CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is the US national stage of PCT/EP2017/050209, filed Jan. 5, 2017, incorporated by reference in its entirety for all purposes, which claims the benefit of EP 16150625.8 filed Jan. 8, 2016; EP 16179288.2 filed Jul. 13, 2016; and EP 16191460.1 filed Sep. 29, 2016.

REFERENCE TO SEQUENCE LISTING

This application includes an electronic sequence listing in a file named 515651_SEQLST.TXT, created on Jun. 28, 2018 and containing 48,205 bytes, which is incorporated by reference.

The present invention relates to CNP prodrugs in which the carrier is covalently and reversibly attached to the ring moiety of a CNP moiety, to pharmaceutical compositions comprising such CNP prodrugs, to their uses and to methods of treating diseases that can be treated with the CNP prodrugs of the present invention.

Gain-of-function mutations in FGFR3 lead to achondroplasia (ACH), hypochondroplasia (HCH), and thanatophoric dysplasia (TD). These conditions, all due to increased signaling of fibroblast-growth-factor-receptor 3 (FGFR3), are characterized by a disproportionate rhizomelic dwarfism and differ in severity, which ranges from mild (HCH) to severe (ACH) and lethal (TD). FGFR3 is a key regulator of endochondral bone growth and signals through several intracellular pathways, including those of the signal transducer and activator of transcription (STAT) and mitogen-activated protein kinase (MAPK). FGFR3 constitutive activation impairs proliferation and terminal differentiation of the growth-plate chondrocytes and synthesis of the extracellular matrix. FGFR3 activation is associated with increased phosphorylation of the STAT and MAPK pathways. The MAPK signaling pathway is regulated by C-type natriuretic peptide (CNP). Binding of CNP to its receptor, natriuretic-peptide receptor B (NPR-B), inhibits FGFR3 downstream signaling and thus triggers endochondral growth and skeletal overgrowth, as observed in both mice and humans overexpressing CNP. Overproduction of CNP in the cartilage or continuous delivery of CNP through intravenous (iv) infusion normalizes the dwarfism of achondroplasic mice, suggesting that administration of CNP at supraphysiological levels is a strategy for treating ACH.

However, given its short half-life of CNP-22 (2 min after intravenous (iv) administration) CNP as a therapeutic agent is challenging in a pediatric population because it would require continuous infusion. Furthermore, as CNP is extensively inactivated in the subcutaneous tissue iv infusion is required.

Potter (FEBS Journal 278 (2011) 1808-1817) describes the clearance of CNP to occur by two degradation routes: receptor-mediated degradation and degradation by extracellular proteases. CNP is degraded by the action of neutral endopeptidase 24.11 (NEP) and is removed from systemic circulation by natriuretic peptide clearance receptor, NPR-C, that binds to and deposits CNP into lysosomes, where CNP is degraded.

Reducing degradation by one or both of these clearance routes would serve to prolong the half-life of CNP.

Due to the limited size of its active site cavity, NEP preferably recognizes substrates smaller than about 3 kDa. U.S. Pat. No. 8,377,884 B2 describe variants of CNP which optionally are permanently conjugated to PEG polymer to increases resistance to NEP cleavage. However, addition of PEG, even as small as 0.6 kDa, to wild-type CNP was found to reduce CNP activity, and addition of greater than about 2 or 3 kDa of PEG to CNP or variants thereof reduce CNP activity in a size-dependent manner. Therefore, attachment of PEG molecules larger than 2 to 3 kDa to reduce NEP degradation is accompanied by a loss of activity, which may reduce the therapeutic potential of such molecules.

In addition to negatively impacting activity of the peptide, conjugation of PEG or another macromolecule to CNP may also prevent effective distribution to the growth plate. Farnum et al. (Anat Rec A Discov Mol Cell Evol Biol. 2006 January; 288(1): 91-103) demonstrated that distribution of molecules from the systemic vasculature to the growth plate was size dependent, and that small molecules (up to 10 kDa) could distribute to the growth plate, whereas a molecular size of 40 kDa and larger prevented entry to the growth plate.

International application WO 2009/156481 A1 relates to reversible PEG-conjugates of BNP which term was defined as including all members of the family of natriuretic peptides. This application only focuses on the cardiovascular effects of this class of peptides, which are mediated through the natriuretic peptide receptor A (NPR-A). WO 2009/156481 A1 fails to disclose CNP's specific properties regarding the regulating of growth, proliferation and differentiation of cartilaginous growth plate chondrocytes, mediated via activation of the natriuretic peptide receptor B (NPR-B).

A different approach to create a NEP resistant CNP molecule and enable subcutaneous administration was described in The American Journal of Human Genetics 91, 1108-1114. BMN-111 is a modified recombinant human C-type Natriuretic Peptide (CNP) where 17 amino acids have been added to form a 39 amino acid CNP pharmacological analog. BMN-111 mimics CNP pharmacological activity at the growth plate and has an extended half-life as a result of neutral-endopeptidase (NEP) resistance that allows once-daily subcutaneous (SC) administration. As BMN-111 is a non-natural occurring peptide, the risk of inducing an immunological response is increased compared to the native peptide, and as described by Martz in “sFGFR for achondroplasia” (SciBx, Biocentury October 2013), an immunological response to BMN-111 has been observed in animal studies, with the presence of antibodies not affecting the pharmacological activity of the drug. However, BMN-111 only has a half-life of 20 minutes, which when dosed daily is associated with a short duration of exposure to efficacious drug levels.

To increase exposure to efficacious drug levels the dose of the drug having CNP activity may be increased. As natriuretic peptides are a family of hormones that may affect blood volume and blood pressure, an increase in dose may be associated with cardiovascular adverse effects. Studies of BMN-111 in animals and man have demonstrated that as the dose increases, arterial blood pressure drops and heart rate increases. Doses of BMN-111 up to 15 μg/kg were associated with mild hypotension in healthy volunteers. Therefore increasing the dose of a drug having CNP activity to increase drug exposure may be associated with unacceptable cardiovascular side effects.

In WO2009/0676639A2 and WO2010/135541A2 the use of PEGylated CNP is contemplated. However, the authors consider retaining the functionality of such a PEGylated CNP an essential property of their conjugates. Therefore, the PEGylated CNP conjugates disclosed therein maintain their ability to bind to NPR-B and thus still pose the problem of hypotension.

In summary, there is a need for a more convenient and safer CNP treatment with a reduced risk of hypotension.

It is therefore an object of the present invention to at least partially overcome the shortcomings described above.

This object is achieved with a CNP prodrug or a pharmaceutically acceptable salt thereof comprisinga CNP moiety -D comprising a ring moiety; anda carrier moiety —Z that is conjugated through a moiety -L2- to a reversible prodrug linker moiety -L1-, which reversible prodrug linker moiety -L1- is covalently and reversibly conjugated to a side chain of an amino acid residue of said ring moiety of -D or to the backbone of said ring moiety of -D; and wherein -L2- is a chemical bond or a spacer.

It was surprisingly found that attachment of a carrier —Z through a reversible prodrug linker moiety to the ring of a CNP moiety significantly reduces binding of the prodrug to the NPR-B receptor compared to, for example, N-terminal attachment or attachment outside the ring moiety of CNP. This allows the administration of a CNP compound with low or nearly no residual activity, i.e. with reduced NPR-B binding, which in turn reduces the risk of hypotension. As the attachment of the polymer —Z to the CNP moiety is reversible, NPR-B binding is restored upon release from the CNP prodrug and the released CNP can be effectively distributed to the growth plate.

This combination of initial inactivity towards NPR-B and subsequent controlled release from the prodrug conjugate which converts the CNP moiety into a fully active form of the drug with regard to NPR-B binding provides several advantages, such as a reduction in the risk of hypotension, the option of administering higher doses and a decrease in the administration frequency which enhances compliance by and convenience for the patient.

It was furthermore surprisingly found that a continuous release of CNP, such as from a controlled release system, such as from the prodrugs of the present invention, is more efficacious than a once-daily bolus injection.

Within the present invention the terms are used having the meaning as follows.

As used herein the term “CNP” refers to all CNP polypeptides, preferably from mammalian species, more preferably from human and mammalian species, more preferably from human and murine species, as well as their variants, analogs, orthologs, homologs, and derivatives and fragments thereof, that are characterized by regulating the growth, proliferation and differentiation of cartilaginous growth plate chondrocytes. Preferably, the term “CNP” refers to the CNP polypeptide of SEQ ID NO:24 as well as its variants, homologs and derivatives exhibiting essentially the same biological activity, i.e. regulating the growth, proliferation and differentiation of cartilaginous growth plate chondrocytes. More preferably, the term “CNP” refers to the polypeptide of SEQ ID NO:24.

In another preferred embodiment the term “CNP” refers to the polypeptide of SEQ ID NO:20.

In another preferred embodiment the term “CNP” refers to the polypeptide of SEQ ID NO:21.

In another preferred embodiment the term “CNP” refers to the polypeptide of SEQ ID NO:22.

In another preferred embodiment the term “CNP” refers to the polypeptide of SEQ ID NO:23.

In another preferred embodiment the term “CNP” refers to the polypeptide of SEQ ID NO:30.

Naturally occurring CNP-22 (SEQ ID NO:1) has the following sequence: GLSKGCFGLKLDRIGSMSGLGC,

wherein the cysteines at position 6 and 22 are connected through a disulfide-bridge, as illustrated inFIG. 1.

SEQ ID NO:24 has the following sequence:

LQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC,

wherein the cysteines at position 22 and 38 are connected through a disulfide-bride.

The term “CNP” also includes all CNP variants, analogs, orthologs, homologs and derivatives and fragments thereof as disclosed in WO 2009/067639 A2 and WO 2010/135541 A2, which are herewith incorporated by reference.

Accordingly, the term “CNP” also refers preferably to the following peptide sequences:

SEQ ID NO: 2 (CNP-53):DLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 3 (G-CNP-53):GDLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 4 (M-CNP-53):MDLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 5 (P-CNP-53):PDLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 6 (CNP-53 M48N):DLRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSNSGLGC;SEQ ID NO: 7 (CNP-53 415-31):DLRVDTKSRAAWARGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 8 (CNP-52):LRVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 9 (CNP-51):RVDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 10 (CNP-50):VDTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 11 (CNP-49):DTKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 12 (CNP-48):TKSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 13 (CNP-47):KSRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 14 (CNP-46):SRAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 15 (CNP-45):RAAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 16 (CNP-44):AAWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 17 (CNP-44 Δ14-22):AAWARLLQEHPNAGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 18 (CNP-44 Δ15-22):AAWARLLQEHPNARGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 19 (CNP-43):AWARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 20 (CNP-42):WARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 21 (CNP-41):ARLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 22 (CNP-40):RLLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 23 (CNP-39):LLQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 24 (CNP-38):LQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 25 (CNP-37):QEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 26 (CNP-37 Q1pQ, wherein pQ =pyroglutamate):pQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 27 (G-CNP-37):GQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 28 (P-CNP-37):PQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 29 (M-CNP-37):MQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 30 (PG-CNP-37):PGQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 31 (MG-CNP-37):MGQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 32 (CNP-37 M32N):QEHPNARKYKGANKKGLSKGCFGLKLDRIGSNSGLGC;SEQ ID NO: 33 (G-CNP-37 M32N):GQEHPNARKYKGANKKGLSKGCFGLKLDRIGSNSGLGC;SEQ ID NO: 34 (G-CNP-37 K14Q):GQEHPNARKYKGANQKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 35 (G-CNP-37 K14P):GQEHPNARKYKGANPKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 36 (G-CNP-37 K14Q, Δ15):GQEHPNARKYKGANQGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 37 (G-CNP-37 K14Q, K15Q):GQEHPNARKYKGANQQGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 38 (CNP-36):EHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 39 (CNP-35):HPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 40 (CNP-34):PNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 41 (CNP-33):NARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 42 (CNP-32):ARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 43 (CNP-31):RKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 44 (CNP-30):KYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 45 (CNP-29):YKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 46 (CNP-28):KGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 47 (GHKSEVAHRF-CNP-28):GHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 48 (CNP-27):GANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 49 (CNP-27 K4Q, K5Q):GANQQGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 50 (CNP-27 K4R, K5R):GANRRGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 51 (CNP-27 K4P, K5R):GANPRGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 52 (CNP-27 K4S, K5S):GANSSGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 53 (CNP-27 K4P, K5R):GANGANPRGLSRGCFGLKLDRIGSMSGLGC;SEQ ID NO: 54 (CNP-27 K4R, K5R, K9R):GANRRGLSRGCFGLKLDRIGSMSGLGC;SEQ ID NO: 55 (CNP-27 K4R, K5R, K9R, M22N):GANRRGLSRGCFGLKLDRIGSNSGLGC;SEQ ID NO: 56 (P-CNP-27 K4R, K5R, K9R):PGANRRGLSRGCFGLKLDRIGSMSGLGC;SEQ ID NO: 57 (M-CNP-27 K4R, K5R, K9R):MGANRRGLSRGCFGLKLDRIGSMSGLGC;SEQ ID NO: 58 (HSA fragment-CNP-27):GHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLG;SEQ ID NO: 59 (HSA fragment-CNP-27 M22N):GHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSNSGLGC;SEQ ID NO: 60 (M-HSA fragment-CNP-27):MGHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 61 (P-HSA fragment-CNP-27):PGHKSEVAHRFKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 62 (CNP-26):ANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 63 (CNP-25):NKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 64 (CNP-24):KKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 65 (CNP-23):KGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 66 (R-CNP-22):RGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 67 (ER-CNP-22):ERGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 68 (R-CNP-22 K4R):RGLSRGCFGLKLDRIGSMSGLGC;SEQ ID NO: 69 (ER-CNP-22 4KR):ERGLSRGCFGLKLDRIGSMSGLGC;SEQ ID NO: 70 (RR-CNP-22):RRGLSRGCFGLKLDRIGSMSGLGC;SEQ ID NO: 71 (HRGP fragment-CNP-22):GHHSHEQHPHGANQQGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 72 (HRGP fragment-CNP-22):GAHHPHEHDTHGANQQGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 73 (HRGP fragment-CNP-22):GHHSHEQHPHGANPRGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 74 (IgG1(Fc) fragment-CNP-22):GQPREPQVYTLPPSGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 75 (HSA fragment-CNP-22):GQHKDDNPNLPRGANPRGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 76 (HSA fragment-CNP-22):GERAFKAWAVARLSQGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 77 (osteocrin NPR C inhibitor fragment-CNP22):FGIPMDRIGRNPRGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 78 (FGF2 heparin-binding domainfragment-CNP22):GKRTGQYKLGSKTGPGPKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 79 (IgG1(Fc) fragment-CNP-22 K4R):GQPREPQVYTGANQQGLSRGCFGLKLDRIGSMSGLGC;SEQ ID NO: 80 (HSA fragment-CNP-22 K4R):GVPQVSTSTGANQQGLSRGCFGLKLDRIGSMSGLGC;SEQ ID NO: 81 (fibronectin fragment-CNP-22 K4R):GQPSSSSQSTGANQQGLSRGCFGLKLDRIGSMSGLGC;SEQ ID NO: 82 (fibronectin fragment-CNP-22 K4R):GQTHSSGTQSGANQQGLSRGCFGLKLDRIGSMSGLGC;SEQ ID NO: 83 (fibronectin fragment-CNP-22 K4R):GSTGQWHSESGANQQGLSRGCFGLKLDRIGSMSGLGC;SEQ ID NO: 84 (zinc finger fragment-CNP-22 K4R):GSSSSSSSSSGANQQGLSRGCFGLKLDRIGSMSGLGC;SEQ ID NO: 85 (CNP-21):LSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 86 (CNP-20):SKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 87 (CNP-19):KGCFGLKLDRIGSMSGLGC;SEQ ID NO: 88 (CNP-18):GCFGLKLDRIGSMSGLGC;SEQ ID NO: 89 (CNP-17):CFGLKLDRIGSMSGLGC;SEQ ID NO: 90 (BNP fragment-CNP-17-BNP fragment):SPKMVQGSGCFGLKLDRIGSMSGLGCKVLRRH;SEQ ID NO: 91 (CNP-38 L1G):GQEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC;SEQ ID NO: 92 (Ac-CNP-37; wherein Ac = acetyl):Ac-QEHPNARKYKGANKKGLSKGCFGLKLDRIGSMSGLGC.

More preferably, the term “CNP” refers to the sequence of SEQ ID NOs: 2, 19, 20, 21, 22, 23, 24, 25, 26, 30, 32, 38, 39, 40, 41, 42, 43, 91, 92. Even more preferably, the term “CNP” refers to the sequence of SEQ ID NOs: 23, 24, 25, 26, 38, 39, 91 and 92. In a particularly preferred embodiment the term “CNP” refers to the sequence of SEQ ID NO:24. In an equally preferred embodiment the term “CNP” refers to the sequence of SEQ ID NO:30. In an equally preferred embodiment the term “CNP” refers to the sequence of SEQ ID NO:20. In an equally preferred embodiment the term “CNP” refers to the sequence of SEQ ID NO:21. In an equally preferred embodiment the term “CNP” refers to the sequence of SEQ ID NO:22. In an equally preferred embodiment the term “CNP” refers to the sequence of SEQ ID NO:23.

In one embodiment the term “CNP” refers to a sequence of SEQ ID NO:93 QEHPNARX1YX2GANX3X4GLSX5GCFGLX6LDRIGSMSGLGC,

wherein X1, X2, X3, X4, X5and X6are independently of each other selected from the group consisting of K, R, P, S and Q, with the provision that at least one of X1, X2, X3, X4, X5and X6is selected from the group consisting of R, P, S and Q; preferably X1, X2, X3, X4, X5and X6are selected from the group consisting of K and R, with the provision that at least one of X1, X2, X3, X4, X5and X6is R;

more preferably to a sequence of SEQ ID NO:94

QEHPNARKYKGANX1X2GLSX3GCFGLX4LDRIGSMSGLGC, wherein X1, X2, X3and X4are independently of each other selected from the group consisting of K, R, P, S and Q, with the provision that at least one of X1, X2, X3and X4is selected from the group consisting of R, P, S and Q; preferably X1, X2, X3and X4are selected from K and R, with the provision that at least one of X1, X2, X3and X4is R;

and even more preferably to a sequence of SEQ ID NO:95 QEHPNARKYKGANX1X2GLSKGCFGLKLDRIGSMSGLGC,

wherein X1X2are selected from the group consisting of KR, RK, KP, PK, SS, RS, SR, QK, QR, KQ, RQ, RR and QQ.

It is understood that also the equivalents of the cysteines in positions 22 and 38 of SEQ ID NO:24 are connected through a disulfide-bridge in SEQ ID NOs: 2 to 95.

It is understood that the present invention also encompasses CNP variants in which any one or more, up to all, residues susceptible to deamidation or a deamidation-like reaction (e.g., isomerization) may be converted to other residues via deamidation or a deamidation-like reaction to any extent, up to 100% conversion per converted residue. In certain embodiments, the disclosure encompasses CNP variants in which:

(1) any one or more, up to all, asparagine (Asn/N) residues may be converted to aspartic acid or aspartate, and/or to isoaspartic acid or isoaspartate, via deamidation up to about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% conversion per converted residue; or

(2) any one or more, up to all, glutamine (Gln/Q) residues may be converted to glutamic acid or glutamate, and/or to isoglutamic acid or isoglutamate, via deamidation up to about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% conversion per converted residue; or

(3) any one or more, up to all, aspartic acid or aspartate (Asp/D) residues may be converted to isoaspartic acid or isoaspartate via a deamidation-like reaction (also called isomerization) up to about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% conversion per converted residue; or

(4) any one or more, up to all, glutamic acid or glutamate (Glu/E) residues may be converted to isoglutamic acid or isoglutamate via a deamidation-like reaction (also called isomerization) up to about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% conversion per converted residue; or

(5) the N-terminal glutamine (if present) may be converted into pyroglutamate up to about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% conversion; or

(6) a combination of the above.

As used herein, the term “CNP polypeptide variant” refers to a polypeptide from the same species that differs from a reference CNP polypeptide. Preferably, such reference CNP polypeptide sequence is the sequence of SEQ ID NO:24. Generally, differences are limited so that the amino acid sequence of the reference and the variant are closely similar overall and, in many regions, identical. Preferably, CNP polypeptide variants are at least 70%, 80%, 90%, or 95% identical to a reference CNP polypeptide, preferably the CNP polypeptide of SEQ ID NO:24. By a polypeptide having an amino acid sequence at least, for example, 95% “identical” to a query amino acid sequence, it is intended that the amino acid sequence of the subject polypeptide is identical to the query sequence except that the subject polypeptide sequence may include up to five amino acid alterations per each 100 amino acids of the query amino acid sequence. These alterations of the reference sequence may occur at the amino (N-terminal) or carboxy terminal (C-terminal) positions of the reference amino acid sequence or anywhere between those terminal positions, interspersed either individually among residues in the reference sequence or in one or more contiguous groups within the reference sequence. The query sequence may be an entire amino acid sequence of the reference sequence or any fragment specified as described herein. Preferably, the query sequence is the sequence of SEQ ID NO:24.

Such CNP polypeptide variants may be naturally occurring variants, such as naturally occurring allelic variants encoded by one of several alternate forms of a CNP occupying a given locus on a chromosome or an organism, or isoforms encoded by naturally occurring splice variants originating from a single primary transcript. Alternatively, a CNP polypeptide variant may be a variant that is not known to occur naturally and that can be made by mutagenesis techniques known in the art.

It is known in the art that one or more amino acids may be deleted from the N-terminus or C-terminus of a bioactive peptide or protein without substantial loss of biological function. Such N- and/or C-terminal deletions are also encompassed by the term CNP polypeptide variant.

It is also recognized by one of ordinary skill in the art that some amino acid sequences of CNP polypeptides can be varied without significant effect of the structure or function of the peptide. Such mutants include deletions, insertions, inversions, repeats, and substitutions selected according to general rules known in the art so as to have little effect on activity. For example, guidance concerning how to make phenotypically silent amino acid substitutions is provided in Bowie et al. (1990), Science 247:1306-1310, which is hereby incorporated by reference in its entirety, wherein the authors indicate that there are two main approaches for studying the tolerance of the amino acid sequence to change.

The term CNP polypeptide also encompasses all CNP polypeptides encoded by CNP analogs, orthologs, and/or species homologs. As used herein, the term “CNP analog” refers to CNP of different and unrelated organisms which perform the same functions in each organism, but which did not originate from an ancestral structure that the organisms' ancestors had in common. Instead, analogous CNPs arose separately and then later evolved to perform the same or similar functions. In other words, analogous CNP polypeptides are polypeptides with quite different amino acid sequences that perform the same biological activity, namely regulating the growth, proliferation and differentiation of cartilaginous growth plate chondrocytes.

As used herein the term “CNP ortholog” refers to CNP within two different species which sequences are related to each other via a common homologous CNP in an ancestral species, but which have evolved to become different from each other.

As used herein, the term “CNP homolog” refers to CNP of different organisms which perform the same functions in each organism and which originate from an ancestral structure that the organisms' ancestors had in common. In other words, homologous CNP polypeptides are polypeptides with quite similar amino acid sequences that perform the same biological activity, namely regulating the growth, proliferation and differentiation of cartilaginous growth plate chondrocytes. Preferably, CNP polypeptide homologs may be defined as polypeptides exhibiting at least 40%, 50%, 60%, 70%, 80%, 90% or 95% identity to a reference CNP polypeptide, preferably the CNP polypeptide of SEQ ID NO:24.

Thus, a CNP polypeptide according to the invention may be, for example: (i) one in which at least one of the amino acid residues is substituted with a conserved or non-conserved amino acid residue, preferably a conserved amino acid residue, and such substituted amino acid residue may or may not be one encoded by the genetic code; and/or (ii) one in which at least one of the amino acid residues includes a substituent group; and/or (iii) one in which the CNP polypeptide is fused with another compound, such as a compound to increase the half-life of the polypeptide (for example, polyethylene glycol); and/or (iv) one in which additional amino acids are fused to the CNP polypeptide, such as an IgG Fc fusion region peptide or leader or secretory sequence or a sequence which is employed for purification of the above form of the polypeptide or a pre-protein sequence.

As used herein, the term “CNP polypeptide fragment” refers to any peptide comprising a contiguous span of a part of the amino acid sequence of a CNP polypeptide, preferably the polypeptide of SEQ ID NO:24.

More specifically, a CNP polypeptide fragment comprises at least 6, such as at least 8, at least 10 or at least 17 consecutive amino acids of a CNP polypeptide, more preferably of the polypeptide of SEQ ID NO:24. A CNP polypeptide fragment may additionally be described as sub-genuses of CNP polypeptides comprising at least 6 amino acids, wherein “at least 6” is defined as any integer between 6 and the integer representing the C-terminal amino acid of a CNP polypeptide, preferably of the polypeptide of SEQ ID No:24. Further included are species of CNP polypeptide fragments at least 6 amino acids in length, as described above, that are further specified in terms of their N-terminal and C-terminal positions. Also encompassed by the term “CNP polypeptide fragment” as individual species are all CNP polypeptide fragments, at least 6 amino acids in length, as described above, that may be particularly specified by a N-terminal and C-terminal position. That is, every combination of a N-terminal and C-terminal position that a fragment at least 6 contiguous amino acid residues in length could occupy, on any given amino acid sequence of a CNP polypeptide, preferably the CNP polypeptide of SEQ ID NO:24, is included in the present invention.

The term “CNP” also includes poly(amino acid) conjugates which have a sequence as described above, but have a backbone that comprises both amide and non-amide linkages, such as ester linkages, like for example depsipeptides. Depsipeptides are chains of amino acid residues in which the backbone comprises both amide (peptide) and ester bonds. Accordingly, the term “side chain” as used herein refers either to the moiety attached to the alpha-carbon of an amino acid moiety, if the amino acid moiety is connected through amine bonds such as in polypeptides, or to any carbon atom-comprising moiety attached to the backbone of a poly(amino acid) conjugate, such as for example in the case of depsipeptides. Preferably, the term “CNP” refers to polypeptides having a backbone formed through amide (peptide) bonds.

As the term CNP includes the above-described variants, analogs, orthologs, homologs, derivatives and fragments of CNP, all references to specific positions within a reference sequence also include the equivalent positions in the variants, analogs, orthologs, homologs, derivatives and fragments of a CNP moiety, even if not explicitly mentioned.

As used herein, the term “random coil” refers to a peptide or protein adopting/having/forming, preferably having, a conformation which substantially lacks a defined secondary and tertiary structure as determined by circular dichroism spectroscopy performed in aqueous buffer at ambient temperature, and pH 7.4. Preferably, ambient temperature is about 20° C., i.e. between 18° C. and 22° C., most preferably ambient temperature is 20° C.

As used herein, the term “ring moiety” refers to the stretch of consecutive amino acid residues of the CNP drug or moiety that is located between two cysteine residues that form an intramolecular disulfide bridge or between homologous amino acid residues which are connected through a chemical linker. Preferably, the ring moiety is located between two cysteine residues that form an intramolecular disulfide bridge. These two cysteines correspond to the cysteines at position 22 and position 38 in the sequence of CNP-38 (SEQ ID NO:24). Accordingly, amino acids 23 to 37 are located in said ring moiety, if the CNP drug or moiety has the sequence of CNP-38.

Independently of the length of the CNP moiety, the sequence of the ring moiety of wild-type CNP is FGLKLDRIGSMSGLG (SEQ ID NO:96).

As described above, the term “CNP” relates to CNP drugs or moieties having different numbers of amino acids. The person skilled in the art understands that in CNP drugs or moieties of different lengths the positions of equivalent amino acids vary and the skilled artisan will have no difficulty identifying the two cysteines forming the disulfide bridge or their two homologous amino acid residues connected to each other through a chemical linker in longer, shorter and/or otherwise modified CNP versions.

As the term CNP includes the above-described variants, analogs, orthologs, homologs, derivatives and fragments of CNP, the term “ring moiety” also includes the corresponding variants, analogs, orthologs, homologs, derivatives and fragments of the sequence of SEQ ID NO:96. Accordingly, all references to specific positions within a reference sequence also include the equivalent positions in variants, analogs, orthologs, homologs, derivatives and fragments of a CNP moiety, even if not explicitly mentioned.

As used herein the term “liquid composition” refers to a mixture comprising water-soluble CNP prodrug and one or more solvents, such as water.

As used herein, the term “dry composition” means that a pharmaceutical composition is provided in a dry form. Suitable methods for drying are spray-drying and lyophilization, i.e. freeze-drying. Such dry composition of the prodrug of the present invention has a residual water content of a maximum of 10%, preferably less than 5% and more preferably less than 2%, determined according to Karl Fischer. Preferably, the pharmaceutical composition of the present invention is dried by lyophilization.

The term “drug” as used herein refers to a substance used in the treatment, cure, prevention, or diagnosis of a disease or used to otherwise enhance physical or mental well-being. If a drug D-H is conjugated to another moiety, the moiety -D of the resulting product that originated from the drug is referred to as “biologically active moiety”.

As used herein the term “prodrug” refers to a biologically active moiety reversibly and covalently connected to a specialized protective group through a reversible prodrug linker moiety which is a linker moiety comprising a reversible linkage with the biologically active moiety and wherein the specialized protective group alters or eliminates undesirable properties in the parent molecule. This also includes the enhancement of desirable properties in the drug and the suppression of undesirable properties. The specialized non-toxic protective group is referred to as “carrier”. A prodrug releases the reversibly and covalently bound biologically active moiety -D in the form of its corresponding drug D-H. In other words, a prodrug is a conjugate comprising a biologically active moiety which is covalently and reversibly conjugated to a carrier moiety via a reversible prodrug linker moiety, which covalent and reversible conjugation of the carrier to the reversible prodrug linker moiety is either directly or through a spacer, such as -L2-. Such conjugate releases the formerly conjugated biologically active moiety in the form of a free drug.

A “biodegradable linkage” or a “reversible linkage” is a linkage that is hydrolytically degradable, i.e. cleavable, in the absence of enzymes under physiological conditions (aqueous buffer at pH 7.4, 37° C.) with a half-life ranging from one hour to six months, preferably from one hour to four months, even more preferably from one hour to three months, even more preferably from one hour to two months, even more preferably from one hour to one month. Accordingly, a “stable linkage” is a linkage having a half-life under physiological conditions (aqueous buffer at pH 7.4, 37° C.) of more than six months.

Accordingly, a “reversible prodrug linker moiety” is a moiety which is covalently conjugated to a biologically active moiety, such as CNP, through a reversible linkage and is also covalently conjugated to a carrier moiety, such as —Z, wherein the covalent conjugation to said carrier moiety is either directly or through a spacer moiety, such as -L2-. Preferably the linkage between —Z and -L2- is a stable linkage.

As used herein, the term “traceless prodrug linker” means a reversible prodrug linker which upon cleavage releases the drug in its free form. As used herein, the term “free form” of a drug means the drug in its unmodified, pharmacologically active form.

As used herein the term “pharmaceutical composition” refers to a composition containing one or more active ingredients, for example a drug or a prodrug, here specifically the CNP prodrugs of the present invention, and optionally one or more excipients, as well as any product which results, directly or indirectly, from combination, complexation or aggregation of any two or more of the ingredients of the composition, or from dissociation of one or more of the ingredients, or from other types of reactions or interactions of one or more of the ingredients. Accordingly, the pharmaceutical compositions of the present invention encompass any composition made by admixing one or more CNP prodrugs of the present invention and optionally a pharmaceutically acceptable excipient.

As used herein, the term “excipient” refers to a diluent, adjuvant, or vehicle with which the therapeutic, such as a drug or prodrug, is administered. Such pharmaceutical excipient can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, including but not limited to peanut oil, soybean oil, mineral oil, sesame oil and the like. Water is a preferred excipient when the pharmaceutical composition is administered orally. Saline and aqueous dextrose are preferred excipients when the pharmaceutical composition is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions are preferably employed as liquid excipients for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, mannitol, trehalose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like. The pharmaceutical composition, if desired, can also contain minor amounts of wetting or emulsifying agents, pH buffering agents, like, for example, acetate, succinate, tris, carbonate, phosphate, HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), MES (2-(N-morpholino)ethanesulfonic acid), or can contain detergents, like Tween, poloxamers, poloxamines, CHAPS, Igepal, or amino acids like, for example, glycine, lysine, or histidine. These pharmaceutical compositions can take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained-release formulations and the like. The pharmaceutical composition can be formulated as a suppository, with traditional binders and excipients such as triglycerides. Oral formulation can include standard excipients such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, etc. Such compositions will contain a therapeutically effective amount of the drug or biologically active moiety, together with a suitable amount of excipient so as to provide the form for proper administration to the patient. The formulation should suit the mode of administration.

As used herein the term “micelle” means an aggregate of amphiphilic molecules dispersed in a liquid colloid. In aqueous solution a typical micelle forms an aggregate with the hydrophilic moiety of the surfactant molecules facing the surrounding solvent and the hydrophobic moiety of the surfactant molecule facing inwards, also called “normal-phase micelle”. “Invers micelles” have the hydrophilic moiety facing inwards and the hydrophobic moiety facing the surrounding solvent.

As used herein the term “liposome” refers to a vesicle, preferably a spherical vesicle, having at least one lipid bilayer. Preferably, liposomes comprise phospholipids, even more preferably phosphatidylcholine. The term “liposome” refers to various structures and sizes, such as, for example, to multilamellar liposome vesicles (MLV) having more than one concentric lipid bilayer with an average diameter of 100 to 1000 nm, small unilamellar liposome vesicles (SUV) having one lipid bilayer and an average diameter of 25 to 100 nm, large unilamellar liposome vesicles (LUV) having one lipid bilayer and an average diameter of about 1000 μm and giant unilamellar vesicles (GUV) having one lipid bilayer and an average diameter of 1 to 100 μm. The term “liposome” also includes elastic vesicles such as transferosomes and ethosomes, for example.

As used herein the term “aquasome” refers to spherical nanoparticles having a diameter of 60 to 300 nm that comprise at least three layers of self-assembled structure, namely a solid phase nanocrystalline core coated with an oligomeric film to which drug molecules are adsorbed with or without modification of the drug.

As used herein the term “ethosome” refers to lipid vesicles comprising phospholipids and ethanol and/or isopropanol in relatively high concentration and water, having a size ranging from tens of nanometers to micrometers.

As used herein the term “LeciPlex” refers to positively charged phospholipid-based vesicular system which comprises soy PC, a cationic agent, and a bio-compatible solvent like PEG 300, PEG 400, diethylene glycol monoethyl ether, tetrahydrofurfuryl alcohol polyethylene glycol ether or 2-pyrrolidoneor N-methyl-2-pyrrolidone.

As used herein the term “niosome” refers to unilamellar or multilamellar vesicles comprising non-ionic surfactants.

As used herein the term “pharmacosome” refers to ultrafine vesicular, micellar or hexagonal aggregates from lipids covalently bound to biologically active moieties.

As used herein the term “proniosome” refers to dry formulations of surfactant-coated carrier which on rehydration and mild agitation gives niosomes.

As used herein the term “polymersome” refers to an artificial spherical vesicle comprising a membrane formed from amphiphilic synthetic block copolymers and may optionally comprise an aqueous solution in its core. A polymersome has a diameter ranging from 50 nm to 5 μm and larger. The term also includes syntosomes, which are polymersomes engineered to comprise channels that allow certain chemicals to pass through the membrane into or out of the vesicle.

As used herein the term “sphingosome” refers to a concentric, bilayered vesicle in which an aqueous volume is entirely enclosed by a membranous lipid bilayer mainly composed of natural or synthetic sphingolipid.

As used herein the term “transferosome” refers to ultraflexible lipid vesicles comprising an aqueous core that are formed from a mixture of common polar and suitable edge-activated lipids which facilitate the formation of highly curved bilayers which render the transferosome highly deformable.

As used herein the term “ufasome” refers to a vesicle comprising unsaturated fatty acids.

As used herein, the term “reagent” means a chemical compound which comprises at least one functional group for reaction with the functional group of another chemical compound or drug. It is understood that a drug comprising a functional group (such as a primary or secondary amine or hydroxyl functional group) is also a reagent.

As used herein, the term “moiety” means a part of a molecule, which lacks one or more atoms compared to the corresponding reagent. If, for example, a reagent of the formula “H—X—H” reacts with another reagent and becomes part of the reaction product, the corresponding moiety of the reaction product has the structure “H—X—” or “—X—”, whereas each “—” indicates attachment to another moiety. Accordingly, a biologically active moiety is released from a prodrug as a drug.

It is understood that if the sequence or chemical structure of a group of atoms is provided which group of atoms is attached to two moieties or is interrupting a moiety, said sequence or chemical structure can be attached to the two moieties in either orientation, unless explicitly stated otherwise. For example, a moiety “—C(O)N(R1)—” can be attached to two moieties or interrupting a moiety either as “—C(O)N(R1)—” or as “—N(R1)C(O)—”. Similarly, a moiety


can be attached to two moieties or can interrupt a moiety either as



As used herein, the term “functional group” means a group of atoms which can react with other groups of atoms. Functional groups include but are not limited to the following groups: carboxylic acid (—(C═O)OH), primary or secondary amine (—NH2, —NH—), maleimide, thiol (—SH), sulfonic acid (—(O═S═O)OH), carbonate, carbamate (—O(C═O)N<), hydroxyl (—OH), aldehyde (—(C═O)H), ketone (—(C═O)—), hydrazine (>N—N<), isocyanate, isothiocyanate, phosphoric acid (—O(P═O)OHOH), phosphonic acid (—O(P═O)OHH), haloacetyl, alkyl halide, acryloyl, aryl fluoride, hydroxylamine, disulfide, sulfonamides, sulfuric acid, vinyl sulfone, vinyl ketone, diazoalkane, oxirane, and aziridine.

In case the prodrugs of the present invention comprise one or more acidic or basic groups, the invention also comprises their corresponding pharmaceutically or toxicologically acceptable salts, in particular their pharmaceutically utilizable salts. Thus, the prodrugs of the present invention comprising acidic groups can be used according to the invention, for example, as alkali metal salts, alkaline earth metal salts or as ammonium salts. More precise examples of such salts include sodium salts, potassium salts, calcium salts, magnesium salts or salts with ammonia or organic amines such as, for example, ethylamine, ethanolamine, triethanolamine or amino acids. Prodrugs of the present invention comprising one or more basic groups, i.e. groups which can be protonated, can be present and can be used according to the invention in the form of their addition salts with inorganic or organic acids. Examples for suitable acids include hydrogen chloride, hydrogen bromide, phosphoric acid, sulfuric acid, nitric acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acids, oxalic acid, acetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, formic acid, propionic acid, pivalic acid, diethylacetic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, malic acid, sulfaminic acid, phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic acid, citric acid, adipic acid, and other acids known to the person skilled in the art. For the person skilled in the art further methods are known for converting the basic group into a cation like the alkylation of an amine group resulting in a positively-charge ammonium group and an appropriate counterion of the salt. If the prodrugs of the present invention simultaneously comprise acidic and basic groups, the invention also includes, in addition to the salt forms mentioned, inner salts or betaines (zwitterions). The respective salts can be obtained by customary methods which are known to the person skilled in the art like, for example by contacting these prodrugs with an organic or inorganic acid or base in a solvent or dispersant, or by anion exchange or cation exchange with other salts. The present invention also includes all salts of the prodrugs of the present invention which, owing to low physiological compatibility, are not directly suitable for use in pharmaceuticals but which can be used, for example, as intermediates for chemical reactions or for the preparation of pharmaceutically acceptable salts.

The term “pharmaceutically acceptable” means a substance that does cause harm when administered to a patient and preferably means approved by a regulatory agency, such as the EMA (Europe) and/or the FDA (US) and/or any other national regulatory agency for use in animals, preferably for use in humans.

As used herein the term “about” in combination with a numerical value is used to indicate a range ranging from and including the numerical value plus and minus no more than 10% of said numerical value, more preferably no more than 8% of said numerical value, even more preferably no more than 5% of said numerical value and most preferably no more than 2% of said numerical value. For example, the phrase “about 200” is used to mean a range ranging from and including 200+/−10%, i.e. ranging from and including 180 to 220; preferably 200+/−8%, i.e. ranging from and including 184 to 216; even more preferably ranging from and including 200+/−5%, i.e. ranging from and including 190 to 210; and most preferably 200+/−2%, i.e. ranging from and including 196 to 204. It is understood that a percentage given as “about 20%” does not mean “20%+/−10%”, i.e. ranging from and including 10 to 30%, but “about 20%” means ranging from and including 18 to 22%, i.e. plus and minus 10% of the numerical value which is 20.

As used herein, the term “polymer” means a molecule comprising repeating structural units, i.e. the monomers, connected by chemical bonds in a linear, circular, branched, crosslinked or dendrimeric way or a combination thereof, which may be of synthetic or biological origin or a combination of both. It is understood that a polymer may also comprise one or more other chemical groups and/or moieties, such as, for example, one or more functional groups. Preferably, a soluble polymer has a molecular weight of at least 0.5 kDa, e.g. a molecular weight of at least 1 kDa, a molecular weight of at least 2 kDa, a molecular weight of at least 3 kDa or a molecular weight of at least 5 kDa. If the polymer is soluble, it preferable has a molecular weight of at most 1000 kDa, such as at most 750 kDa, such as at most 500 kDa, such as at most 300 kDa, such as at most 200 kDa, such as at most 100 kDa. It is understood that also a protein is a polymer in which the amino acids are the repeating structural units, even though the side chains of each amino acid may be different.

As used herein, the term “polymeric” means a reagent or a moiety comprising one or more polymers or polymer moieties. A polymeric reagent or moiety may optionally also comprise one or more other moieties, which are preferably selected from the group consisting of:C1-50alkyl, C2-50alkenyl, C2-50alkynyl, C3-10cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, phenyl, naphthyl, indenyl, indanyl, and tetralinyl; andlinkages selected from the group comprising

whereindashed lines indicate attachment to the remainder of the moiety or reagent, and —R and —Raare independently of each other selected from the group consisting of —H, methyl, ethyl, propyl, butyl, pentyl and hexyl.

The person skilled in the art understands that the polymerization products obtained from a polymerization reaction do not all have the same molecular weight, but rather exhibit a molecular weight distribution. Consequently, the molecular weight ranges, molecular weights, ranges of numbers of monomers in a polymer and numbers of monomers in a polymer as used herein, refer to the number average molecular weight and number average of monomers, i.e. to the arithmetic mean of the molecular weight of the polymer or polymeric moiety and the arithmetic mean of the number of monomers of the polymer or polymeric moiety.

Accordingly, in a polymeric moiety comprising “x” monomer units any integer given for “x” therefore corresponds to the arithmetic mean number of monomers. Any range of integers given for “x” provides the range of integers in which the arithmetic mean numbers of monomers lies. An integer for “x” given as “about x” means that the arithmetic mean numbers of monomers lies in a range of integers of x+/−10%, preferably x+/−8%, more preferably x+/−5% and most preferably x+/−2%.

As used herein, the term “number average molecular weight” means the ordinary arithmetic mean of the molecular weights of the individual polymers.

As used herein the term “water-soluble” with reference to a carrier means that when such carrier is part of the CNP prodrug of the present invention at least 1 g of the CNP prodrug comprising such water-soluble carrier can be dissolved in one liter of water at 20° C. to form a homogeneous solution. Accordingly, the term “water-insoluble” with reference to a carrier means that when such carrier is part of the CNP prodrug of the present invention less than 1 g of the CNP prodrug comprising such water-insoluble carrier can be dissolved in one liter of water at 20° C. to form a homogeneous solution.

As used herein, the term “PEG-based” in relation to a moiety or reagent means that said moiety or reagent comprises PEG. Preferably, a PEG-based moiety or reagent comprises at least 10% (w/w) PEG, such as at least 20% (w/w) PEG, such as at least 30% (w/w) PEG, such as at least 40% (w/w) PEG, such as at least 50% (w/w), such as at least 60 (w/w) PEG, such as at least 70% (w/w) PEG, such as at least 80% (w/w) PEG, such as at least 90% (w/w) PEG, such as at least 95% (w/w) PEG. The remaining weight percentage of the PEG-based moiety or reagent are other moieties preferably selected from the following moieties and linkages:C1-50alkyl, C2-50alkenyl, C2-50alkynyl, C3-10cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, phenyl, naphthyl, indenyl, indanyl, and tetralinyl; andlinkages selected from the group comprising

whereindashed lines indicate attachment to the remainder of the moiety or reagent, and —R and —Raare independently of each other selected from the group consisting of —H, methyl, ethyl, propyl, butyl, pentyl and hexyl.

As used herein, the term “PEG-based comprising at least X % PEG” in relation to a moiety or reagent means that said moiety or reagent comprises at least X % (w/w) ethylene glycol units (—CH2CH2O), wherein the ethylene glycol units may be arranged blockwise, alternating or may be randomly distributed within the moiety or reagent and preferably all ethylene glycol units of said moiety or reagent are present in one block; the remaining weight percentage of the PEG-based moiety or reagent are other moieties preferably selected from the following moieties and linkages:C1-50alkyl, C2-50alkenyl, C2-50alkynyl, C3-10cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, phenyl, naphthyl, indenyl, indanyl, and tetralinyl; andlinkages selected from the group comprising

whereindashed lines indicate attachment to the remainder of the moiety or reagent, and —R and —Raare independently of each other selected from the group consisting of —H, methyl, ethyl, propyl, butyl, pentyl and hexyl.

The term “hyaluronic acid-based comprising at least X % hyaluronic acid” is used accordingly.

The term “substituted” as used herein means that one or more —H atoms of a molecule or moiety are replaced by a different atom or a group of atoms, which are referred to as “substituent”.

Preferably, the one or more further optional substituents are independently of each other selected from the group consisting of halogen, —CN, —COORx1, —ORx1, —C(O)Rx1, —C(O)N(Rx1Rx1a), —S(O)2N(Rx1Rx1a), —S(O)N(Rx1Rx1a), —S(O)2Rx1, —S(O)Rx1, —N(Rx1)S(O)2N(Rx1aRx1b), —SRx1, —N(Rx1Rx1a), —NO2—, —OC(O)Rx1, —N(Rx1)C(O)Rx1a, —N(Rx1)S(O)2Rx1a, —N(Rx1)S(O)Rx1a, —N(Rx1)C(O)ORx1a, —N(Rx1)C(O)N(Rx1aRx1b), —OC(O)N(Rx1Rx1a), -T0, C1-50alkyl, C2-50alkenyl, and C2-50alkynyl; wherein -T0, C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally substituted with one or more —Rx2, which are the same or different and wherein C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T0-, —C(O)O—, —O—, —C(O)—, —C(O)N(Rx3), —S(O)2N(Rx3)—, —S(O)N(Rx3)—, —S(O)2—, —S(O)—, —N(Rx3)S(O)2N(Rx3a)—, —S—, —N(Rx3)—, —OC(ORx3)(Rx3a)—, —N(Rx3)C(O)N(Rx3a)—, and —OC(O)N(Rx3)—;

—Rx1, —Rx1a, —Rx1bare independently of each other selected from the group consisting of —H, -T0, C1-50alkyl, C2-50alkenyl, and C2-50alkynyl; wherein -T0, C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally substituted with one or more —Rx2, which are the same or different and wherein C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T0-, —C(O)O—, —O—, —C(O)—, —C(O)N(Rx3)—, —S(O)2N(Rx3)—, —S(O)N(Rx3)—, —S(O)2—, —S(O)—, —N(Rx3)S(O)2N(Rx3a)—, —S—, —N(Rx3)—, —OC(ORx3)(Rx3a)—, —N(Rx3)C(O)N(Rx3a)—, and —OC(O)N(Rx3)—;

each T0is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C3-10cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; wherein each T0is independently optionally substituted with one or more —Rx2, which are the same or different;

each —Rx2is independently selected from the group consisting of halogen, —CN, oxo (═O), —COORx4, —ORx4, —C(O)Rx4, —C(O)N(Rx4Rx4a), —S(O)2N(Rx4Rx4a), —S(O)N(Rx4Rx4a), —S(O)2Rx4, —S(O)Rx4, —N(Rx4)S(O)2N(Rx4aRx4b), —SRx4, —N(Rx4Rx4a), —NO2—, —OC(O)Rx4, —N(Rx4)C(O)Rx4a, —N(Rx4)S(O)2Rx4a, —N(Rx4)S(O)Rx4a, —N(Rx4)C(O)ORx4a, —N(Rx4)C(O)N(Rx4aRx4b), —OC(O)N(Rx4Rx4a), and C1-6alkyl; wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different;

each —Rx3, —Rx3a, —Rx4, —Rx4a, —Rx4bis independently selected from the group consisting of —H and C1-6alkyl; wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different.

More preferably, the one or more further optional substituents are independently of each other selected from the group consisting of halogen, —CN, —COORx1, —ORx1, —C(O)Rx1, —C(O)N(Rx1Rx1a), —S(O)2N(Rx1Rx1a), —S(O)N(Rx1Rx1a), —S(O)2Rx1, —S(O)Rx1, —N(Rx1)S(O)2N(Rx1aRx1b), —SRx1, —N(Rx1Rx1a), —NO2—, —OC(O)Rx4, —N(Rx1)C(O)Rx1a, —N(Rx1)S(O)2Rx1a, —N(Rx1)S(O)Rx1a, —N(Rx1)C(O)ORx1a, —N(Rx1)C(O)N(Rx1aRx1b), —OC(O)N(Rx4Rx1a), -T0, C1-10alkyl, C2-10alkenyl, and C2-10alkynyl; wherein -T0, C1-10alkyl, C2-10alkenyl, and C2-10alkynyl are optionally substituted with one or more —Rx2, which are the same or different and wherein C1-10alkyl, C2-10alkenyl, and C2-10alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T0-, —C(O)O—, —O—, —C(O)—, —C(O)N(Rx3), —S(O)2N(Rx3)—, —S(O)N(Rx3)—, —S(O)2—, —S(O)—, —N(Rx3)S(O)2N(Rx3a)—, —S—, —N(Rx3)—, —OC(ORx3)(Rx3a)—, —N(Rx3)C(O)N(Rx3a)—, and —OC(O)N(Rx3)—;

each —Rx1, —Rx1a, —Rx1b, —Rx3, —Rx3ais independently selected from the group consisting of —H, halogen, C1-6alkyl, C2-6alkenyl, and C2-6alkynyl;

each T0is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C3-10cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; wherein each T0is independently optionally substituted with one or more —Rx2, which are the same or different;

each —Rx2is independently selected from the group consisting of halogen, —CN, oxo (═O), —COORx4, —ORx4, —C(O)Rx4, —C(O)N(Rx4Rx4a), —S(O)2N(Rx4Rx4a), —S(O)N(Rx4Rx4a), —S(O)2Rx4, —S(O)Rx4, —N(Rx4)S(O)2N(Rx4aRx4b), —SRx4, —N(Rx4Rx4a), —NO2—, —OC(O)Rx4, —N(Rx4)C(O)Rx4a, —N(Rx4)S(O)2Rx4a, —N(Rx4)S(O)Rx4a, —N(Rx4)C(O)ORx4a, —N(Rx4)C(O)N(Rx4aRx4b), —OC(O)N(Rx4Rx4a), and C1-6alkyl; wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different;

each —Rx4, —Rx4a, —Rx4bis independently selected from the group consisting of —H, halogen, C1-6alkyl, C2-6alkenyl, and C2-6alkynyl;

Even more preferably, the one or more further optional substituents are independently of each other selected from the group consisting of halogen, —CN, —COORx1, —ORx1, —C(O)Rx1, —C(O)N(Rx1Rx1a), —S(O)2N(Rx1Rx1a), —S(O)N(Rx1Rx1a), —S(O)2Rx1, —S(O)Rx1, —N(Rx1)S(O)2N(Rx1aRx1b), —SRx1, —N(Rx1Rx1a), —NO2—, —OC(O)Rx1, —N(Rx1)C(O)Rx1a, —N(Rx1)S(O)2Rx1a, —N(Rx1)S(O)Rx1a, —N(Rx1)C(O)ORx1a, —N(Rx1)C(O)N(Rx1aRx1b), —OC(O)N(Rx1Rx1a), -T0, C1-6alkyl, C2-6alkenyl, and C2-6alkynyl; wherein -T0, C1-6alkyl, C2-6alkenyl, and C2-6alkynyl are optionally substituted with one or more —Rx2, which are the same or different and wherein C1-6alkyl, C2-6alkenyl, and C2-6alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T0-, —C(O)O—, —O—, —C(O)—, —C(O)N(Rx3), —S(O)2N(Rx3)—, —S(O)N(Rx3)—, —S(O)N(Rx3)—, —S(O)2—, —S(O)—, —N(Rx3)S(O)2N(Rx3a)—, —S—, —N(Rx3)—, —OC(ORx3)(Rx3a)—, —N(Rx3)C(O)N(Rx3a)—, and —OC(O)N(Rx3)—;

each —Rx1, —Rx1a, —Rx1b, —Rx2, —Rx3, —Rx3ais independently selected from the group consisting of —H, halogen, C1-6alkyl, C2-6alkenyl, and C2-6alkynyl;

each T0is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C3-10cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; wherein each T0is independently optionally substituted with one or more —Rx2, which are the same or different.

Preferably, a maximum of 6 —H atoms of an optionally substituted molecule are independently replaced by a substituent, e.g. 5 —H atoms are independently replaced by a substituent, 4 —H atoms are independently replaced by a substituent, 3 —H atoms are independently replaced by a substituent, 2 —H atoms are independently replaced by a substituent, or 1 —H atom is replaced by a substituent.

The term “interrupted” means that a moiety is inserted between two carbon atoms or if the insertion is at one of the moiety's ends—between a carbon or heteroatom and a hydrogen atom.

As used herein, the term “C1-4alkyl” alone or in combination means a straight-chain or branched alkyl moiety having 1 to 4 carbon atoms. If present at the end of a molecule, examples of straight-chain or branched C1-4alkyl are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl. When two moieties of a molecule are linked by the C1-4alkyl, then examples for such C1-4alkyl groups are —CH2—, —CH2—CH2—, —CH(CH3)—, —CH2—CH2—CH2—, —CH(C2H5)—, —C(CH3)2—. Each hydrogen of a C1-4alkyl carbon may optionally be replaced by a substituent as defined above. Optionally, a C1-4alkyl may be interrupted by one or more moieties as defined below.

As used herein, the term “C1-6alkyl” alone or in combination means a straight-chain or branched alkyl moiety having 1 to 6 carbon atoms. If present at the end of a molecule, examples of straight-chain and branched C1-6alkyl groups are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl and 3,3-dimethylpropyl. When two moieties of a molecule are linked by the C1-6alkyl group, then examples for such C1-6alkyl groups are —CH2—, —CH2—CH2—, —CH(CH3)—, —CH2—CH2—CH2—, —CH(C2H5)— and —C(CH3)2—. Each hydrogen atom of a C1-6carbon may optionally be replaced by a substituent as defined above. Optionally, a C1-6alkyl may be interrupted by one or more moieties as defined below.

Accordingly, “C1-10alkyl”, “C1-20alkyl” or “C1-50alkyl” means an alkyl chain having 1 to 10, 1 to 20 or 1 to 50 carbon atoms, respectively, wherein each hydrogen atom of the C1-10, C1-20or C1-50carbon may optionally be replaced by a substituent as defined above. Optionally, a C1-10or C1-50alkyl may be interrupted by one or more moieties as defined below.

As used herein, the term “C2-6alkenyl” alone or in combination means a straight-chain or branched hydrocarbon moiety comprising at least one carbon-carbon double bond having 2 to 6 carbon atoms. If present at the end of a molecule, examples are —CH═CH2, —CH═CH—CH3, —CH2—CH═CH2, —CH═CHCH2—CH3and —CH═CH—CH═CH2. When two moieties of a molecule are linked by the C2-6alkenyl group, then an example for such C2-6alkenyl is —CH═CH—. Each hydrogen atom of a C2-6alkenyl moiety may optionally be replaced by a substituent as defined above. Optionally, a C2-6alkenyl may be interrupted by one or more moieties as defined below.

Accordingly, the term “C2-10alkenyl”, “C2-20alkenyl” or “C2-50alkenyl” alone or in combination means a straight-chain or branched hydrocarbon moiety comprising at least one carbon-carbon double bond having 2 to 10, 2 to 20 or 2 to 50 carbon atoms. Each hydrogen atom of a C2-10alkenyl, C2-20alkenyl or C2-50alkenyl group may optionally be replaced by a substituent as defined above. Optionally, a C2-10alkenyl, C2-20alkenyl or C2-50alkenyl may be interrupted by one or more moieties as defined below.

As used herein, the term “C2-6alkynyl” alone or in combination means a straight-chain or branched hydrocarbon moiety comprising at least one carbon-carbon triple bond having 2 to 6 carbon atoms. If present at the end of a molecule, examples are —C≡CH, —CH2—C≡CH, CH2—CH2—C≡CH and CH2—C≡C—CH3. When two moieties of a molecule are linked by the alkynyl group, then an example is —C≡C—. Each hydrogen atom of a C2-6alkynyl group may optionally be replaced by a substituent as defined above. Optionally, one or more double bonds may occur. Optionally, a C2-6alkynyl may be interrupted by one or more moieties as defined below.

Accordingly, as used herein, the term “C2-10alkynyl”, “C2-20alkynyl” and “C2-50alkynyl” alone or in combination means a straight-chain or branched hydrocarbon moiety comprising at least one carbon-carbon triple bond having 2 to 10, 2 to 20 or 2 to 50 carbon atoms, respectively. Each hydrogen atom of a C2-10alkynyl, C2-20alkynyl or C2-50alkynyl group may optionally be replaced by a substituent as defined above. Optionally, one or more double bonds may occur. Optionally, a C2-10alkynyl, C2-20alkynyl or C2-50alkynyl may be interrupted by one or more moieties as defined below.

As mentioned above, a C1-4alkyl, C1-6alkyl, C1-10alkyl, C1-20alkyl, C1-50alkyl, C2-6alkenyl, C2-10alkenyl, C2-20alkenyl, C2-50alkenyl, C2-6alkynyl, C2-10alkynyl, C2-20alkenyl or C2-50alkynyl may optionally be interrupted by one or more moieties which are preferably selected from the group consisting of


whereindashed lines indicate attachment to the remainder of the moiety or reagent; and—R and —Raare independently of each other selected from the group consisting of —H, methyl, ethyl, propyl, butyl, pentyl and hexyl.

As used herein, the term “C3-10cycloalkyl” means a cyclic alkyl chain having 3 to 10 carbon atoms, which may be saturated or unsaturated, e.g. cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl. Each hydrogen atom of a C3-10cycloalkyl carbon may be replaced by a substituent as defined above. The term “C3-10cycloalkyl” also includes bridged bicycles like norbornane or norbornene.

The term “8- to 30-membered carbopolycyclyl” or “8- to 30-membered carbopolycycle” means a cyclic moiety of two or more rings with 8 to 30 ring atoms, where two neighboring rings share at least one ring atom and that may contain up to the maximum number of double bonds (aromatic or non-aromatic ring which is fully, partially or un-saturated). Preferably a 8- to 30-membered carbopolycyclyl means a cyclic moiety of two, three, four or five rings, more preferably of two, three or four rings.

As used herein, the term “3- to 10-membered heterocyclyl” or “3- to 10-membered heterocycle” means a ring with 3, 4, 5, 6, 7, 8, 9 or 10 ring atoms that may contain up to the maximum number of double bonds (aromatic or non-aromatic ring which is fully, partially or un-saturated) wherein at least one ring atom up to 4 ring atoms are replaced by a heteroatom selected from the group consisting of sulfur (including —S(O)—, —S(O)2—), oxygen and nitrogen (including ═N(O)—) and wherein the ring is linked to the rest of the molecule via a carbon or nitrogen atom. Examples for 3- to 10-membered heterocycles include but are not limited to aziridine, oxirane, thiirane, azirine, oxirene, thiirene, azetidine, oxetane, thietane, furan, thiophene, pyrrole, pyrroline, imidazole, imidazoline, pyrazole, pyrazoline, oxazole, oxazoline, isoxazole, isoxazoline, thiazole, thiazoline, isothiazole, isothiazoline, thiadiazole, thiadiazoline, tetrahydrofuran, tetrahydrothiophene, pyrrolidine, imidazolidine, pyrazolidine, oxazolidine, isoxazolidine, thiazolidine, isothiazolidine, thiadiazolidine, sulfolane, pyran, dihydropyran, tetrahydropyran, imidazolidine, pyridine, pyridazine, pyrazine, pyrimidine, piperazine, piperidine, morpholine, tetrazole, triazole, triazolidine, tetrazolidine, diazepane, azepine and homopiperazine. Each hydrogen atom of a 3- to 10-membered heterocyclyl or 3- to 10-membered heterocyclic group may be replaced by a substituent as defined below.

As used herein, the term “8- to 11-membered heterobicyclyl” or “8- to 11-membered heterobicycle” means a heterocyclic moiety of two rings with 8 to 11 ring atoms, where at least one ring atom is shared by both rings and that may contain up to the maximum number of double bonds (aromatic or non-aromatic ring which is fully, partially or un-saturated) wherein at least one ring atom up to 6 ring atoms are replaced by a heteroatom selected from the group consisting of sulfur (including —S(O)—, —S(O)2—), oxygen and nitrogen (including ═N(O)—) and wherein the ring is linked to the rest of the molecule via a carbon or nitrogen atom. Examples for an 8- to 11-membered heterobicycle are indole, indoline, benzofuran, benzothiophene, benzoxazole, benzisoxazole, benzothiazole, benzisothiazole, benzimidazole, benzimidazoline, quinoline, quinazoline, dihydroquinazoline, quinoline, dihydroquinoline, tetrahydroquinoline, decahydroquinoline, isoquinoline, decahydroisoquinoline, tetrahydroisoquinoline, dihydroisoquinoline, benzazepine, purine and pteridine. The term 8- to 11-membered heterobicycle also includes spiro structures of two rings like 1,4-dioxa-8-azaspiro[4.5]decane or bridged heterocycles like 8-aza-bicyclo[3.2.1]octane. Each hydrogen atom of an 8- to 11-membered heterobicyclyl or 8- to 11-membered heterobicycle carbon may be replaced by a substituent as defined below.

Similary, the term “8- to 30-membered heteropolycyclyl” or “8- to 30-membered heteropolycycle” means a heterocyclic moiety of more than two rings with 8 to 30 ring atoms, preferably of three, four or five rings, where two neighboring rings share at least one ring atom and that may contain up to the maximum number of double bonds (aromatic or non-aromatic ring which is fully, partially or unsaturated), wherein at least one ring atom up to 10 ring atoms are replaced by a heteroatom selected from the group of sulfur (including S(O)—, —S(O)2—), oxygen and nitrogen (including ═N(O)—) and wherein the ring is linked to the rest of a molecule via a carbon or nitrogen atom.

It is understood that the phrase “the pair Rx/Ryis joined together with the atom to which they are attached to form a C3-10cycloalkyl or a 3- to 10-membered heterocyclyl” in relation with a moiety of the structure


means that Rx and Ry form the following structure:


wherein R is C3-10cycloalkyl or 3- to 10-membered heterocyclyl.

It is also understood that the phrase “the pair Rx/Ryis joint together with the atoms to which they are attached to form a ring A” in relation with a moiety of the structure



means that Rxand Ryform the following structure:



As used herein, the term “terminal alkyne” means a moiety



As used herein, “halogen” means fluoro, chloro, bromo or iodo. It is generally preferred that halogen is fluoro or chloro.

In general, the term “comprise” or “comprising” also encompasses “consist of” or “consisting of”.

In the present invention -L1- is covalently and reversibly conjugated to the side chain of an amino acid residue of said ring moiety of -D or to the backbone of said ring moiety of -D. Preferably, -L1- is covalently and reversibly conjugated to the side chain of an amino acid residue of said ring moiety of -D.

It was also surprisingly found that an increase in the lengths of the CNP sequence is beneficial with regard to NEP-stability: CNP-22 was more susceptible towards NEP-degradation than CNP-34 which in turn was more susceptible than CNP-38.

Preferably -D has the sequence of SEQ ID NO:24, SEQ ID NO:25 or SEQ ID NO:30.

In one embodiment -D has the sequence of SEQ ID NO:25.

In another embodiment -D has the sequence of SEQ ID NO:20.

In another embodiment -D has the sequence of SEQ ID NO:21.

In another embodiment -D has the sequence of SEQ ID NO:22.

In another embodiment -D has the sequence of SEQ ID NO:23.

In one embodiment -D has the sequence of SEQ ID NO:30.

In a preferred embodiment -D has the sequence of SEQ ID NO:24.

Said amino acid residue located in the ring moiety of -D is preferably any amino acid having a functional group.

Preferably, the moiety -L1- is conjugated to the ring moiety of -D through a functional group of the corresponding CNP drug D-H selected from the group consisting of carboxylic acid, primary and secondary amine, maleimide, thiol, sulfonic acid, carbonate, carbamate, hydroxyl, aldehyde, ketone, hydrazine, isocyanate, isothiocyanate, phosphoric acid, phosphonic acid, haloacetyl, alkyl halide, acryloyl, aryl fluoride, hydroxylamine, sulfate, disulfide, vinyl sulfone, vinyl ketone, diazoalkane, oxirane, guanidine and aziridine. Most preferably the moiety -L1- is conjugated to the ring moiety of -D through a functional group of the corresponding CNP drug D-H selected from the group consisting hydroxyl, primary and secondary amine and guanidine. In a particular preferred embodiment the moiety -L1- is conjugated to the ring moiety of D through a primary or secondary amine group, preferably through a primary amine group.

The moiety -L1- can be connected to the ring moiety of -D through any type of linkage, provided that it is reversible. Preferably, -L1- is connected to the ring moiety of -D through a linkage selected from the group consisting of amide, ester, carbamate, acetal, aminal, imine, oxime, hydrazone, disulfide and acylguanidine. Even more preferably -L1- is connected to the ring moiety of -D through a linkage selected from the group consisting of amide, ester, carbamate and acylguanidine. It is understood that these linkages may not be per se reversible, such as for example the amide linkage, but that suitably chosen neighboring groups comprised in -L1- may render such linkages reversible.

In one embodiment -L1- is connected to the ring moiety of -D through an ester linkage.

In another embodiment -L1- is connected to the ring moiety of -D through a carbamate linkage.

In another embodiment -L1- is connected to the ring moiety of -D through an acylguanidine.

In a preferred embodiment -L1- is connected to the ring moiety of -D through an amide linkage.

The amino acid residue of the ring moiety of -D to which -L1- is conjugated is selected from the group consisting of proteinogenic amino acid residues and non-proteinogenic amino acid residues.

In one embodiment the amino acid residue of the ring moiety of -D to which -L1- is conjugated is a non-proteinogenic amino acid.

In a preferred embodiment the amino acid residue of the ring moiety of -D to which -L1- is conjugated is a proteinogenic amino acid. Even more preferably said amino acid is selected from the group consisting of histidine, lysine, tryptophan, serine, threonine, tyrosine, aspartic acid, glutamic acid and arginine. Even more preferably said amino acid is selected from the group consisting of lysine, aspartic acid, arginine and serine. Even more preferably said amino acid is selected from the group consisting of lysine, arginine and serine.

In one embodiment the amino acid residue of the ring moiety of -D to which -L1- is conjugated is a histidine. It is understood that such histidine does not occur in the sequence of SEQ ID NO:96 and that it may only be present in variants, analogs, orthologs, homologs and derivatives thereof.

In one embodiment the amino acid residue of the ring moiety of -D to which -L1- is conjugated is a tryptophan. It is understood that such tryptophan does not occur in the sequence of SEQ ID NO:96 and that it may only be present in variants, analogs, orthologs, homologs and derivatives thereof.

In one embodiment the amino acid residue of the ring moiety of -D to which -L1- is conjugated is a threonine. It is understood that such threonine does not occur in the sequence of SEQ ID NO:96 and that it may only be present in variants, analogs, orthologs, homologs and derivatives thereof.

In one embodiment the amino acid residue of the ring moiety of -D to which -L1- is conjugated is a tyrosine. It is understood that such tyrosine does not occur in the sequence of SEQ ID NO:96 and that it may only be present in variants, analogs, orthologs, homologs and derivatives thereof.

In one embodiment the amino acid residue of the ring moiety of -D to which -L1- is conjugated is a glutamic acid. It is understood that such glutamic acid does not occur in the sequence of SEQ ID NO:96 and that it may only be present in variants, analogs, orthologs, homologs and derivatives thereof.

In one embodiment the amino acid residue of the ring moiety of -D to which -L1- is conjugated is a lysine. Preferably, said amino acid is the lysine at position 4 of SEQ ID NO:96 which corresponds to the lysine at position 26 of SEQ ID NO:24.

In another embodiment the amino acid residue of the ring moiety of -D to which -L1- is conjugated is an aspartic acid. Preferably, said amino acid is the aspartic acid at position 6 of SEQ ID NO:96 which corresponds to the aspartic acid at position 28 of SEQ ID NO:24.

In another embodiment the amino acid residue of the ring moiety of -D to which -L1- is conjugated is an arginine. Preferably, said amino acid is the arginine at position 7 of SEQ ID NO:96 which corresponds to the arginine at position 29 of SEQ ID NO:24.

In another embodiment the amino acid residue of the ring moiety of -D to which -L1- is conjugated a serine. Preferably, said amino acid is the serine at position 10 or 12 of SEQ ID NO:96. In one embodiment said amino acid is the serine at position 10 of SEQ ID NO:96 which corresponds to the serine at position 32 of SEQ ID NO:24. In another embodiment said amino acid is the serine at position 12 of SEQ ID NO:96 which corresponds to the serine at position 34 of SEQ ID NO:24.

In a preferred embodiment the amino acid residue of the ring moiety of -D to which -L1- is conjugated is a lysine. Most preferably, -D has the sequence of SEQ ID NO:24 and -L1- is conjugated to the lysine at position 26.

Preferably, the CNP prodrug of the present invention is of formula (I)
Z-L2-L1D  (I)wherein-D is a CNP moiety;-L1- is a reversible prodrug linker moiety which is covalently and reversibly conjugated to a side chain of an amino acid residue of the ring moiety of -D or to the backbone of said ring moiety of -D;-L2- is a single chemical bond or a spacer moiety; and—Z is a carrier moiety.

The moiety -L1- is a reversible prodrug linker from which the drug, i.e. CNP, is released in its free form, i.e. it is a traceless prodrug linker. Suitable prodrug linkers are known in the art, such as for example the reversible prodrug linker moieties disclosed in WO 2005/099768 A2, WO 2006/136586 A2, WO 2011/089216 A1 and WO 2013/024053 A1, which are incorporated by reference herewith.

In another embodiment -L1- is a reversible prodrug linker as described in WO 2011/012722 A1, WO 2011/089214 A1, WO 2011/089215 A1, WO 2013/024052 A1 and WO 2013/160340 A1 which are incorporated by reference herewith.

A particularly preferred moiety -L1- is disclosed in WO 2009/095479 A2. Accordingly, in one preferred embodiment the moiety -L1- is of formula (II):

wherein the dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond;—X— is —C(R4R4a)—; —N(R4)—; —O—; —C(R4R4a)—C(R5R5a)—; —C(R5R5a)—C(R4R4a)—; —C(R4R4a)—N(R6)—; —N(R6)—C(R4R4a)—; —C(R4R4a)—O—; —O—C(R4R4a)—; or —C(R7R7a)—;X1is C; or S(O);—X2— is —C(R8R8a)—; or —C(R8R8a)—C(R9R9a)—;═X3is ═O; ═S; or ═N—CN;—R1, —R1a, —R2, —R2a, —R4, —R4a, —R5, —R5a, —R6, —R8, —R8a, —R9, —R9a, are independently selected from the group consisting of —H; and C1-6alkyl;—R3, —R3aare independently selected from the group consisting of —H; and C1-6alkyl, provided that in case one of —R3, —R3aor both are other than —H they are connected to N to which they are attached through an SP3-hybridized carbon atom;—R7is —N(R10R10a) or —NR10—(C═O)—R11;—R7a, —R10, —R10a, —R11are independently of each other —H; or C1-6alkyl;optionally, one or more of the pairs —R1a/—R4a, —R1a/—R5a, —R1a/—R7a, —R4a/—R5a, —R8a/—R9aform a chemical bond;optionally, one or more of the pairs —R1/—R1a, —R2/—R2a, —R4/—R4a, —R5/—R5a, —R8/—R8a, —R9/—R9aare joined together with the atom to which they are attached to form a C3-10cycloalkyl; or 3- to 10-membered heterocyclyl;optionally, one or more of the pairs —R1/—R4, —R1/—R5, —R1/—R6, —R1/—R7a, —R4/—R5, —R4/—R6, —R8/—R9, —R2/—R3are joined together with the atoms to which they are attached to form a ring A;optionally, R3/R3aare joined together with the nitrogen atom to which they are attached to form a 3- to 10-membered heterocycle;A is selected from the group consisting of phenyl; naphthyl; indenyl; indanyl; tetralinyl; C3-10cycloalkyl; 3- to 10-membered heterocyclyl; and 8- to 11-membered heterobicyclyl; andwherein -L1- is substituted with -L2-Z and wherein -L1- is optionally further substituted, provided that the hydrogen marked with the asterisk in formula (II) is not replaced by -L2-Z or a substituent;whereinL2- is a single chemical bond or a spacer;—Z is a carrier moiety.

Preferably -L1- of formula (II) is substituted with one moiety -L2-Z.

In one embodiment -L1- of formula (II) is not further substituted.

It is understood that if —R3/—R3aof formula (II) are joined together with the nitrogen atom to which they are attached to form a 3- to 10-membered heterocycle, only such 3- to 10-membered heterocycles may be formed in which the atoms directly attached to the nitrogen are SP3-hybridized carbon atoms. In other words, such 3- to 10-membered heterocycle formed by —R3/—R3atogether with the nitrogen atom to which they are attached has the following structure:

whereinthe dashed line indicates attachment to the rest of -L1-;the ring comprises 3 to 10 atoms comprising at least one nitrogen; andR#and R##represent an SP3-hydridized carbon atom.

It is also understood that the 3- to 10-membered heterocycle may be further substituted.

Exemplary embodiments of suitable 3- to 10-membered heterocycles formed by —R3/—R3aof formula (II) together with the nitrogen atom to which they are attached are the following:

whereindashed lines indicate attachment to the rest of the molecule; and—R is selected from the group consisting of —H and C1-6alkyl.

-L1- of formula (II) may optionally be further substituted. In general, any substituent may be used as far as the cleavage principle is not affected, i.e. the hydrogen marked with the asterisk in formula (II) is not replaced and the nitrogen of the moiety


of formula (II) remains part of a primary, secondary or tertiary amine, i.e. —R3and —R3aare independently of each other —H or are connected to —N< through an SP3-hybridized carbon atom.

In one embodiment —R1or —R1aof formula (II) is substituted with -L2-Z. In another embodiment —R2or —R2aof formula (II) is substituted with -L2-Z. In another embodiment —R3or —R3aof formula (II) is substituted with -L2-Z. In another embodiment —R4of formula (II) is substituted with -L2-Z. In another embodiment —R5or —R5aof formula (II) is substituted with -L2-Z. In another embodiment —R6of formula (II) is substituted with -L2-Z. In another embodiment —R7or —R7aof formula (II) is substituted with -L2-Z. In another embodiment —R8or —R8aof formula (II) is substituted with -L2-Z. In another embodiment —R9or —R9aof formula (II) is substituted with -L2-Z.

Most preferably —R4of formula (II) is substituted with -L2-Z.

Preferably, —X— of formula (II) is —C(R4R4a)— or —N(R4)—. Most preferably, —X— of formula (II) is —C(R4R4a)—.

Preferably, X1of formula (II) is C.

Preferably, —X3of formula (II) is ═O.

Preferably, —X2— of formula (II) is —C(R8R8a)—.

Preferably —R8and —R8aof formula (II) are independently selected from the group consisting of —H, methyl and ethyl. More preferably at least one of —R8and —R8aof formula (II) is —H. Even more preferably both —R8and —R8aof formula (II) are —H.

Preferably, —R1and —R1aof formula (II) are independently selected from the group consisting of —H, methyl and ethyl. More preferably, at least one of —R1and —R1aof formula (II) is —H. Even more preferably both —R1and —R1aof formula (II) are —H.

Preferably, —R2and —R2aof formula (II) are independently selected from the group consisting of —H, methyl and ethyl. More preferably, at least one of —R2and —R2aof formula (II) is —H. Even more preferably both —R2and —R2aof formula (II) are H.

Preferably, —R3and —R3aof formula (II) are independently selected from the group consisting of —H, methyl, ethyl, propyl and butyl. Even more preferably at least one of —R3and —R3aof formula (II) is methyl. In an equally preferred embodiment —R3and —R3aof formula (II) are both —H. In another equally preferred embodiment —R3and —R3aof formula (II) are both methyl.

Preferably, —R3of formula (II) is —H and —R3aof formula (II) is methyl.

Preferably, —R4and —R4aof formula (II) are independently selected from the group consisting of —H, methyl and ethyl. More preferably, at least one of —R4and —R4aof formula (II) is —H. Even more preferably both —R4and —R4aof formula (II) are —H.

Preferably the moiety -L1- is of formula (IIa):

whereinthe dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond;—R1, —R1a, —R2a, —R3, —R3a, —R4, —R4aand —X2— are used as defined in formula (II); andwherein -L1- is substituted with -L2-Z and wherein -L1- is optionally further substituted, provided that the hydrogen marked with the asterisk in formula (IIa) is not replaced by -L2-Z or a substituent.

Preferably -L1- of formula (IIa) is substituted with one moiety -L2-Z.

Preferably the moiety -L1- of formula (IIa) is not further substituted.

In one embodiment —R1or —R1aof formula (IIa) is substituted with -L2-Z. In another embodiment —R2or —R2aof formula (IIa) is substituted with -L2-Z. In another embodiment —R3or —R3aof formula (IIa) is substituted with -L2-Z. In another embodiment —R4of formula (IIa) is substituted with -L2-Z. In another embodiment —R8or —R8aof formula (IIa) is substituted with -L2-Z. In another embodiment —R9or —R9aof formula (IIa) is substituted with -L2-Z.

Most preferably —R4of formula (IIa) is substituted with -L2-Z.

Preferably, —R1and —R1aof formula (IIa) are independently selected from the group consisting of —H, methyl and ethyl. More preferably, at least one of —R1and —R1aof formula (IIa) is —H. Even more preferably both —R1and —R1aof formula (IIa) are —H.

Preferably, —R4and —R4aof formula (IIa) are independently selected from the group consisting of —H, methyl and ethyl. More preferably, at least one of —R4and —R4aof formula (IIa) is —H. Even more preferably both —R4and —R4aof formula (IIa) are —H.

Preferably, —X2— of formula (IIa) is —C(R8R8a)—.

Preferably —R8and —R8aof formula (IIa) are independently selected from the group consisting of —H, methyl and ethyl. More preferably at least one of —R8and —R8aof formula (IIa) is —H. Even more preferably both —R8and —R8aof formula (IIa) are —H.

Preferably, —R2and —R2aof formula (IIa) are independently selected from the group consisting of —H, methyl and ethyl. More preferably, at least one of —R2and —R2aof formula (IIa) is —H. Even more preferably both —R2and —R2aof formula (IIa) are H.

Preferably, —R3and —R3aof formula (IIa) are independently selected from the group consisting of —H, methyl, ethyl, propyl and butyl. Even more preferably at least one of —R3and —R3aof formula (IIa) is methyl. In an equally preferred embodiment —R3and —R3aof formula (IIa) are both —H. In another equally preferred embodiment —R3and —R3aof formula (IIa) are both methyl.

Preferably, —R3of formula (IIa) is —H and —R3aof formula (IIa) is methyl.

Preferably the moiety -L1- is of formula (IIb):

whereinthe dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond;—R2, —R2a, —R3, —R3aand —X2— are used as defined in formula (II); andwherein -L1- is substituted with -L2-Z and wherein -L1- is optionally further substituted, provided that the hydrogen marked with the asterisk in formula (IIb) is not replaced by -L2-Z or a substituent.

Preferably -L1- of formula (IIb) is substituted with one moiety -L2-Z.

Preferably the moiety -L1- of formula (IIb) is not further substituted.

Preferably, —X2— of formula (IIb) is —C(R8R8a)—.

Preferably —R8and —R8aof formula (IIb) are independently selected from the group consisting of —H, methyl and ethyl. More preferably at least one of —R8and —R8aof formula (IIb) is —H. Even more preferably both —R8and —R8aof formula (IIb) are —H.

Preferably, —R2and —R2aof formula (IIb) are independently selected from the group consisting of —H, methyl and ethyl. More preferably, at least one of —R2and —R2aof formula (IIb) is —H. Even more preferably both —R2and —R2aof formula (IIb) are H.

Preferably, —R3and —R3aof formula (IIb) are independently selected from the group consisting of —H, methyl, ethyl, propyl and butyl. Even more preferably at least one of —R3and —R3aof formula (IIb) is methyl. In an equally preferred embodiment —R3and —R3aof formula (IIb) are both —H. In another equally preferred embodiment —R3and —R3aof formula (IIb) are both methyl.

Most preferably, —R3of formula (IIb) is —H and —R3aof formula (IIb) is methyl.

Even more preferably the moiety -L1- is of formula (IIb′):

whereinthe unmarked dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; the dashed line marked with the asterisk indicates attachment to -L2-;—R2, —R2a, —R3, —R3aand —X2— are used as defined in formula (II); andwherein -L1- is optionally further substituted, provided that the hydrogen marked with the asterisk in formula (IIb′) is not replaced by a substituent.

Preferably the moiety -L1- of formula (IIb′) is not further substituted.

Preferably, —X2— of formula (IIb′) is —C(R8R8a)—.

Preferably —R8and —R8aof formula (IIb′) are independently selected from the group consisting of —H, methyl and ethyl. More preferably at least one of —R8and —R8aof formula (IIb′) is —H. Even more preferably both —R8and —R8aof formula (IIb′) are —H.

Preferably, —R2and —R2aof formula (IIb′) are independently selected from the group consisting of —H, methyl and ethyl. More preferably, at least one of —R2and —R2aof formula (IIb′) is —H. Even more preferably both —R2and —R2aof formula (IIb′) are H.

Preferably, —R3and —R3aof formula (IIb′) are independently selected from the group consisting of —H, methyl, ethyl, propyl and butyl. Even more preferably at least one of —R3and —R3aof formula (IIb′) is methyl.

Most preferably, —R3of formula (IIb′) is —H and —R3aof formula (IIb′) is methyl.

Preferably the moiety -L1- is of formula (IIc):

whereinthe dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andwherein -L1- is substituted with -L2-Z and wherein -L1- is optionally further substituted, provided that the hydrogen marked with the asterisk in formula (IIc) is not replaced by -L2-Z or a substituent.

Preferably -L1- of formula (IIc) is substituted with one moiety -L2-Z.

Preferably the moiety -L1- of formula (IIc) is not further substituted.

In another preferred embodiment the moiety -L1- is of formula (IIc-a):

the dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andwherein -L1- is substituted with -L2-Z and wherein -L1- is optionally further substituted, provided that the hydrogen marked with the asterisk in formula (IIc-a) is not replaced by -L2-Z or a substituent.

Preferably -L1- of formula (IIc-a) is substituted with one moiety -L2-Z.

Preferably the moiety -L1- of formula (IIc-a) is not further substituted.

In another preferred embodiment the moiety -L1- is of formula (IIc-b):

the dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andwherein -L1- is substituted with -L2-Z and wherein -L1- is optionally further substituted, provided that the hydrogen marked with the asterisk in formula (IIc-b) is not replaced by -L2-Z or a substituent.

Preferably -L1- of formula (IIc-b) is substituted with one moiety -L2-Z.

Preferably the moiety -L1- of formula (IIc-b) is not further substituted.

Even more preferably the moiety -L1- is selected from the group consisting of formula (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv) and (IIc-v):

whereinthe unmarked dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond;the dashed line marked with the asterisk indicates attachment to -L2-Z; and-L1- is optionally further substituted, provided that the hydrogen marked with the asterisk in formula (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv) and (IIc-v) is not replaced by a substituent.

Preferably, the moiety -L1- of formula (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv) and (IIc-v) is not further substituted.

In a particularly preferred embodiment the moiety -L1- is of formula (IIc-ii)

whereinthe unmarked dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to -L2-Z.

Preferably -L1- of formula (IIc-ii) is substituted with one moiety -L2-Z.

In an equally preferred embodiment the moiety -L1- is selected from the group consisting of formula (IIc-i′), (IIc-ii′), (IIc-iii′), (IIc-iv′) and (IIc-v′):

whereinthe unmarked dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond;the dashed line marked with the asterisk indicates attachment to -L2-Z; and-L1- is optionally further substituted, provided that the hydrogen marked with the asterisk in formula (IIc-i′), (IIc-ii′), (IIc-iii′), (IIc-iv′) and (IIc-v′) is not replaced by a substituent.

Preferably, the moiety -L1- of formula (IIc-i′), (IIc-ii′), (IIc-iii′), (IIc-iv′) and (IIc-v′) is not further substituted.

In another particularly preferred embodiment the moiety -L1- is of formula (IIc-ii′)

whereinthe unmarked dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to -L2-Z.

Preferably -L1- of formula (IIc-ii) is substituted with one moiety -L2-Z.

In an equally preferred embodiment the moiety -L1- is selected from the group consisting of formula (IIc-i″), (IIc-ii″), (IIc-iii″) and (IIc-iv″):

whereinthe unmarked dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond;the dashed line marked with the asterisk indicates attachment to -L2-Z; and-L1- is optionally further substituted, provided that the hydrogen marked with the asterisk in formula (IIc-i″), (IIc-ii″), (IIc-iii″) and (IIc-iv″) is not replaced by a substituent.

Preferably, the moiety -L1- of formula (IIc-i″), (IIc-ii″), (IIc-iii″) and (IIc-iv″) is not further substituted.

In another particularly preferred embodiment the moiety -L1- is of formula (IIc-ii″)

whereinthe unmarked dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to -L2-Z.

Preferably -L1- of formula (IIc-ii″) is substituted with one moiety -L2-Z.

The optional further substituents of -L1- of formula (II), (IIa), (IIb), (IIb′), (IIc), (IIc-a), (III-b), (IIc-i), (IIc-ii), (IIc-iii), (IIc-iv), (IIc-v), (IIc-i′), (IIc-ii′), (IIc-iii′), (IIc-iv′), (IIc-v′), (IIc-i″), (IIc-ii″), (IIc-iii″), (IIc-iv″), are preferably as described above.

Another particularly preferred moiety -L1- is disclosed in WO2012/020373A1. Accordingly, in another preferred embodiment the moiety -L1- is of formula (III):

whereinthe dashed line indicates attachment to a primary or secondary amine or hydroxyl of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide or ester linkage, respectively;—R1, —R1a, —R2, —R2a, —R3and —R3aare independently of each other selected from the group consisting of —H, —C(R8R8aR8b), —C(═O)R8, —C≡N, —C(—NR8)R8a, —CR8(═CR8aR8b), —C≡CR8and -T;—R4, —R5and —R5aare independently of each other selected from the group consisting of —H, —C(R9R9aR9b) and -T;a1 and a2 are independently of each other 0 or 1;each —R6, —R6a, —R7, —R7a, —R8, —R8a, —R9, —R9a, —R9bare independently of each other selected from the group consisting of —H, halogen, —CN, —COOR10, —OR10, —C(O)R10, —C(O)N(R10R10a), —S(O)2N(R10R10a), —S(O)N(R10R10a), —S(O)2R10, —S(O)R10, —N(R10)S(O)2N(R10aR10b), —SR10, —N(R10R10a), —NO2—, —OC(O)R10, —N(R10)C(O)R10a, —N(R10)S(O)2N(R10a, —N(R10)S(O)R10a, —N(R10)C(O)OR10a, —N(R10)C(O)N(R10aR10b), —OC(O)N(R10R10a), -T, C1-20alkyl, C2-20alkenyl, and C2-20alkynyl; wherein -T, C1-20alkyl, C2-20alkenyl, and C2-20alkynyl are optionally substituted with one or more —R11, which are the same or different and wherein C1-20alkyl, C2-20alkenyl, and C2-20alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T-, —C(O)O—, —O—, —C(O)—, —C(O)N(R12)—, —S(O)2N(R12)—, —S(O)N(R12)—, —S(O)2—, —S(O)—, —N(R12)S(O)2N(R12a)—, —S—, —N(R12)—, —OC(OR12)(R12a)—, —N(R12)C(O)N(R12a)—, and —OC(O)N(R12)—;each —R10, —R10a, —R10bis independently selected from the group consisting of —H, -T, C1-20alkyl, C2-20alkenyl, and C2-20alkynyl; wherein -T, C1-20alkyl, C2-20alkenyl, and C2-20alkynyl are optionally substituted with one or more —R11, which are the same or different and wherein C1-20alkyl, C2-20alkenyl, and C2-20alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T-, —C(O)O—, —O—, —C(O)—, —C(O)N(R12)—, —S(O)2N(R12)—, —S(O)N(R12)—, —S(O)2—, —S(O)—, —N(R12)S(O)2R12a)—, —S—, —N(R12)—, —OC(OR12)(R12a)—, —N(R12)C(O)N(R12a)—, and —OC(O)N(R12)—;each T is independently of each other selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C3-10cycloalkyl, 3- to 10-membered heterocyclyl, and 8- to 11-membered heterobicyclyl; wherein each T is independently optionally substituted with one or more —R11, which are the same or different;each —R11is independently of each other selected from halogen, —CN, oxo (═O), —COOR13, —OR13, —C(O)R13, —C(O)N(R13R13a), —S(O)2N(R13R13a), —S(O)N(R13R13a), —S(O)2R13, —S(O)R13, —N(R13)S(O)2N(R13aR13b), —SR13, —N(R13R13a), —NO2—, —OC(O)R13, —N(R13)C(O)R13a, —N(R13)S(O)2R13a, —N(R13)S(O)R13a, —N(R13)C(O)OR13a, —N(R13)C(O)N(R13aR13b), —OC(O)N(R13R13a), and C1-6alkyl; wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different;each —R12, —R12a, —R13, —R13a, —R13bis independently selected from the group consisting of —H, and C1-6alkyl; wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different;optionally, one or more of the pairs —R1/—R1a, —R2/—R2a, —R3/—R3a, —R6/—R6a, —R7/—R7aare joined together with the atom to which they are attached to form a C3-10cycloalkyl or a 3- to 10-membered heterocyclyl;optionally, one or more of the pairs —R1/—R2, —R1/—R3, —R1/—R4, —R1/—R5, —R1/—R6, —R1/—R7, —R2/—R3, —R2/—R4, —R2/—R5, —R2/—R6, —R2/—R7, —R3/—R4, —R3/—R5, —R3/—R6, —R3/—R7, —R4/—R5, —R4/—R6, —R4/—R7, —R5/—R6, —R5/—R7, —R6/—R7are joint together with the atoms to which they are attached to form a ring A;A is selected from the group consisting of phenyl; naphthyl; indenyl; indanyl; tetralinyl; C3-10cycloalkyl; 3- to 10-membered heterocyclyl; and 8- to 11-membered heterobicyclyl;wherein -L1- is substituted with -L2-Z and wherein -L1- is optionally further substituted;wherein-L2- is a single chemical bond or a spacer; and—Z is a carrier moiety.

The optional further substituents of -L1- of formula (III) are preferably as described above.

Preferably -L1- of formula (III) is substituted with one moiety -L2-Z.

In one embodiment -L1- of formula (III) is not further substituted.

Additional preferred embodiments for -L1- are disclosed in EP 1536334B1, WO2009/009712A1, WO2008/034122A1, WO2009/143412A2, WO2011/082368A2, and U.S. Pat. No. 8,618,124B2, which are herewith incorporated by reference in their entirety.

Additional preferred embodiments for -L1- are disclosed in U.S. Pat. No. 8,946,405B2 and U.S. Pat. No. 8,754,190B2, which are herewith incorporated by reference in their entirety. Accordingly, a preferred moiety -L1- is of formula (IV):


whereinthe dashed line indicates attachment to the CNP moiety and wherein attachment is through a functional group of an amino acid residue of the ring moiety of -D selected from the group consisting of —OH, —SH and —NH2;m is 0 or 1;at least one or both of —R1and —R2is/are independently of each other selected from the group consisting of —CN, —NO2, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted alkenyl, optionally substituted alkynyl, —C(O)R3, —S(O)R3, —S(O)2R3, and —SR4,one and only one of —R1and —R2is selected from the group consisting of —H, optionally substituted alkyl, optionally substituted arylalkyl, and optionally substituted heteroarylalkyl;—R3is selected from the group consisting of —H, optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, —OR9and —N(R9)2;R4is selected from the group consisting of optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl;each —R5is independently selected from the group consisting of —H, optionally substituted alkyl, optionally substituted alkenylalkyl, optionally substituted alkynylalkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl and optionally substituted heteroarylalkyl;R9is selected from the group consisting of —H and optionally substituted alkyl;—Y— is absent and —X— is —O— or —S—; or—Y— is —N(Q)CH2— and —X— is —O—;Q is selected from the group consisting of optionally substituted alkyl, optionally substituted aryl, optionally substituted arylalkyl, optionally substituted heteroaryl and optionally substituted heteroarylalkyl;optionally, —R1and —R2may be joined to form a 3 to 8-membered ring; andoptionally, both —R9together with the nitrogen to which they are attached form a heterocyclic ring;wherein -L1- is substituted with -L2-Z and wherein -L1- is optionally further substituted;wherein-L2- is a single chemical bond or a spacer; and—Z is a carrier moiety.

The optional further substituents of -L1- of formula (IV) are preferably as described above.

Preferably -L1- of formula (IV) is substituted with one moiety -L2-Z.

In one embodiment -L1- of formula (IV) is not further substituted.

Only in the context of formula (IV) the terms used have the following meaning:

The term “alkyl” as used herein includes linear, branched or cyclic saturated hydrocarbon groups of 1 to 8 carbons, or in some embodiments 1 to 6 or 1 to 4 carbon atoms.

The term “alkoxy” includes alkyl groups bonded to oxygen, including methoxy, ethoxy, isopropoxy, cyclopropoxy, cyclobutoxy, and similar.

The term “alkenyl” includes non-aromatic unsaturated hydrocarbons with carbon-carbon double bonds.

The term “alkynyl” includes non-aromatic unsaturated hydrocarbons with carbon-carbon triple bonds.

The term “aryl” includes aromatic hydrocarbon groups of 6 to 18 carbons, preferably 6 to 10 carbons, including groups such as phenyl, naphthyl, and anthracenyl. The term “heteroaryl” includes aromatic rings comprising 3 to 15 carbons containing at least one N, O or S atom, preferably 3 to 7 carbons containing at least one N, O or S atom, including groups such as pyrrolyl, pyridyl, pyrimidinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, quinolyl, indolyl, indenyl, and similar.

In some instance, alkenyl, alkynyl, aryl or heteroaryl moieties may be coupled to the remainder of the molecule through an alkylene linkage. Under those circumstances, the substituent will be referred to as alkenylalkyl, alkynylalkyl, arylalkyl or heteroarylalkyl, indicating that an alkylene moiety is between the alkenyl, alkynyl, aryl or heteroaryl moiety and the molecule to which the alkenyl, alkynyl, aryl or heteroaryl is coupled.

The term “halogen” includes bromo, fluoro, chloro and iodo.

The term “heterocyclic ring” refers to a 4 to 8 membered aromatic or non-aromatic ring comprising 3 to 7 carbon atoms and at least one N, O, or S atom. Examples are piperidinyl, piperazinyl, tetrahydropyranyl, pyrrolidine, and tetrahydrofuranyl, as well as the exemplary groups provided for the term “heteroaryl” above.

When a ring system is optionally substituted, suitable substituents are selected from the group consisting of alkyl, alkenyl, alkynyl, or an additional ring, each optionally further substituted. Optional substituents on any group, including the above, include halo, nitro, cyano, —OR, —SR, —NR2, —OCOR, —NRCOR, —COOR, —CONR2, —SOR, —SO2R, —SONR2, —SO2NR2, wherein each R is independently alkyl, alkenyl, alkynyl, aryl or heteroaryl, or two R groups taken together with the atoms to which they are attached form a ring.

An additional preferred embodiment for -L1- is disclosed in WO2013/036857A1, which is herewith incorporated by reference in its entirety. Accordingly, a preferred moiety -L1- is of formula (V):

whereinthe dashed line indicates attachment to an amine functional group of a side chain of an amino acid residue of the ring moiety of the CNP moiety;—R1is selected from the group consisting of optionally substituted C1-C6linear, branched, or cyclic alkyl; optionally substituted aryl; optionally substituted heteroaryl; alkoxy; and —NR52;—R2is selected from the group consisting of —H; optionally substituted C1-C6alkyl; optionally substituted aryl; and optionally substituted heteroaryl;—R3is selected from the group consisting of —H; optionally substituted C1-C6alkyl; optionally substituted aryl; and optionally substituted heteroaryl;—R4is selected from the group consisting of —H; optionally substituted C1-C6alkyl; optionally substituted aryl; and optionally substituted heteroaryl;each —R5is independently of each other selected from the group consisting of —H; optionally substituted C1-C6alkyl; optionally substituted aryl; and optionally substituted heteroaryl; or when taken together two —R5can be cycloalkyl or cyclo hetero alkyl;wherein -L1- is substituted with -L2-Z and wherein -L1- is optionally further substituted;wherein-L2- is a single chemical bond or a spacer; and—Z is a carrier moiety.

The optional further substituents of -L1- of formula (V) are preferably as described above.

Preferably -L1- of formula (V) is substituted with one moiety -L2-Z.

In one embodiment -L1- of formula (V) is not further substituted.

Only in the context of formula (V) the terms used have the following meaning:

“Alkyl”, “alkenyl”, and “alkynyl” include linear, branched or cyclic hydrocarbon groups of 1-8 carbons or 1-6 carbons or 1-4 carbons wherein alkyl is a saturated hydrocarbon, alkenyl includes one or more carbon-carbon double bonds and alkynyl includes one or more carbon-carbon triple bonds. Unless otherwise specified these contain 1-6 C.

“Aryl” includes aromatic hydrocarbon groups of 6-18 carbons, preferably 6-10 carbons, including groups such as phenyl, naphthyl, and anthracene “Heteroaryl” includes aromatic rings comprising 3-15 carbons containing at least one N, O or S atom, preferably 3-7 carbons containing at least one N, O or S atom, including groups such as pyrrolyl, pyridyl, pyrimidinyl, imidazolyl, oxazolyl, isoxazolyl, thiszolyl, isothiazolyl, quinolyl, indolyl, indenyl, and similar.

The term “substituted” means an alkyl, alkenyl, alkynyl, aryl, or heteroaryl group comprising one or more substituent groups in place of one or more hydrogen atoms. Substituents may generally be selected from halogen including F, Cl, Br, and I; lower alkyl including linear, branched, and cyclic; lower haloalkyl including fluoroalkyl, chloroalkyl, bromoalkyl, and iodoalkyl; OH; lower alkoxy including linear, branched, and cyclic; SH; lower alkylthio including linear, branched and cyclic; amino, alkylamino, dialkylamino, silyl including alkylsilyl, alkoxysilyl, and arylsilyl; nitro; cyano; carbonyl; carboxylic acid, carboxylic ester, carboxylic amide, aminocarbonyl; aminoacyl; carbamate; urea; thiocarbamate; thiourea; ketne; sulfone; sulfonamide; aryl including phenyl, naphthyl, and anthracenyl; heteroaryl including 5-member heteroaryls including as pyrrole, imidazole, furan, thiophene, oxazole, thiazole, isoxazole, isothiazole, thiadiazole, triazole, oxadiazole, and tetrazole, 6-member heteroaryls including pyridine, pyrimidine, pyrazine, and fused heteroaryls including benzofuran, benzothiophene, benzoxazole, benzimidazole, indole, benzothiazole, benzisoxazole, and benzisothiazole.

A further preferred embodiment for -L1- is disclosed in U.S. Pat. No. 7,585,837B2, which is herewith incorporated by reference in its entirety. Accordingly, a preferred moiety -L1- is of formula (VI):

whereinthe dashed line indicates attachment to an amine functional group of a side chain of an amino acid residue of the ring moiety of a CNP moiety;R1and R2are independently selected from the group consisting of hydrogen, alkyl, alkoxy, alkoxyalkyl, aryl, alkaryl, aralkyl, halogen, nitro, —SO3H, —SO2NHR5, amino, ammonium, carboxyl, PO3H2, and OPO3H2;R3, R4, and R5are independently selected from the group consisting of hydrogen, alkyl, and aryl;wherein -L1- is substituted with -L2-Z and wherein -L1- is optionally further substituted;wherein-L2- is a single chemical bond or a spacer; and—Z is a carrier moiety.

Suitable substituents for formulas (VI) are alkyl (such as C1 -6alkyl), alkenyl (such as C2-6alkenyl), alkynyl (such as C2-6alkynyl), aryl (such as phenyl), heteroalkyl, heteroalkenyl, heteroalkynyl, heteroaryl (such as aromatic 4 to 7 membered heterocycle) or halogen moieties.

Preferably -L1- of formula (VI) is substituted with one moiety -L2-Z.

In one embodiment -L1- of formula (VI) is not further substituted.

Only in the context of formula (VI) the terms used have the following meaning:

The terms “alkyl”, “alkoxy”, “alkoxyalkyl”, “aryl”, “alkaryl” and “aralkyl” mean alkyl radicals of 1-8, preferably 1-4 carbon atoms, e.g. methyl, ethyl, propyl, isopropyl and butyl, and aryl radicals of 6-10 carbon atoms, e.g. phenyl and naphthyl. The term “halogen” includes bromo, fluoro, chloro and iodo.

A further preferred embodiment for -L1- is disclosed in WO2002/089789A1, which is herewith incorporated by reference in its entirety. Accordingly, a preferred moiety -L1- is of formula (VII):

whereinthe dashed line indicates attachment to an amine functional group of a side chain of an amino acid residue of the ring moiety of a CNP moiety;L1is a bifunctional linking group,Y1and Y2are independently O, S or NR7;R2, R3, R4, R5, R6and R7are independently selected from the group consisting of hydrogen, C1-6alkyls, C3-12branched alkyls, C3-8cycloalkyls, C1-6substituted alkyls, C3-8substituted cycloalkyls, aryls, substituted aryls, aralkyls, C1-6heteroalkyls, substituted C1-6heteroalkyls, C1-6alkoxy, phenoxy, and C1-6heteroalkoxy;Ar is a moiety which when included in formula (VII) forms a multisubstituted aromatic hydrocarbon or a multi-substituted heterocyclic group;X is a chemical bond or a moiety that is actively transported into a target cell, a hydrophobic moiety, or a combination thereof,y is 0 or 1;wherein -L1- is substituted with -L2-Z and wherein -L1- is optionally further substituted;wherein-L2- is a single chemical bond or a spacer; and—Z is a carrier moiety.

The optional further substituents of -L1- of formula (VII) are preferably as described above.

Preferably -L1- of formula (VII) is substituted with one moiety -L2-Z.

In one embodiment -L1- of formula (VII) is not further substituted.

Only in the context of formula (VII) the terms used have the following meaning:

The term “alkyl” shall be understood to include, e.g. straight, branched, substituted C1-12alkyls, including alkoxy, C3-8cycloalkyls or substituted cycloalkyls, etc.

The term “substituted” shall be understood to include adding or replacing one or more atoms contained within a functional group or compounds with one or more different atoms.

Substituted alkyls include carboxyalkyls, amino alkyls, dialkylaminos, hydroxyalkyls and mercaptoalkyls; substituted cycloalkyls include moieties such as 4-chlorocyclohexyl; aryls include moieties such as napthyl; substituted aryls include moieties such as 3-bromo-phenyl; aralkyls include moieties such as toluyl; heteroalkyls include moieties such as ethylthiophene; substituted heteroalkyls include moieties such as 3-methoxythiophone; alkoxy includes moieities such as methoxy; and phenoxy includes moieties such as 3-nitrophenoxy. Halo-shall be understood to include fluoro, chloro, iodo and bromo.

In another preferred embodiment -L1- comprises a substructure of formula (VIII)

whereinthe dashed line marked with the asterisk indicates attachment to an amine functional group of a side chain of an amino acid residue of the ring moiety of a CNP moiety; the unmarked dashed lines indicate attachment to the remainder of -L1-; andwherein -L1- is substituted with -L2-Z and wherein -L1- is optionally further substituted;wherein-L2- is a single chemical bond or a spacer; and—Z is a carrier moiety.

The optional further substituents of -L1- of formula (VIII) are preferably as described above.

Preferably -L1- of formula (VIII) is substituted with one moiety -L2-Z.

In one embodiment -L1- of formula (VIII) is not further substituted.

In another preferred embodiment -L1- comprises a substructure of formula (IX)

whereinthe dashed line marked with the asterisk indicates attachment to a nitrogen of a side chain of an amino acid residue of the ring moiety of a CNP moiety by forming a carbamate bond;the unmarked dashed lines indicate attachment to the remainder of -L1-; andwherein -L1- is substituted with -L2-Z and wherein -L1- is optionally further substituted;wherein-L2- is a single chemical bond or a spacer; and—Z is a carrier moiety.

The optional further substituents of -L1- of formula (IX) are preferably as described above.

Preferably -L1- of formula (IX) is substituted with one moiety -L2-Z.

In one embodiment -L1- of formula (IX) is not further substituted.

The moiety -L2- can be attached to -L1- by replacing any —H present, except where explicitly excluded.

In the prodrugs of the present invention -L2- is a chemical bond or a spacer moiety.

In one embodiment -L2- is a chemical bond.

In another embodiment -L2- is a spacer moiety.

Preferably, one to five, preferably one, of the hydrogens given by —R1, —R1a, —R2, —R2a, —R3, —R3a, —R4, —R4a, —R5, —R5a, —R6, —R7, —R7a, —R8, —R8a, —R9, —R9a, —R10, ——R10aand/or —R11of formula (II) are replaced by -L2-. Preferably, one to five, preferably one, of the hydrogens given by —R1, —R1a, —R2, —R2a, —R3, —R3a, —R4, —R5, —R5a, —R6, —R6a, —R7, —R7a, —R8, —R8a, —R8b, —R9, —R9a, —R9b, —R10, —R10a, —R10b, —R11, —R12, —R12a, —R13, —R13a, and/or —R13bof formula (III) are replaced by -L2-.

In the prodrugs of the present invention -L2- is a chemical bond or a spacer moiety.

In one embodiment -L2- is a chemical bond.

In another embodiment -L2- is a spacer moiety.

When -L2- is other than a single chemical bond, -L2- is preferably selected from the group consisting of -T-, —C(O)O—, —O—, —C(O)—, —C(O)N(Ry1)—, —S(O)2N(Ry1)—, —S(O)N(Ry1)—, —S(O)2—, —S(O)—, —N(Ry1)S(O)2N(Ry1a)—, —S—, —N(Ry1)—, —OC(ORy1)(Ry1a)—, —N(Ry1)C(O)N(Ry1a)—, —OC(O)N(Ry1)—; C1-50alkyl, C2-50alkenyl, and C2-50alkynyl; wherein -T-, C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally substituted with one or more —Ry2, which are the same or different and wherein C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T-, —C(O)O—, —O—, —C(O)—, —C(O)N(Ry3)—, —S(O)2N(Ry3)—, —S(O)N(Ry3)—, —S(O)2—, —S(O)—, —N(Ry3)S(O)2N(Ry3a)—, —S—, —N(Ry3)—, —OC(ORy3)(Ry3a)—, —N(Ry3)C(O)N(Ry3a)—, and —OC(O)N(Ry3)—;

Ry1and —Ry1aare independently of each other selected from the group consisting of —H, -T, C1-50alkyl, C2-50alkenyl, and C2-50alkynyl; wherein -T, C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally substituted with one or more —Ry2which are the same or different, and wherein C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T-, —C(O)O—, —O—, —C(O)—, —C(O)N(Ry4)—, —S(O)2N(Ry4)—, —S(O)N(Ry4)—, —S(O)2—, —S(O)—, —N(Ry4)S(O)2N(Ry4a)—, —S—, —N(Ry4)—, —OC(ORy4)(Ry4a)—, —N(Ry4)C(O)N(Ry4a)—, and —OC(O)N(Ry4)—;

each T is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C3-10cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl;

wherein each T is independently optionally substituted with one or more —Ry2, which are the same or different;

each —Ry2is independently selected from the group consisting of halogen, —CN, oxo (═O), —COORy5, —ORy5, —C(O)Ry5, —C(O)N(Ry5Ry5a), —S(O)2N(Ry5Ry5a), —S(O)N(Ry5Ry5a), —S(O)2Ry5, —S(O)Ry5, —N(Ry5)S(O)2N(Ry5aRy5b), —SRy5, —N(Ry5Ry5a), —NO2—, —OC(O)Ry5, —N(Ry5)C(O)Ry5a, —N(Ry5)S(O)2Ry5a, —N(Ry5)S(O)Ry5a, —N(Ry5)C(O)ORy5a, —N(Ry5)C(O)N(Ry5aRy5a), —OC(O)N(Ry5Ry5a), and C1-6alkyl; wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different; and

each —Ry3, Ry3a, —Ry4, —Ry4a, —Ry5, —Ry5aand —Ry5bis independently selected from the group consisting of —H, and C1-6alkyl, wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different.

When -L2- is other than a single chemical bond, -L2- is even more preferably selected from -T-, —C(O)O—, —O—, —C(O)—, —C(O)N(Ry1)—, —S(O)2N(Ry1)—, —S(O)N(Ry1)—, —S(O)2—, —S(O)—, —N(Ry1)S(O)2N(Ry1a)—, —S—, —N(Ry1)—, —OC(ORy1)(Ry1a)—, —N(Ry1)C(O)N(Ry1a)—, —OC(O)N(Ry1)—, C1-50alkyl, C2-50alkenyl, and C2-50alkynyl; wherein -T-, C1-20alkyl, C2-20alkenyl, and C2-20alkynyl are optionally substituted with one or more —Ry2, which are the same or different and wherein C1-20alkyl, C2-20alkenyl, and C2-20alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T-, —C(O)O—, —O—, —C(O)—, —C(O)N(Ry3)—, —S(O)2N(Ry3)—, —S(O)N(Ry3)—, —S(O)2—, —S(O)—, —N(Ry3)S(O)2N(Ry3a)—, —S—, —N(Ry3)—, —OC(ORy3)(Ry3a)—, —N(Ry3)C(O)N(Ry3a)—, and —OC(O)N(Ry3)—;

—Ry1and —Ry1aare independently of each other selected from the group consisting of —H, -T, C1-10alkyl, C2-10alkenyl, and C2-10alkynyl; wherein -T, C1-10alkyl, C2-10alkenyl, and C2-10alkynyl are optionally substituted with one or more —Ry2, which are the same or different, and wherein C1-10alkyl, C2-10alkenyl, and C2-10alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T-, —C(O)O—, —O—, —C(O)—, —C(O)N(Ry4)—, —S(O)2N(Ry4)—, —S(O)N(Ry4)—, —S(O)2—, —S(O)—, —N(Ry4)S(O)2N(Ry4a)—, —S—, —N(Ry4)—, —OC(ORy4)(Ry4a)—, —N(Ry4)C(O)N(Ry4a)—, and —OC(O)N(Ry4)—;

each T is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C3-10cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl; wherein each T is independently optionally substituted with one or more —Ry2, which are the same or different;

—RY2is selected from the group consisting of halogen, —CN, oxo (═O), —COORy5, —ORy5, —C(O)Ry5, —C(O)N(Ry5Ry5a), —S(O)2N(Ry5Ry5a), —S(O)N(Ry5Ry5a), —S(O)2Ry5, —S(O)Ry5, —N(Ry5)S(O)2N(Ry5aRy5b), —SRy5, —N(Ry5Ry5a), —NO2—, —OC(O)Ry5, —N(Ry5)C(O)Ry5a, —N(Ry5)S(O)2Ry5a, —N(Ry5)S(O)Ry5a, —N(Ry5)C(O)ORy5a, —N(Ry5)C(O)N(Ry5aRy5a), —OC(O)N(Ry5Ry5a), and C1-6alkyl; wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different; andeach —Ry3, —Ry3a, —Ry4, —Ry4a, —Ry5, —Ry5aand —Ry5bis independently of each other selected from the group consisting of —H, and C1-6alkyl; wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different.

When -L2- is other than a single chemical bond, -L2- is even more preferably selected from the group consisting of -T-, —C(O)O—, —O—, —C(O)—, —C(O)N(Ry1)—, —S(O)2N(Ry1)—, —S(O)N(Ry1)—, —S(O)2—, —S(O)—, —N(Ry1)S(O)2N(Ry1a)—, —S—, —N(Ry1)—, —OC(ORy1)(Ry1a)—, —N(Ry1)C(O)N(Ry1a)—, and —OC(O)N(Ry1)—, C1-50alkyl, C2-50alkenyl, and C2-50alkynyl; wherein -T-, C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally substituted with one or more —Ry2, which are the same or different and wherein C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T-, —C(O)O—, —O—, —C(O)—, —C(O)N(Ry3)—, —S(O)2N(Ry3)—, —S(O)N(Ry3)—, —S(O)2—, —S(O)—, —N(Ry3)S(O)2N(Ry3a)—, —S—, —N(Ry3)—, —OC(ORy3)(Ry3a)—, —N(Ry3)C(O)N(Ry3a)—, and —OC(O)N(Ry3)—;

—Ry1and —Ry1aare independently selected from the group consisting of —H, -T, C1-10alkyl, C2-10alkenyl, and C2-10alkynyl;

each T is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C3-10cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl;

each —Ry2is independently selected from the group consisting of halogen, and C1-6alkyl; and each —Ry3, —Ry3a, —Ry4, Ry4a, —Ry5, —Ry5aand —Ry5bis independently of each other selected from the group consisting of —H, and C1-6alkyl; wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different.

Even more preferably, -L2- is a C1-20alkyl chain, which is optionally interrupted by one or more groups independently selected from —O—, -T- and —C(O)N(Ry1)—; and which C1-20alkyl chain is optionally substituted with one or more groups independently selected from —OH, -T and —C(O)N(Ry6Ry6a); wherein —Ry1, —Ry6, —Ry6aare independently selected from the group consisting of H and C1-4alkyl and wherein T is selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C3-10cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl.

Preferably, -L2- has a molecular weight in the range of from 14 g/mol to 750 g/mol.

Preferably, -L2- comprises a moiety selected from



wherein

dashed lines indicate attachment to the rest of -L2-, -L1- and/or —Z and/or, respectively; and —R and —Raare independently of each other selected from the group consisting of —H, methyl, ethyl, propyl, butyl, pentyl and hexyl.

In one preferred embodiment -L2- has a chain lengths of 1 to 20 atoms.

As used herein the term “chain length” with regard to the moiety -L2- refers to the number of atoms of -L2- present in the shortest connection between -L1- and —Z.

Preferably, -L2- is of formula (i)

whereinthe dashed line marked with the asterisk indicates attachment to -L1-;the unmarked dashed line indicates attachment to —Z;—R1is selected from the group consisting of —H, C1-6alkyl, C2-6alkenyl and C2-6alkynyl;n is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 and 18; andwherein the moiety of formula (i) is optionally further substituted.

Preferably —R1of formula (i) is selected from the group consisting of —H, methyl, ethyl, propyl, and butyl. Even more preferably —R1of formula (i) is selected from the group consisting of —H, methyl, ethyl and propyl. Even more preferably —R1of formula (i) is selected from the group consisting of —H and methyl. Most preferably —R1of formula (i) is methyl.

Preferably n of formula (i) is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10. Even more preferably n of formula (i) is selected from the group consisting of 0, 1, 2, 3, 4 and 5. Even more preferably n of formula (i) is selected from the group consisting of 0, 1, 2 and 3. Even more preferably n of formula (i) is selected from the group consisting of 0 and 1. Most preferably n of formula (i) is 0.

In one preferred embodiment -L2- is a moiety selected from the group consisting of

whereinthe dashed line marked with the asterisk indicates attachment to -L1-;the unmarked dashed line indicates attachment to —Z; andwherein the moieties (ii), (iii), (iv), (v), (vi), (vii), (viii), (ix), (x), (xi), (xii), (xiii), (xiv), (xv), (xvi) and (xvii) are optionally further substituted.

In a preferred embodiment -L2- is selected from the group consisting of

whereinthe dashed line marked with the asterisk indicates attachment to -L1-; andthe unmarked dashed line indicates attachment to —Z.

Even more preferred -L2- is selected from the group consisting of

whereinthe dashed line marked with the asterisk indicates attachment to -L1-; andthe unmarked dashed line indicates attachment to —Z.

Even more preferably -L2- is of formula (xvi)

whereinthe dashed line marked with the asterisk indicates attachment to -L1-; andthe unmarked dashed line indicates attachment to —Z.

In one preferred embodiment the moiety -L1-L2- is selected from the group consisting of

whereinthe unmarked dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z.

In an even more preferred embodiment the moiety -L1-L2- is of formula (IId-ii)

whereinthe unmarked dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z.

In a most preferred embodiment the moiety -L1-L2- is of formula (IId-ii′)

whereinthe unmarked dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z.

In another preferred embodiment the moiety -L1-L2- is selected from the group consisting of

whereinthe unmarked dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z.

In an even more preferred embodiment the moiety -L1-L2- is of formula (IId-iia)

whereinthe unmarked dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z.

In a most preferred embodiment the moiety -L1-L2- is of formula (IId-iia′)

whereinthe unmarked dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z.

In another preferred embodiment the moiety -L1-L2- is selected from the group consisting of

whereinthe unmarked dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z.

In an even more preferred embodiment the moiety -L1-L2- is of formula (IId-iib)

whereinthe unmarked dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z.

In a most preferred embodiment the moiety -L1-L2- is of formula (IId-iib′)

whereinthe unmarked dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z.

Preferably, —Z is water-soluble.

Preferably, —Z has a molecular weight ranging from 5 to 500 kDa. Even more preferably, —Z has a molecular weight ranging from 10 to 250 kDa, even more preferably ranging from 10 to 150 kDa, even more preferably from 12 to 100 and most preferably from 15 to 80. In one preferred embodiment —Z has a molecular weight of about 20 kDa. In another preferred embodiment —Z has a molecular weight of about 40 kDa.

In one embodiment —Z comprises a C8-24alkyl moiety.

In a preferred embodiment —Z comprises a polymer, preferably a polymer selected from the group consisting of 2-methacryloyl-oxyethyl phosphoyl cholins, poly(acrylic acids), poly(acrylates), poly(acrylamides), poly(alkyloxy) polymers, poly(amides), poly(amidoamines), poly(amino acids), poly(anhydrides), poly(aspartamides), poly(butyric acids), poly(glycolic acids), polybutylene terephthalates, poly(caprolactones), poly(carbonates), poly(cyanoacrylates), poly(dimethylacrylamides), poly(esters), poly(ethylenes), poly(ethyleneglycols), poly(ethylene oxides), poly(ethyl phosphates), poly(ethyloxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl-oxazolines), wpoly(hydroxymethacrylates), poly(hydroxypropylmethacrylamides), poly(hydroxypropyl methacrylates), poly(hydroxypropyloxazo lines), poly(iminocarbonates), poly(lactic acids), poly(lactic-co-glycolic acids), poly(methacrylamides), poly(methacrylates), poly(methyloxazolines), poly(organophosphazenes), poly(ortho esters), poly(oxazo lines), poly(propylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinyl amines), poly(vinylmethylethers), poly(vinylpyrrolidones), silicones, celluloses, carbomethyl celluloses, hydroxypropyl methylcelluloses, chitins, chitosans, dextrans, dextrins, gelatins, hyaluronic acids and derivatives, functionalized hyaluronic acids, mannans, pectins, rhamnogalacturonans, starches, hydroxyalkyl starches, hydroxyethyl starches and other carbohydrate-based polymers, xylans, and copolymers thereof.

The moiety —Z of formula (I) may be a linear, branched, multi-arm or dendritic polymeric moiety.

In one embodiment —Z of formula (I) is a linear polymeric moiety.

In another embodiment —Z of formula (I) is a multi-arm polymeric moiety.

In another embodiment —Z of formula (I) is a dendritic polymeric moiety.

In a preferred embodiment —Z of formula (I) is a branched polymeric moiety.

In one embodiment —Z of formula (I) comprises a protein. Preferred proteins are selected from the group consisting of carboxyl-terminal peptide of the chorionic gonadotropin as described in US 2012/0035101 A1 which are herewith incorporated by reference; albumin; XTEN sequences as described in WO 2011123813 A2 which are herewith incorporated by reference; proline/alanine random coil sequences as described in WO 2011/144756 A1 which are herewith incorporated by reference; proline/alanine/serine random coil sequences as described in WO 2008/155134 A1 and WO 2013/024049 A1 which are herewith incorporated by reference; and Fc fusion proteins.

In one embodiment —Z of formula (I) is a polysarcosine.

In another preferred embodiment —Z of formula (I) comprises a poly(N-methylglycine).

In a particularly preferred embodiment —Z of formula (I) comprises a random coil protein moiety.

In one preferred embodiment —Z of formula (I) comprises one random coil protein moiety.

In another preferred embodiment —Z of formula (I) comprises two random coil protein moieties.

In another preferred embodiment —Z of formula (I) comprises three random coil protein moieties.

In another preferred embodiment —Z of formula (I) comprises four random coil protein moieties.

In another preferred embodiment —Z of formula (I) comprises five random coil protein moieties.

In another preferred embodiment —Z of formula (I) comprises six random coil protein moieties.

In another preferred embodiment —Z of formula (I) comprises seven random coil protein moieties.

In another preferred embodiment —Z of formula (I) comprises eight random coil protein moieties.

Preferably such random coil protein moiety comprises at least 25 amino acid residues and at most 2000 amino acids. Even more preferably such random coil protein moiety comprises at least 30 amino acid residues and at most 1500 amino acid residues. Even more preferably such random coil protein moiety comprises at least 50 amino acid residues and at most 500 amino acid residues.

In a preferred embodiment, —Z of formula (I) comprises a random coil protein moiety of which at least 80%, preferably at least 85%, even more preferably at least 90%, even more preferably at least 95%, even more preferably at least 98% and most preferably at least 99% of the total number of amino acids forming said random coil protein moiety are selected from alanine and proline. Even more preferably, at least 10%, but less than 75%, preferably less than 65%, of the total number of amino acid residues of such random coil protein moiety are proline residues. Preferably, such random coil protein moiety is as described in WO 2011/144756 A1 which is hereby incorporated by reference in its entirety. Even more preferably —Z comprises at least one moiety selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:51 and SEQ ID NO:61 as disclosed in WO2011/144756 which are hereby incorporated by reference. A moiety comprising such random coil protein comprising alanine and proline will be referred to as “PA” or “PA moiety”.

Accordingly, —Z of formula (I) comprises a PA moiety.

In an equally preferred embodiment, —Z of formula (I) comprises a random coil protein moiety of which at least 80%, preferably at least 85%, even more preferably at least 90%, even more preferably at least 95%, even more preferably at least 98% and most preferably at least 99% of the total number of amino acids forming said random coil protein moiety are selected from alanine, serine and proline. Even more preferably, at least 4%, but less than 40% of the total number of amino acid residues of such random coil protein moiety are proline residues. Preferably, such random coil protein moiety is as described in WO 2008/155134 A1 which is hereby incorporated by reference in its entirety. Even more preferably —Z comprises at least one moiety selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:46, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54 and SEQ ID NO:56 as disclosed in WO 2008/155134 A1, which are hereby incorporated by reference. A moiety comprising such random coil protein moiety comprising alanine, serine and proline will be referred to as “PAS” or “PAS moiety”.

Accordingly, —Z of formula (I) comprises a PAS moiety.

In an equally preferred embodiment, —Z of formula (I) comprises a random coil protein moiety of which at least 80%, preferably at least 85%, even more preferably at least 90%, even more preferably at least 95%, even more preferably at least 98% and most preferably at least 99% of the total number of amino acids forming said random coil protein moiety are selected from alanine, glycine and proline. A moiety comprising such random coil protein moiety comprising alanine, glycine and proline will be referred to as “PAG” or “PAG moiety”.

Accordingly, —Z of formula (I) comprises a PAG moiety.

In an equally preferred embodiment, —Z of formula (I) comprises a random coil protein moiety of which at least 80%, preferably at least 85%, even more preferably at least 90%, even more preferably at least 95%, even more preferably at least 98% and most preferably at least 99% of the total number of amino acids forming said random coil protein moiety are selected from proline and glycine. A moiety comprising such random coil protein moiety comprising proline and glycine will be referred to as “PG” or “PG moiety”.

Preferably, such PG moiety comprises a moiety of formula (a-0)
[(Gly)p-Pro-(Gly)q]r(a-0);whereinp is selected from the group consisting of 0, 1, 2, 3, 4 and 5;q is selected from the group consisting of 0, 1, 2, 3, 4 and 5;r is an integer ranging from and including 10 to 1000;provided that at least one of p and q is at least 1;

Preferably, p of formula (a-0) is selected from the group consisting of 1, 2 and 3.

Preferably, q of formula (a-0) is selected from 0, 1 and 2.

Even more preferably the PG moiety comprises the sequence of SEQ ID NO:97: GGPGGPGPGGPGGPGPGGPG

Even more preferably, the PG moiety comprises the sequence of formula (a-0-a)(GGPGGPGPGGPGGPGPGGPG)V(a-0-a),whereinv is an integer ranging from and including 1 to 50.

It is understood that the sequence of formula (a-0-a) comprises v replicates of the sequence of SEQ ID NO:97.

Accordingly, —Z of formula (I) comprises a PG moiety.

In an equally preferred embodiment, —Z of formula (I) comprises a random coil protein moiety of which at least 80%, preferably at least 85%, even more preferably at least 90%, even more preferably at least 95%, even more preferably at least 98% and most preferably at least 99% of the total number of amino acids forming said random coil protein moiety are selected from alanine, glycine, serine, threonine, glutamate and proline. Preferably, such random coil protein moiety is as described in WO 2010/091122 A1 which is hereby incorporated by reference. Even more preferably —Z comprises at least one moiety selected from the group consisting of SEQ ID NO:182, SEQ ID NO:183, SEQ ID NO:184; SEQ ID NO:185, SEQ ID NO:186, SEQ ID NO:187, SEQ ID NO:188, SEQ ID NO:189, SEQ ID NO:190, SEQ ID NO:191, SEQ ID NO:192, SEQ ID NO:193, SEQ ID NO:194, SEQ ID NO:195, SEQ ID NO:196, SEQ ID NO:197, SEQ ID NO:198, SEQ ID NO:199, SEQ ID NO:200, SEQ ID NO:201, SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:205, SEQ ID NO:206, SEQ ID NO:207, SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:211, SEQ ID NO:212, SEQ ID NO:213, SEQ ID NO:214, SEQ ID NO:215, SEQ ID NO:216, SEQ ID NO:217, SEQ ID NO:218, SEQ ID NO:219, SEQ ID NO:220, SEQ ID NO:221, SEQ ID NO:759, SEQ ID NO:760, SEQ ID NO:761, SEQ ID NO:762, SEQ ID NO:763, SEQ ID NO:764, SEQ ID NO:765, SEQ ID NO:766, SEQ ID NO:767, SEQ ID NO:768, SEQ ID NO:769, SEQ ID NO:770, SEQ ID NO:771, SEQ ID NO:772, SEQ ID NO:773, SEQ ID NO:774, SEQ ID NO:775, SEQ ID NO:776, SEQ ID NO:777, SEQ ID NO:778, SEQ ID NO:779, SEQ ID NO:1715, SEQ ID NO:1716, SEQ ID NO:1718, SEQ ID NO:1719, SEQ ID NO:1720, SEQ ID NO:1721 and SEQ ID NO:1722 as disclosed in WO2010/091122A1, which are hereby incorporated by reference. A moiety comprising such random coil protein moiety comprising alanine, glycine, serine, threonine, glutamate and proline will be referred to as “XTEN” or “XTEN moiety” in line with its designation in WO 2010/091122 A1.

Accordingly, —Z of formula (I) comprises an XTEN moiety.

In another preferred embodiment, —Z of formula (I) comprises a fatty acid derivate. Preferred fatty acid derivatives are those disclosed in WO 2005/027978 A2 and WO 2014/060512 A1 which are herewith incorporated by reference.

In another preferred embodiment —Z of formula (I) is a hyaluronic acid-based polymer.

In one embodiment —Z of formula (I) is a carrier as disclosed in WO 2012/02047 A1 which is herewith incorporated by reference.

In another embodiment —Z of formula (I) is a carrier as disclosed in WO 2013/024048 A1 which is herewith incorporated by reference.

In another preferred embodiment —Z of formula (I) is a PEG-based polymer.

In a particularly preferred embodiment —Z of formula (I) is a branched polymer. In one embodiment —Z of formula (I) is a branched polymer having one, two, three, four, five or six branching points. Preferably, —Z of formula (I) is a branched polymer having one, two or three branching points. In one embodiment —Z of formula (I) is a branched polymer having one branching point. In another embodiment —Z of formula (I) is a branched polymer having two branching points. In another embodiment —Z of formula (I) is a branched polymer having three branching points.

A branching point is preferably selected from the group consisting of —N<, —CH< and >C<.

Preferably such branched moiety —Z of formula (I) is PEG-based.

In one embodiment such branched moiety —Z of formula (I) has a molecular weight ranging from and including 5 kDa to 500 kDa, more preferably ranging from and including 10 kDa to 250 Da, even more preferably ranging from and including 10 kDa to 150 kDa, even more preferably ranging from and including 12 kDa to 100 kDa and most preferably ranging from and including 15 kDa to 80 kDa.

Preferably, such branched moiety —Z of formula (I) has a molecular weight ranging from and including 10 kDa to 80 kDa. In one embodiment the molecular weight of —Z of formula (I) is about 10 kDa. In another embodiment the molecular weight of such branched moiety —Z of formula (I) is about 20 kDa. In another embodiment the molecular weight of such branched moiety —Z of formula (I) is about 30 kDa. In another embodiment the molecular weight of such a branched moiety —Z of formula (I) is about 40 kDa. In another embodiment the molecular weight of such a branched moiety —Z of formula (I) is about 50 kDa. In another embodiment the molecular weight of such a branched moiety —Z of formula (I) is about 60 kDa. In another embodiment the molecular weight of such a branched moiety —Z of formula (I) is about 70 kDa. In another embodiment the molecular weight of such a branched moiety —Z of formula (I) is about 80 kDa. Most preferably, such branched moiety —Z of formula (I) has a molecular weight of about 40 kDa.

Applicants found that an N-terminal attachment of a moiety -L1-L2-Z is significantly more efficient with regard to NEP-stability than attachment at an internal site and that the least efficient attachment site with regard to NEP-stability is at the ring part of a CNP moiety. However, applicants surprisingly found that this disadvantage of attachment to the ring with regard to NEP-stability can be compensated by using a branched moiety —Z having a molecular weight of at least 10 kDa, such as at least 12 kDa, such as at least 15 kDa, such as at least 18 kDa, such as at least 20 kDa, such as at least 24 kDa, such as at least 25 kDa, such as at least 27 kDa, such as at least 30 kDa. Preferably, such branched moiety —Z has a molecular weight of no more than 500 kDa, preferably of no more than 250 kDa, preferably of no more than 200 Da, preferably of no more than 150 kDa and most preferably no more than 100 kDa. Most preferably such branched moiety —Z has a molecular weight of about 40 kDa. Consequently, the use of such branched moiety —Z at the ring part of the CNP moiety does not only lead to increased NEP-stability, but combines increased NEP-stability with reduced NPR-B binding associated with attachment to the ring.

It was surprisingly found that even though the ring moiety is involved in NPR-C binding, attachment of a 5 kDa carrier to the ring moiety did not have a significant effect on NPR-C affinity. Furthermore, it was surprisingly found that a 4× 10 kDa carrier, i.e. a branched carrier having four 10 kDa arms, attached to the ring moiety is more efficient in reducing NPR-C affinity than a 2× 20 kDa carrier, i.e. a branched carrier having two 20 kDa arms, even though the total molecular weight was the same. It is thus not only the total molecular weight of the carrier attached to the ring moiety, but the particular branching pattern of the carrier that influences NPR-C binding affinity.

This finding is also supported by the NPR-C affinity measured with a 4-arm 40 kDa carrier having a different branching pattern which still exhibited a high NPR-C affinity.

In summary, it was surprisingly found that NPR-C affinity can be efficiently reduced with a multi-branched carrier attached to the ring moiety having a first branching point close to the CNP moiety, such as less than 300 atoms from the CNP moiety, preferably 200 atoms from the CNP moiety, even more preferably 100 atoms from the CNP moiety, even more preferably less than 50 atoms from the CNP moiety, even more preferably less than 25 atoms from the CNP moiety and most preferably less than 10 atoms from the CNP moiety.

Even more preferably, one or more further branching point(s) is/are located within less than 500 atoms from the CNP moiety, even more preferably 300 atoms from the CNP moiety, even more preferably less than 200 atoms from the CNP moiety, even more preferably less than 100 atoms from the CNP moiety, even more preferably less than 75 atoms from the CNP moiety, even more preferably less than 50 atoms from the CNP moiety, even more preferably less than 40 atoms from the CNP moiety and most preferably less than 35 atoms from the CNP moiety.

It was in addition also found that such branching pattern is beneficial for in vivo stability of the CNP moiety, i.e. protection against proteolytic degradation. It was surprisingly found that N-terminal degradation was stronger when using a 2× 20 kDa carrier compared to 4× 10 kDa carrier. Likewise, using a 4-arm 40 kDa carrier having a different branching pattern exhibited even stronger N-terminal degradation.

Preferably, —Z of formula (I) comprises a moiety



In an equally preferred embodiment —Z of formula (I) comprises an amide bond.

In one embodiment —Z of formula (I) comprises a moiety of formula (a)

whereinthe dashed line indicates attachment to -L2- or to the remainder of —Z;BPais a branching point selected from the group consisting of —N<, —CR< and >C<;—R is selected from the group consisting of —H and C1-6alkyl;a is 0 if BPais —N< or —CR< and a is 1 if BPais >C<;—Sa—, —Sa′, —Sa″— and —Sa′″— are independently of each other a chemical bond or are selected from the group consisting of C1-50alkyl, C2-50alkenyl, and C2-50alkynyl;wherein C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally substituted with one or more —R1, which are the same or different and wherein C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T-, —C(O)O—, —O—, —C(O)—, —C(O)N(R2)—, —S(O)2N(R2)—, —S(O)N(R2)—, —S(O)2—, —S(O)—, —N(R2)S(O)2N(R2a)—, —S—, —N(R2)—, —OC(OR2)(R2a)—, —N(R2)C(O)N(R2a)—, and —OC(O)N(R2)—;each -T- is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C3-10cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl; wherein each -T- is independently optionally substituted with one or more —R15which are the same or different;each —R1is independently selected from the group consisting of halogen, —CN, oxo (═O), —COOR3, —OR3, —C(O)R3, —C(O)N(R3R3a), —S(O)2N(R3R3a), —S(O)N(R3R3a), —S(O)2R3, —S(O)R3, —N(R3)S(O)2N(R3aR3b), —SR3, —N(R3R3a), —NO2—, —OC(O)R3, —N(R3)C(O)R3a, —N(R3)S(O)2R3a, —N(R3)S(O)R3a, —N(R3)C(O)OR3a, —N(R3)C(O)N(R3aR3a), —OC(O)N(R3R3a), and C1-6alkyl; wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different;each —R2, —R2a, —R3, —R3aand —R3bis independently selected from the group consisting of —H, and C1-6alkyl, wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different; andPa′, —Pa″and —Pa′″are independently a polymeric moiety.

Optionally, the moiety of formula (a) is substituted with one or more substituents.

In one embodiment BPaof formula (a) is —N<.

In another embodiment BPaof formula (a) is —CR<. Preferably, —R is —H. Accordingly, a of formula (a) is preferably 0.

In another embodiment BPaof formula (a) is >C<.

In one embodiment —Sa— of formula (a) is a chemical bond.

In another embodiment —Sa— of formula (a) is selected from the group consisting of C1-10alkyl, C2-10alkenyl and C2-10alkynyl, which C1-10alkyl, C2-10alkenyl and C2-10alkynyl are optionally interrupted by one or more chemical groups selected from the group consisting of —C(O)O—, —O—, —C(O)—, —C(O)N(R4)—, —S(O)2N(R4)—, —S(O)N(R4)—, —S(O)2—, —S(O)—, —N(R4)S(O)2N(R4a)—, —S—, —N(R4)—, —OC(OR4)(R4a)—, —N(R4)C(O)N(R4a)—, and —OC(O)N(R4)—; wherein —R4and —R4aare independently selected from the group consisting of —H, methyl, ethyl, propyl and butyl. Preferably —S1— of formula (a) is selected from the group consisting of methyl, ethyl, propyl, butyl, which are optionally interrupted by one or more chemical groups selected from the group consisting of —O—, —C(O)— and —C(O)N(R4)—.

In one embodiment —Sa′— of formula (a) is a chemical bond.

In another embodiment —Sa′— of formula (a) is selected from the group consisting of C1-10alkyl, C2-10alkenyl and C2-10alkynyl, which C1-10alkyl, C2-10alkenyl and C2-10alkynyl are optionally interrupted by one or more chemical groups selected from the group consisting of —C(O)O—, —O—, —C(O)—, —C(O)N(R4)—, —S(O)2N(R4)—, —S(O)N(R4)—, —S(O)2—, —S(O)—, —N(R4)S(O)2N(R4a)—, —S—, —N(R4)—, —OC(OR4)(R4a)—, —N(R4)C(O)N(R4a)—, and —OC(O)N(R4)—; wherein —R4and —R4aare independently selected from the group consisting of —H, methyl, ethyl, propyl and butyl. Preferably —S2— of formula (a) is selected from the group consisting of methyl, ethyl, propyl, butyl, which are optionally interrupted by one or more chemical groups selected from the group consisting of —O—, —C(O)— and —C(O)N(R4)—.

In one embodiment —Sa″— of formula (a) is a chemical bond.

In another embodiment —Sa″ of formula (a) is selected from the group consisting of C1-10alkyl, C2-10alkenyl and C2-10alkynyl, which C1-10alkyl, C2-10alkenyl and C2-10alkynyl are optionally interrupted by one or more chemical groups selected from the group consisting of —C(O)O—, —O—, —C(O)—, —C(O)N(R4)—, —S(O)2N(R4)—, —S(O)N(R4)—, —S(O)2—, —S(O)—, —N(R4)S(O)2N(R4a)—, —S—, —N(R4)—, —OC(OR4)(R4a)—, —N(R4)C(O)N(R4a)—, and —OC(O)N(R4)—; wherein —R4and —R4aare independently selected from the group consisting of —H, methyl, ethyl, propyl and butyl. Preferably —S3— of formula (a) is selected from the group consisting of methyl, ethyl, propyl, butyl, which are optionally interrupted by one or more chemical groups selected from the group consisting of —O—, —C(O)— and —C(O)N(R4)—.

In one embodiment —Sa′″— of formula (a) is a chemical bond.

In another embodiment —Sa′″of formula (a) is selected from the group consisting of C1-10alkyl, C2-10alkenyl and C2-10alkynyl, which C1-10alkyl, C2-10alkenyl and C2-10alkynyl are optionally interrupted by one or more chemical groups selected from the group consisting of —C(O)O—, —O—, —C(O)—, —C(O)N(R4)—, —S(O)2N(R4)—, —S(O)N(R4)—, —S(O)2—, —S(O)—, —N(R4)S(O)2N(R4a)—, —S—, —N(R4)—, —OC(OR4)(R4a)—, —N(R4)C(O)N(R4a)—, and —OC(O)N(R4)—; wherein —R4and —R4aare independently selected from the group consisting of —H, methyl, ethyl, propyl and butyl. Preferably —S4— of formula (a) is selected from the group consisting of methyl, ethyl, propyl, butyl, which are optionally interrupted by one or more chemical groups selected from the group consisting of —O—, —C(O)— and —C(O)N(R4)—.

Preferably, —Pa′, —Pa″and —Pa′″of formula (a) independently comprise a polymer selected from the group consisting of 2-methacryloyl-oxyethyl phosphoyl cholins, poly(acrylic acids), poly(acrylates), poly(acrylamides), poly(alkyloxy) polymers, poly(amides), poly(amidoamines), poly(amino acids), poly(anhydrides), poly(aspartamides), poly(butyric acids), poly(glycolic acids), polybutylene terephthalates, poly(caprolactones), poly(carbonates), poly(cyanoacrylates), poly(dimethylacrylamides), poly(esters), poly(ethylenes), poly(ethyleneglycols), poly(ethylene oxides), poly(ethyl phosphates), poly(ethyloxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl-oxazolines), poly(hydroxymethacrylates), poly(hydroxypropylmethacrylamides), poly(hydroxypropyl methacrylates), poly(hydroxypropyloxazo lines), poly(iminocarbonates), poly(lactic acids), poly(lactic-co-glycolic acids), poly(methacrylamides), poly(methacrylates), poly(methyloxazolines), poly(organophosphazenes), poly(ortho esters), poly(oxazo lines), polypropylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinyl amines), poly(vinylmethylethers), poly(vinylpyrrolidones), silicones, celluloses, carbomethyl celluloses, hydroxypropyl methylcelluloses, chitins, chitosans, dextrans, dextrins, gelatins, hyaluronic acids and derivatives, functionalized hyaluronic acids, mannans, pectins, rhamnogalacturonans, starches, hydroxyalkyl starches, hydroxyethyl starches and other carbohydrate-based polymers, xylans, and copolymers thereof.

Preferably, —Pa′, —Pa″and —Pa′″of formula (a) independently have a molecular weight ranging from and including 5 kDa to 50 kDa, more preferably have a molecular weight ranging from and including 5 kDa to 40 kDa, even more preferably ranging from and including 7.5 kDa to 35 kDa, even more preferably ranging from and 7.5 to 30 kDa, even more preferably ranging from and including 10 to 30 kDa.

In one embodiment —Pa′, —Pa″and —Pa′″of formula (a) have a molecular weight of about 5 kDa.

In another embodiment —Pa′, —Pa″and —Pa′″of formula (a) have a molecular weight of about 7.5 kDa.

In another embodiment —Pa′, —Pa″and —Pa′″of formula (a) have a molecular weight of about 10 kDa.

In another embodiment —Pa′, —Pa″and —Pa′″of formula (a) have a molecular weight of about 12.5 kDa.

In another embodiment —Pa′, —Pa″and —Pa′″of formula (a) have a molecular weight of about 15 kDa.

In another embodiment —Pa′, —Pa″and —Pa′″of formula (a) have a molecular weight of about 20 kDa.

More preferably, —Pa′, —Pa″and —Pa′″of formula (a) independently comprise a PEG-based moiety. Even more preferably, —Pa′, —Pa″and —Pa′″of formula (a) independently comprise a PEG-based moiety comprising at least 20% PEG, even more preferably at least 30%, even more preferably at least 40% PEG, even more preferably at least 50% PEG, even more preferably at least 60% PEG, even more preferably at least 70% PEG, even more preferably at least 80% PEG and most preferably at least 90% PEG.

In an equally preferred embodiment —Pa′, —Pa″and —Pa′″of formula (a) independently comprise a protein moiety, more preferably a random coil protein moiety and most preferably a random coil protein moiety selected from the group consisting of PA, PAS, PAG, PG and XTEN moieties.

In one embodiment —Pa′, —Pa″and —Pa′″of formula (a) are a PA moiety.

In another embodiment —Pa′, —Pa″and —Pa′″of formula (a) are a PAS moiety.

In another embodiment —Pa′, —Pa″and —Pa′″of formula (a) are a PAG moiety.

In another embodiment —Pa′, —Pa″and —Pa′″of formula (a) are a PG moiety.

In another embodiment —Pa′, —Pa″and —Pa′″of formula (a) are an XTEN moiety.

In one embodiment —Z of formula (I) comprises one moiety of formula (a).

In another embodiment —Z of formula (I) comprises two moieties of formula (a).

In another embodiment —Z of formula (I) comprises three moieties of formula (a).

In another embodiment —Z of formula (I) comprises four moieties of formula (a).

In another embodiment —Z of formula (I) comprises five moieties of formula (a).

In another embodiment —Z of formula (I) comprises six moieties of formula (a).

In a preferred embodiment —Z of formula (I) comprises two moieties of formula (a).

In a preferred embodiment —Z of formula (I) comprises a moiety of formula (b)

whereinthe dashed line indicates attachment to -L2- or to the remainder of —Z;b1 is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7 and 8;b2 is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7 and 8;b3 is an integer ranging from and including 150 to 1000; preferably ranging from and including 150 to 500; and most preferably ranging from and including 200 to 460; andb4 is an integer ranging from and including 150 to 1000; preferably ranging from and including 150 to 500; and most preferably ranging from and including 200 to 460.

Optionally, the moiety of formula (b) is substituted with one or more substituents.

Preferably, b3 and b4 of formula (b) are the same integer.

In one preferred embodiment b3 and b4 both an integer ranging from 200 to 250 and most preferably b3 and b4 of formula (b) are about 225.

In another preferred embodiment b3 and b4 are both an integer ranging from 400 to 500 and most preferably b3 and b4 of formula (b) are about 450.

Preferably, b1 of formula (b) is selected from the group consisting of 0, 1, 2, 3 and 4. More preferably b1 of formula (b) is selected from the group consisting of 1, 2 and 3. Most preferably b1 of formula (b) is 2.

Preferably, b2 of formula (b) is selected from the group consisting of 1, 2, 3, 4 and 5. More preferably b2 of formula (b) is selected from the group consisting of 2, 3 and 4. Most preferably b2 of formula (b) is 3.

In one particularly preferred embodiment b1 of formula (b) is 2, b2 of formula (b) is 3, and b3 and b4 are both about 450.

In another particularly preferred embodiment b1 of formula (b) is 2, b2 of formula (b) is 3, and b3 and b4 are both about 225.

In one embodiment —Z comprises one moiety of formula (b).

In another embodiment —Z comprises two moieties of formula (b).

In another embodiment —Z comprises three moieties of formula (b).

In another embodiment —Z comprises four moieties of formula (b).

In another embodiment —Z comprises five moieties of formula (b).

In another embodiment —Z comprises six moieties of formula (b).

In a preferred embodiment —Z comprises two moieties of formula (b).

In an even more preferred embodiment —Z comprises a moiety of formula (c)

whereinthe dashed line indicates attachment to -L2- or to the remainder of —Z;c1 and c2 are independently an integer ranging from and including 150 to 500;preferably ranging from and including 200 to 460.

Preferably both c1 and c2 of formula (c) are the same integer.

In one preferred embodiment c1 and c2 of formula (c) range from and include 200 to 250 and most preferably are about 225. In another preferred embodiment c1 and c2 of formula (c) range from and include 400 to 500 and most preferably are about 450.

In a preferred embodiment the moiety —Z is a branched PEG-based polymer comprising at least 10% PEG, has one branching point and two PEG-based polymer arms and has a molecular weight of about 40 kDa. Accordingly, each of the two PEG-based polymer arms has a molecular weight of about 20 kDa. Preferably the branching point is —CH<.

In one embodiment —Z of formula (I) comprises one moiety of formula (c).

In another embodiment —Z of formula (I) comprises two moieties of formula (c).

In another embodiment —Z of formula (I) comprises three moieties of formula (c).

In another embodiment —Z of formula (I) comprises four moieties of formula (c).

In another embodiment —Z of formula (I) comprises five moieties of formula (c).

In another embodiment —Z of formula (I) comprises six moieties of formula (c).

In a preferred embodiment —Z of formula (I) comprises two moieties of formula (c).

In one preferred embodiment the moiety —Z of formula (I) is of formula (d)

whereinthe dashed line indicates attachment to -L2-;—Zb— is selected from the group consisting of C1-50alkyl, C2-50alkenyl, and C2-50alkynyl; wherein C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally substituted with one or more —R1, which are the same or different and wherein C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T-, —C(O)O—, —O—, —C(O)—, —C(O)N(R2)—, —S(O)2N(R2)—, —S(O)N(R2)—, —S(O)2—, —S(O)—, —N(R2)S(O)2N(R2a)—, —S—, —N(R2)—, —OC(OR2)(R2a)—, —N(R2)C(O)N(R2a)—, and —OC(O)N(R2)—;each -T- is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C3-10cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl; wherein each -T- is independently optionally substituted with one or more —R1, which are the same or different;each —R1is independently selected from the group consisting of halogen, —CN, oxo (═O), —COOR3, —OR3, —C(O)R3, —C(O)N(R3R3a), —S(O)2N(R3R3a), —S(O)N(R3R3a), —S(O)2R3, —S(O)R3, —N(R3)S(O)2N(R3aR3b), —SR3, —N(R3R3a), —NO2—, —OC(O)R3, —N(R3)C(O)R3a, —N(R3)S(O)2R3a, —N(R3)S(O)R3a, —N(R3)C(O)OR3a, —N(R3)C(O)N(R3aR3a), —OC(O)N(R3R3a), and C1-6alkyl;wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different;each —R2, —R2a, —R3, —R3aand —R3bis independently selected from the group consisting of —H, and C1-6alkyl, wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different;and—Zais

whereinBPa, —Sa—, —Sa′—, —Sa″—, —Sa′″—, —Pa′, —Pa″, —Pa′″and a are used as defined for formula (a).

Optionally, the moiety of formula (d) is substituted with one or more substituents.

Preferred embodiments of BPa, —Sa—, —Sa′—, —Sa″—, —Sa′″—, —Pa′, —Pa″, —Pa′″of formula (d) are as defined above for formula (a).

Preferably, —Zaof formula (d) is of formula (b). Preferred embodiments of b1, b2, b3 and b4 are as described for formula (b).

Even more preferably, —Zaof formula (d) is of formula (c). Preferred embodiments for c1 and c2 are as described for formula (c).

In an even more preferred embodiment the moiety —Z of formula (I) is of formula (e)

whereinthe dashed line indicates attachment to -L2-;e is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 and 15; and—Zais

whereinb1, b2, b3 and b4 are used as defined for formula (b).

Optionally, the moiety of formula (e) is substituted with one or more substituents.

Preferred embodiments for b1, b2, b3 and b4 of formula (e) are as defined above for formula (b).

In one embodiment e of formula (e) is 1. In another embodiment e of formula (e) is 2. In another embodiment e of formula (e) is 3. In another embodiment e of formula (e) is 4. In another embodiment e of formula (e) is 5. In another embodiment e of formula (e) is 6. In another embodiment e of formula (e) is 7. In another embodiment e of formula (e) is 8. In another embodiment e of formula (e) is 9. In another embodiment e of formula (e) is 10. In another embodiment e of formula (e) is 11. In another embodiment e of formula (e) is 12. In another embodiment e of formula (e) is 13. In another embodiment e of formula (e) is 14. In another embodiment e of formula (e) is 15.

Preferably e of formula (e) is selected from the group consisting of 2, 3, 4, 5, 6, 7, 8 and 9.

Even more preferably, e of formula (e) is selected from 3, 4, 5 and 6. Most preferably e of formula (e) is 5.

Preferably e of formula (e) is 5, b1 of formula (e) is 2, b2 of formula (e) is 3 and b3 and b4 of formula (e) are both about 450.

In an equally preferred embodiment the moiety —Z of formula (I) is of formula (e-i) or (e-i′):

whereinthe dashed line indicates attachment to -L2-,e is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 and 15;—Zais

whereinb1, b2, b3 and b4 are used as defined for formula (b).

Preferred embodiments for b1, b2, b3 and b4 of formula (e-i) and (e-i′) are as defined above for formula (b).

Preferred embodiments for e of formula (e-i) and (e-i′) are as described for formula (e).

Preferably, b1 of formula (e-i) and (e-i′) is 2, b2 of formula (e-i) and (e-i′) is 3 and b3 and b4 of formula (e-i) and (e-i′) are both about 450.

In a preferred embodiment —Z of formula (I) is of formula (e-i).

In another preferred embodiment the moiety —Z is a branched PEG-based polymer comprising at least 10% PEG, has three branching points and four PEG-based polymer arms and has a molecular weight of about 40 kDa. Accordingly, each of the four PEG-based polymer arms has a molecular weight of about 10 kDa. Preferably each of the three branching points is —CH<.

In a preferred embodiment the moiety —Z is of formula (f)

whereinthe dashed line indicates attachment to -L2-;BPfis a branching point selected from the group consisting of —N<, —CR< and >C<;—R is selected from the group consisting of —H and C1-6alkyl;f is 0 if BPfis —N< or —CR< and f is 1 if BPfis >C<;—Sf—, —Sf—, —Sf′— and —Sf″— are independently either a chemical bond or are independently selected from the group consisting of C1-50alkyl, C2-50alkenyl, and C2-50alkynyl; wherein C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally substituted with one or more —R1, which are the same or different and wherein C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T-, —C(O)O—, —O—, —C(O)—, —C(O)N(R2)—, —S(O)2N(R2)—, —S(O)N(R2)—, —S(O)2—, —S(O)—, —N(R2)S(O)2N(R2a)—, —S—, —N(R2)—, —OC(OR2)(R2a)—, —N(R2)C(O)N(R2a)—, and —OC(O)N(R2)—;each -T- is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C3-10cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl; wherein each -T- is independently optionally substituted with one or more —R1, which are the same or different;each R1is independently selected from the group consisting of halogen, —CN, oxo (═O), —COOR3, —OR3, —C(O)R3, —C(O)N(R3R3a), —S(O)2N(R3R3a), —S(O)N(R3R3a), —S(O)2R3, —S(O)R3, —N(R3)S(O)2N(R3aR3b), —SR3, —N(R3R3a), —NO2—, —OC(O)R3, —N(R3)C(O)R3a, —N(R3)S(O)2R3a, —N(R3)S(O)R3a, —N(R3)C(O)OR3a, —N(R3)C(O)N(R3aR3a), —OC(O)N(R3R3a), and C1-6alkyl; wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different;each —R2, —R2a, —R3, —R3aand —R3bis independently selected from the group consisting of —H, and C1-6alkyl, wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different;and—Za′, —Za″and —Za′″are independently

whereinBPa, —Sa—, —Sa′—, —Sa″—, —Sa′″—, —Pa′, —Pa″, —Pa′″and a are used as defined for formula (a).

Optionally, the moiety of formula (f) is substituted with one or more substituents.

Preferred embodiments of BPa, —Sa—, —Sa′—, —Sa″—, —Sa′″—, —Pa′, —Pa″, —Pa′″of formula (f) are as defined above for formula (a).

Preferably BPfof formula (f) is —CR< and r is 0. Preferably —R is —H.

Preferably —Sf— of formula (f) is a chemical bond.

Preferably, —Za′, —Za″and —Za′″of formula (f) have the same structure. Preferably, —Za′, —Za″and —Za′″of formula (f) are of formula (b).

Preferred embodiments of b1, b2, b3 and b4 are as described for formula (b).

Preferably —Sf— of formula (f) is a chemical bond, BPaof formula (f) is —CR< with —R being —H. Even more preferably —Sf— of formula (f) is a chemical bond, BPaof formula (f) is —CR< with —R being —H and —Za′, —Za″and —Za′″of formula (f) are of formula (b).

Even more preferably —Z is of formula (g)

whereinthe dashed line indicates attachment to -L2-;—S8—, —S8′— and S8″— are independently selected from the group consisting of C1-50alkyl, C2-50alkenyl, and C2-50alkynyl; wherein C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally substituted with one or more —R1, which are the same or different and wherein C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T-, —C(O)O—, —O—, —C(O)—, —C(O)N(R2)—, —S(O)2N(R2)—, —S(O)N(R2)—, —S(O)2—, —S(O)—, —N(R2)S(O)2N(R2a)—, —S—, —N(R2)—, —OC(OR2)(R2a)—, —N(R2)C(O)N(R2a)—, and —OC(O)N(R2)—;each -T- is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C3-10cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl; wherein each -T- is independently optionally substituted with one or more —R1, which are the same or different; each R1is independently selected from the group consisting of halogen, —CN, oxo (═O), —COOR3, —OR3, —C(O)R3, —C(O)N(R3R3a), —S(O)2N(R3R3a), —S(O)N(R3R3a), —S(O)2R3, —S(O)R3, —N(R3)S(O)2N(R3aR3b), —SR3, —N(R3R3a), —NO2—, —OC(O)R3, —N(R3)C(O)R3a, —N(R3)S(O)2R3a, —N(R3)S(O)R3a, —N(R3)C(O)OR3a, —N(R3)C(O)N(R3aR3a), —OC(O)N(R3R3a), and C1-6alkyl;wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different;each —R2, —R2a, —R3, —R3aand —R3bis independently selected from the group consisting of —H, and C1-6alkyl, wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different;and—Zaand —Za′are independently

whereinBPa, —Sa—, —Sa′—, —Sa″—, —Sa′″—, —Pa′, —Pa″, —Pa′″and a are used as defined for formula (a).

Optionally, the moiety of formula (g) is substituted with one or more substituents.

Preferred embodiments of BPa, —Sa—, —Sa′—, —Sa″—, —Sa′″—, —Pa′, —Pa″, —Pa′″of formula (g) are as defined above for formula (a).

Preferably, —Sg— of formula (g) is selected from the group consisting of C1-6alkyl, C2-6alkenyl and C2-6alkynyl, which are optionally substituted with one or more —R1, which is the same or different,wherein—R1is selected from the group consisting of halogen, oxo (═O), —COOR3, —OR3, —C(O)R3, —C(O)N(R3R3a), —S(O)2N(R3R3a), —S(O)N(R3R3a), —S(O)2R3, —S(O)R3, —N(R3)S(O)2N(R3aR3b), —SR3, —N(R3R3a), —NO2—, —OC(O)R3, —N(R3)C(O)R3a, —N(R3)S(O)2R3a, —N(R3)S(O)R3a, —N(R3)C(O)OR3a, —N(R3)C(O)N(R3aR3a), —OC(O)N(R3R3a), and C1-6alkyl; wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different; and —R3, —R3aand —R3bare independently selected from —H, methyl, ethyl, propyl and butyl.

Even more preferably —Sg— of formula (g) is selected from C1-6alkyl.

Preferably, —Sg′— of formula (g) is selected from the group consisting of C1-6alkyl, C2-6alkenyl and C2-6alkynyl, which are optionally substituted with one or more —R1, which is the same or different,wherein—R1is selected from the group consisting of halogen, oxo (═O), —COOR3, —OR3, —C(O)R3, —C(O)N(R3R3a), —S(O)2N(R3R3a), —S(O)N(R3R3a), —S(O)2R3, —S(O)R3, —N(R3)S(O)2N(R3aR3b), —SR3, —N(R3R3a), —NO2—, —OC(O)R3, —N(R3)C(O)R3a, —N(R3)S(O)2R3a, —N(R3)S(O)R3a, —N(R3)C(O)OR3a, —N(R3)C(O)N(R3aR3a), —OC(O)N(R3R3a), and C1-6alkyl; wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different; and —R3, —R3aand —R3bare independently selected from —H, methyl, ethyl, propyl and butyl.

Even more preferably —Sg′— of formula (g) is selected from C1-6alkyl.

Preferably, —S8″— of formula (g) is selected from the group consisting of C1-6alkyl, C2-6alkenyl and C2-6alkynyl, which are optionally substituted with one or more —R1, which is the same or different,wherein—R1is selected from the group consisting of halogen, oxo (═O), —COOR3, —OR3, —C(O)R3, —C(O)N(R3R3a), —S(O)2N(R3R3a), —S(O)N(R3R3a), —S(O)2R3, —S(O)R3, —N(R3)S(O)2N(R3aR3b), —SR3, —N(R3R3a), —NO2—, —OC(O)R3, —N(R3)C(O)R3a, —N(R3)S(O)2R3a, —N(R3)S(O)R3a, —N(R3)C(O)OR3a, —N(R3)C(O)N(R3aR3a), —OC(O)N(R3R3a), and C1-6alkyl; wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different; and —R3, —R3aand —R3bare independently selected from —H, methyl, ethyl, propyl and butyl.

Even more preferably —Sg″of formula (g) is selected from C1-6alkyl.

Preferably, —Zaand —Za′of formula (g) have the same structure. Preferably, —Zaand —Za′of formula (g) are of formula (b).

In an alternative even more preferred embodiment —Z is of formula (g-i)

whereinthe dashed line indicates attachment to -L2-;—Sg—, —Sg′— and —Sg″are independently selected from the group consisting of C1-50alkyl, C2-50alkenyl, and C2-50alkynyl; wherein C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally substituted with one or more —R1, which are the same or different and wherein C1-50alkyl, C2-50alkenyl, and C2-50alkynyl are optionally interrupted by one or more groups selected from the group consisting of -T-, —C(O)O—, —O—, —C(O)—, —C(O)N(R2)—, —S(O)2N(R2)—, —S(O)N(R2)—, —S(O)2—, —S(O)—, —N(R2)S(O)2N(R2a)—, —S—, —N(R24)—, —OC(OR24)(R2a)—, —N(R24)C(O)N(R2a)—, and —OC(O)N(R24)—;each -T- is independently selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C3-10cycloalkyl, 3- to 10-membered heterocyclyl, 8- to 11-membered heterobicyclyl, 8- to 30-membered carbopolycyclyl, and 8- to 30-membered heteropolycyclyl; wherein each -T- is independently optionally substituted with one or more —R1, which are the same or different;each R1is independently selected from the group consisting of halogen, —CN, oxo (═O), —COOR3, —OR3, —C(O)R3, —C(O)N(R3R3a), —S(O)2N(R3R3a), —S(O)N(R3R3a), —S(O)2R3, —S(O)R3, —N(R3)S(O)2N(R3aR3b), —SR3, —N(R3R3a), —NO2—, —OC(O)R3, —N(R3)C(O)R3a, —N(R3)S(O)2R3a, —N(R3)S(O)R3a, —N(R3)C(O)OR3a, —N(R3)C(O)N(R3aR3a), —OC(O)N(R3R3a), and C1-6alkyl;wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different;each —R2, —R2a, —R3, —R3aand —R3bis independently selected from the group consisting of —H, and C1-6alkyl, wherein C1-6alkyl is optionally substituted with one or more halogen, which are the same or different;—Ya1— and —Ya1′— are

and—Zaand —Za′are independently

whereinBPa, —Sa—, —Sa′—, —Sa″—, —Sa′″—, —Pa′, —Pa″, —Pa′″and a are used as defined for formula (a).

Optionally, the moiety of formula (g-i) is substituted with one or more substituents.

Preferably, —Ya1— and —Ya1′— of formula (g-i) are both


wherein the dashed line marked with the asterisk is attached to —Zaor —Za′, respectively.

Preferred embodiments of BPa, —Sa—, —Sa′—, —Sa″—, —Sa′″—, —Pa′, —Pa″, —Pa′″of formula (g-i) are as defined above for formula (a).

Preferred embodiments of —Sg—, —Sg′— and —Sg″— of formula (g-i) are as defined for formula (g).

Preferably, —Zaand —Za′of formula (g-i) have the same structure. Preferably, —Zaand —Za′of formula (g-i) are of formula (b). Preferred embodiments for b1, b2, b3 and b4 are as described for formula (b).

Even more preferably —Z is of formula (h)

whereinthe dashed line indicates attachment to -L2-; andeach —Zcis a moiety

whereineach c1 is an integer independently ranging from about 200 to 250.

Optionally, the moiety of formula (h) is substituted with one or more substituents.

Preferably both c1 of formula (h) are the same.

Preferably both c1 of formula (h) are about 225.

Even more preferably —Z is of formula (h-a)

whereinthe dashed line indicates attachment to -L2-;each k is independently of each other selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12;—Ya1— and —Ya1′— are

andeach —Zcis a moiety

whereineach c1 is an integer independently ranging from about 200 to 250.

Optionally, the moiety of formula (h-a) is substituted with one or more substituents.

Preferably, each k of formula (h-a) is independently selected from the group consisting of 2, 3, 4, 5, 6 and 7. Preferably, both k of formula (h-a) are identical.

Preferably both c1 of formula (h-a) are the same.

Preferably both c1 of formula (h-a) are about 225.

Preferably, —Ya1— and —Ya1′— of formula (h-a) are both


wherein the dashed line marked with the asterisk is attached to —Zc.

In an even more preferred embodiment the moiety —Z is of formula (h-i)

whereinthe dashed line indicates attachment to -L2-; andeach —Zcis a moiety

each c1 is an integer independently ranging from 200 to 250.

Optionally, the moiety of formula (h-i) is substituted with one or more substituents.

Preferably both c1 of formula (h-i) are the same.

Preferably both c1 of formula (h-i) are about 225.

In an alternative even more preferred embodiment the moiety —Z is of formula (h-ia)

whereinthe dashed line indicates attachment to -L2-;each k is independently of each other selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12;—Ya1— and —Ya1′— are

andeach —Zcis a moiety



each c1 is an integer independently ranging from 200 to 250.

Preferably, each k of formula (h-ia) is independently selected from the group consisting of 2, 3, 4, 5, 6 and 7. Preferably, both k of formula (h-ia) are identical.

Preferably both c1 of formula (h-ia) are the same.

Preferably both c1 of formula (h-ia) are about 225.

Preferably, —Ya1— and —Ya1′— of formula (h-ia) are both


wherein the dashed line marked with the asterisk is attached to —Zc.

In an equally preferred the embodiment —Z comprises a moiety selected from the group consisting of


whereinthe dashed line indicates attachment to -L2-;s1, s2, s3, s4, s5, s6, s7, s8, s9, s10, s11, s12, s13, s14 and s15 are independently of each other selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10;

—Xd1, —Xd2, —Xd3and —Xd4are independently of each other selected from the group consisting of —OH, —SH and —NRg1Rg2; preferably —OH;

—Xe1, —Xe2, —Xe3and —Xe4are independently of each other selected from the group consisting of —H, C1-6alkyl, C2-6alkenyl and C2-6alkynyl;

Rg1and —Rg2are independently of each other selected from the group consisting of —H, C1-6alkyl, C2-6alkenyl and C2-6alkynyl;

—Xf1, —Xf2, —Xf3, —Xf4, —Xf5, —Xf6, —Xf7, —Xf8, —Xf9, —Xf10, —Xf11, —Xf12, —Xf13and —Xf14, are independently of each other selected from the group consisting of —H, C1-6alkyl, C2-6alkenyl and C2-6alkynyl; preferably —H;

—Yd1—, —Yd2—, —Yd3— and —Yd4— are independently of each other selected from the group consisting of


and —Zd1, —Zd2, —Zd3and —Zd4are independently of each other a protein, more preferably a random coil protein and most preferably a random coil protein selected from the group consisting of PA, PAS, PAG, PG and XTEN.

In one preferred embodiment, —Yd1— and —Yd2— of formula (j-iv), (j-v) and (j-vi) and —Yd1—, —Xd2—, —Xd3— and —Xd4— of formula (j-vii) are



In another preferred embodiment, —Yd1— and —Yd2— of formula (j-iv), (j-v) and (j-vi) and —Xd1—, —Xd2—, —Xd3— and —Xd4— of formula (j-vii) are


wherein the dashed line marked with the asterisk is oriented towards —Zd1, —Zd2, —Zd3and —Zd4, respectively, and the unmarked dashed line is oriented towards -L2-.

Preferably, —Xf1, —Xf2, —Xf3, —Xf4, —Xf5, —Xf6, —Xf7and —Xf8, of formula (j-i) are —H; —Xd1and —Xd2of formula (j-i) are —OH; —Xe1and —Xe2of formula (j-i) are selected from the group consisting of —H and methyl; and s1, s2, s3 and s4 of formula (j-i) are selected from the group consisting of 2, 3, 4, 5 and 6. Even more preferably —Xf1, —Xf2, —Xf3, —Xf4, —Xf5, —Xf6, —Xf7and —Xf8of formula (j-i) are —H; —Xd1and —Xd2of formula (j-i) are —OH; —Xe1and —Xe2of formula (j-i) are —H; and s1, s2, s3 and s4 of formula (j-i) are 4.

Preferably, —Xf1, —Xf2, —Xf3and —Xf4of formula (j-ii) are —H; —Xd1, —Xd2, —Xd3and —Xd2of formula (j-ii) are —OH; —Xe1, —Xe2, —Xe3and —Xe4of formula (j-ii) are selected from the group consisting of —H and methyl; s1, s2, s3, s4 and s5 of formula (j-ii) are selected from the group consisting of 1, 2, 3, 4, 5 and 6. Even more preferably —Xf1, —Xf2, —Xf3and —Xf4of formula (j-ii) are —H; —Xd1, —Xd2, —Xd3and —Xd2of formula (j-ii) are —OH; —Xe1, —Xe2, —Xe3and —Xe4of formula (j-ii) are —H; s1 is 4 of formula (j-ii) and s2, s3, s4 and s5 of formula (j-ii) are 1.

Preferably, —Xf1, —Xf2, —Xf3, —Xf4, —Xf5, —Xf6, —Xf7, —Xf8, —Xf9and —Xf10of formula (j-iii) are —H; —Xd1, —Xd2, —Xd3and —Xd4of formula (j-iii) are —OH; —Xe1, —Xe2, —Xe3and —Xe4of formula (j-iii) are selected from the group consisting of —H and methyl; and s1, s2 and s3 of formula (j-iii) are selected from the group consisting of 2, 3, 4, 5 and 6. Even more preferably —Xf1, —Xf2, —Xf3, —Xf4, —Xf5, —Xf6, —Xf7, —Xf8, —Xf9and —Xf10of formula (j-iii) are —H; —Xd1, —Xd2, —Xd3and —Xd4of formula (j-iii) are —OH; —Xe1, —Xe2, —Xe3and —Xe4of formula (j-iii) are —H; and s1, s2 and s3 of formula (j-iii) are 4.

Preferably, —Xf1, —Xf2, —Xf3, —Xf4, —Xf5and —Xf6of formula (j-iv) are —H; s1, s2, s3, s4, s5, s6 and s7 of formula (j-iv) are selected from the group consisting of 1, 2, 3, 4, 5, 6 and 7; —Yd1— and —Yd2— are selected from the group consisting of



In an even more preferred embodiment —Xf1, —Xf2, —Xf3, —Xf4, —Xf5and —Xf6of formula (j-iv) are —H; s1 of formula (j-iv) is 3, s2 of formula (j-iv) is 5, s3 of formula (j-iv) is 2, s4 of formula (j-iv) is 4, s5 of formula (j-iv) is 5, s6 of formula (j-iv) is 2 and s7 of formula (j-iv) is 4; and —Yd1— and —Yd2— of formula (j-iv) are



In an equally preferred embodiment —Xf1, —Xf2, —Xf3, —Xf4, —Xf5and —Xf6of formula (j-iv) are —H; s1 of formula (j-iv) is 3, s2 of formula (j-iv) is 5, s3 of formula (j-iv) is 2, s4 of formula (j-iv) is 4, s5 of formula (j-iv) is 5, s6 of formula (j-iv) is 2 and s7 of formula (j-iv) is 4; and —Yd1— and —Yd2— of formula (j-iv) are



wherein the dashed line marked with the asterisk is oriented towards —Zd1, —Zd2, —Zd3and —Zd4respectively, and the unmarked dashed line is oriented towards -L2-.

Preferably, —Xf1, —Xf2, —Xf3and —Xf4of formula (j-v) are —H; s1, s2, s3, s4 and s5 of formula (j-v) are selected from the group consisting of 1, 2, 3, 4, 5, 6 and 7; —Yd1— and —Yd2— of formula (j-v) are selected from the group consisting of



In an even more preferred embodiment —Xf1, —Xf2, —Xf3and —Xf4of formula (j-v) are —H; s1 of formula (j-v) is 3, s2 of formula (j-v) is 2, s3 of formula (j-v) is 1, s4 of formula (j-v) is 2 and s5 of formula (j-v) is 1; and —Yd1— and —Yd2— of formula (j-v) are


In an equally preferred embodiment —Xf1, —Xf2, —Xf3and —Xf4of formula (j-v) are —H; s1 of formula (j-v) is 3, s2 of formula (j-v) is 2, s3 of formula (j-v) is 1, s4 of formula (j-v) is 2 and s5 of formula (j-v) is 1; and —Yd1— and —Yd2— of formula (j-v) are



wherein the dashed line marked with the asterisk is oriented towards —Zd1, —Zd2, —Zd3and —Zd4, respectively, and the unmarked dashed line is oriented towards -L2-.

Preferably, —Xf1, —Xf2, —Xf3, —Xf4, —Xf5, —Xf6, —Xf7, —Xf8, —Xf9and —Xf10of formula (j-vi) are —H s1, s2, s3, s4, s5, s6, s7, s8 and s9 of formula (j-vi) are selected from the group consisting of 1, 2, 3, 4, 5, 6 and 7; —Yd1— and —Yd2— of formula (j-vi) are selected from the group consisting of



In an even more preferred embodiment —Xf1, —Xf2, —Xf3, —Xf4, —Xf5, —Xf6, —Xf7, —Xf8, —Xf9and —Xf10of formula (j-vi) are —H; s1 of formula (j-vi) is 4, s2 of formula (j-vi) is 5, s3 of formula (j-vi) is 2, s4 of formula (j-vi) is 4, s5 of formula (j-vi) is 4, s6 of formula (j-vi) is 5, s7 of formula (j-vi) is 2, s8 of formula (j-vi) is 4 and s9 of formula (j-vi) is 4; and —Yd1— and —Yd2— of formula (j-v) are



In an equally preferred embodiment —Xf1, —Xf2, —Xf3, —Xf4, —Xf5, —Xf6, —Xf7, —Xf8, —Xf9, —Xf10of formula (j-vi) are —H; s1 of formula (j-vi) is 4, s2 of formula (j-vi) is 5, s3 of formula (j-vi) is 2, s4 of formula (j-vi) is 4, s5 of formula (j-vi) is 4, s6 of formula (j-vi) is 5, s7 of formula (j-vi) is 2, s8 of formula (j-vi) is 4 and s9 of formula (j-vi) is 4; and —Yd1— and —Yd2— of formula (j-v) are


wherein the dashed line marked with the asterisk is oriented towards —Zd1, —Zd2, —Zd3and —Zd4, respectively, and the unmarked dashed line is oriented towards -L2-.

Preferably, —Xf1, —Xf2, —Xf3, —Xf4, —Xf5, —Xf6, —Xf7, —Xf8, —Xf9, —Xf10, —Xf11, —Xf12, —Xf13and —Xf14of formula (j-vii) are —H; s1, s2, s3, s4, s5, s6, s7, s8, s9, s10, s11, s12, s13, s14 and s15 of formula (j-vii) are selected from the group consisting of 1, 2, 3, 4, 5, 6 and 7; —Yd1—, —Yd2—, —Yd3— and —Yd4— of formula (j-vii) are selected from the group consisting of



In an even more preferred embodiment —Xf1, —Xf2, —Xf3, —Xf4, —Xf5, —Xf6, —Xf7, —Xf8, —Xf9, —Xf10, —Xf11, —Xf12, —Xf13and —Xf14of formula (j-vii) are —H; are —H; s1 of formula (j-vii) is 4, s2 of formula (j-vii) is 4, s3 of formula (j-vii) is 5, s4 of formula (j-vii) is 2, s5 of formula (j-vii) is 4, s6 of formula (j-vii) is 5, s7 of formula (j-vii) is 2, s8 of formula (j-vii) is 4, s9 of formula (j-vii) is 4, s10 of formula (j-vii) is 5, s11 of formula (j-vii) is 2, s12 of formula (j-vii) is 4, s13 of formula (j-vii) is 5, s14 of formula (j-vii) is 2 and s15 of formula (j-vii) is 4; and —Yd1—, —Yd2—, —Yd3— and —Yd4— of formula (j-vii) are


In an equally preferred embodiment —Xf1, —Xf2, —Xf3, —Xf4, —Xf5, —Xf6, —Xf7, —Xf8, —Xf9, —Xf10, —Xf11, —Xf12, —Xf13and —Xf14of formula (j-vii) are —H; are —H; s1 of formula (j-vii) is 4, s2 of formula (j-vii) is 4, s3 of formula (j-vii) is 5, s4 of formula (j-vii) is 2, s5 of formula (j-vii) is 4, s6 of formula (j-vii) is 5, s7 of formula (j-vii) is 2, s8 of formula (j-vii) is 4, s9 of formula (j-vii) is 4, s10 of formula (j-vii) is 5, s11 of formula (j-vii) is 2, s12 of formula (j-vii) is 4, s13 of formula (j-vii) is 5, s14 of formula (j-vii) is 2 and s15 of formula (j-vii) is 4; and —Yd1—, —Yd2—, —Yd3— and —Yd4— of formula (j-vii) are



wherein the dashed line marked with the asterisk is oriented towards —Zd1, —Zd2, —Zd3and —Zd4, respectively, and the unmarked dashed line is oriented towards -L2-.

Preferably —Zd1, —Zd2, —Zd3and —Zd4of formula (j-i), (j-ii), (j-iii), (j-iv), (j-v), (j-vi) and (j-vii) have the same structure.

In one embodiment —Zd1, —Zd2, —Zd3and —Zd4of formula (j-i), (j-ii), (j-iii), (j-iv), (j-v), (j-vi) and (j-vii) are a PA moiety.

In another embodiment —Zd1, —Zd2, —Zd3and —Zd4of formula (j-i), (j-ii), (j-iii), (j-iv), (j-v), (j-vi) and (j-vii) are a PAS moiety.

In another embodiment —Zd1, —Zd2, —Zd3and —Zd4of formula (j-i), (j-ii), (j-iii), (j-iv), (j-v), (j-vi) and (j-vii) are a PAG moiety.

In another embodiment —Zd1, —Zd2, —Zd3and —Zd4of formula (j-i), (j-ii), (j-iii), (j-iv), (j-v), (j-vi) and (j-vii) are a PG moiety.

In another embodiment —Zd1, —Zd2, —Zd3and —Zd4of formula (j-i), (j-ii), (j-iii), (j-iv), (j-v), (j-vi) and (j-vii) are a XTEN moiety.

In a preferred embodiment the CNP prodrug of the present invention is of formula (IIe)

whereinthe unmarked dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to a moiety

whereineach c1 is an integer independently ranging from 400 to 500.

Preferably, c1 of formula (IIe) is about 450.

In an equally preferred embodiment the CNP prodrug of the present invention is of formula (IIe-i)

whereinthe unmarked dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to a moiety

whereineach c1 is an integer independently ranging from 400 to 500.

Preferably, c1 of formula (IIe-i) is about 450.

In another equally preferred embodiment the CNP prodrug of the present invention is of formula (IIe-ii)

whereinthe unmarked dashed line indicates attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to a moiety

whereineach c1 is an integer independently ranging from 400 to 500.

Preferably, c1 of formula (IIe-ii) is about 450.

In one embodiment the CNP moiety of the CNP prodrug of formula (IIe), (IIe-i) and (IIe-ii) has the sequence of SEQ ID NO:25.

In another embodiment the CNP moiety of the CNP prodrug of formula (IIe), (IIe-i) and (IIe-ii) has the sequence of SEQ ID NO:30.

In another embodiment the CNP moiety of the CNP prodrug of formula (IIe), (IIe-i) and (IIe-ii) has the sequence of SEQ ID NO:20.

In another embodiment the CNP moiety of the CNP prodrug of formula (IIe), (IIe-i) and (IIe-ii) has the sequence of SEQ ID NO:21.

In another embodiment the CNP moiety of the CNP prodrug of formula (IIe), (IIe-i) and (IIe-ii) has the sequence of SEQ ID NO:22.

In another embodiment the CNP moiety of the CNP prodrug of formula (IIe), (IIe-i) and (IIe-ii) has the sequence of SEQ ID NO:23.

In a preferred embodiment the CNP moiety of the CNP prodrug of formula (IIe), (IIe-i) and (IIe-ii) has the sequence of SEQ ID NO:24.

In a preferred embodiment the CNP prodrug of the present invention is of formula (IIe), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:24 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 26.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIe-i), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:24 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 26.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIe-ii), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:24 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 26.

In a preferred embodiment the CNP prodrug of the present invention is of formula (IIe), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:20 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 30.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIe-i), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:20 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 30.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIe-ii), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:20 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 30.

In a preferred embodiment the CNP prodrug of the present invention is of formula (IIe), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:21 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 29.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIe-i), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:21 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 29.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIe-ii), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:21 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 29.

In a preferred embodiment the CNP prodrug of the present invention is of formula (IIe), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:22 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 28.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIe-i), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:22 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 28.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIe-ii), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:22 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 28.

In a preferred embodiment the CNP prodrug of the present invention is of formula (IIe), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:23 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 27.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIe-i), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:23 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 27.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIe-ii), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:23 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 27.

In a preferred embodiment the CNP prodrug of the present invention is of formula (IIe), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:30 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 27.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIe-i), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:30 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 27.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIe-ii), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:30 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 27.

Accordingly, in a preferred embodiment the CNP prodrug of the present invention is of formula (IIe′)

whereinthe unmarked dashed line indicates attachment to a nitrogen provided by the side chain of the lysine at position 26 of the CNP moiety of SEQ ID NO:24 by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to a moiety

whereineach c1 is an integer independently ranging from 400 to 500.

Preferably, each c1 of formula (IIe′) is about 450.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIe-i′)

whereinthe unmarked dashed line indicates the attachment to a nitrogen provided by the side chain of the lysine at position 26 of the CNP moiety of SEQ ID NO:24 by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to a moiety

whereineach c1 is an integer independently ranging from 400 to 500.

Preferably, each c1 of formula (IIe-i′) is about 450.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIe-ii′)

whereinthe unmarked dashed line indicates the attachment to a nitrogen provided by the side chain of the lysine at position 26 of the CNP moiety of SEQ ID NO:24 by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to a moiety

whereineach c1 is an integer independently ranging from 400 to 500.

Preferably, each c1 of formula (IIe-ii) is about 450.

In another preferred embodiment the CNP prodrug of the present invention is of formula (III)

whereinthe unmarked dashed line indicates the attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z having the structure

whereineach —Zais

whereineach c1 is an integer independently ranging from 200 to 250.

Preferably, each c1 of formula (IIf) is about 225.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIf-i)

whereinthe unmarked dashed line indicates the attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z having the structure

whereineach —Zais

whereineach c1 is an integer independently ranging from 200 to 250.

Preferably, each c1 of formula (IIf-i) is about 225.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIf-ii)

whereinthe unmarked dashed line indicates the attachment to a nitrogen of an amino acid side chain of the ring moiety of the CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z having the structure

whereineach —Zais

whereineach c1 is an integer independently ranging from 200 to 250.

Preferably, each c1 of formula (IIf-ii) is about 225.

In one embodiment the CNP moiety of the CNP prodrug of formula (IIf), (IIf-i) and (IIf-ii) has the sequence of SEQ ID NO:25.

In one embodiment the CNP moiety of the CNP prodrug of formula (IIf), (IIf-i) and (IIf-ii) has the sequence of SEQ ID NO:30.

In another embodiment the CNP moiety of the CNP prodrug of formula (IIf), (IIf-i) and (IIf-ii) has the sequence of SEQ ID NO:20.

In another embodiment the CNP moiety of the CNP prodrug of formula (IIf), (IIf-i) and (IIf-ii) has the sequence of SEQ ID NO:21.

In another embodiment the CNP moiety of the CNP prodrug of formula (IIf), (IIf-i) and (IIf-ii) has the sequence of SEQ ID NO:22.

In another embodiment the CNP moiety of the CNP prodrug of formula (IIf), (IIf-i) and (IIf-ii) has the sequence of SEQ ID NO:23.

In a preferred embodiment the CNP moiety of the CNP prodrug of formula (IIf), (IIf-i) and (IIf-ii) has the sequence of SEQ ID NO:24.

In a preferred embodiment the CNP prodrug of the present invention is of formula (IIf), wherein c1 is about 225, the CNP moiety has the sequence of SEQ ID NO:24 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 26.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIf-i), wherein c1 is about 225, the CNP moiety has the sequence of SEQ ID NO:24 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 26.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIf-ii), wherein c1 is about 225, the CNP moiety has the sequence of SEQ ID NO:24 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 26.

In a preferred embodiment the CNP prodrug of the present invention is of formula (IIf), wherein c1 is about 225, the CNP moiety has the sequence of SEQ ID NO:30 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 27.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIf-i), wherein c1 is about 225, the CNP moiety has the sequence of SEQ ID NO:30 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 27.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIf-ii), wherein c1 is about 225, the CNP moiety has the sequence of SEQ ID NO:30 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 27.

In a preferred embodiment the CNP prodrug of the present invention is of formula (IIf), wherein c1 is about 225, the CNP moiety has the sequence of SEQ ID NO:20 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 30.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIf-i), wherein c1 is about 225, the CNP moiety has the sequence of SEQ ID NO:20 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 30.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIf-ii), wherein c1 is about 225, the CNP moiety has the sequence of SEQ ID NO:20 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 30.

In a preferred embodiment the CNP prodrug of the present invention is of formula (IIf), wherein c1 is about 225, the CNP moiety has the sequence of SEQ ID NO:21 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 29.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIf-i), wherein c1 is about 225, the CNP moiety has the sequence of SEQ ID NO:21 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 29.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIf-ii), wherein c1 is about 225, the CNP moiety has the sequence of SEQ ID NO:21 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 29.

In a preferred embodiment the CNP prodrug of the present invention is of formula (IIf), wherein c1 is about 225, the CNP moiety has the sequence of SEQ ID NO:22 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 28.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIf-i), wherein c1 is about 225, the CNP moiety has the sequence of SEQ ID NO:22 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 28.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIf-ii), wherein c1 is about 225, the CNP moiety has the sequence of SEQ ID NO:22 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 28.

In a preferred embodiment the CNP prodrug of the present invention is of formula (IIf), wherein c1 is about 225, the CNP moiety has the sequence of SEQ ID NO:23 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 27.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIf-i), wherein c1 is about 225, the CNP moiety has the sequence of SEQ ID NO:23 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 27.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIf-ii), wherein c1 is about 225, the CNP moiety has the sequence of SEQ ID NO:23 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 27.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIf′)

whereinthe unmarked dashed line indicates the attachment to a nitrogen provided by the side chain of the lysine at position 26 of the CNP moiety of SEQ ID NO:24 by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z having the structure

whereineach Zais

whereineach c1 is an integer independently ranging from 200 to 250.

Preferably, each c1 of formula (IIf′) is about 225.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIf-i′)

whereinthe unmarked dashed line indicates the attachment to a nitrogen provided by the side chain of the lysine at position 26 of the CNP moiety of SEQ ID NO:24 by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z having the structure

whereineach Zais

whereineach c1 is an integer independently ranging from 200 to 250.

Preferably, each c1 of formula (IIf-i′) is about 225.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIf-ii′)

whereinthe unmarked dashed line indicates the attachment to a nitrogen provided by the side chain of the lysine at position 26 of the CNP moiety of SEQ ID NO:24 by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z having the structure

whereineach Zais

whereineach c1 is an integer independently ranging from 200 to 250.

Preferably, each c1 of formula (IIf-ii) is about 225.

In an equally preferred embodiment the CNP prodrug of the present invention is of formula (IIea)

whereinthe unmarked dashed line indicates the attachment to a nitrogen of -D which is a CNP moiety by forming an amide bond;k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12; and the dashed line marked with the asterisk indicates attachment to a moiety

whereineach c1 is an integer independently ranging from 400 to 500.

Preferably, c1 of formula (IIea) is about 450.

Preferably, k of formula (IIea) is selected from the group consisting of 2, 3, 4, 5, 6 and 7.

In an equally preferred embodiment the CNP prodrug of the present invention is of formula (IIea-i)

whereinthe unmarked dashed line indicates the attachment to a nitrogen of -D which is a CNP moiety by forming an amide bond;k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12; andthe dashed line marked with the asterisk indicates attachment to a moiety

whereineach c1 is an integer independently ranging from 400 to 500.

Preferably, k of formula (IIea-i) is selected from the group consisting of 2, 3, 4, 5, 6 and 7.

Preferably, c1 of formula (IIea-i) is about 450.

In another equally preferred embodiment the CNP prodrug of the present invention is of formula (IIea-ii)

whereinthe unmarked dashed line indicates the attachment to a nitrogen of -D which is a CNP moiety by forming an amide bond;k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12; andthe dashed line marked with the asterisk indicates attachment to a moiety

whereineach c1 is an integer independently ranging from 400 to 500.

Preferably, k of formula (IIea-ii) is selected from the group consisting of 2, 3, 4, 5, 6 and 7.

Preferably, c1 of formula (IIea-ii) is about 450.

In one embodiment the CNP moiety of the CNP prodrug of formula (IIea), (IIea-i) and (IIea-ii) has the sequence of SEQ ID NO:25.

In another embodiment the CNP moiety of the CNP prodrug of formula (IIea), (IIea-i) and (IIea-ii) has the sequence of SEQ ID NO:30.

In another embodiment the CNP moiety of the CNP prodrug of formula (IIea), (IIea-i) and (IIea-ii) has the sequence of SEQ ID NO:20.

In another embodiment the CNP moiety of the CNP prodrug of formula (IIea), (IIea-i) and (IIea-ii) has the sequence of SEQ ID NO:21.

In another embodiment the CNP moiety of the CNP prodrug of formula (IIea), (IIea-i) and (IIea-ii) has the sequence of SEQ ID NO:22.

In another embodiment the CNP moiety of the CNP prodrug of formula (IIea), (IIea-i) and (IIea-ii) has the sequence of SEQ ID NO:23.

In a preferred embodiment the CNP moiety of the CNP prodrug of formula (IIea), (IIea-i) and (IIea-ii) has the sequence of SEQ ID NO:24.

In one embodiment the CNP moiety is attached to -L1- in the CNP prodrug of formula (IIea), (IIea-i) and (IIea-ii) through the nitrogen of the N-terminal amine functional group of CNP.

In a preferred embodiment the CNP prodrug of the present invention is of formula (IIea), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:24 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 26.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIea-i), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:24 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 26.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIea-ii), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:24 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 26.

In a preferred embodiment the CNP prodrug of the present invention is of formula (IIea), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:20 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 30.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIea-i), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:20 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 30.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIea-ii), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:20 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 30.

In a preferred embodiment the CNP prodrug of the present invention is of formula (IIea), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:21 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 29.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIea-i), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:21 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 29.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIea-ii), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:21 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 29.

In a preferred embodiment the CNP prodrug of the present invention is of formula (IIea), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:22 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 28.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIea-i), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:22 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 28.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIea-ii), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:22 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 28.

In a preferred embodiment the CNP prodrug of the present invention is of formula (IIea), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:23 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 27.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIea-i), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:23 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 27.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIea-ii), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:23 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 27.

In a preferred embodiment the CNP prodrug of the present invention is of formula (IIea), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:30 and is attached to -L1-through the amine functional group provided by the side chain of the lysine at position 27.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIea-i), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:30 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 27.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIea-ii), wherein c1 is about 450, the CNP moiety has the sequence of SEQ ID NO:30 and is attached to -L1- through the amine functional group provided by the side chain of the lysine at position 27.

Accordingly, in a preferred embodiment the CNP prodrug of the present invention is of formula (IIea′)

whereinthe unmarked dashed line indicates the attachment to the nitrogen provided by the side chain of the lysine at position 26 of the CNP moiety of SEQ ID NO:24 by forming an amide bond;k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12; andthe dashed line marked with the asterisk indicates attachment to a moiety

whereineach c1 is an integer independently ranging from 400 to 500.

Preferably, k of formula (Ika′) is selected from the group consisting of 2, 3, 4, 5, 6 and 7.

Preferably, each c1 of formula (IIea′) is about 450.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIea-i′)

whereinthe unmarked dashed line indicates the attachment to a nitrogen provided by the side chain of the lysine at position 26 of the CNP moiety of SEQ ID NO:24 by forming an amide bond;k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12; andthe dashed line marked with the asterisk indicates attachment to a moiety

whereineach c1 is an integer independently ranging from 400 to 500.

Preferably, k of formula (IIea-i′) is selected from the group consisting of 2, 3, 4, 5, 6 and 7.

Preferably, each c1 of formula (IIea-i′) is about 450.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIea-ii′)

whereinthe unmarked dashed line indicates the attachment to a nitrogen provided by the side chain of the lysine at position 26 of the CNP moiety of SEQ ID NO:24 by forming an amide bond;k is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12; andthe dashed line marked with the asterisk indicates attachment to a moiety

whereineach c1 is an integer independently ranging from 400 to 500.

Preferably, k of formula (IIea-ii′) is selected from the group consisting of 2, 3, 4, 5, 6 and 7.

Preferably, each c1 of formula (IIea-ii′) is about 450.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIfa)

whereinthe unmarked dashed line indicates the attachment to a nitrogen of -D which is a CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z having the structure

whereink is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12;each —Zais

whereineach c1 is an integer independently ranging from 200 to 250.

Preferably, k of formula (IIfa) is selected from the group consisting of 2, 3, 4, 5, 6 and 7.

Preferably, each c1 of formula (IIfa) is about 225.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIfa-i)

whereinthe unmarked dashed line indicates the attachment to a nitrogen of -D which is a CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z having the structure

whereink is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12;each —Zais

whereineach c1 is an integer independently ranging from 200 to 250.

Preferably, k of formula (IIfa-i) is selected from the group consisting of 2, 3, 4, 5, 6 and 7.

Preferably, each c1 of formula (IIfa-i) is about 225.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIfa-ii)

whereinthe unmarked dashed line indicates the attachment to a nitrogen of -D which is a CNP moiety by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z having the structure

whereink is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12;each —Zais

whereineach c1 is an integer independently ranging from 200 to 250.

Preferably, each c1 of formula (IIfa-ii) is about 225.

In one embodiment the CNP moiety of the CNP prodrug of formula (IIfa), (IIfa-i) and (IIfa-ii) has the sequence of SEQ ID NO:25.

In another embodiment the CNP moiety of the CNP prodrug of formula (IIfa), (IIfa-i) and (IIfa-ii) has the sequence of SEQ ID NO:30.

In another embodiment the CNP moiety of the CNP prodrug of formula (IIfa), (IIfa-i) and (IIfa-ii) has the sequence of SEQ ID NO:20.

In another embodiment the CNP moiety of the CNP prodrug of formula (IIfa), (IIfa-i) and (IIfa-ii) has the sequence of SEQ ID NO:21.

In another embodiment the CNP moiety of the CNP prodrug of formula (IIfa), (IIfa-i) and (IIfa-ii) has the sequence of SEQ ID NO:22.

In another embodiment the CNP moiety of the CNP prodrug of formula (IIfa), (IIfa-i) and (IIfa-ii) has the sequence of SEQ ID NO:23.

In a preferred embodiment the CNP moiety of the CNP prodrug of formula (IIfa), (IIfa-i) and (IIfa-ii) has the sequence of SEQ ID NO:24.

In one embodiment the CNP moiety is attached to -L1- in the CNP prodrug of formula (IIfa), (IIfa-i) and (IIfa-ii) through the nitrogen of the N-terminal amine functional group of CNP.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIfa′)

whereinthe unmarked dashed line indicates the attachment to the nitrogen provided by the side chain of the lysine at position 26 of the CNP moiety of SEQ ID NO:24 by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z having the structure

whereink is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12;each Zais

whereineach c1 is an integer independently ranging from 200 to 250.

Preferably, k of formula (IIfa′) is selected from the group consisting of 2, 3, 4, 5, 6 and 7.

Preferably, each c1 of formula (IIfa′) is about 225.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIfa-i′)

whereinthe unmarked dashed line indicates the attachment to a nitrogen provided by the side chain of the lysine at position 26 of the CNP moiety of SEQ ID NO:24 by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z having the structure

whereink is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12;each Zais

whereineach c1 is an integer independently ranging from 200 to 250.

Preferably, k of formula (IIfa-i′) is selected from the group consisting of 2, 3, 4, 5, 6 and 7.

Preferably, each c1 of formula (IIfa-i′) is about 225.

In another preferred embodiment the CNP prodrug of the present invention is of formula (IIfa-ii′)

whereinthe unmarked dashed line indicates the attachment to a nitrogen provided by the side chain of the lysine at position 26 of the CNP moiety of SEQ ID NO:24 by forming an amide bond; andthe dashed line marked with the asterisk indicates attachment to —Z having the structure

whereink is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12;each Zais

whereineach c1 is an integer independently ranging from 200 to 250.

Preferably, k of formula (IIfa-ii′) is selected from the group consisting of 2, 3, 4, 5, 6 and 7.

Preferably, each c1 of formula (IIfa-ii′) is about 225.

Preferably, the total mass of the CNP prodrug of the present invention is at least 10 kDa, such as at least 12 kDa, such as at least 15 kDa, such as at least 20 kDa or such as at least 30 kDa. It is preferred that the total mass of the CNP prodrug of the present invention is at most 250 kDa, such as at most 200 kDa, 180 kDa, 150 kDa or 100 kDa.

In a preferred embodiment the residual activity of the CNP prodrug of the present invention is less than 10%, more preferably less than 1%, even more preferably less than 0.1%, even more preferably less than 0.01%, even more preferably less than 0.001% and most preferably less than 0.0001%.

As used herein the term “residual activity” refers to the activity exhibited by the CNP prodrug of the present invention with the CNP moiety bound to a carrier in relation to the activity exhibited by the corresponding free CNP. In this context the term “activity” refers to NPR-B binding. It is understood that measuring the residual activity of the CNP prodrug of the present invention takes time during which a certain amount of CNP will be released the CNP prodrug of the present invention and that such released CNP will distort the results measured for the CNP prodrug. It is thus accepted practice to test the residual activity of a prodrug with a conjugate in which the drug moiety, in this case CNP, is non-reversibly, i.e. stably, bound to a carrier, which as closely as possible resembles the structure of the CNP prodrug for which residual activity is to be measured.

A suitable assay for measuring CNP activity and the residual activity of the CNP prodrug of the present invention, preferably in the form of a stable analog, is described in WO 2010/135541 A1, example 4, page 143/144.

Another aspect of the present invention is a pharmaceutical composition comprising at least one CNP prodrug or a pharmaceutically acceptable salt thereof of the present invention and at least one excipient.

In one embodiment the pharmaceutical composition comprising CNP prodrug molecules of the present invention comprises a mixture of CNP prodrugs in which the CNP moieties are attached to -L1- through functional groups, preferably through amine functional groups, of different amino acid residues of the ring moiety of the CNP moiety.

In a preferred embodiment the CNP moieties of all CNP prodrug molecules comprised in the pharmaceutical composition are attached to -L1- through a functional group of the same amino acid residue of the ring moiety of the CNP moiety, preferably through an amine functional group of the same amino acid residue of the ring moiety of the CNP moiety. In a preferred embodiment the CNP moieties of all CNP prodrug molecules comprised in the pharmaceutical composition are attached to -L1- through the amine functional group of the side chain of lysine 26, if the CNP moiety has the sequence of SEQ ID:NO 24.

Preferably, the pharmaceutical composition comprising at least one CNP prodrug or a pharmaceutically acceptable salt thereof of the present invention has a pH ranging from and including pH 3 to pH 8. More preferably, the pharmaceutical composition has a pH ranging from and including pH 4 to pH 6. Most preferably, the pharmaceutical composition has a pH ranging from and including pH 4 to pH 5.

In one embodiment the pharmaceutical composition comprising at least one CNP prodrug or a pharmaceutically acceptable salt thereof of the present invention and at least one excipient is a liquid formulation.

In another embodiment the pharmaceutical composition comprising at least one CNP prodrug or a pharmaceutically acceptable salt thereof of the present invention and at least one excipient is a dry formulation.

Such liquid or dry pharmaceutical composition comprises at least one excipient. Excipients used in parenteral formulations may be categorized as, for example, buffering agents, isotonicity modifiers, preservatives, stabilizers, anti-adsorption agents, oxidation protection agents, viscosifiers/viscosity enhancing agents, or other auxiliary agents. However, in some cases, one excipient may have dual or triple functions. Preferably, the at least one excipient comprised in the pharmaceutical composition of the present invention is selected from the group consisting of(i) Buffering agents: physiologically tolerated buffers to maintain pH in a desired range, such as sodium phosphate, bicarbonate, succinate, histidine, citrate and acetate, sulphate, nitrate, chloride, pyruvate; antacids such as Mg(OH)2or ZnCO3may be also used;(ii) Isotonicity modifiers: to minimize pain that can result from cell damage due to osmotic pressure differences at the injection depot; glycerin and sodium chloride are examples; effective concentrations can be determined by osmometry using an assumed osmolality of 285-315 mOsmol/kg for serum;(iii) Preservatives and/or antimicrobials: multidose parenteral formulations require the addition of preservatives at a sufficient concentration to minimize risk of patients becoming infected upon injection and corresponding regulatory requirements have been established; typical preservatives include m-cresol, phenol, methylparaben, ethylparaben, propylparaben, butylparaben, chlorobutanol, benzyl alcohol, phenylmercuric nitrate, thimerosol, sorbic acid, potassium sorbate, benzoic acid, chlorocresol, and benzalkonium chloride;(iv) Stabilizers: Stabilisation is achieved by strengthening of the protein-stabilising forces, by destabilisation of the denatured state, or by direct binding of excipients to the protein; stabilizers may be amino acids such as alanine, arginine, aspartic acid, glycine, histidine, lysine, proline, sugars such as glucose, sucrose, trehalose, polyols such as glycerol, mannitol, sorbitol, salts such as potassium phosphate, sodium sulphate, chelating agents such as EDTA, hexaphosphate, ligands such as divalent metal ions (zinc, calcium, etc.), other salts or organic molecules such as phenolic derivatives; in addition, oligomers or polymers such as cyclodextrins, dextran, dendrimers, PEG or PVP or protamine or HSA may be used;(v) Anti-adsorption agents: Mainly ionic or non-ionic surfactants or other proteins or soluble polymers are used to coat or adsorb competitively to the inner surface of the formulation's container; e.g., poloxamer (Pluronic F-68), PEG dodecyl ether (Brij 35), polysorbate 20 and 80, dextran, polyethylene glycol, PEG-polyhistidine, BSA and HSA and gelatins; chosen concentration and type of excipient depends on the effect to be avoided but typically a monolayer of surfactant is formed at the interface just above the CMC value;(vi) Oxidation protection agents: antioxidants such as ascorbic acid, ectoine, methionine, glutathione, monothioglycerol, morin, polyethylenimine (PEI), propyl gallate, and vitamin E; chelating agents such as citric acid, EDTA, hexaphosphate, and thioglycolic acid may also be used;(vii) Viscosifiers or viscosity enhancers: retard settling of the particles in the vial and syringe and are used in order to facilitate mixing and resuspension of the particles and to make the suspension easier to inject (i.e., low force on the syringe plunger); suitable viscosifiers or viscosity enhancers are, for example, carbomer viscosifiers like Carbopol 940, Carbopol Ultrez 10, cellulose derivatives like hydroxypropylmethylcellulose (hypromellose, HPMC) or diethylaminoethyl cellulose (DEAE or DEAE-C), colloidal magnesium silicate (Veegum) or sodium silicate, hydroxyapatite gel, tricalcium phosphate gel, xanthans, carrageenans like Satia gum UTC 30, aliphatic poly(hydroxy acids), such as poly(D,L- or L-lactic acid) (PLA) and poly(glycolic acid) (PGA) and their copolymers (PLGA), terpolymers of D,L-lactide, glycolide and caprolactone, poloxamers, hydrophilic poly(oxyethylene) blocks and hydrophobic poly(oxypropylene) blocks to make up a triblock of poly(oxyethylene)-poly(oxypropylene)-poly(oxyethylene) (e.g. Pluronic®), polyetherester copolymer, such as a polyethylene glycol terephthalate/polybutylene terephthalate copolymer, sucrose acetate isobutyrate (SAIB), dextran or derivatives thereof, combinations of dextrans and PEG, polydimethylsiloxane, collagen, chitosan, polyvinyl alcohol (PVA) and derivatives, polyalkylimides, poly (acrylamide-co-diallyldimethyl ammonium (DADMA)), polyvinylpyrrolidone (PVP), glycosaminoglycans (GAGs) such as dermatan sulfate, chondroitin sulfate, keratan sulfate, heparin, heparan sulfate, hyaluronan, ABA triblock or AB block copolymers composed of hydrophobic A-blocks, such as polylactide (PLA) or poly(lactide-co-glycolide) (PLGA), and hydrophilic B-blocks, such as polyethylene glycol (PEG) or polyvinyl pyrrolidone; such block copolymers as well as the abovementioned poloxamers may exhibit reverse thermal gelation behavior (fluid state at room temperature to facilitate administration and gel state above sol-gel transition temperature at body temperature after injection);(viii) Spreading or diffusing agent: modifies the permeability of connective tissue through the hydrolysis of components of the extracellular matrix in the intrastitial space such as but not limited to hyaluronic acid, a polysaccharide found in the intercellular space of connective tissue; a spreading agent such as but not limited to hyaluronidase temporarily decreases the viscosity of the extracellular matrix and promotes diffusion of injected drugs; and(ix) Other auxiliary agents: such as wetting agents, viscosity modifiers, antibiotics, hyaluronidase; acids and bases such as hydrochloric acid and sodium hydroxide are auxiliary agents necessary for pH adjustment during manufacture.

Another aspect of the present invention is CNP prodrug or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising at least one CNP prodrug of the present invention for use as a medicament.

Preferably, said medicament is used in the treatment of a disease selected from the group consisting of achondroplasia, hypochondroplasia, short stature, dwarfism, osteochondrodysplasias, thanatophoric dysplasia, osteogenesis imperfecta, achondrogenesis, chondrodysplasia punctata, homozygous achondroplasia, camptomelic dysplasia, congenital lethal hypophosphatasia, perinatal lethal type of osteogenesis imperfecta, short-rib polydactyly syndromes, rhizomelic type of chondrodysplasia punctata, Jansen-type metaphyseal dysplasia, spondyloepiphyseal dysplasia congenita, atelosteogenesis, diastrophic dysplasia, congenital short femur, Langer-type mesomelic dysplasia, Nievergelt-type mesomelic dysplasia, Robinow syndrome, Reinhardt syndrome, acrodysostosis, peripheral dysostosis, Kniest dysplasia, fibrochondrogenesis, Roberts syndrome, acromesomelic dysplasia, micromelia, Morquio syndrome, Kniest syndrome, metatrophic dysplasia, spondyloepimetaphyseal dysplasia, neurofibromatosis, Legius syndrome, LEOPARD syndrome, Noonan syndrome, hereditary gingival fibromatosis, neurofibromatosis type 1, Legius syndrome, cardiofaciocutaneous syndrome, Costello syndrome, SHOX deficiency, idiopathic short stature, growth hormone deficiency, osteoarthritis, cleidocranial dysostosis, craniosynostosis (e.g., Muenke syndrome, Crouzon syndrome, Apert syndrome, Jackson-Weiss syndrome, Pfeiffer syndrome, or Crouzonodermoskeletal syndrome), dactyly, brachydactyly, camptodactyly, polydactyly, syndactyly, dyssegmental dysplasia, enchondromatosis, fibrous dysplasia, hereditary multiple exostoses, hypophosphatemic rickets, Jaffe-Lichtenstein syndrome, Marfan syndrome, McCune-Albright syndrome, osteopetrosis and osteopoikilosis.

In another embodiment said medicament is used in the treatment of an ophthalmic disorder, such as glaucoma and/or elevated intraocular pressure.

In another embodiment said medicament is used in the treatment of a cancer disease associated with overactivation of FGFR3, e.g., multiple myeloma, myeloproliferative syndrome, leukemia, plasma cell leukemia, lymphoma, glioblastoma, prostate cancer, bladder cancer, or mammary cancer.

In another embodiment said medicament is used in the treatment of a vascular smooth muscle disorder, preferably selected from the group consisting of hypertension, restenosis, arteriosclerosis, acute decompensated heart failure, congestive heart failure, cardiac edema, nephredema, hepatic edema, acute renal insufficiency, and chronic renal insufficiency.

In another embodiment said medicament is used in the treatment of hemorrhagic shock.

Preferably said medicament is used in the treatment of an achondroplasia phenotype selected from the group consisting of growth retardation, skull deformities, orthodontic defects, cervical cord compression, spinal stenosis, hydrocephalus, hearing loss due to chronic otitis, cardiovascular disease, neurological disease, and obesity.

Most preferably said medicament is used in the treatment of achondroplasia.

Another aspect of the present invention is the CNP prodrug or a pharmaceutically acceptable salt thereof or the pharmaceutical composition comprising at least one CNP prodrug of the present invention for use in a method of treatment of a disease which can be treated with CNP.

Preferably, said disease is selected from the group consisting of achondroplasia, hypochondroplasia, short stature, dwarfism, osteochondrodysplasias, thanatophoric dysplasia, osteogenesis imperfecta, achondrogenesis, chondrodysplasia punctata, homozygous achondroplasia, camptomelic dysplasia, congenital lethal hypophosphatasia, perinatal lethal type of osteogenesis imperfecta, short-rib polydactyly syndromes, rhizomelic type of chondrodysplasia punctata, Jansen-type metaphyseal dysplasia, spondyloepiphyseal dysplasia congenita, atelosteogenesis, diastrophic dysplasia, congenital short femur, Langer-type mesomelic dysplasia, Nievergelt-type mesomelic dysplasia, Robinow syndrome, Reinhardt syndrome, acrodysostosis, peripheral dysostosis, Kniest dysplasia, fibrochondrogenesis, Roberts syndrome, acromesomelic dysplasia, micromelia, Morquio syndrome, Kniest syndrome, metatrophic dysplasia, spondyloepimetaphyseal dysplasia, neurofibromatosis, Legius syndrome, LEOPARD syndrome, Noonan syndrome, hereditary gingival fibromatosis, neurofibromatosis type 1, Legius syndrome, cardiofaciocutaneous syndrome, Costello syndrome, SHOX deficiency, idiopathic short stature, growth hormone deficiency, osteoarthritis, cleidocranial dysostosis, craniosynostosis (e.g., Muenke syndrome, Crouzon syndrome, Apert syndrome, Jackson-Weiss syndrome, Pfeiffer syndrome, or Crouzonodermoskeletal syndrome), dactyly, brachydactyly, camptodactyly, polydactyly, syndactyly, dyssegmental dysplasia, enchondromatosis, fibrous dysplasia, hereditary multiple exostoses, hypophosphatemic rickets, Jaffe-Lichtenstein syndrome, Marfan syndrome, McCune-Albright syndrome, osteopetrosis and osteopoikilosis.

In another embodiment the disease is an ophthalmic disorder, such as glaucoma and/or elevated intraocular pressure.

In another embodiment said disease is associated with overactivation of FGFR3 in cancer, e.g., multiple myeloma, myeloproliferative syndrome, leukemia, plasma cell leukemia, lymphoma, glioblastoma, prostate cancer, bladder cancer, or mammary cancer.

In another embodiment said disease is a vascular smooth muscle disorder, preferably selected from the group consisting of hypertension, restenosis, arteriosclerosis, acute decompensated heart failure, congestive heart failure, cardiac edema, nephredema, hepatic edema, acute renal insufficiency, and chronic renal insufficiency.

In another embodiment said disease is hemorrhagic shock.

Preferably said disease is an achondroplasia phenotype selected from the group consisting of growth retardation, skull deformities, orthodontic defects, cervical cord compression, spinal stenosis, hydrocephalus, hearing loss due to chronic otitis, cardiovascular disease, neurological disease, and obesity.

Most preferably said disease is achondroplasia.

In one embodiment the patient undergoing the method of treatment of the present invention is a mammalian patient, preferably a human patient. In one embodiment this human patient is an adult. In a preferred embodiment the human patient is a pediatric patient.

Another aspect of the present invention is the use of the CNP prodrug or a pharmaceutically acceptable salt thereof or the pharmaceutical composition comprising at least one CNP prodrug of the present invention for the manufacture of a medicament for treating a disease which can be treated with CNP.

Preferably, said disease is selected from the group consisting of achondroplasia, hypochondroplasia, short stature, dwarfism, osteochondrodysplasias, thanatophoric dysplasia, osteogenesis imperfecta, achondrogenesis, chondrodysplasia punctata, homozygous achondroplasia, camptomelic dysplasia, congenital lethal hypophosphatasia, perinatal lethal type of osteogenesis imperfecta, short-rib polydactyly syndromes, rhizomelic type of chondrodysplasia punctata, Jansen-type metaphyseal dysplasia, spondyloepiphyseal dysplasia congenita, atelosteogenesis, diastrophic dysplasia, congenital short femur, Langer-type mesomelic dysplasia, Nievergelt-type mesomelic dysplasia, Robinow syndrome, Reinhardt syndrome, acrodysostosis, peripheral dysostosis, Kniest dysplasia, fibrochondrogenesis, Roberts syndrome, acromesomelic dysplasia, micromelia, Morquio syndrome, Kniest syndrome, metatrophic dysplasia, spondyloepimetaphyseal dysplasia, neurofibromatosis, Legius syndrome, LEOPARD syndrome, Noonan syndrome, hereditary gingival fibromatosis, neurofibromatosis type 1, Legius syndrome, cardiofaciocutaneous syndrome, Costello syndrome, SHOX deficiency, idiopathic short stature, growth hormone deficiency, osteoarthritis, cleidocranial dysostosis, craniosynostosis (e.g., Muenke syndrome, Crouzon syndrome, Apert syndrome, Jackson-Weiss syndrome, Pfeiffer syndrome, or Crouzonodermoskeletal syndrome), dactyly, brachydactyly, camptodactyly, polydactyly, syndactyly, dyssegmental dysplasia, enchondromatosis, fibrous dysplasia, hereditary multiple exostoses, hypophosphatemic rickets, Jaffe-Lichtenstein syndrome, Marfan syndrome, McCune-Albright syndrome, osteopetrosis and osteopoikilosis.

In another embodiment the disease is an ophthalmic disorder, such as glaucoma and/or elevated intraocular pressure.

In another embodiment said disease is associated with overactivation of FGFR3 in cancer, e.g., multiple myeloma, myeloproliferative syndrome, leukemia, plasma cell leukemia, lymphoma, glioblastoma, prostate cancer, bladder cancer, or mammary cancer.

In another embodiment said disease is a vascular smooth muscle disorder, preferably selected from the group consisting of hypertension, restenosis, arteriosclerosis, acute decompensated heart failure, congestive heart failure, cardiac edema, nephredema, hepatic edema, acute renal insufficiency, and chronic renal insufficiency.

In another embodiment said disease is hemorrhagic shock.

Preferably said disease is an achondroplasia phenotype selected from the group consisting of growth retardation, skull deformities, orthodontic defects, cervical cord compression, spinal stenosis, hydrocephalus, hearing loss due to chronic otitis, cardiovascular disease, neurological disease, and obesity.

Most preferably said disease is achondroplasia.

In one embodiment the disease to be treated with the CNP prodrug or a pharmaceutically acceptable salt thereof or the pharmaceutical composition comprising at least one CNP prodrug of the present invention occurs in a mammalian patient, preferably in a human patient. In one embodiment this human patient is an adult. In a preferred embodiment the human patient is a pediatric patient.

A further aspect of the present invention is a method of treating, controlling, delaying or preventing in a mammalian patient, preferably a human patient, in need of the treatment of one or more diseases which can be treated with CNP, comprising the step of administering to said patient in need thereof a therapeutically effective amount of CNP prodrug or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising CNP prodrug of the present invention. In one embodiment the human patient is an adult. In a preferred embodiment the human patient is a pediatric patient.

Preferably, the one or more diseases which can be treated with CNP is selected from the group consisting of achondroplasia, hypochondroplasia, short stature, dwarfism, osteochondrodysplasias, thanatophoric dysplasia, osteogenesis imperfecta, achondrogenesis, chondrodysplasia punctata, homozygous achondroplasia, camptomelic dysplasia, congenital lethal hypophosphatasia, perinatal lethal type of osteogenesis imperfecta, short-rib polydactyly syndromes, rhizomelic type of chondrodysplasia punctata, Jansen-type metaphyseal dysplasia, spondyloepiphyseal dysplasia congenita, atelosteogenesis, diastrophic dysplasia, congenital short femur, Langer-type mesomelic dysplasia, Nievergelt-type mesomelic dysplasia, Robinow syndrome, Reinhardt syndrome, acrodysostosis, peripheral dysostosis, Kniest dysplasia, fibrochondrogenesis, Roberts syndrome, acromesomelic dysplasia, micromelia, Morquio syndrome, Kniest syndrome, metatrophic dysplasia, spondyloepimetaphyseal dysplasia, neurofibromatosis, Legius syndrome, LEOPARD syndrome, Noonan syndrome, hereditary gingival fibromatosis, neurofibromatosis type 1, Legius syndrome, cardiofaciocutaneous syndrome, Costello syndrome, SHOX deficiency, idiopathic short stature, growth hormone deficiency, osteoarthritis, cleidocranial dysostosis, craniosynostosis (e.g., Muenke syndrome, Crouzon syndrome, Apert syndrome, Jackson-Weiss syndrome, Pfeiffer syndrome, or Crouzonodermoskeletal syndrome), dactyly, brachydactyly, camptodactyly, polydactyly, syndactyly, dyssegmental dysplasia, enchondromatosis, fibrous dysplasia, hereditary multiple exostoses, hypophosphatemic rickets, Jaffe-Lichtenstein syndrome, Marfan syndrome, McCune-Albright syndrome, osteopetrosis and osteopoikilosis.

In another embodiment the one or more diseases which can be treated with CNP is an ophthalmic disorder, such as glaucoma and/or elevated intraocular pressure.

In another embodiment the one or more diseases which can be treated with CNP is associated with overactivation of FGFR3 in cancer, e.g., multiple myeloma, myeloproliferative syndrome, leukemia, plasma cell leukemia, lymphoma, glioblastoma, prostate cancer, bladder cancer, or mammary cancer.

In another embodiment the one or more diseases which can be treated with CNP is a vascular smooth muscle disorder, preferably selected from the group consisting of hypertension, restenosis, arteriosclerosis, acute decompensated heart failure, congestive heart failure, cardiac edema, nephredema, hepatic edema, acute renal insufficiency, and chronic renal insufficiency.

In another embodiment the one or more disease which can be treated with CNP is hemorrhagic shock.

Preferably the one or more diseases which can be treated with CNP is an achondroplasia phenotype selected from the group consisting of growth retardation, skull deformities, orthodontic defects, cervical cord compression, spinal stenosis, hydrocephalus, hearing loss due to chronic otitis, cardiovascular disease, neurological disease, and obesity.

Most preferably the one or more diseases which can be treated with CNP is achondroplasia.

An additional aspect of the present invention is a method of administering the CNP prodrug or a pharmaceutically acceptable salt thereof or pharmaceutical composition of the present invention, wherein the method comprises the step of administering the CNP prodrug or the pharmaceutical composition of the present invention via topical, enteral or parenteral administration and by methods of external application, injection or infusion, including intraarticular, periarticular, intradermal, subcutaneous, intramuscular, intravenous, intraosseous, intraperitoneal, intrathecal, intracapsular, intraorbital, intravitreal, intratympanic, intravesical, intracardiac, transtracheal, subcuticular, subcapsular, subarachnoid, intraspinal, intraventricular, intrasternal injection and infusion, direct delivery to the brain via implanted device allowing delivery of the invention or the like to brain tissue or brain fluids (e.g., Ommaya Reservoir), direct intracerebroventricular injection or infusion, injection or infusion into brain or brain associated regions, injection into the subchoroidal space, retro-orbital injection and ocular instillation, preferably via subcutaneous injection.

In a preferred embodiment, the present invention relates to a CNP prodrug or pharmaceutically acceptable salt thereof or a pharmaceutical composition of the present invention, for use in the treatment of achondroplasia via subcutaneous injection.

In a further aspect the present invention relates to a pharmaceutical composition comprising at least one CNP prodrug of the present invention or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition comprises at least one further biologically active moiety or drug.

The at least one further biologically active moiety or drug may be in its free form (i.e in the form of a free drug), may be in the form of a stable conjugate or may be in the form of a controlled-release compound.

In one embodiment, the at least one further biologically active moiety or drug is a drug in its free form, i.e. the pharmaceutical composition of the present invention comprises at least one CNP prodrug and at least one further drug.

Preferably, the at least one further drug is selected from the group consisting of antihistamins; human anti-FGFR3 antibodies; soluble forms of human fibroblast growth factor receptor 3;

tyrosine kinase inhibitors; statins; CNP agonists; growth hormone; IGF-1; ANP; BNP; inhibitors of peptidases and proteases; and inhibitors of NPR-C.

A preferred antihistamin is meclozine.

A preferred tyrosine kinase inhibitor is NVP-BGJ398.

A preferred statin is rosuvastatin.

A preferred CNP agonist for the at least one further drug is vosoritide.

Preferred inhibitors of peptidases and proteases are NEP and furin inhibitors.

A preferred inhibitor for NEP are thiorphan and candoxatril.

Preferred inhibitors of NPR-C are the fragment of SEQ ID NO:98 (FGIPMDRIGRNPR) and antibody B701.

Preferred inhibitors of tyrosine kinases are as disclosed in U.S. Pat. Nos. 6,329,375 and 6,344,459, which are herewith incorporated by reference.

In one embodiment the at least one further drug is an antihistamin.

In another embodiment the at least one further drug is a human anti-FGFR3 antibody.

In another embodiment the at least one further drug is a soluble forms of human fibroblast growth factor receptor 3 (sFGFR3).

In another embodiment the at least one further drug is a tyrosine kinase inhibitor.

In another embodiment the at least one further drug is a statin.

In another embodiment the at least one further drug is a growth hormone.

In another embodiment the at least one further drug is a CNP agonist.

In another embodiment the at least one further drug is IGF-1.

In another embodiment the at least one further drug is ANP.

In another embodiment the at least one further is BNP.

In another embodiment the at least one further drug is an inhibitor of peptidases and proteases.

In another embodiment the at least one further drug is an inhibitor of NPR-C.

In another embodiment, the at least one further biologically active moiety or drug is in the form of a stable conjugate.

In one embodiment the at least one further biologically active moiety in the form of a stable conjugate comprises at least one biologically active moiety covalently conjugated through a stable linkage to a polymeric moiety, preferably to a water-soluble polymeric moiety, either directly or through a spacer moiety.

Preferably, such polymeric moiety, even more preferably water-soluble polymeric moiety, comprises a polymer selected from the group consisting of 2-methacryloyl-oxyethyl phosphoyl cholins, poly(acrylic acids), poly(acrylates), poly(acrylamides), poly(alkyloxy) polymers, poly(amides), poly(amidoamines), poly(amino acids), poly(anhydrides), poly(aspartamides), poly(butyric acids), poly(glycolic acids), polybutylene terephthalates, poly(caprolactones), poly(carbonates), poly(cyanoacrylates), poly(dimethylacrylamides), poly(esters), poly(ethylenes), poly(ethyleneglycols), poly(ethylene oxides), poly(ethyl phosphates), poly(ethyloxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl-oxazo lines), poly(hydroxymethacrylates), poly(hydroxypropylmethacrylamides), poly(hydroxypropyl methacrylates), poly(hydroxypropyloxazolines), poly(imino carbonates), poly(lactic acids), poly(lactic-co-glycolic acids), poly(methacrylamides), poly(methacrylates), poly(methyloxazolines), poly(organophosphazenes), poly(ortho esters), poly(oxazolines), polypropylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinyl amines), poly(vinylmethylethers), poly(vinylpyrrolidones), silicones, celluloses, carbomethyl celluloses, hydroxypropyl methylcelluloses, chitins, chitosans, dextrans, dextrins, gelatins, hyaluronic acids and derivatives, functionalized hyaluronic acids, mannans, pectins, rhamnogalacturonans, starches, hydroxyalkyl starches, hydroxyethyl starches and other carbohydrate-based polymers, xylans, and copolymers thereof.

In another embodiment the at least one further biologically active moiety in the form of a stable conjugate is covalently conjugated through a stable linkage to an albumin-binding moiety. Preferably, said albumin-binding moiety is a C8-24alkyl moiety or fatty acid derivative. Preferred fatty acid derivatives are those disclosed in WO 2005/027978 A2 and WO 2014/060512 A1 which are herewith incorporated by reference.

Preferably, the at least one further biologically active moiety in the form of a stable conjugate comprises a biologically active moiety selected from the group consisting of antihistamins; human anti-FGFR3 antibodies; soluble forms of human fibroblast growth factor receptor 3 (sFGFR3); tyrosine kinase inhibitors; statins; CNP agonists; growth hormone; IGF-1; ANP; BNP; inhibitors of peptidases and proteases; and inhibitors of NPR-C.

A preferred antihistamin is meclozine.

A preferred tyrosine kinase inhibitor is NVP-BGJ398.

A preferred statin is rosuvastatin.

A preferred CNP agonist for the at least one further biologically active moiety is vosoritide.

Preferred inhibitors of peptidases and proteases are NEP and furin inhibitors.

A preferred inhibitor for NEP are thiorphan and candoxatril.

Preferred inhibitors of NPR-C are the fragment of SEQ ID NO:98 (FGIPMDRIGRNPR) and antibody B701.

Preferred inhibitors of tyrosine kinases are as disclosed in U.S. Pat. Nos. 6,329,375 and 6,344,459, which are herewith incorporated by reference.

In one embodiment the at least one further biologically active moiety in the form of a stable conjugate comprises an antihistamin moiety.

In another embodiment the at least one further biologically active moiety in the form of a stable conjugate comprises a human anti-FGFR3 antibody moiety.

In another embodiment the at least one further biologically active moiety in the form of a stable conjugate comprises a soluble forms of human fibroblast growth factor receptor 3 (sFGFR3) moiety.

In another embodiment the at least one further biologically active moiety in the form of a stable conjugate comprises a tyrosine kinase inhibitor moiety.

In another embodiment the at least one further biologically active moiety in the form of a stable conjugate comprises a statin moiety.

In another embodiment the at least one further biologically active moiety in the form of a stable conjugate comprises a growth hormone moiety.

In another embodiment the at least one further biologically active moiety in the form of a stable conjugate comprises a CNP agonist moiety.

In another embodiment the at least one further biologically active moiety in the form of a stable conjugate comprises an IGF-1 moiety.

In another embodiment the at least one further biologically active moiety in the form of a stable conjugate comprises an ANP moiety.

In another embodiment the at least one further biologically active moiety in the form of a stable conjugate comprises a BNP moiety.

In another embodiment the at least one further biologically active moiety in the form of a stable conjugate comprises an inhibitor of peptidases and proteases moiety.

In another embodiment the at least one further biologically active moiety in the form of a stable conjugate comprises an inhibitor of NPR-C moiety.

In another embodiment the at least one further biologically active moiety or drug is in the form of a controlled-release compound.

Preferably, the at least one further biologically active moiety or drug in the form of a controlled-release compound comprises at least one biologically active moiety or drug selected from the group consisting of antihistamins; human anti-FGFR3 antibodies; soluble forms of human fibroblast growth factor receptor 3; statins; CNP agonists; growth hormone; IGF-1; ANP; BNP; inhibitors of peptidases and proteases; inhibitors of tyrosine kinases; and inhibitors of NPR-C.

A preferred antihistamin is meclozine.

A preferred tyrosine kinase inhibitor is NVP-BGJ398.

A preferred statin is rosuvastatin.

A preferred CNP agonist for the at least one further drug is vosoritide.

Preferred inhibitors of peptidases and proteases are NEP and furin inhibitors.

A preferred inhibitor for NEP are thiorphan and candoxatril.

Preferred inhibitors of NPR-C are the fragment of SEQ ID NO:98 (FGIPMDRIGRNPR) and antibody B701.

Preferred inhibitors of tyrosine kinases are as disclosed in U.S. Pat. Nos. 6,329,375 and 6,344,459, which are herewith incorporated by reference.

In one embodiment the at least one further biologically active moiety or drug in the form of a controlled-release comprises an antihistamin moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a controlled-release comprises a human anti-FGFR3 antibody moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a controlled-release comprises a soluble forms of human fibroblast growth factor receptor 3 (sFGFR3) moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a controlled-release comprises a tyrosine kinase inhibitor moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a controlled-release comprises a statin moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a controlled-release comprises a growth hormone moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a controlled-release comprises a CNP agonist moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a controlled-release comprises an IGF-1 moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a controlled-release comprises an ANP moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a controlled-release comprises a BNP moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a controlled-release comprises an inhibitor of peptidases and proteases moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a controlled-release comprises an inhibitor of NPR-C moiety or drug.

In one embodiment the at least one further biologically active moiety or drug in the form of a controlled-release compound is water-insoluble.

Preferably, such water-insoluble controlled-release compound is selected from the group consisting of crystals, nanoparticles, microparticles, nanospheres and microspheres.

In one embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release compound is a crystal comprising at least one drug or biologically active moiety.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release compound is a nanoparticle comprising at least one drug or biologically active moiety.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release compound is a microparticle comprising at least one drug or biologically active moiety.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release compound is a nanosphere comprising at least one drug or biologically active moiety.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release compound is a microsphere comprising at least one drug or biologically active moiety.

In one embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release compound is a vesicle comprising at least one drug or biologically active moiety. Preferably, such vesicle comprising at least one drug or biologically active moiety is a micelle, liposome or polymersome.

In one embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release compound is a micelle comprising at least one drug or biologically active moiety.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release compound is a liposome comprising at least one drug or biologically active moiety. Preferably, such liposome is selected from the group consisting of aquasomes; non-ionic surfactant vesicles, such as niosomes and proniosomes; cationic liposomes, such as LeciPlex; transfersomes; ethosomes; ufasomes; sphingosomes; and pharmacosomes.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release compound is a polymersome at least one drug or biologically active moiety.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release compound comprises at least one biologically active moiety or drug non-covalently embedded in a water-insoluble polymer. Preferably, such water-insoluble polymer comprises a polymer selected from the group consisting of 2-methacryloyl-oxyethyl phosphoyl cholins, poly(acrylic acids), poly(acrylates), poly(acrylamides), poly(alkyloxy) polymers, poly(amides), poly(amidoamines), poly(amino acids), poly(anhydrides), poly(aspartamides), poly(butyric acids), poly(glycolic acids), polybutylene terephthalates, poly(caprolactones), poly(carbonates), poly(cyanoacrylates), poly(dimethylacrylamides), poly(esters), poly(ethylenes), poly(ethyleneglycols), poly(ethylene oxides), poly(ethyl phosphates), poly(ethyloxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl-oxazolines), poly(hydroxymethacrylates), poly(hydroxypropylmethacrylamides), poly(hydroxypropyl methacrylates), poly(hydroxypropyloxazo lines), poly(imino carbonates), poly(lactic acids), poly(lactic-co-glycolic acids), poly(methacrylamides), poly(methacrylates), poly(methyloxazo lines), poly(organophosphazenes), poly(ortho esters), poly(oxazolines), polypropylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinyl amines), poly(vinylmethylethers), poly(vinylpyrrolidones), silicones, celluloses, carbomethyl celluloses, hydroxypropyl methylcelluloses, chitins, chitosans, dextrans, dextrins, gelatins, hyaluronic acids and derivatives, functionalized hyaluronic acids, mannans, pectins, rhamnogalacturonans, starches, hydroxyalkyl starches, hydroxyethyl starches and other carbohydrate-based polymers, xylans, and copolymers thereof.

In a preferred embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release compound comprises at least one drug or biologically active moiety non-covalently embedded in poly(lactic-co-glycolic acid) (PLGA).

In another embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release compound comprises at least one biologically active moiety covalently and reversibly conjugated to a water-insoluble polymer. Preferably such water-insoluble polymer comprises a polymer selected from the group consisting of 2-methacryloyl-oxyethyl phosphoyl cholins, poly(acrylic acids), poly(acrylates), poly(acrylamides), poly(alkyloxy) polymers, poly(amides), poly(amidoamines), poly(amino acids), poly(anhydrides), poly(aspartamides), poly(butyric acids), poly(glycolic acids), polybutylene terephthalates, poly(caprolactones), poly(carbonates), poly(cyanoacrylates), poly(dimethylacrylamides), poly(esters), poly(ethylenes), poly(ethyleneglycols), poly(ethylene oxides), poly(ethyl phosphates), poly(ethyloxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl-oxazolines), poly(hydroxymethacrylates), poly(hydroxypropylmethacrylamides), poly(hydroxypropyl methacrylates), poly(hydroxypropyloxazo lines), poly(imino carbonates), poly(lactic acids), poly(lactic-co-glycolic acids), poly(methacrylamides), poly(methacrylates), poly(methyloxazo lines), poly(organophosphazenes), poly(ortho esters), poly(oxazolines), polypropylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinyl amines), poly(vinylmethylethers), poly(vinylpyrrolidones), silicones, celluloses, carbomethyl celluloses, hydroxypropyl methylcelluloses, chitins, chitosans, dextrans, dextrins, gelatins, hyaluronic acids and derivatives, functionalized hyaluronic acids, mannans, pectins, rhamnogalacturonans, starches, hydroxyalkyl starches, hydroxyethyl starches and other carbohydrate-based polymers, xylans, and copolymers thereof.

Preferably, the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release compound comprises at least one biologically active moiety or drug selected from the group consisting of antihistamins; human anti-FGFR3 antibodies; soluble forms of human fibroblast growth factor receptor 3; tyrosine kinase inhibitors; statins; CNP agonists; growth hormone; IGF-1; ANP; BNP; inhibitors of peptidases and proteases; and inhibitors of NPR-C.

A preferred antihistamin is meclozine.

A preferred tyrosine kinase inhibitor is NVP-BGJ398.

A preferred statin is rosuvastatin.

A preferred CNP agonist for the at least one further drug is vosoritide.

Preferred inhibitors of peptidases and proteases are NEP and furin inhibitors.

A preferred inhibitor for NEP are thiorphan and candoxatril.

Preferred inhibitors of NPR-C are the fragment of SEQ ID NO:98 (FGIPMDRIGRNPR) and antibody B701.

Preferred inhibitors of tyrosine kinases are as disclosed in U.S. Pat. Nos. 6,329,375 and 6,344,459, which are herewith incorporated by reference.

In one embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release comprises an antihistamin moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release comprises a human anti-FGFR3 antibody moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release comprises a soluble forms of human fibroblast growth factor receptor 3 (sFGFR3) moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release comprises a tyrosine kinase inhibitor moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release comprises a statin moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release comprises a growth hormone moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release comprises a CNP agonist moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release comprises an IGF-1 moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release comprises an ANP moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release comprises a BNP moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release comprises an inhibitor of peptidases and proteases moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-insoluble controlled-release comprises an inhibitor of NPR-C moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a controlled-release compound is water-soluble.

In one embodiment the at least one further biologically active moiety or drug in the form of a water-soluble controlled-release compound comprises at least one biologically active moiety covalently conjugated through a reversible linkage to a water-soluble polymeric moiety, either directly or through a spacer moiety.

Preferably, such water-soluble polymeric moiety comprises a polymer selected from the group consisting of 2-methacryloyl-oxyethyl phosphoyl cholins, poly(acrylic acids), poly(acrylates), poly(acrylamides), poly(alkyloxy) polymers, poly(amides), poly(amidoamines), poly(amino acids), poly(anhydrides), poly(aspartamides), poly(butyric acids), poly(glycolic acids), polybutylene terephthalates, poly(caprolactones), poly(carbonates), poly(cyanoacrylates), poly(dimethylacrylamides), poly(esters), poly(ethylenes), poly(ethyleneglycols), poly(ethylene oxides), poly(ethyl phosphates), poly(ethyloxazolines), poly(glycolic acids), poly(hydroxyethyl acrylates), poly(hydroxyethyl-oxazolines), poly(hydroxymethacrylates), poly(hydroxypropylmethacrylamides), poly(hydroxypropyl methacrylates), poly(hydroxypropyloxazo lines), poly(iminocarbonates), poly(lactic acids), poly(lactic-co-glycolic acids), poly(methacrylamides), poly(methacrylates), poly(methyloxazolines), poly(organophosphazenes), poly(ortho esters), poly(oxazolines), poly(propylene glycols), poly(siloxanes), poly(urethanes), poly(vinyl alcohols), poly(vinyl amines), poly(vinylmethylethers), poly(vinylpyrrolidones), silicones, celluloses, carbomethyl celluloses, hydroxypropyl methylcelluloses, chitins, chitosans, dextrans, dextrins, gelatins, hyaluronic acids and derivatives, functionalized hyaluronic acids, mannans, pectins, rhamnogalacturonans, starches, hydroxyalkyl starches, hydroxyethyl starches and other carbohydrate-based polymers, xylans, and copolymers thereof.

In another embodiment the at least one further biologically active moiety in the form of a water-soluble controlled-release compound is covalently conjugated through a stable linkage to an albumin-binding moiety. Preferably, said albumin-binding moiety is a C8-24alkyl moiety or fatty acid derivative. Preferred fatty acid derivatives are those disclosed in WO 2005/027978 A2 and WO 2014/060512 A1 which are herewith incorporated by reference.

Preferably, the at least one further biologically active moiety in the form of a water-soluble controlled-release comprises a biologically active moiety selected from the group consisting of antihistamins; human anti-FGFR3 antibodies; soluble forms of human fibroblast growth factor receptor 3; tyrosine kinase inhibitors; statins; CNP agonists; growth hormone; IGF-1; ANP; BNP; inhibitors of peptidases and proteases; and inhibitors of NPR-C.

A preferred antihistamin is meclozine.

A preferred tyrosine kinase inhibitor is NVP-BGJ398.

A preferred statin is rosuvastatin.

A preferred CNP agonist for the at least one further drug is vosoritide.

Preferred inhibitors of peptidases and proteases are NEP and furin inhibitors.

A preferred inhibitor for NEP are thiorphan and candoxatril.

Preferred inhibitors of NPR-C are the fragment of SEQ ID NO:98 (FGIPMDRIGRNPR) and antibody B701.

Preferred inhibitors of tyrosine kinases are as disclosed in U.S. Pat. Nos. 6,329,375 and 6,344,459, which are herewith incorporated by reference.

In one embodiment the at least one further biologically active moiety or drug in the form of a water-soluble controlled-release comprises an antihistamin moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-soluble controlled-release comprises a human anti-FGFR3 antibody moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-soluble controlled-release comprises a soluble forms of human fibroblast growth factor receptor 3 (sFGFR3) moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-soluble controlled-release comprises a tyrosine kinase inhibitor moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-soluble controlled-release comprises a statin moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-soluble controlled-release comprises a growth hormone moiety or drug. A preferred water-soluble controlled-release growth hormone compound is compound 2 of example 2 of WO2016/079114A1. Accordingly, a preferred water-soluble controlled-release growth hormone compound has the following structure:



In another embodiment the at least one further biologically active moiety or drug in the form of a water-soluble controlled-release comprises a CNP agonist moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-soluble controlled-release comprises an IGF-1 moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-soluble controlled-release comprises an ANP moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-soluble controlled-release comprises a BNP moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-soluble controlled-release comprises an inhibitor of peptidases and proteases moiety or drug.

In another embodiment the at least one further biologically active moiety or drug in the form of a water-soluble controlled-release comprises an inhibitor of NPR-C moiety or drug.

Another aspect of the present invention is the pharmaceutical composition comprising at least one CNP prodrug and at least one further biologically active moiety or drug of the present invention for use as a medicament.

Another aspect of the present invention is the pharmaceutical composition comprising at least one CNP prodrug and at least one further biologically active moiety or drug of the present invention for use in the treatment of a patient suffering from a disorder that benefits from stimulating growth.

Preferably, the patient is a mammalian patient, more preferably a human patient.

Preferably, such disorders that benefit from stimulating growth are selected from the group comprising achondroplasia, hypochondroplasia, short stature, dwarfism, osteochondrodysplasias, thanatophoric dysplasia, osteogenesis imperfecta, achondrogenesis, chondrodysplasia punctata, homozygous achondroplasia, camptomelic dysplasia, congenital lethal hypophosphatasia, perinatal lethal type of osteogenesis imperfecta, short-rib polydactyly syndromes, rhizomelic type of chondrodysplasia punctata, Jansen-type metaphyseal dysplasia, spondyloepiphyseal dysplasia congenita, atelosteogenesis, diastrophic dysplasia, congenital short femur, Langer-type mesomelic dysplasia, Nievergelt-type mesomelic dysplasia, Robinow syndrome, Reinhardt syndrome, acrodysostosis, peripheral dysostosis, Kniest dysplasia, fibrochondrogenesis, Roberts syndrome, acromesomelic dysplasia, micromelia, Morquio syndrome, Kniest syndrome, metatrophic dysplasia, and spondyloepimetaphyseal dysplasia. Most preferably, the disorder that benefits from stimulating growth is achondroplasia.

Another aspect of the present invention is a method of treating a patient suffering from a disorder that benefits from stimulating growth by administering the pharmaceutical composition of the present invention.

Preferably, the patient is a mammalian patient, more preferably a human patient.

Preferably, such disorders that benefit from stimulating growth are selected from the group comprising achondroplasia, hypochondroplasia, short stature, dwarfism, osteochondrodysplasias, thanatophoric dysplasia, osteogenesis imperfecta, achondrogenesis, chondrodysplasia punctata, homozygous achondroplasia, camptomelic dysplasia, congenital lethal hypophosphatasia, perinatal lethal type of osteogenesis imperfecta, short-rib polydactyly syndromes, rhizomelic type of chondrodysplasia punctata, Jansen-type metaphyseal dysplasia, spondyloepiphyseal dysplasia congenita, atelosteogenesis, diastrophic dysplasia, congenital short femur, Langer-type mesomelic dysplasia, Nievergelt-type mesomelic dysplasia, Robinow syndrome, Reinhardt syndrome, acrodysostosis, peripheral dysostosis, Kniest dysplasia, fibrochondrogenesis, Roberts syndrome, acromesomelic dysplasia, micromelia, Morquio syndrome, Kniest syndrome, metatrophic dysplasia, and spondyloepimetaphyseal dysplasia. Most preferably, the disorder that benefits from stimulating growth is achondroplasia.

The polypeptide moiety of the CNP prodrug may be prepared by standard solid-phase peptide synthesis methods, e.g. by Boc chemistry (R. B. Merrifield, J. Am. Chem. Soc., 85(14): 2149-2154 (1963)). Alternatively, Fmoc (fluorenylmethoxycarbonyl) chemistry may be employed.

Methods known in the art can be employed to improve purity and/or yield, including the use of pseudoproline or other dipeptide building blocks, fragment coupling and others (J. Wade et al., Lett. Pept. Sci., 7(2):107-112 (2000); Y. Fujiwara et al., Chem. Pharm. Bull., 44(7):1326-1331 (1996); P. Cherkupally et al., Eur. J. Org. Chem., 6372-6378 (2013)).

Alternatively, the polypeptide moiety of the CNP prodrug may be produced by recombinant synthesis processes.

FIG. 1: Structure of CNP according to SEQ ID NO:1.

