REFERENCE TO A SEQUENCE LISTING

This application incorporates-by-reference nucleotide and/or amino acid sequences which are present in the file named “190723_91046 Sequence_Listing_CAS.txt”, which is 701 bytes in size, and which was created Jan. 23, 2019 in the IBM-PC machine format, having an operating system compatibility with MS-Windows, which is contained in the text file filed Jan. 23, 2019 as part of this application.

FIELD

The present disclosure is generally related to novel, potent and selective 2-arylsulfonamido-N-arylacetamide derivatized Stat3 inhibitors of Formulae I-VIII, and pharmaceutically acceptable salts thereof. The present disclosure also relates to pharmaceutical compositions containing the inhibitors and their use in the treatment or prevention of cancer, and other pathogenic conditions in which Stat3 activation is implicated. As an example, the disclosure provides methods and compositions for the treatment of cancer by modulating Stat3.

BACKGROUND

The following includes information that may be useful in understanding various aspects and embodiments of the present disclosure. It is not an admission that any of the information provided herein is prior art, or relevant, to the presently described or claimed inventions, or that any publication or document that is specifically or implicitly referenced is prior art.

The signal transducer and activator of transcription (Stat) family of cytoplasmic transcription factors have important roles in many cellular processes, including cell growth and differentiation, inflammation and immune responses. (Bromberg, et al.,Breast Cancer Res.2:86-90 (2000); Darnell, J., et al.,Nat. Rev. Cancer2:740-749 (2002)). STAT proteins are classically activated by tyrosine (Tyr) kinases, such as Janus kinases (JAKs) and Src family kinases, in response to the binding of cytokine and growth factors to their cognate receptors. The Tyr phosphorylation (pTyr) promotes dimerization between two activated STAT:STAT monomers through a reciprocal pTyr-Src homology SH2 domain interactions. Active STAT:STAT dimers translocate to the nucleus to induce gene transcription by binding to specific DNA-response elements in the promoters of target genes to regulate gene expression. By contrast, aberrantly-active Stat3, one of the Stat family members, has been implicated in many human tumors and represents an attractive target for drug discovery. The aberrant activation of Stat3 occurs in glioma, breast, prostate, ovarian, and many other human cancers, whereby it promotes malignant progression (Yu & Jove,Nat. Rev. Cancer4:97-105 (2004)). Mechanisms by which constitutively-active Stat3 mediates tumorigenesis include dysregulation of gene expression that leads to uncontrolled growth and survival of tumor cells, enhanced tumor angiogenesis, and metastasis and the suppression of tumor immune surveillance (Yu & Jove (2004); Bromberg & Darnell,Oncogene19:2468-2473 (2000); Bowman et al.,Oncogene19:2474-2488 (2000); Turkson J & Jove,Oncogene19:6613-6626 (2000); Turkson,Expert Opin Ther Targets8:409-422 (2004); Wang et al.,Nat Med10:48-54 (2004)).

Stat3 modulates mitochondrial functions and Stat3 crosstalk with other proteins, such as NF-κB, that promotes the malignant phenotype. Many human tumors harbor aberrantly-active signal transducer and activator of transcription Stat3 signaling.

SUMMARY OF THE INVENTION

In one aspect, this invention relates to novel, selective and potent 2-arylsulfonamido-N-arylacetamide derivatized Stat3 inhibitors, useful as cancer therapeutics. In some aspects, the compounds of this invention are useful for inhibiting malignant transformation, tumor development and progression.

In one aspect, this invention relates to compounds of Formula I, which selectively inhibit Stat3.



wherein R1can include or exclude aryl or a 5 or 6-membered heteroaryl, where the heteroatoms are one or more of O, N, and S(A)2, where S is sulfur and A can include or exclude oxygen or an electron pair, the aryl or the 5 or 6-membered heteroaryl are optionally substituted with halo, CF3, C1-C6alkyl, C1-C6branched alkyl, aryl, heteroaryl, C3-C7cycloalkyl, C3-C7cycloalkenyl, three- to six-membered heterocycle, three- to seven-membered saturated heterocycle, fused C2-C5alkylene, where one or more CH2groups can be replaced with O, NR9, and S(A)2, where S is sulfur and A can include or exclude oxygen or an electron pair, the aryl or the 5 or 6-membered heteroaryl are optionally substituted naphthalene, optionally substituted indole, benzofuran, benzothiophene; R2and R3are independently selected from H or C1-C6alkyl; where R2and R3can form a C3-C6cycloalkane ring, where this C3-C6cycloalkane ring can be substituted with 1 or more of C1-C6alkyl, hydroxyl, NR9R10, or C1-C6alkoxy; R4can include or exclude H, C1-C6alkyl, (CH2)fNR9R10, (CH2)fOR9, (CH2)fCO2R9, (CH2)fCO2NR9R10; R5can include or exclude C1-C6alkyl, (CH2)fNR9R10, (CH2)fOR9, (CH2)fCO2R9, (CH2)fCO2NR9R10, where R4and R5can form a C3-C6cycloalkane ring, where this C3-C6cycloalkane ring can be substituted with 1 or more of C1-C6alkyl, hydroxyl, NR9R10, or C1-C6alkoxy, where one or more CH2groups can be replaced with O, NR9, and S(A)2, where S is sulfur and A can include or exclude oxygen or an electron pair; R6can include or exclude C1-C6alkyl; where R4and R6can form a C3-C6cycloalkane ring, where this C3-C6cycloalkane ring can be substituted with 1 or more of C1-C6alkyl, hydroxyl, NR9R10and where one or more CH2groups can be replaced with O, NR9, and S(A)2, where S is sulfur and A can include or exclude oxygen or an electron pair; wherein R4and R6can form an optionally substituted pyrrole ring, wherein one or more CH groups of said pyrrole ring can be replaced with O, N, and S(A)2, where S is sulfur and A can include or exclude oxygen or an electron pair; R7can include or exclude CF3, aryl or heteroaryl group where the aryl or heteroaryl group is substituted with 1-5 substituents selected from C1-C6alkyl, halo, hydroxyl, CN, or CF3; R8is substitution selected from one or more of H, C1-C6alkyl, C1-C6alkoxy, hydroxyl, halogen, OC(O)CH3, NR9R10, CN, CF3, CO2R9, CO2NR9R10, (CH2)fNR9R10, (CH2)fOR9, or (CH2)fCO2R9; R9can include or exclude H or C1-C6alkyl; R10can include or exclude H, C1-C6alkyl; W can include or exclude CO2H, tetrazole, benzyl, C(O)NHOR10and CF2OH; Q is C, CH, N, O, S; where Q and R8can form a heterocyclic ring; p is selected from 0 or 1; y is selected from 0 or 1, f is selected from 0 to 4; t is selected from 0 or 1;

and solvates, hydrates, or pharmaceutically acceptable salts thereof.

In one aspects, this invention relates to pharmaceutical compositions comprising a therapeutically effective amount of a compound of Formula I and a pharmaceutically acceptable excipient.

In one aspect, this invention relates to compounds of Formula II, which selectively inhibit Stat3.



wherein R11can include or exclude optionally substituted aryl or optionally substituted heteroaryl,

wherein the aryl or heteroaryl can be substituted with one or more of hydroxyl, carboxylic acid, carboxylate, benzohydroxamic acid, hydroxyl-substituted alkyl;

wherein R12can include or exclude (R)—CH(CH3), (S)—CHCH3, (S)—CH(CH2CH3), (R)—CH(CH2CH3), bridging cylcopropyl


C(CH3)2, (S)—CH(CH2OH), (R)—CH(CH2OH), (R)—CH(CH2CH2OH), (S)—CH(CH2CH2OH), (S)—CH[(R)—CH(CH3)OH], (R)—CH[(S)—CH(CH3)OH], (S)—CH(CH2NH2), (R)—CH(CH2NH2), (S)—CH(CH2CO2H), and (R)—CH(CH2CO2H);

and solvates, hydrates, or pharmaceutically acceptable salts thereof.

In one aspect, this invention relates to compounds of Formula III, which selectively inhibit Stat3.



wherein R11can include or exclude aryl or heteroaryl,

wherein the aryl or heteroaryl can be substituted with one or more of hydroxyl, carboxylic acid, carboxylate, benzohydroxamic acid, nitroso, alkyl carboxylic acid, C1-C6alkyl, and halo;

wherein R12is (R)—CH(CH3);

and solvates, hydrates, or pharmaceutically acceptable salts thereof.

In some aspects, R11can include or exclude:



and salts thereof.

In one aspect, this invention relates to compounds of Formula IV, which selectively inhibit Stat3.



wherein R14can include or exclude a five- to seven-membered cycloalkyl or heteroxycloalkyl group, wherein the cycloalkyl group can be substituted with one or more of halo;

wherein R15can include or exclude substituted aryl, wherein the substitution is one or more of hydroxy, caroboxylic acid, and benzohydroxamic acid;

wherein R16is (R)—CH(CH3);

wherein Y can include or exclude CH and N,

and solvates, hydrates, or pharmaceutically acceptable salts thereof.

In some aspects, R14can include or exclude:



In some aspects, R15can include or exclude:



In one aspect, this invention relates a compound selected from:



and solvates, hydrates, or pharmaceutically acceptable salts thereof.

In one aspect, this invention relates to compounds of Formula VI, which selectively inhibit Stat3.



wherein n is selected from 1 to 3

wherein X can include or exclude CH and N;

wherein R17can include or exclude CH2and O;

wherein R18can include or exclude hydrogen and halo;

wherein R19can include or exclude hydrogen and hydroxyl;

wherein R20can include or exclude —OH, NHOH, and —(O−, Na+),

and solvates, hydrates, or pharmaceutically acceptable salts thereof.

In one aspect, this invention relates a compound selected from:



and solvates, hydrates, or pharmaceutically acceptable salts thereof.

In some aspects, this invention relates to compounds of Formula VII, which selectively inhibit Stat3.



wherein W, X, Y, and Z each independently can include or exclude CH or N,

wherein R21can include or exclude optionally substituted aryl, optionally substituted heteroaryl, or an optionally substituted aryl fused with a heterocyclic ring,

wherein the substituted aryl or substituted heteroaryl or the substituted aryl fused with a heterocyclic ring can be substituted with one or more of hydroxyl, carboxylic acid, carboxylate, hydroxamic acid, amide, alkyl amide, dialkylamide, alkoxyamino, alkyl carboxylic acid, C1-C6 alkyl, and halo,

wherein n is selected from 1 to 3;

and solvates, hydrates, or pharmaceutically acceptable salts thereof.

In some aspects, R21can include or exclude any of the following moieties:



In some aspects, the compounds of this disclosure can include or exclude:



In some aspects, this inventions relates to compounds of Formula VIII.



wherein W, X, Y, Z, and Q each independently can include or exclude CH or N,

wherein d is selected from 0, 1, 2, 3, or 4,

wherein R21is as described herein, and

wherein R22is halogen.

In some aspects, any R group described herein can include or exclude the recited options.

In one aspect, the compounds of this invention inhibit Stat3 while exhibiting little or no inhibition of Stat1 at concentrations of at least twice the IC50for Stat3 inhibition. The compounds of this invention uniquely interact with three sub-pockets on the stat3:stat3 dimer interface, in contrast to other previously described Stat3 inhibitors, which interacts with only two sub-pockets. As a result of the unique and specific mechanism by which the 2-arylsulfonamido-N-arylacetamide derivatized Stat3 inhibitors of this invention exert their effects, the compounds are more potent and less toxic. The compounds of this invention also suprisingly selectively bind inhibit the activated form of Stat3, consequently attenuating Stat3 functions in cancer cells. The compounds of this invention are useful, for example, for inhibiting cancer cell growth, survival, migration and/or metastasis.

In one aspect, this invention relates to compounds which preferentially inhibit Stat3 DNA-binding activity with IC50's of 10 μM or less, but exhibit little or no disruption of Stat5 DNA-binding activity. In one aspect, this invention relates to compositions and formulations useful for inhibiting cancer growth. In some aspects, the anti-cancer activity of the compounds is determined by the ability to inhibit growth of mouse xenografts of human breast and non-small cell lung cancers.

Dimerization of Stat3 occurs through SH2-phosphotyrosyl peptide interactions. See Shuai et al., Interferon activation of the transcription factor Stat91 involves dimerization through SH2-phosphotyrosyl peptide interactions (Cell,76:821-828 (1994); Miklossy et al.Nat Rev Drug Discov12:611-629 (2013); Turkson et al.,Mol Cancer Ther3:261-269 (2004); Turkson et al.,J. Biol. Chem.276:45443-45455 (2001); Siddiquee et al.,Proc Natl Acad Sci USA.104:7391-7396 (2007); Coleman et al.,J Med Chem.48(6661-70) (2005)).

In one aspect, the invention relates to the inventors' design of 2-arylsulfonamido-N-arylacetamide derivatized Stat3 inhibitors which interfere with the dimerization between two monomers, and the inventors' recognition that this represents an attractive strategy to develop drugs that inhibit Stat3 activation and functions.

The inventors have appreciated that glycine-based compounds such as BP-1-102 inhibit Stat3 activity (Zhang et al.,Proc Natl Acad Sci USA109:9623-8 (2012)). In the present invention, the inventors have developed analogues of glycine based compounds with increased potency, reduced lipophilicity, and lower molecular weight compared to the glycine based compounds such as BP-1-102.

The compounds of this invention surprisingly show improved Stat3-inhibitory activity. For example, the compounds of this invention have IC50's or activities that are two to ten times lower than (and therefore more potent than) the compound BP-1-102.

The present disclosure provides novel, selective 2-arylsulfonamido-N-arylacetamide derivatized Stat3 inhibitors, and pharmaceutical formulations and kits comprising the inhibitors. The compounds and pharmaceutical formulations are useful as therapeutics for cancer and other conditions mediated by aberrantly active Stat3, a substrate for growth factor receptor tyrosine kinases, or cytoplasmic tyrosine kinases, including Janus kinases or the Src family kinases. In some aspects, the processes inhibited by the compounds and compositions of this invention include proliferation, survival, angiogenesis, migration/metastasis/invasion, and immunity.

The compounds of this invention are useful for inhibiting activities resulting from constitutive Stat3 activation, which include: a) stimulating proliferation by increasing the expression of c-Myc and/or cyclin D1/D2, and/or decreasing expression of p53; b) increasing survival by increasing the expression of survivin, Bcl-x/Bcl-2, Mcl-1 and/or Akt-2; stimulating angiogenesis by increasing expression of VEGF; and/or increasing migration/metastasis or invasion by increasing the expression MMP-2 or MMP-9.

In some aspects, the compounds of Formulae I-VIII including compounds of Examples 25-31, 50, 52, 53, 60-64, 79, 85, 102, 104, 110, 113, 119, 122, 125, 134-137, 143, 148, 149, 151 and 152 are used in the compositions and methods of this invention as described herein.

In one aspect, the present disclosure provides the use of a compound of any of Formulae I-VIII including compounds of Examples 25-31, 50, 52, 53, 60-64, 79, 85, 102, 104, 110, 113, 119, 122, 125, 134-137, 143, 148, 149, 151 and 152 for the preparation of a medicament for the treatment of a condition selected from the group consisting of cancer, hyperplasia, autoimmune indications, and neoplasia. In one aspect, the tumor progression, including metastasis and/or growth is thereby inhibited and/or reduced. In one aspect, multi-drug resistance is thereby inhibited and/or reduced.

In another aspect the present disclosure provides a method of treating cancer comprising administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a compound of any of Formulae I-VIII including compounds of Examples 25-31, 50, 52, 53, 60-64, 79, 85, 102, 104, 110, 113, 119, 122, 125, 134-137, 143, 148, 149, 151 and 152 whereby the cancer is treated, cancer progression is stopped or slowed, and/or Stat3 is inhibited.

In one aspect, the level of Stat3 activity is reduced in cancer cells. In one aspect, the effective dose of the 2-arylsulfonamido-N-arylacetamide derivatized Stat3 inhibitor compound is administered at a dose ranging from 0.05 mg/kg to 4 mg/kg. In some aspects, the therapeutically effective dose is about 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8. 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, or about 4.0 mg/kg, or any range in between any two of the recited doses. In some aspects, the dose will be 0.08 mg/kg to about 0.5 mg/kg, from about 0.08 to about 0.24 mg/kg, or from about 0.24 to about 0.5 mg/kg. In another aspect, the effective dose of the Stat3 inhibitor is given in one or more doses. In some aspects, the therapeutically effective amount of the Stat3 inhibitor dose is selected from: 0.08, 0.24, and 0.5 mg/kg for each dose. In one aspect, the dose is administered by a delivery route selected from the group consisting of intraperitoneal, intradermal, intramuscular, intraperitoneal, intravenous, topical, subcutaneous, intranasal, or epidural routes. In one aspect, the one or more effective doses of the 2-arylsulfonamido-N-arylacetamide derivatized Stat3 inhibitor are administered orally, intravenously, intramuscularly, or subcutaneously. In one aspect, the one or more effective doses of the Stat3 inhibitor are administered orally. In one aspect, the one or more effective doses of the Stat3 inhibitor are administered intravenously. In some aspects, the one or more effective doses of the Stat3 inhibitor are administered subcutaneously. In one aspect, the one or more effective doses of the Stat3 inhibitor are administered intramuscularly.

In one aspect, this disclosure provides a method of treatment comprising administering to a subject in need thereof an effective amount of a pharmaceutical composition comprising a 2-arylsulfonamido-N-arylacetamide derivatized Stat3 inhibitor of this invention. In one aspect, the subject has a glioma, breast cancer, or pancreatic cancer. In some aspects, the subject has a solid tumor cancer. In one aspect, the solid tumor comprises sarcomas, carcinomas or lymphomas. In one aspect, the cancer can include or exclude: brain tumors, such as gliomas, medulloblastomas, cerebral menangiomas, breast, prostate, pancreatic, ovarian, bladder, head and neck, malignant melanoma, multiple myeloma, lymphomas, including anaplastic large T cell lymphoma, sezary syndrome, EBV-related Burkitt's Lymphoma, HSV Saimiri-dependent (T Cell), cutaneous T cell lymphoma, mycosis fungoides, leukemia, including HTLV-I dependent leukemia, erythroleukemia, acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myelogenous leukemia (AML), chronic myelogenous lekemia (CML), megakaryocytic leukemia, and large granula lymphocyte (LGL) leukemia, or thyroid, skin, lung, or kidney cancer. In some aspects the cancer can include or exclude: renal cell carcinoma, pancreatic adenocarcinoma, ovarian carcinoma, suamous cell carcinoma of the head and neck, or Hodgkin's Lymphoma.

According to one aspect of the present invention, there are provided novel compositions comprising compounds represented by Formulae I-VIII including compounds of Examples 25-31, 50, 52, 53, 60-64, 79, 85, 102, 104, 110, 113, 119, 122, 125, 134-137, 143, 148, 149, 151 and 152 their pharmaceutically acceptable salts, and pharmaceutical compositions containing them, or mixture thereof.

The inventions described and claimed herein have many attributes and embodiments, including, but not limited to, those set forth, or described, or referenced, in this Brief Summary. It is not intended to be all-inclusive and the inventions described and claimed herein are not limited to, or by the features or embodiments identified in, this Brief Summary, which is included for purposes of illustration only and not restriction. Additional embodiments may be disclosed in the Detailed Description below.

