Biomolecule conjugatesThe present invention relates to biomolecule conjugates which comprise a biomolecule wherein at least one non-natural amino acid (NNAA) is integral to the structure of the biomolecule and wherein the NNAA is a point of attachment of a linker to which a payload, particularly a cytotoxic agent, is attached. More specifically, this invention relates to conjugates of cell-binding agents and active release products comprising cytotoxic agents wherein the conjugates are produced by means of a cycloaddition reaction. Methods of production, pharmaceutical compositions and methods of use are provided.A kind of 1. compound of the separation with Formula IWhereinA is strain alkynes ring or strain alkene ring, and the wherein ring is carbocylic radical or heterocyclic radical；D is pay(useful) load；Each E is made up of independently selected from the following group, the group the following：-CO-、-CR1R2-、-NR3-、-S-S-、-S-、-SO-、- SO2-、-O-、-CR3=N-NR3-、-CR3=N-O- ,-CR3=N-NR3- CO- ,-N=N-CO-, alkyl, C3-C8 carbocylic radicals ,-O- (CR1R2)a-, aryl ,-(CR1R2)a- aryl, heteroaryl ,-(CR1R2)a- heteroaryl ,-(CR1R2)a- C3-C8 carbocylic radicals, heterocycle Base ,-(CR1R2)a- heterocyclic radical ,-(CH2CH2O)a-、-(CH2CH2O)a-(CH2)b、-(CH2)aC (O)-, amino acid and peptide；R1And R2The following group is each independently selected from, the group is made up of the following：H、F、Cl、Br、I、OH、-N(R3)2、-N(R3)3+、 C1-C8 alkyl halides, carboxylate radical, sulfate radical, sulfamic acid root, sulfonate radical ,-SO2R3,-S (=O) R3、-SR3、-SO2N(R3)2、-C (=O) R3、-CO2R3,-C (=O) N (R3)2、-CN、-N3、-NO2, C1-C8 alkoxyl, poly- ethyleneoxy group, phosphonate radical, phosphoric acid Root, C1-C8 alkyl, C2-C8 alkenyls, C2-C8 alkynyls, aryl, heteroaryl, C3-C8 carbocylic radicals and C1-C20 heterocyclic radicals；Or, When taken together, R1And R2Form the spiral shell carbocyclic ring of carbonyl (=O) or 3 to 7 carbon atoms；Also,R3It is selected from the group, the group is made up of the following：H, C1-C8 alkyl, C2-C8 alkenyls, C2-C8 alkynyls, C6-C20 aryl, C6-C20 heteroaryls, C3-C8 carbocylic radicals and C1-C20 heterocyclic radicals；The one or more that alkyl, alkenyl, alkynyl, aryl, heteroaryl, carbocylic radical and heterocyclic radical are optionally independently selected from the group Substituent substitutes, and the group is made up of the following：F、Cl、Br、I、OH、-N(R3)2、-N(R3)3+, C1-C8 alkyl halides, carboxylate radical, Sulfate radical, sulfamic acid root, sulfonate radical, C1-C8 alkyl azochlorosulfonates, C1-C8 alkyl aminos, 4- dialkylaminopyridiniums, C1- C8 alkyl hydroxies, C1-C8 alkylthio groups ,-SO2R ,-S (=O) R3、-SR3、-SO2N(R3)2,-C (=O) R3、-CO2R3,-C (=O) N (R3)2、-CN、-N3、-NO2, C1-C8 alkoxyl, the fluoro-alkyls of C1-C8 tri-, C1-C8 alkyl, C3-C12 carbocyclic rings, C6-C20 virtue Base, C6-C20 heteroaryls, C3-C8 carbocylic radicals, C2-C20 heterocyclic radicals, poly- ethyleneoxy group, phosphonate radical and phosphate radical；M is the integer from 1 to 100；Also,A and b is individually the integer from 1 to 100.
생체분자 컨쥬게이트본 발명은 생체분자를 포함하는 생체분자 컨쥬게이트에 관한 것으로, 여기서 적어도 하나의 비-천연 아미노산(NNAA)이 생체분자의 구조에 필수적이며, NNAA는 페이로드, 특히 세포독성제가 부착되는 링커의 부착 지점이다. 보다 상세하게는, 본 발명은 세포결합제, 및 세포독성제를 포함하는 활성 방출 생성물의 컨쥬게이트에 관한 것으로, 컨쥬게이트는 고리첨가 반응에 의해 생성된다. 생성 방법, 약학적 조성물 및 사용 방법이 제공된다.하기 화학식 I의 분리된 화합물:[화학식 I][이미지](상기 식에서,A는 변형된 알킨 고리 또는 변형된 알켄 고리이고, 여기서 고리는 카르보사이클릴 또는 헤테로사이클릴이고;D는 페이로드(payload)이고;각각의 E는 독립적으로 -CO-, -CR1R2-, -NR3-, -S-S-, -S-, -SO-, -SO2-, -O-, -CR3=N-NR3-, -CR3=N-O-, -CR3=N-NR3-CO-, -N=N-CO-, 알킬, C3-C8 카르보사이클릴, -O-(CR1R2)a-, 아릴, -(CR1R2)a-아릴, 헤테로아릴, -(CR1R2)a-헤테로아릴, -(CR1R2)a-C3-C8 카르보사이클릴, 헤테로사이클릴, -(CR1R2)a-헤테로사이클릴, -(CH2CH2O)a-, -(CH2CH2O)a-(CH2)b, ―(CH2)aC(O)-, 아미노산, 및 펩티드로 구성된 군으로부터 선택되고;R1 및 R2는 각각 독립적으로 H, F, Cl, Br, I, OH, -N(R3)2, -N(R3)3+, C1-C8 알킬할라이드, 카르복실레이트, 설페이트, 설파메이트, 설포네이트, -SO2R3, -S(=O)R3, -SR3, -SO2N(R3)2, -C(=O)R3, -CO2R3, -C(=O)N(R3)2, ―CN, ―N3, ―NO2, C1-C8 알콕시, 폴리에틸렌옥시, 포스포네이트, 포스페이트, C1-C8 알킬, C2-C8 알케닐, C2-C8 알키닐, 아릴, 헤테로아릴, C3-C8 카르보사이클릴, 및 C1-C20 헤테로사이클릴로 구성된 군으로부터 선택되거나; 함께 취해지는 경우, R1 및 R2는 카르보닐(=O), 또는 3 내지 7개의 탄소 원자의 스피로 카르보사이클릭 고리를 형성하고;R3는 H, C1-C8 알킬, C2-C8 알케닐, C2-C8 알키닐, C6-C20 아릴, C6-C20 헤테로아릴, C3-C8 카르보사이클릴, 및 C1-C20 헤테로사이클릴로 구성된 군으로부터 선택되고;알킬, 알케닐, 알키닐, 아릴, 헤테로아릴, 카르보사이클릴, 및 헤테로사이클릴은 임의로 F, Cl, Br, I, OH, ―N(R3)2, ―N(R3)3+, C1-C8 알킬할라이드, 카르복실레이트, 설페이트, 설파메이트, 설포네이트, C1-C8 알킬설포네이트, C1-C8 알킬아미노, 4-디알킬아미노피리디늄, C1-C8 알킬하이드록실, C1-C8 알킬티올, -SO2R, -S(=O)R3, ―SR3, ―SO2N(R3)2, -C(=O)R3, -CO2R3, -C(=O)N(R3)2, ―CN, ―N3, ―NO2, C1-C8 알콕시, C1-C8 트리플루오로알킬, C1-C8 알킬, C3-C12 카르보사이클, C6-C20 아릴, C6-C20 헤테로아릴, C3-C8 카르보사이클릴, C2-C20 헤테로사이클릴, 폴리에틸렌옥시, 포스포네이트, 및 포스페이트로 구성된 군으로부터 선택되는 하나 이상의 치환기로 독립적으로 치환되고;m은 1 내지 100의 정수이고;a 및 b는 각각 1 내지 100의 정수임).