Compositions containing, methods involving, and uses of non-natural amino acid linked dolastatin derivatives

An antibody drug conjugate comprising a trastuzumab antibody conjugated to a dolastatin, wherein the conjugation occurs via a non-naturally occurring amino acid in the antibody as depicted in Formula (VIII) or (IX), wherein the non-naturally occurring amino acid is located at position 121 of the antibody:

• wherein:
• A is optional, and when present is lower alkylene, substituted lower alkylene, lower cycloalkylene, substituted lower cycloalkylene, lower alkenylene, substituted lower alkenylene, alkynylene, lower heteroalkylene, substituted heteroalkylene, lower heterocycloalkylene, substituted lower heterocycloalkylene, arylene, substituted arylene, heteroarylene, substituted heteroarylene, alkarylene, substituted alkarylene, aralkylene, or substituted aralkylene;
• B is optional, and when present is a linker selected from the group consisting of lower alkylene, substituted lower alkylene, lower alkenylene, substituted lower alkenylene, lower heteroalkylene, substituted lower heteroalkylene, —O—, —O-(alkylene or substituted alkylene)-, —S—, —S-(alkylene or substituted alkylene)-, —S(O)k— where k is 1, 2, or 3, —S(O)k-(alkylene or substituted alkylene)-, —C(O)—, —C(O)-(alkylene or substituted alkylene)-, —C(S)—, —C(S)-(alkylene or substituted alkylene)-, —N(R′)—, —NR′-(alkylene or substituted alkylene)-, —C(O) N(R′)—, —CON(R′)-(alkylene or substituted alkylene)-, —CSN(R′)—, —CSN(R′)-(alkylene or substituted alkylene)-, —N(R′)CO-(alkylene or substituted alkylene)-, —N(R′)C(O)O—, —S(O)kN(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(S) N(R′)—, —N(R′)S(O)kN(R′)—, —N(R′)—N═, —C(R′)═N—, —C(R′)═N—N(R′)—, —C(R′)═N—N=, —C(R′)2—N═N—, and —C(R′)2—N(R′)—N(R′)—,
• wherein each R′ is independently H, alkyl, or substituted alkyl;
• R is H, alkyl, substituted alkyl, cycloalkyl, or substituted cycloalkyl;
• R1 is H, an amino protecting group, resin, at least one amino acid, polypeptide, or polynucleotide;
• R2 is OH, an ester protecting group, resin, at least one amino acid, polypeptide, or polynucleotide;
• wherein at least one of R1 and R2 is a polypeptide, wherein the polypeptide is the trastuzumab antibody, wherein amino acid position 121 of the trastuzumab antibody is substituted with the non-naturally encoded amino acid;
• R3 and R4 are each independently H, halogen, lower alkyl, or substituted lower alkyl, or R3 and R4 or two R3 groups optionally form a cycloalkyl or a heterocycloalkyl;
• Z has the structure of:
• R5 is H, COR8, C1-C6 alkyl or thiazole;
• R8 is OH;
• R6 is OH or H;
• Ar is phenyl or pyridine;
• R7 is C1-C6 alkyl or hydrogen;
• and
• L is a linker selected from the group consisting of -alkylene-, -alkylene-C(O)—, -(alkylene-O)n-alkylene-, -(alkylene-O)n-alkylene-C(O)—, -(alkylene-O)n—(CH2)n′—NHC(O)—(CH2)n″—C(Me)2-S—S—(CH2)n″—NHC(O)-(alkylene-O)n″-alkylene-, -(alkylene-O)n-alkylene-W—, -alkylene-C(O)—W—, -(alkylene-O)n-alkylene-U-alkylene-C(O)—, and -(alkylene-O)n-alkylene-U-alkylene-;
• wherein:
• W has the structure of:
• U has the structure of:
• and
• each n, n′, n″, n′″ and n″ is independently an integer from 0 to 20;
• wherein substituted means substituted with one or more substituents independently selected from the group consisting of halo, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C1-C10 alkoxy, C5-C12 aralkyl, C3-C12 cycloalkyl, C4-C12 cycloalkenyl, phenyl, toluolyl, xylenyl, biphenyl, C2-C12 alkoxyalkyl, C5-C12 alkoxyaryl, C5-C12 aryloxyalkyl, C7-C12 oxyaryl, C1-C6 alkylsulfonyl, C1-C10 alkylsulfonyl, —(CH2)m—O—(C1-C10 alkyl), aryl, fluoroalkyl, heterocyclic radical, nitroalkyl, —NO2, —CN, —NR″C(O)—(C1-C10 alkyl), —C(O)—(C1-C10 alkyl), C2-C10 alkthioalkyl, —C(O)O—(C1-C10 alkyl), —OH, —SO2, ═S, —COOH, —NR″2, carbonyl, —C(O)—(C1-C10 alkyl)-CF3, —C(O)—CF3, —C(O)NR″2, —(C1-C10 aryl)-S—(C6-C10 aryl), —C(O)—(C6-C10 aryl), —(CH2)m—O—(CH2)m—O—(C1-C10 alkyl), —C(O)NR″2, —C(S)NR″2, —SO2NR″2, —NR″C(O)NR″2 and —NR″C(S)NR″2;
• wherein each R″ group is independently H, alkyl, aryl or alkaryl;
• and each m is from 1 to 8;
• or a pharmaceutically acceptable salt or solvate thereof.