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

Patent metadata

Field	Value
Publication number	US-2018009845-A1
Application number	US-201715702682-A
Country	US
Kind code	A1
Filing date	Sep 12, 2017
Priority date	May 27, 2011
Publication date	Jan 11, 2018
Grant date	—

Abstract

Disclosed herein are non-natural amino acids and dolastatin analogs that include at least one non-natural amino acid, and methods for making such non-natural amino acids and polypeptides. The dolastatin analogs can include a wide range of possible functionalities, but typically have at least one oxime, carbonyl, dicarbonyl, and/or hydroxylamine group. Also disclosed herein are non-natural amino acid dolastatin analogs that are further modified post-translationally, methods for effecting such modifications, and methods for purifying such dolastatin analogs. Typically, the modified dolastatin analogs include at least one oxime, carbonyl, dicarbonyl, and/or hydroxylamine group. Further disclosed are methods for using such non-natural amino acid dolastatin analogs and modified non-natural amino acid dolastatin analogs, including therapeutic, diagnostic, and other biotechnology use.

First claim

What is claimed:

1 - 24 . (canceled)

25 . A compound comprising Formula (VIII) or (IX), wherein the compound is a trastuzumab antibody conjugated to a dolastatin, wherein the conjugation occurs via a non-naturally encoded amino acid in the antibody, wherein Formula (VIII) or (IX) correspond to:

wherein:
A is optional, and when present is lower alkylene, substituted lower alkylene, lower alkenylene, substituted lower alkenylene, 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, —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) k N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(S)N(R′)—, —N(R′)S(O) k N(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′)—, where each R′ is independently H, alkyl, or substituted alkyl;
each R′ is independently H, alkyl, or substituted alkyl;

R is H, alkyl, substituted alkyl, cycloalkyl, or substituted cycloalkyl;
R 1 is H, an amino protecting group, resin, at least one amino acid, polypeptide or polynucleotide;
R 2 is OH, an ester protecting group, resin, at least one amino acid, polypeptide or polynucleotide;
R 3 and R 4 are each independently H, halogen, lower alkyl, or substituted lower alkyl, or R 3 and R 4 or two R 3 groups optionally form a cycloalkyl or a heterocycloalkyl.
Z has the structure of:

R 5 is H, COR 8 , C 1 -C 6 alkyl, or thiazole;
R 8 is OH;
R 6 is OH or H;
Ar is phenyl or pyridine;

R 7 is C 1 -C 6 alkyl or hydrogen;
L is a linker selected from the group consisting of a bond, -alkylene-, -alkylene-C(O)—, -(alkylene-O) n -alkylene-, -(alkylene-O) n -alkylene-C(O)—, -(alkylene-O) n —(CH 2 ) n′ —NHC(O)—(CH 2 ) n″ —C(Me) 2 -S—S—(CH 2 ) n′″ —NHC(O)-(alkylene-O) n″″ -alkylene-, -(alkylene-O) n -alkylene-W—, -alkylene-C(O)—W—, -(alkylene-O) n -alkylene-J-, -alkylene′-J-(alkylene-O) n -alkylene-, -(alkylene-O) n -alkylene-J-alkylene′, -J-(alkylene-O) n -alkylene-, -(alkylene-O) n -alkylene-J-(alkylene-O) n ′-alkylene-J′-, —W—, -alkylene-W—, alkylene′-J-(alkylene-NMe) n -alkylene-W—, and J-(alkylene-NMe) n -alkylene-W—, -(alkylene-O) n -alkylene-U-alkylene-C(O)—, -(alkylene-O) n -alkylene-U-alkylene-; -J-alkylene-NMe-alkylene′—NMe-alkylene″-W—, and -alkylene-J-alkylene′-NMe-alkylene″-NMe-alkylene′″—W—;
W has the structure of:

U has the structure of:

each n, n′, n″, n′″ and n″″ are independently integers from 0 to 20; wherein substituted means substituted with one or more substituents independently selected from: halo, C 1 -C 10 alkyl, C 2 -C 10 alkenyl, C 2 -C 10 alkynyl, C 1 -C 10 alkoxy, C 5 -C 12 aralkyl, C 3 -C 12 cycloalkyl, C 4 -C 12 cycloalkenyl, phenyl, toluoyl, xylenyl, biphenyl, C 2 -C 12 alkoxyalkyl, C 5 -C 12 alkoxyaryl, C 5 -C 12 aryloxyalkyl, C 7 -C 12 oxyaryl, C 1 -C 6 alkylsulfinyl, C 1 -C 10 alkylsulfonyl, —(CH 2 ) m —O—(C 1 -C 10 alkyl) wherein m is from 1 to 8, aryl, fluoroalkyl, heterocyclic radical, nitroalkyl, —NO 2 , —CN, —NR″C(O)—(C 1 -C 10 alkyl), —C(O)—(C 1 -C 10 alkyl), C 2 -C 10 alkthioalkyl, —C(O)O—(C 1 -C 10 alkyl), —OH, —SO 2 , ═S, —COOH, —NR″ 2 , carbonyl, —C(O)—(C 1 -C 10 alkyl)-CF 3 , —C(O)—CF 3 , —C(O)NR″ 2 , —(C 1 -C 10 aryl)-S—(C 6 -C 10 aryl), —C(O)—(C 6 -C 10 aryl), —(CH2) m —O—(CH 2 ) m -O—(C 1 -C 10 alkyl) wherein each m is from 1 to 8, —C(O)NR″ 2 , —C(S)NR″ 2 , —SO 2 NR″ 2 , —NR″C(O)NR″ 2 , —NR″C(S)NR″ 2 ; wherein each R″ group is independently selected from H, alkyl, aryl, or alkaryl,

or an active metabolite, or a pharmaceutically acceptable prodrug or solvate thereof.

26 - 57 . (canceled)

58 . A method for derivatizing a dolastatin analog comprising Formula (I), (III), (IV), (V), or (VI), wherein the derivatized dolastatin analog is a trastuzumab antibody conjugated to a dolastatin, wherein the conjugation occurs via a non-naturally encoded amino acid in the antibody, wherein the method comprising contacting the dolastatin analog with a reagent of Formula (XXXVII), wherein Formula (I), (III), (IV), (V), or (VI) correspond to:

wherein:
Z has the structure of:

R 5 is H, COR 8 , C 1 -C 6 alkyl, or thiazole;
R 8 is OH or —NH-(alkylene-O) n —NH 2 ;

R 6 is OH or H;
Ar is phenyl or pyridine;

R 7 is C 1 -C 6 alkyl or hydrogen;
Y is NH 2 —O— or methyl and V is NH 2 —O—;
L, L 1 , L 2 , L 3 , and L 4 are each linkers selected from the group consisting of a bond, -alkylene-, -alkylene-C(O)—, -(alkylene-O) n -alkylene-, -(alkylene-O) n -alkylene-C(O)—, -(alkylene-O) n —(CH 2 ) n′ , —NHC(O)—(CH 2 ) n″ —C(Me) 2 -S—S—(CH 2 ) n′″ —NHC(O)-(alkylene-O) n″″ -alkylene-, -(alkylene-O) n -alkylene-W—, -alkylene-C(O)—W—, -(alkylene-O) n -alkylene-J-, -alkylene′-J-(alkylene-O) n -alkylene-, -(alkylene-O) n -alkylene-J-alkylene′, -J-(alkylene-O) n -alkylene-, -(alkylene-O) n -alkylene-J-(alkylene-O) n ′-alkylene-J′-, —W—, -alkylene-W—, alkylene′-J-(alkylene-NMe) n -alkylene-W—, and J-(alkylene-NMe) n -alkylene-W—, -(alkylene-O) n -alkylene-U-alkylene-C(O)—, -(alkylene-O) n -alkylene-U-alkylene-; -J-alkylene-NMe-alkylene′-NMe-alkylene″-W—, and -alkylene-J-alkylene′-NMe-alkylene″-NMe-alkylene′″—W—;
W has the structure of:

U has the structure of:

each J and J′ independently have the structure of:

or L is absent, Y is methyl, R 5 is COR 8 , and R 5 is —NH-(alkylene-O) n —NH 2 ; and
each n, n′, n″, n′″ and n″″ are independently integers from 0 to 20;

wherein Formula (XXXVII) corresponds to:

wherein:
A is optional, and when present is lower alkylene, substituted lower alkylene, lower alkenylene, substituted lower alkenylene, arylene, substit

Assignees

• Ambrx Inc

Inventors

• Maio Zhenwei
• Atkinson Kyle
• Biroc Sandra
• Buss Timothy
• Neal Melissa
• Kraynov Vadim
• Marsden Robin
• Pinkstaff Jaosn
• Skidmore Lillian
• Sun Ying
• Szydlik Agnieszka
• Lopez De Valenta Delia Lanina

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• C07D417/12

  linked by a chain containing hetero atoms as chain links · CPC title

• C07K5/0205 Primary

  containing the structure -NH-(X)3-C(=0)-, e.g. statine or derivatives thereof · CPC title

• C07D207/08

  with hydrocarbon radicals, substituted by hetero atoms, attached to ring carbon atoms · CPC title

• A61K47/6861

  the tumour determinant being from kidney or bladder cancer cell · CPC title