Method of targeting specific cell populations using cell-binding agent maytansinoid conjugates linked via a non-cleavable linker, said conjugates and methods of making said

Patent metadata

Field	Value
Publication number	US-10844135-B2
Application number	US-201715413153-A
Country	US
Kind code	B2
Filing date	Jan 23, 2017
Priority date	Oct 10, 2003
Publication date	Nov 24, 2020
Grant date	Nov 24, 2020

Abstract

The present invention discloses a method for targeting maytansinoids to a selected cell population, the method comprising contacting a cell population or tissue suspected of containing the selected cell population with a cell-binding agent maytansinoid conjugate, wherein one or more maytansinoids is covalently linked to the cell-binding agent via a non-cleavable linker and the cell-binding agent binds to cells of the selected cell population.

First claim

What is claimed is:

1. A cell-binding agent maytansinoid conjugate comprising at least one thiol-containing maytansinoid linked to a cell-binding agent via a non-cleavable linker,
wherein the cell-binding agent is an antibody, a single chain antibody or an antibody fragment that specifically binds to a target cell,
wherein the antibody, single chain antibody or antibody fragment comprises a human constant region,
wherein the non-cleavable linker is a linker that is substantially resistant to acid-induced cleavage, light-induced cleavage, peptidase-induced cleavage, esterase-induced cleavage, and disulfide bond cleavage,
wherein the conjugate is highly cytotoxic, and
wherein the maximum tolerated dose (MTD) in rodents of the cell-binding maytansinoid conjugate is at least about 150 mg/kg,
provided that the cell-binding agent is not an anti-erbB antibody.

2. The cell-binding agent maytansinoid conjugate of claim 1 ,
wherein the non-cleavable linker is derived from a maleimido-based moiety.

3. The cell-binding agent maytansinoid conjugate of claim 2 , wherein the non-cleavable linker is derived from a maleimido-based moiety selected from N-succinimidyl 4-(maleimidomethyl)cyclohexanecarboxylate (SMCC), N-succinimidyl-4-(N-maleimidomethyl)-cyclohexane-1-carboxy-(6-amidocaproate) (LC-SMCC), k-maleimidoundecanoic acid N-succinimidyl ester (KMUA), γ-maleimidobutyric acid N-succinimidyl ester (GMBS), ε-maleimidcaproic acid N-hydroxysuccinimide ester (EMCS), m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS), N-αa-maleimidoacetoxy)-succinimide ester (AMAS), succinimidyl-6-(β-maleimidopropionamido)hexanoate (SMPH), N-succinimidyl 4-(p-maleimidophenyl)-butyrate (SMPB), N-(p-maleimidophenyl)isocyanate (PMPI), or a sulfo-succinimidyl variant or an analog thereof.

4. The cell-binding agent maytansinoid conjugate of claim 3 , wherein the non-cleavable linker is derived from SMCC.

5. The cell-binding agent maytansinoid conjugate of claim 1 , wherein the linker is at any one of the C-3 hydroxyl, C-14 hydroxymethyl, C-15 hydroxyl or C-20 desmethyl groups of the at least one maytansinoid.

6. The cell-binding agent maytansinoid conjugate of claim 1 , wherein the at least one maytansinoid is an N-methyl-alanine-containing ester of maytansinol.

7. The cell-binding agent maytansinoid conjugate of claim 1 , wherein the at least one maytansinoid is an N-methyl-cysteine-containing ester of maytansinol.

8. The cell-binding agent maytansinoid conjugate of claim 1 , wherein the at least one maytansinoid is represented by formula (II′-L), (II′-D) or (II′-D,L):

wherein:
Y 1 ′ represents (CR 7 R 8 ) 1 (CR 9 =CR 10 ) p (C=C) q A 0 (CR 5 R 6 ) m D u (CR 11 =CR 12 ) r (C=C) s B t (CR 3 R 4 ) n CR 1 R 2 S,
wherein:
R 1 to R 12 are each independently linear alkyl or alkenyl having 1 to 10 carbon atoms, branched or cyclic alkyl or alkenyl having 3 to 10 carbon atoms, phenyl, substituted phenyl or heterocyclic aromatic or heterocycloalkyl radical, and in addition, R 2 to R 12 can be H;
A, B, and D, each independently is cyclic alkyl or cyclic alkenyl having 3 to 10 carbon atoms, simple or substituted aryl, or heterocyclic aromatic or heterocycloalkyl radical;
1, m, n, o, p, q, r, s, t and u are each independently 0 or an integer of 1 to 5, provided that at least two of 1, m, n, o, p, q, r, s, t and u are not both zero; and
May represents a maytansinoid that bears a side chain at C-3 hydroxyl, C-14 hydroxymethyl, C-15 hydroxyl or C-20 desmethyl.

9. The cell-binding agent maytansinoid conjugate of claim 8 , wherein R 1 is methyl and R 2 is H or R 1 and R 2 are methyl.

10. The cell-binding agent maytansinoid conjugate of claim 8 , wherein R 1 is methyl, R 2 is H, R 5 , R 6 , R 7 and R 8 are each H, 1 and m are each 1, and n is 0; or
wherein R 1 and R 2 are methyl, R 5 , R 6 , R 7 , R 8 are each H, 1 and m are each 1, and n is 0.

11. The cell-binding agent maytansinoid conjugate of claim 1 , wherein the at least one maytansinoid is represented by formula (II-L), (II-D), or (II-D,L):

wherein:
Y 1 represents (CR 7 R 8 ) l (CR 5 R 6 ) m (CR 3 R 4 ) n CR 1 R 2 S, wherein:
R 1 to R 8 are each independently, linear alkyl or alkenyl having 1 to 10 carbon atoms, branched or cyclic alkyl or alkenyl having 3 to 10 carbon atoms, phenyl, substituted phenyl or heterocyclic aromatic or heterocycloalkyl radical, and in addition R 2 to R 8 can be H;
l, m and n are each independently an integer of 1 to 5, and in addition n can be 0; and
May represents a maytansinoid that bears the side chain at C-3 hydroxyl, C-14 hydroxymethyl, C-15 hydroxyl or C-20 desmethyl.

12. The cell-binding agent maytansinoid conjugate of claim 11 , wherein R 1 is methyl and R 2 is H or R 1 and R 2 are methyl.

13. The cell-binding agent maytansinoid conjugate of claim 11 , wherein R 1 is methyl, R 2 is H, R 5 , R 6 , R 7 and R 8 are each H, 1 and m are each 1, and n is 0; or
wherein R 1 and R 2 are methyl, R 5 , R 6 , R 7 , R 8 are each H, 1 and m are each 1, and n is 0.

14. The cell-binding agent maytansinoid conjugate of claim 1 , wherein the at least one maytansinoid is represented by formula 4 1 ′:

wherein:
Y 1 ′ represents (CR 7 R 8 ) l (CR 9 =CR 10 ) p (C=C) q A 0 (CR 5 R 6 ) m (CR 11 =CR 12 ) r (C=C) 8 B t (CR 3 R 4 ) n CR 1 R 2 S,
wherein:
R 1 to R 12 are each independently linear alkyl or alkenyl having 1 to 10 carbon atoms, branched or cyclic alkyl or alkenyl having 3 to 10 carbon atoms, phenyl, substituted phenyl or heterocyclic aromatic or heterocycloalkyl radical, and in addition, R 2 to R 12 can be H;
A, B, and D, each independently is cyclic alkyl or cyclic alkenyl having 3 to 10 carbon atoms, simple or substituted aryl, or heterocyclic aromatic or heterocycloalkyl radical;
l, m, n, o, p, q, r, s, t and u are each independently 0 or an integer of 1 to 5, provided that at least two of l, m, n, o, p, q, r, s, t and u are not both zero; and
May represents a maytansinoid that bears a side chain at C-3 hydroxyl, C-14 hydroxymethyl, C-15 hydroxyl or C-20 desmethyl.

15. The cell-binding agent maytansinoid conjugate of claim 14 , wherein R 1 is methyl and R 2 is H or R 1 and R 2 are methyl.

16. The cell-binding agent maytansinoid conjugate of claim 14 , wherein R 1 is methyl, R 2 is H, R 5 , R 6 , R 7 and R 8 are each H, 1 and m are each 1, and n is 0; or
wherein R 1 and R 2 are methyl, R 5 , R 6 , R 7 , R 8 are each H, 1 and m are each 1, and n is 0.

17. The cell-binding agent maytansinoid conjugate of claim 1 , wherein the at least one maytansinoid is represented by formula 4 1 :

wherein:
Y 1 represents (CR 7 R 8 ) l (CR 5 R 6 ) m (CR 3 R 4 ) n CR 1 R 2 S, wherein:
R 1 to R 5 are each independently, linear alkyl or alkenyl having 1 to 10 carbon atoms, branched or cyclic alkyl or alkenyl having 3 to 10 carbon atoms, phenyl, substituted phenyl or heterocyclic aromatic or heterocycloalkyl radical, and in addition R 2 to R 8 can be H; and
l, m and n are each independently an integer of

Assignees

• Immunogen Inc

Inventors

• Chari Ravi V J
• Blattler Walter A

Classifications

• C07K16/3092 Primary

  against tumour-associated mucins · CPC title

• A61K47/68033

  the drug being a maytansine · CPC title

• C07K16/32 Primary

  against translation products of oncogenes · CPC title

• C12N1/00 Primary

  Microorganisms; Compositions thereof (medicinal preparations containing material from protozoa, bacteria or viruses A61K35/66, from algae A61K36/02, from fungi A61K36/06; preparing medicinal bacterial antigen or antibody compositions, e.g. bacterial vaccines, A61K39/00); Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor · CPC title

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title