Methods of making DLL3 antibody drug conjugates

Patent metadata

Field	Value
Publication number	US-9931420-B2
Application number	US-201715694602-A
Country	US
Kind code	B2
Filing date	Sep 1, 2017
Priority date	Feb 24, 2012
Publication date	Apr 3, 2018
Grant date	Apr 3, 2018

Abstract

Novel modulators, including antibodies and derivatives thereof, and methods of using such modulators to treat proliferative disorders are provided.

First claim

The invention claimed is:

1. A method of making an antibody drug conjugate comprising an anti-DLL3 antibody, a linker and a drug, the method comprising the steps of:
(a) providing a linker-drug conjugate, wherein the drug comprises a pyrrolobenzodiazepine (PBD) comprising the formula AC:

wherein:
the dotted lines indicate the optional presence of a double bond, and wherein only one of the dotted lines in a given ring can be a double bond;
R 2 is selected from H, OH, ═O, ═CH 2 , CN, R, OR, ═CH—R D , ═C(R D ) 2 , OSO 2 R, CO 2 R, COR, and halo, where R D is selected from R, CO 2 R, COR, CHO, CO 2 H, and halo;
R 6 and R 9 are each independently selected from H, R, OH, OR, SH, SR, NH 2 , NHR, NRR′, NO 2 , Me 3 Sn and halo;
R 7 is selected from H, R, OH, OR, SH, SR, NH 2 , NHR, NRR′, NO 2 , Me 3 Sn and halo;
R 10 is the linker connected to the anti-DLL3 antibody;
Q is selected from O, S and NH;
R 11 is either H, or R or, where Q is O, SO 3 M, where M is a metal cation;
R and R′ are each independently selected from optionally substituted C 1-12 alkyl, C 3-20 heterocyclyl and C 5-20 aryl groups, and optionally in relation to the group NRR′, R and R′ together with the nitrogen atom to which they are attached form an optionally substituted 4-, 5-, 6- or 7-membered heterocyclic ring;
X is selected from O, S, and N(H);
R 2″ , R 6″ , R 7″ , R 9″ , and X″ are as defined according to R 2 , R 6 , R 7 , R 9 , and X, respectively; and
R″ is a C 3-12 alkylene group, which comprises a chain optionally interrupted by one or more heteroatoms, one or more rings, or both one or more heteroatoms and one or more rings, wherein the optional one or more rings are optionally substituted; and
(b) conjugating the linker-drug conjugate to the anti-DLL3 antibody, wherein the anti-DLL3 antibody comprises a humanized anti-DLL3 antibody comprising a light chain variable region set forth as SEQ ID NO: 212 and a heavy chain variable region set forth as SEQ ID NO: 213.

2. The method of claim 1 , wherein the linker comprises a terminal thiol reactive group.

3. The method of claim 2 , wherein the terminal thiol reactive group comprises a maleimide group.

4. The method of claim 2 , wherein the humanized anti-DLL3 antibody comprises a reducible cysteine that can be made reactive for conjugating with the linker-drug.

5. The method of claim 4 , wherein the humanized anti-DLL3 antibody and the linker-drug conjugate are conjugated by reaction of the reduced cysteine of the humanized anti-DLL3 antibody with the terminal thiol reactive group of the linker.

6. The method of claim 1 , wherein the antibody drug conjugate comprises the structure:

wherein:
CBA is a cell binding agent, which is the humanized anti-DLL3 antibody;
A, L 1 , and L 2 are components of the linker;
A is a connecting group connecting L 1 to the cell binding agent (CBA);
L 1 is optionally a cleavable linker;
L 2 is a covalent bond or together with the —OC(═O)— group forms a self-immolative linker; and
wherein the linker is attached to the pyrrolobenzodiazepine (PBD) at the position of the asterisk (*).

7. The method of claim 6 , wherein the L 1 comprises a cleavable linker.

8. The method of claim 7 , wherein the cleavable linker comprises a dipeptide.

9. The method of claim 8 , wherein the dipeptide is Phe-Lys, Val-Ala, Val-Lys, Ala-Lys, Val-Cit, Phe-Cit, Leu-Cit, Ile-Cit, Phe-Arg, or Trp-Cit.

10. The method of claim 9 , wherein the dipeptide is Val-Ala.

11. The method of claim 6 , wherein the moiety:

comprises the structure:

wherein the wavy line indicates the point of attachment of the structure directly to A or to a remaining portion of L 1 that is further connected to A.

12. The method of claim 1 , wherein R 2 is R, wherein R is a C 5-20 aryl group.

13. The method of claim 1 , wherein R 6 and R 9 are H.

14. The method of claim 1 , wherein R 7 is OR.

15. The method of claim 14 , wherein R is a C 1 alkyl.

16. The method of claim 1 , wherein Q is O.

17. The method of claim 16 , wherein R 11 is H.

18. The method of claim 1 , wherein X and X″ are O.

19. The method of claim 1 , further comprising the step of:
(c) purifying the antibody drug conjugate by ion exchange column chromatography.

20. The method of claim 1 , further comprising the step of:
(d) analyzing drug to antibody ratio (DAR).

21. A method of making an antibody drug conjugate comprising an anti-DLL3 antibody, a linker and a drug, wherein the antibody drug conjugate comprises the structure:

wherein:
CBA is a cell binding agent, which is the anti-DLL3 antibody;
A, L 1 , and L 2 are components of the linker;
A is a connecting group connecting L 1 to the cell binding agent (CBA);
L 1 is optionally a cleavable linker;
L 2 is a covalent bond or together with the —OC(═O)— group forms a self-immolative linker; and
wherein the linker is attached to the drug at the position of the asterisk (*),
the method comprising the steps of:
(a) providing a linker-drug conjugate, wherein the drug comprises a pyrrolobenzodiazepine (PBD) comprising the formula AC:

wherein:
the dotted lines indicate the optional presence of a double bond, and wherein only one of the dotted lines in a given ring can be a double bond;
R 2 is selected from H, OH, ═O, ═CH 2 , CN, R, OR, ═CH—R D , ═C(R D ) 2 , OSO 2 R, CO 2 R, COR, and halo, where R D is selected from R, CO 2 R, COR, CHO, CO 2 H, and halo;
R 6 and R 9 are each independently selected from H, R, OH, OR, SH, SR, NH 2 , NHR, NRR′, NO 2 , Me 3 Sn and halo;
R 7 is selected from H, R, OH, OR, SH, SR, NH 2 , NHR, NRR′, NO 2 , Me 3 Sn and halo;
R 10 is the linker connected to the anti-DLL3 antibody;
Q is selected from O, S and NH;
R 11 is either H, or R or, where Q is O, SO 3 M, where M is a metal cation;
R and R′ are each independently selected from optionally substituted C 1-12 alkyl, C 3-20 heterocyclyl and C 5-20 aryl groups, and optionally in relation to the group NRR′, R and R′ together with the nitrogen atom to which they are attached form an optionally substituted 4-, 5-, 6- or 7-membered heterocyclic ring;
X is selected from O, S, and N(H);
R 2″ , R 6″ , R 7″ , R 9″ , and X″ are as defined according to R 2 , R 6 , R 7 , R 9 , and X, respectively; and
R″ is a C 3-12 alkylene group, which comprises a chain optionally interrupted by one or more heteroatoms, one or more rings, or both one or more heteroatoms and one or more rings, wherein the optional one or more rings are optionally substituted;
wherein the moiety:

Assignees

• Abbvie Stemcentrx Llc

Inventors

• Stull Robert A
• Saunders Laura
• Dylla Scott J
• Foord Orit
• Liu David
• Torgov Michael
• Shao Hui

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• A61P11/00

  Drugs for disorders of the respiratory system · CPC title

• G01N33/5759

  involving compounds localised on the membrane of tumour or cancer cells · CPC title

• G01N33/5752

  of the lungs · CPC title

• A61K47/6809

  Antibiotics, e.g. antitumor antibiotics anthracyclins, adriamycin, doxorubicin or daunomycin · CPC title