Diels-alder conjugation methods

Patent metadata

Field	Value
Publication number	US-11701427-B2
Application number	US-202117232854-A
Country	US
Kind code	B2
Filing date	Apr 16, 2021
Priority date	Apr 16, 2020
Publication date	Jul 18, 2023
Grant date	Jul 18, 2023

Abstract

Described herein are protein-payload conjugates and compositions thereof that are useful, for example, for target-specific delivery of therapeutic and/or imaging agent moieties. In certain embodiments, provided are specific and efficient methods for producing protein-payload constructs (e.g., antibody-drug conjugates) utilizing a combination of transglutaminase and Diels-Alder techniques. Antibody-drug conjugates and compositions which comprise glutaminyl-modified antibodies, Diels-Alder adducts, and reactive payloads and are provided.

First claim

What is claimed is:

1. A compound having a structure according to Formula (II):

wherein:
BA is a binding agent;
Gln is a glutamine residue of said BA;
SP is absent or a spacer;
Q is CHR, (CHR) 2 , or O bridge;
R is independently at each occurrence H, C 1-3 alkyl, —OC 1-3 alkyl, or —NHC 1-3 alkyl;
L is a linker;
D is a therapeutic moiety, wherein the therapeutic moiety is a maytansinoid, a tubulysin, an auristatin, a dolastatin, a camptothesin, a pyrrolobenzodiazepine, an antibiotic, an antiviral agent, an anti-inflammatory agent, an immunomodulator, an antifungal agent, a steroid, or D is an imaging agent moiety;
n is an integer from 1 to 3; and
d is an integer from 1 to 6.

2. The compound of claim 1 according to Formula (IIA):

3. The compound of claim 1 according to Formula (IIB):

4. The compound of claim 3 , wherein the two moieties D are the same.

5. The compound of claim 3 , wherein the two moieties D are different.

6. The compound of claim 1 according to Formula (IIIA):

7. The compound of claim 1 according to Formula (IIIB):

8. The compound of claim 1 according to Formula (IIIC):

9. The compound of claim 1 , wherein R is H at each occurrence.

10. The compound of claim 1 , wherein at least one R is not H.

11. The compound of claim 10 , wherein at least one R is C 1-3 alkyl.

12. The compound of claim 10 , wherein at least one R is methyl.

13. The compound of claim 1 having a structure according to Formula (IV):

14. The compound according to claim 1 , wherein BA is an antibody, or an antigen-binding fragment thereof.

15. The compound according to claim 14 , wherein BA is a monoclonal humanized antibody.

16. The compound according to claim 14 , wherein BA is a human epidermal growth factor receptor 2 (HER-2) antibody, an antigen-binding fragment thereof, a macrophage scavenger receptor 1 (MSR1) antibody, or an antigen-binding fragment thereof.

17. The compound according to claim 1 , wherein Gln is independently at each occurrence Q295 or N297Q.

18. The compound according to claim 1 , wherein d is 2, 4, or 6.

19. The compound according to claim 18 , wherein d is 2.

20. The compound according to claim 18 , wherein d is 4.

21. The compound according to claim 1 , wherein D is a therapeutic moiety, wherein the therapeutic moiety is a maytansinoid, a tubulysin, an auristatin, a dolastatin, a camptothesin, a pyrrolobenzodiazepine (PBD), an antibiotic, an antiviral agent, an anti-inflammatory agent, an immunomodulator, or an antifungal agent.

22. The compound according to claim 1 , wherein D is a therapeutic moiety, wherein the therapeutic moiety is an auristatin, a pyrrolobenzodiazepine (PBD), or an ansamysin antibiotic.

23. The compound according to claim 1 , wherein D is a therapeutic moiety, wherein the therapeutic moiety is monomethyl auristatin E (MMAE), PBD-1, or rifamycin.

24. The compound according to claim 1 , wherein D is an imaging agent moiety selected from the group consisting of a dye, a chelator, a radionuclide, and an oligonucleotide.

25. The compound of claim 24 , wherein D is an Alexa Fluor fluorescent dye selected from the group consisting of Alexa Fluor 647, Alexa Fluor 488, Alexa Fluor 594, Alexa Fluor 555, and Alexa Fluor 568.

26. The compound according to claim 1 , wherein SP is one or more of —(CH 2 ) u —, —((CH 2 ) u —O—) v —, —NH—, or —C(O)— and combinations thereof, wherein subscripts u and v are independently at each occurrence an integer from 1 to 20.

27. The compound according to claim 26 , wherein SP is one or more of: —(CH 2 ) u —, C(O)—, —NH—, —(CH 2 ) u —NH—C(O)—, —(CH 2 ) u —C(O)—NH—, —(CH 2 ) u —C(O)—NH—(CH 2 ) v —, —(CH 2 —CH 2 —O) v —, —(CH 2 ) u —(O—CH 2 —CH 2 ) v —C(O)—NH—, —(CH 2 —CH 2 —O) v —(CH 2 ) u —C(O)—NH—(CH 2 ) u —, —NH—(CH 2 ) u —, —NH—(CH 2 ) u —C(O)—, —NH—(CH 2 ) u —C(O)—NH—(CH 2 ) v —, —NH—(CH 2 —CH 2 —O) v —, —NH—(CH 2 —CH 2 —O) v C(O)—, —NH—(CH 2 —CH 2 —O) v —(CH 2 ) u —, —NH—(CH 2 —CH 2 —O) v —(CH 2 ) u —C(O)—, —NH—(CH 2 —CH 2 —O) v —(CH 2 ) u —C(O)—NH—(CH 2 ) u —, —(CH 2 ) u —NH—C(O)—, —(CH 2 ) u —C(O)—NH—(CH 2 —CH 2 —O) v —C(O)—NH—, —NH—(CH 2 ) u —C(O)—NH—,

or combinations thereof; and
wherein subscripts u, v, w, and x are independently an integer from 1 to 20.

28. The compound according to claim 26 , wherein SP is —(CH 2 ) u —; and
wherein the subscript u is an integer from 1 to 5.

29. The compound according to claim 26 , wherein SP is

wherein subscripts u, v, w, and x are independently an integer from 1 to 12.

30. The compound according to claim 26 , wherein SP is

31. The compound according to claim 1 , wherein L is:

wherein l, m and n are independently at each occurrence an integer from 0 to 20; and p is an integer from 0 to 4.

32. The compound according to claim 31 , wherein the one or more amino acid is independently at each occurrence selected from the group consisting of glycine, serine, alanine, valine, phenylalanine, proline, and citrulline.

33. The compound according to claim 31 , wherein the one or more amino acid is valine-citrulline (-VC-), or valine-alanine (-VA-).

34. The compound according to claim 1 , wherein L is:

wherein n is an integer from 1 to 6; m is an integer from 2 to 10; p is an integer from 1 to 3, and the amino acid is independently at each occurrence valine, citrulline or alanine.

35. The compound according to claim 1 , wherein the moiety D is attached to linker L via a tertiary amino group.

36. The compound according to claim 1 , wherein

is according to Formula (I-DA):

wherein the

is the bond to the spacer or binding agent, an

Assignees

• Regeneren Pharmaceuticals Inc
• Regeneron Pharma

Inventors

• Nittoli Thomas

Classifications

• A61K51/1093

  conjugates with carriers being antibodies · CPC title

• A61K47/65

  Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers · CPC title

• A61K38/1729

  Cationic antimicrobial peptides, e.g. defensins · CPC title

• A61K31/395

  having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins · CPC title

• A61P35/00

  Antineoplastic agents · CPC title