Antibody compounds with reactive arginine and related antibody drug conjugates
The present invention provides antibody compounds that contain a substitution of arginine for the reactive lysine residue (Lys99) in the hydrophobic cleft (38C2_Arg). The invention also provides antibody drug conjugate compounds (ADCs) that contain cargo moieties that are site-specifically conjugated to the engineered arginine residue in the 38C2_Arg variant antibody. Further provided in the invention are therapeutic applications of the compounds.
1. An antibody compound, comprising a variant of catalytic antibody 38C2 (38C2_Arg), or antigen binding fragment thereof, that contains a substitution of arginine for the reactive lysine residue (Lys99) in the hydrophobic cleft.
2. The antibody compound of claim 1 , wherein the catalytic antibody is humanized 38C2 (h38C2).
3. The antibody compound of claim 1 , wherein the 38C2_Arg or antigen binding fragment thereof comprises heavy chain and light chain variable domain sequences respectively shown in SEQ ID NOs:1 and 2.
4. The antibody compound of claim 1 , which is a dual variable domain (DVD) compound or an antigen-binding fragment thereof comprising (i) the 38C2_Arg or antigen binding fragment thereof, and (ii) a second antibody variable domain recognizing a target of interest.
5. The antibody compound of claim 4 , wherein 38C2_Arg is positioned closer to the C-terminus in the antibody compound than the second variable domain.
6. The antibody compound of claim 4 , which is a homodimeric molecule comprising Lys99Arg substitution in both antibody arms.
7. The antibody compound of claim 4 , which is a heterodimeric molecule comprising Lys99Arg substitution in only one antibody arm.
8. The antibody compound of claim 4 , wherein the dual variable domain compound is a bispecific immunoglobulin molecule.
9. The antibody compound of claim 4 , comprising an antigen-binding fragment of a dual variable domain (DVD) compound that is a Fab, Fab′, F(ab′) 2 , Fv or scFv.
10. The antibody compound of claim 9 , comprising a Fab.
11. The antibody compound of claim 4 , comprising a chimeric immunoglobulin sequence or a humanized immunoglobulin sequence.
12. The antibody compound of claim 4 , where the target of interest is a tumor cell surface antigen.
13. The antibody compound of claim 12 , wherein the tumor cell surface antigen is HER2, FOLR1, FCMR, CD138, CD79B, PSMA, BCMA, CD38, SLAMF7, Siglec-6, CD70, ROR1 or ROR2.
14. An antibody drug conjugate (ADC) comprising at least one drug moiety that is conjugated to an antibody compound via a reactive arginine residue in the antibody compound, wherein the antibody compound comprises a variant of catalytic antibody 38C2 (38C2_Arg), or antigen binding fragment thereof, that contains a substitution of arginine for the reactive lysine residue in the hydrophobic cleft.
15. The antibody drug conjugate of claim 14 , wherein the antibody compound is a dual variable domain (DVD) compound or an antigen-binding fragment thereof comprising (i) the 38C2_Arg or antigen binding fragment thereof, and (ii) a second antibody variable domain recognizing a target of interest.
16. The antibody drug conjugate of claim 15 , wherein the 38C2_Arg or antigen binding fragment thereof comprises heavy chain and light chain variable domain sequences respectively shown in SEQ ID NOs:1 and 2, and the target of interest is HER2.
17. The antibody drug conjugate of claim 16 , wherein the antibody compound is a dual variable domain (DVD)-Fab comprising heavy chain and light chain sequences shown in SEQ ID NOs:5 and 7, respectively.
18. The antibody drug conjugate of claim 16 , wherein the antibody compound is a dual variable domain (DVD)-IgG1 comprising heavy chain and light chain sequences shown in SEQ ID NOs:6 and 7, respectively.
19. A pharmaceutical composition, comprising an effective amount of the antibody drug conjugate of claim 15 and optionally a pharmaceutically acceptable carrier.
20. A method for treating cancer in a subject, comprising administering to the subject in need of treatment the pharmaceutical composition of claim 19 .
21. The antibody drug conjugate of claim 14 , wherein the catalytic antibody is humanized 38C2 (h38C2).
22. The antibody drug conjugate of claim 14 , wherein the drug moiety is conjugated to the antibody compound via a linker moiety.
23. The antibody drug conjugate of claim 22 :
wherein the drug moiety is derivatized with the linker moiety prior to conjugation with the antibody compound; or
wherein the linker moiety is a cleavable linker; or
wherein the linker moiety comprises phenylglyoxal (PGO), glyoxal (GO), or methylglyoxal (MGO).
24. The antibody drug conjugate of claim 15 , wherein the DVD compound or an antigen-binding fragment thereof is:
a homodimeric molecule comprising Lys99Arg substitution in both antibody arms; or
a heterodimeric molecule comprising Lys99Arg substitution in only one antibody arm.
25. The antibody drug conjugate of claim 15 , wherein the antibody compound comprises an antigen-binding fragment of a dual variable domain (DVD) compound that is a Fab, Fab′, F(ab′) 2 , Fv or scFv.
26. The antibody drug conjugate of claim 25 , wherein the antibody compound comprises a Fab.
27. The antibody drug conjugate of claim 15 , where the target of interest is a tumor cell surface antigen.
28. The antibody drug conjugate of claim 27 , wherein the tumor cell surface antigen is HER2, FOLR1, FCMR, CD138, CD79B, PSMA, BCMA, CD38, SLAMF7, Siglec-6, CD70, ROR1 or ROR2.
29. The antibody drug conjugate of claim 14 , wherein the drug moiety is a cytotoxic agent or a siRNA.
30. The antibody drug conjugate of claim 29 , wherein the cytotoxic agent is selected from a toxin, a chemotherapeutic agent, a photoabsorber, an antibiotic, a radioactive isotope, a chelated radioactive isotope and a nucleolytic enzyme.
31. The antibody drug conjugate of claim 14 , wherein the drug moiety is an auristatin, a dolastatin, a cemadotin, a camptothecin, an amanitin, a maytansinoid, a pyrrolobenzodiazepine, an indolinobenzodiazepine, a duocarmycin, an enediyne, a doxorubicin or a polymerized drug.
32. The antibody drug conjugate of claim 14 , wherein the drug moiety is monomethyl auristatin F (MMAF).
33. The antibody drug conjugate of claim 18 , wherein the dual variable domain (DVD)-IgG1 is:
a homodimeric molecule comprising Lys99Arg substitution in both antibody arms; or
a heterodimeric molecule comprising Lys99Arg substitution in only one antibody arm.
34. The antibody drug conjugate of claim 33 , wherein two different drug moieties are conjugated to the two antibody arms of the heterodimeric dual variable domain (DVD)-IgG1 molecule.