Anti-B7-H4 antibodies and immunoconjugates
The disclosure provides anti-B7-H4 antibodies and immunoconjugates and methods of using the same.
1. An isolated antibody that binds to B7-H4, wherein the antibody comprises:
(a) (i) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 7, (ii) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 10, and (iii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 6; or
(b) (i) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 41, (ii) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 44, and (iii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 40.
2. An isolated antibody that binds to B7-H4, wherein the antibody comprises:
(a) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 5, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 6, and (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 7; or
(b) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 39, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 40, and (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 41.
3. The antibody of claim 2 , further comprising a heavy chain framework FR3 sequence of SEQ ID NO: 51, 52 or 53.
4. An isolated antibody that binds to B7-H4, wherein the antibody comprises:
(a) (i) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 7, (ii) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 10, (iii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 6, (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 8, (v) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 9, and (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 10; or
(b) (i) HVR-H1 comprising the amino acid sequence of SEQ ID NO: 39, (ii) HVR-H2 comprising the amino acid sequence of SEQ ID NO: 40, (iii) HVR-H3 comprising the amino acid sequence of SEQ ID NO: 41, (iv) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 42, (v) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 43, and (vi) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 44.
5. The antibody of claim 4 , which is a monoclonal antibody.
6. The antibody of claim 4 , which is a human, humanized, or chimeric antibody.
7. The antibody of claim 4 , which is an antibody fragment that binds B7-H4.
8. The antibody of claim 4 , wherein B7-H4 is human B7-H4 of SEQ ID NO: 73.
9. An isolated antibody that binds to B7-H4, wherein the antibody comprises:
(a) (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 8, (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 9, and (iii) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 10; or
(b) (i) HVR-L1 comprising the amino acid sequence of SEQ ID NO: 42, (ii) HVR-L2 comprising the amino acid sequence of SEQ ID NO: 43, and (iii) HVR-L3 comprising the amino acid sequence of SEQ ID NO: 44.
10. The antibody of claim 9 , further comprising a light chain framework FR3 sequence of SEQ ID NO: 47.
11. The antibody of claim 4 , wherein the antibody comprises
(a) a VH sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 4; or
(b) a VL sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 3; or
(c) a VH sequence as in (a) and a VL sequence as in (b); or
(d) a VH sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 36; or
(e) a VL sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 35; or
(f) a VH sequence as in (d) and a VL sequence as in (e); or
(g) a VH sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 37; or
(h) a VH sequence as in (g) and a VL sequence as in (e); or
(i) a VH sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 38; or
(j) a VH sequence as in (i) and a VL sequence as in (e).
12. The antibody of claim 11 , comprising a VH sequence of SEQ ID NO: 4, a VH sequence of SEQ ID NO: 36, a VH sequence of SEQ ID NO: 37, or a VH sequence of SEQ ID NO: 38.
13. The antibody of claim 11 , comprising a VL sequence of SEQ ID NO: 3 or a VL sequence of SEQ ID NO: 35.
14. An isolated antibody that binds to B7-H4 comprising (a) a VH sequence of SEQ ID NO: 4 and a VL sequence of SEQ ID NO: 3; or (b) a VH sequence of SEQ ID NO: 36 and a VL sequence of SEQ ID NO: 35; or (c) a VH sequence of SEQ ID NO: 37 and a VL sequence of SEQ ID NO: 35; or (d) a VH sequence of SEQ ID NO: 38 and a VL sequence of SEQ ID NO: 35.
15. The antibody of claim 4 , which is an IgG1, IgG2a or IgG2b antibody.
16. Isolated nucleic acid encoding the antibody of claim 4 .
17. A host cell comprising the nucleic acid of claim 16 .
18. A method of producing an antibody comprising culturing the host cell of claim 17 so that the antibody is produced.
19. An immunoconjugate comprising the antibody of claim 4 and a cytotoxic agent.
20. The immunoconjugate of claim 19 having the formula Ab-(L-D)p, wherein:
(a) Ab is the antibody;
(b) L is a linker;
(c) D is a drug selected from a maytansinoid, an auristatin, a calicheamicin, a pyrrolobenzodiazepine, and a nemorubicin derivative; and
(d) p ranges from 1-8.
21. The immunoconjugate of claim 20 , wherein D is an auristatin.
22. The immunoconjugate of claim 21 , wherein D has formula D E
and wherein R 2 and R 6 are each methyl, R 3 and R 4 are each isopropyl, R 5 is H, is sec-butyl, each R 8 is independently selected from CH 3 , O—CH 3 , OH, and H; R 9 is H; and R 1-8 is C(R 8 ) 2 C(R 8 ) 2 -aryl.
23. The immunoconjugate of claim 20 , wherein the drug is MMAE.
24. The immunoconjugate of claim 20 , wherein D is a pyrrolobenzodiazepine of Formula A:
wherein the dotted lines indicate the optional presence of a double bond between C1 and C2 or C2 and C3;
R 2 is independently selected from H, OH, ═O, ═CH 2 , CN, R, OR, ═CH—R D , ═C(R D ) 2 , O—SO 2 —R, CO 2 R and COR, and optionally further selected from halo or dihalo, wherein R D is independently selected from R, CO 2 R, COR, CHO, CO 2 H, and halo;
R 6 and R 9 are independently selected from H, R, OH, OR, SH, SR, NH 2 , NHR, NRR′, NO 2 , Me 3 Sn and halo;
R 7 is independently selected from H, R, OH, OR, SH, SR, NH 2 , NHR, NRR′, NO 2 , Me 3 Sn and halo;
Q is independently 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 1-8 alkyl, C 3-8 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;
R 12 , R 16 , R 19 and R 17 are as defined for R 2 , R 6 , R 9 and R 7 respectively;
R″ is a C3-12 alkylene group, which chain may be interrupted by one or more heteroatoms and/or aromatic rings that are optionally substituted; and
X and X′ are independently selected from O, S and N(H).
25. The immunoconjugate of claim 24 , wherein D has the structure:
wherein n is 0 or 1.
26. The immunoconjugate of claim 20 , wherein D is a nemorubicin derivative.
27. The immunoconjugate of claim 26 , wherein D has a structure selected from:
28. The immunoconjugate of claim 20 , wherein the linker is cleavable by a protease.
29. The immunoconjugate of claim 28 , wherein the linker comprises a val-cit dipeptide or a Phe-homoLys dipeptide.
30. The immunoconjugate of claim 20 , wherein the linker is acid-labile.
31. The immunoconjugate of claim 30 , wherein the linker comprises hydrazone.
32. The immunoconjugate of claim 22 having the formula:
wherein S is a sulfur atom.
33. The immunoconjugate of claim 25 having the formula:
34. The immunoconjugate of claim 27 having a formula selected from:
35. The immunoconjugate of claim 20 , wherein p ranges from 2-5.
36. The immunoconjugate of claim 20 , wherein the antibody is an isolated antibody that binds to B7-H4 comprising (a) a VH sequence of SEQ ID NO: 4 and a VL sequence of SEQ ID NO: 3; or (b) a VH sequence of SEQ ID NO: 36 and a VL sequence of SEQ ID NO: 35; or (c) a VH sequence of SEQ ID NO: 37 and a VL sequence of SEQ ID NO: 35; or (d) a VH sequence of SEQ ID NO: 38 and a VL sequence of SEQ ID NO: 35.
37. A pharmaceutical formulation comprising the immunoconjugate of claim 20 and a pharmaceutically acceptable carrier.
38. The pharmaceutical formulation of claim 37 , further comprising an additional therapeutic agent.
39. The pharmaceutical formulation of claim 38 , wherein the additional therapeutic agent is Avastin® (bevacizumab).
40. A method of treating an individual having a B7-H4-positive cancer, the method comprising administering to the individual an effective amount of the immunoconjugate of claim 20 .
41. The method of claim 40 , wherein the B7-H4-positive cancer is selected from breast cancer, ovarian cancer, and endometrial cancer.
42. The method of claim 41 , further comprising administering an additional therapeutic agent to the individual.
43. The method of claim 42 , wherein the additional therapeutic agent is Avastin® (bevacizumab).
44. A method of inhibiting proliferation of an B7-H4-positive cell, the method comprising exposing the cell to the immunoconjugate of claim 20 under conditions permissive for binding of the immunoconjugate to B7-H4 on the surface of the cell, thereby inhibiting proliferation of the cell.
45. The method of claim 44 , wherein the cell is a breast, ovarian, or endometrial cancer cell.
46. The antibody of claim 4 conjugated to a label.
47. The antibody of claim 46 , wherein the label is a positron emitter.
48. The antibody of claim 47 , wherein the positron emitter is 89 Zr.
49. A method of detecting human B7-H4 in a biological sample comprising contacting the biological sample with the anti-B7-H4 antibody of claim 4 under conditions permissive for binding of the anti-B7-H4 antibody to a naturally occurring human B7-H4, and detecting whether a complex is formed between the anti-B7-H4 antibody and a naturally occurring human B7-H4 in the biological sample.
50. The method of claim 49 , wherein the anti-B7-H4 antibody is an antibody as in claim 4 .
51. The method of claim 49 , wherein the biological sample is a breast cancer sample, ovarian cancer sample, or endometrial cancer sample.
52. A method for detecting an B7-H4-positive cancer comprising (i) administering a labeled anti-B7-H4 antibody to a subject having or suspected of having a B7-H4-positive cancer, wherein the labeled anti-B7-H4 antibody comprises the anti-B7-H4 antibody of claim 4 , and (ii) detecting the labeled anti-B7-H4 antibody in the subject, wherein detection of the labeled anti-B7-H4 antibody indicates a B7-H4-positive cancer in the subject.
53. The method of claim 52 , wherein the labeled anti-B7-H4 antibody is an antibody as in claim 14 that is labeled.
54. The method of claim 52 , wherein the labeled anti-B7-H4 antibody comprises an anti-B7-H4 antibody conjugated to a positron emitter.
55. The method of claim 54 , wherein the positron emitter is 89 Zr.