Combination therapies for treatment of cancer

Abstract

Provided herein are methods of treating cancer in a human subject, comprising administering to the subject: i) an antibody-drug conjugate (ADC) comprising an antibody, a cleavable linker and a cytotoxic agent, ii) a bispecific checkpoint inhibitor, and iii) a PARP1 inhibitor. Further provided herein are kits comprising i) an antibody-drug conjugate (ADC) comprising an antibody, a cleavable linker and a cytotoxic agent, ii) a bispecific checkpoint inhibitor, and iii) a PARP1 inhibitor.

Description

AZ Ref B7H4-102-PCT01-NPCOMBINATION THERAPIES FOR TREATMENT OF CANCER CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the priority benefit of U. S. Provisional Application No.63 / 760,149 filed February 19, 2025, and U. S. Provisional Application No. 63 / 913,368 filed November 7, 2025, both of which are incorporated herein by reference in their entireties.INCORPORATION-BY-REFERENCE OF MATERIAL SUBMITTED ELECTRONICALLY

[0002] Incorporated by reference in its entirety herein is a computer-readable nucleotide / amino acid sequence listing submitted concurrently herewith and identified as follows: one 91,011-byte file named “B7H4-102-PCT01-NP.xml” created on February 16, 2026.FIELD

[0003] The present disclosure provides methods of treating cancer in a human subject, comprising administering to the subject an antibody-drug conjugate (ADC), a bispecific checkpoint inhibitor, and a PARP1 inhibitor. The present disclosure further provides kits comprising an ADC, a bispecific checkpoint inhibitor, and a PARP1 inhibitor.BACKGROUND

[0004] Surgery, radiation therapy and chemotherapy, alone or in combination, are the most traditional and widely used treatment methods for cancer. While these treatment methods are effective in removing or killing cancer cells, they often result in undesirable side effects such as hair loss, anemia, severe nausea and death of healthy cells in patients treated. These limitations present an urgent need for innovative and less harmful treatment methods of cancer. Antibodybased cancer therapies rely on the recognition and binding of antibody-drug conjugates to specific proteins on cancer cells. Antibody drug conjugates (ADCs) can take advantage of the specificity of the antibody portion of the conjugate to deliver a highly toxic agent directly to the cells to be killed. Combination therapies have the potential to improve treatment outcomes by attacking malignant cells and tumors in more than one way.AZ Ref: B7H4-102-PCT01-NPSUMMARY

[0005] The present disclosure provides, inter alia, a method of treating cancer in a human subject in need thereof, comprising administering to the human subject: a) an antibody-drug conjugate (ADC) comprising: i) an antibody or antigen binding fragment thereof which binds to a B7-H4 polypeptide, ii) a cleavable linker, and iii) a cytotoxic agent; b) a bispecific checkpoint inhibitor; and c) aPARPl inhibitor.

[0006] In all aspects, the antibody or antigen binding fragment thereof of the ADC comprises:a) a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively, or a functional variant thereof;b) a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12, respectively, or a functional variant thereof;c) a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, and SEQ ID NO: 18, respectively, or a functional variant thereof;d) a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 24, respectively, or a functional variant thereof; ore) a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, and SEQ ID NO: 30, respectively, or a functional variant thereof.AZ Ref: B7H4-102-PCT01-NP

[0007] In specific embodiments according to all aspects, the antibody or antigen binding fragment thereof of the ADC comprises: a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively, or a functional variant thereof.

[0008] The antibody or antigen binding fragment thereof of the ADC may comprise:a) a variable heavy (VH) chain and a variable light (VL) chain comprising the amino acid sequence of SEQ ID NO: 45 and SEQ ID NO: 34, respectively, or a functional variant thereof;b) a variable heavy (VH) chain and a variable light (VL) chain comprising the amino acid sequence of SEQ ID NO: 33 and SEQ ID NO: 34, respectively, or a functional variant thereof;c) a variable heavy (VH) chain and a variable light (VL) chain comprising the amino acid sequence of SEQ ID NO: 43 and SEQ ID NO: 34, respectively, or a functional variant thereof;d) a variable heavy (VH) chain and a variable light (VL) chain comprising the amino acid sequence of SEQ ID NO: 46 and SEQ ID NO: 34, respectively, or a functional variant thereof;e) a variable heavy (VH) chain and a variable light (VL) chain comprising the amino acid sequence of SEQ ID NO: 47 and SEQ ID NO: 34, respectively, or a functional variant thereof;f) a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 31, and SEQ ID NO: 32, respectively, or a functional variant thereof;g) a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 35 and SEQ ID NO: 36, respectively, or a functional variant thereof;AZ Ref: B7H4-102-PCT01-NPh) a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 37 and SEQ ID NO: 38, respectively, or a functional variant thereof; ori) a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 39 and SEQ ID NO: 40, respectively, or a functional variant thereof.

[0009] In specific embodiments according to all aspects, the antibody or antigen binding fragment thereof of the ADC comprises: a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 45 and SEQ ID NO: 34, respectively, or a functional variant thereof. The antibody or antigen binding fragment thereof of the ADC may bind an OVCAR4 cell line. In specific embodiments according to all aspects, the antibody or antigen binding fragment thereof of the ADC comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 41.

[0010] In specific embodiments according to all aspects, the antibody or antigen binding fragment thereof of the ADC comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 52. In specific embodiments according to all aspects, the antibody or antigen binding fragment thereof of the ADC comprises a light chain constant region comprising the amino acid sequence of SEQ ID NO: 42. In specific embodiments according to all aspects, the antibody or antigen binding fragment thereof of the ADC comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 51; and a light chain comprising the amino acid sequence of SEQ ID NO: 44. In specific embodiments according to all aspects, the antibody or antigen binding fragment thereof of the ADC comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 48; and a light chain comprising the amino acid sequence of SEQ ID NO: 44.

[0011] The antibody or antigen binding fragment thereof of the ADC may be a monoclonal antibody. In particular embodiments, the antibody or antigen binding fragment thereof of the ADC may be a human or humanized monoclonal antibody. In specific embodiments according to all aspects, the cleavable linker of the ADC is an mp-PEG8-val-ala linker. In specific embodiments according to all aspects, the cytotoxic agent of the ADC is a topoisomerase inhibitor (TOPli), a tubulysin derivative, a pyrrolobenzodiazepine, or a combination thereof. In specific embodiments according to all aspects, the cytotoxic agent of the ADC is a topoisomerase inhibitor.AZ Ref: B7H4-102-PCT01-NP

[0012] In specific embodiments according to all aspects, the cytotoxic agent is a topoisomerase inhibitor and wherein the ii) cleavable linker and iii) cytotoxic agent of the ADC comprise a compound of formula I:OH Oand salts and solvates thereof, wherein RLis the cleavable linker, which is selected from:Owherein Q is:Q NH, where Qxis such that Q is an amino-acid residue, a dipeptide residue, a tripeptide residue or a tetrapeptide residue;wherein X is:'S' a N H H Hwhere a = 0 to 5, bl = 0 to 16, b2 = 0 to 16, cl = 0 or 1, c2 = 0 or 1, d = 0 to 5, at least bl or b2 = 0, at least cl or c2 = 0; andwherein GLis a linker for connecting to the antibody or antigen binding fragment;AZ Ref: B7H4-102-PCT01-NPandwherein RL1and RL2are independently selected from H and methyl, or together with the carbon atom to which they are bound form a cyclopropylene or cyclobutylene group, and e is 0 or 1.

[0013] In particular embodiments, RLis of formula la.

[0014] In particular embodiments, Q is:(a) an amino acid residue selected from: Phe, Lys, Vai, Ala, Cit, Leu, He, Arg, and Trp; or(b) a dipeptide residue selected from:NH-Phe-Lys-C=o,NH-Vai-Ala-c=o,NH-Val-Lys-c=o,NHAla-Lys-c NH-Val-Cit-c NH-Phe-Cit-c NH-Leu-Cit-c NH-Ile-Cit-c°,NH-Phe-Arg-C=o, NH_Trp-cit_ c=o,andNH _Gly-Val-c=o; or(c) a tripeptide residue selected from:NH-Glu-Val-Ala-C=o,NH-Glu-Val-Cit-c=o,NH-aGlu-Val-Ala-C=o, andNH-aGlu-Val-Cit-c=o; or(d) a tetrapeptide residue selected from:NH-Gly-Gly-Phe-Glyc°; andNH-Gly-Phe-Gly-Glyc°

[0015] In particular embodiments, a is: (a) 0 to 3; or (b) 0 or 1; or (c) 0.

[0016] In particular embodiments, bl is: (a) 0 to 8; or (b) 0; or (c) 2; or (d) 3; or (e) 4; or (f) 5; or (g) 8.

[0017] In particular embodiments, b2 is: (a) 0 to 8; or (b) 0; or (c) 2; or (d) 3; or (e) 4; or (f) 5; or (g) 8.

[0018] In particular embodiments, (a) cl is 0 or 1; (b) c2 is 0 or 1; and (c) at least one of clAZ Ref: B7H4-102-PCT01-NPand c2 is 0.

[0019] In particular embodiments, d is: (a) 0 to 3; or (b) 1 or 2; or (c) 2; or (d) 5.

[0020] In particular embodiments:(a) a is 0, bl is 0, cl is 1, c2 is 0 and d is 2, and b2 is 0, 2, 3, 4, 5 or 8; or(b) a is 1, b2 is 0, cl is 0, c2 is 0 and d is 0, and bl is 0, 2, 3, 4, 5 or 8; or(c) a is 0, bl is 0, cl is 0, c2 is 0 and d is 1, and b2 is 0, 2, 3, 4, 5 or 8; or(d) b 1 is 0, b2 is 0, cl is 0, c2 is 0, one of a and d is 0, and the other of a and d is 1 or 5; or(e) a is 1, b2 is 0, cl is 0, c2 is 1, d is 2, and bl is 0, 2, 3, 4, 5 or 8.

[0021] In particular embodiments, GLis selected from:(GL1 1) O (G16) 0°xz°^'y-Nxo(G1 1-2) o (tnVt „Br—Ar,xo(GL2) 0 (G“)AV-k o0(G13-1) (CHs— s^*’’0(NO2)AZ Ref: B7H4-102-PCT01-NP where the NO2 group is optional(G13-2) (G110)S— / Q HH (NO2)where the NO2 group is optional(G133) (G“')s—O2N-\2_ / where the NO2 group is optional O(G13-4) (GL12)S—02Ndwhere the NO2 group is optional(GL4) t _ 0 (G143) Y / N-N — / Hal N— I * H5Where Hal = I, Br, Cl(G“) (GL14) H2N XHal—w lere Ar represents a C5-6 arylene group, and X represents C1-4 alkyl.

[0022] In particular embodiments, GLis selected from GL land GL1'2.

[0023] In particular embodiments, RLis of formula lb, and:(a) both RL1and RL2are H; orAZ Ref: B7H4-102-PCT01-NP(b) RL1is H and RL2is methyl; or(c) both RL1and RL2are methyl; or(d) wherein RL1and RL2together with the carbon atom to which they are bound form a cyclopropylene group; or(e) wherein RL1and RL2together with the carbon atom to which they are bound form a cyclobutylene group.

[0024] In specific embodiments according to all aspects, the ADC is of formula IV:L - (DL)P(IV)or a pharmaceutically acceptable salt or solvate thereof,wherein L is the i) antibody or antigen binding fragment thereof, DLis a drug linker unit comprising the ii) cleavable linker and the iii) cytotoxic agent, and DLis of formula III:HNOHwherein RLis the cleavable linker selected from:(ia’):, where Q and X are as defined above and GLLis a linker connected to the antibody or antigen binding fragment; andAZ Ref: B7H4-102-PCT01-NP(ib’):lb', where RL1and RL2are as defined above; andp is an integer of from 1 to 20.

[0025] In particular embodiments, GLLis selected from:(GLL1-1) CBA0 (GLL8-1)CBAjxoo (GLL8-2) N »CBACBAj yxo(G"2) 0 (GLL9-1)CBAj N''Vk 00 CEA(G1 1 3-1) (G132'-2)CBApsX■jr''* CEA (G^3-2) (GLL1°)MZ 3\>*H< / AHAZ Ref B7H4-102-PCT01-NP(GIJ-4) CBA| _ (GI L")1 \ HX— N0 HNZ7 / \H L(G"s) p (GI LI2)CBA| / / °H L(GLL6) 0 (GLL13)CBA| / / AAA / (GIJ 7) CBA^ (GI LI4)where Ar represents a C5-6 arylene group and X represents C1-4 alkyl.

[0026] In particular embodiments, GLLis selected from GLL land GLL1'2.

[0027] In specific embodiments according to all aspects, the drug loading (p) of the cytotoxic agent to the antibody or antibody binding fragment thereof of the ADC is an integer from 1 to about 10. This may also be referred to as the Drug to Antibody Ratio (DAR). In a specific embodiment, the drug loading (p) / DAR is about 8; that is to say, there are about 8 cytotoxic agents conjugated to the antibody or antibody binding fragment thereof.

[0028] In specific embodiments according to all aspects, the topoisomerase inhibitor is a compound of formula A:AZ Ref: B7H4-102-PCT01-NPas a single enantiomer or in an enantiomerically enriched form.

[0029] In specific embodiments according to all aspects, the topoisomerase inhibitor is a compound with the formula VI:where Q is as defined above.

[0030] In specific embodiments according to all aspects, the ii) linker and iii) cytotoxic agent together comprise the following compound:AZ Ref: B7H4-102-PCT01-NP(SG3932).

[0031] In specific embodiments according to all aspects, the bispecific checkpoint inhibitor comprises: a) a first binding domain that specifically binds to Programmed Death-1 (PD-1); and b) a second binding domain that specifically binds to T cell immunoreceptor with Ig and ITIM domains (TIGIT). In a specific embodiments according to all aspects, the first binding domain comprises a heavy chain variable domain comprising a HCDR1 having the amino acid sequence of SEQ ID NO: 56, a HCDR2 having the amino acid sequence of SEQ ID NO: 57, and a HCDR3 having the amino acid sequence of SEQ ID NO: 58, and a light chain variable domain comprising a LCDR1 having the amino acid sequence of SEQ ID NO: 59, a LCDR2 having the amino acid sequence of SEQ ID NO: 60 and a LCDR3 having the amino acid sequence of SEQ ID NO: 61.

[0032] In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 62 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 64. In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 62 and a light chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 64. In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain having the amino acid sequence of SEQ ID NO: 63 and a light chain having the amino acid sequence of SEQ ID NO: 65. In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 63 and a light chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 65.AZ Ref: B7H4-102-PCT01-NP

[0033] In specific embodiments according to all aspects, the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain comprising a HCDR1 having the amino acid sequence of SEQ ID NO: 66, a HCDR2 having the amino acid sequence of SEQ ID NO: 67, and a HCDR3 having the amino acid sequence of SEQ ID NO: 68, and a light chain variable domain comprising a LCDR1 having the amino acid sequence of SEQ ID NO: 69, a LCDR2 having the amino acid sequence of SEQ ID NO: 70, and a LCDR3 having the amino acid sequence of SEQ ID NO: 71. In some embodiments, the second binding domain that specifically binds TIGIT comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 72 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 74. In some embodiments, the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 72 and a light chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 74. In some embodiments, the second binding domain that specifically binds to TIGIT comprises a heavy chain having the amino sequence of SEQ ID NO: 73 and a light chain having the amino acid sequence of SEQ ID NO: 75. In some embodiments, the second binding domain that specifically binds to TIGIT comprises a heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 73 and a light chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 75.

[0034] In specific embodiments according to all aspects, the bispecific binding protein comprises an aglycosylated Fc region. In some embodiments, the bispecific binding protein comprises a deglycosylated Fc region. In some embodiments, the bispecific binding protein comprises an Fc region which has reduced fucosylation or is afucosylated. In some embodiments, the bispecific binding protein is a humanized bispecific antibody or antigen-binding fragment thereof. In some embodiments, the bispecific binding protein comprises an IgG heavy chain constant region. In some embodiments, the constant region includes mutations at L234F, L235E and P331S as defined according to EU numbering convention (Sazinsky et al, PNAS, 105: 20167-20172 (2008)). In some embodiments, the IgG heavy chain constant region is an IgGl heavy chain constant region. In some embodiments, the bispecific binding protein is an antibody. In some embodiments, the bispecific binding protein is a full-length antibody. In some embodiments, theAZ Ref: B7H4-102-PCT01-NPantibody is an IgG antibody. In some embodiments, the antibody is an IgGl antibody. In some embodiments, the antibody is human or humanized. In some embodiments, the bispecific binding protein comprises a kappa light chain constant region. In some embodiments, the bispecific binding protein comprises a lambda light chain constant region.

[0035] In specific embodiments according to all aspects, the PARP1 inhibitor is AZD5305 or a pharmaceutically acceptable salt thereof.

[0036] In specific embodiments according to all relevant aspects, the cancer comprises a cancer cell which expresses B7-H4 polypeptide. In some embodiments, the cancer is selected from endometrial cancer, ovarian cancer, breast cancer, biliary tract cancer, NSCLC (squamous and / or adenocarcinoma), uterine cancer, testicular cancer, bladder cancer, head and neck cancer, melanoma, renal cell carcinoma, pancreatic cancer, prostate cancer, cervical cancer, hematological cancer, cholangiocarcinoma, gastrointestinal cancer such as gastric cancer and colorectal cancer, and lung cancer. In some embodiments, the cancer is a breast cancer selected from hormone receptor-positive (HR+) breast cancer, human epidermal growth factor receptor 2 positive (HER2+) breast cancer, and triple negative breast cancer (TNBC). In some embodiments, the cancer is homologous recombination deficient (HRD) cancer. In some embodiments, the cancer comprises one or more cells having a mutation in an HRD gene selected from BRCA1, BRCA2, ATM, BRIP1, BARD1, CDK12, CHEK1, CHEK2, FANCL, BAI. 2, PPP2R2A, RAD51B, RAD51C, RAD51D, and RAD54L. In some embodiments, the mutated HRD gene is selected from BRCA1, BRCA2, ATM.

[0037] The present disclosure also provides a kit or pharmaceutical composition comprising: 1) an antibody-drug conjugate comprising an antibody or antigen binding fragment thereof which binds to a B7-H4 polypeptide, 2) a bispecific checkpoint inhibitor, and 3) a PARP1 inhibitor, wherein the antibody-drug conjugate, bispecific checkpoint inhibitor and PARP1 inhibitor are each as defined above and elsewhere herein. In specific embodiments, the antibody-drug conjugate comprises a drug-linker represented by the following formula:AZ Ref: B7H4-102-PCT01-NPand wherein the drug-linker is conjugated to the antibody or antigen binding fragment thereof. In some embodiments, the antibody or antigen binding fragment thereof that binds to B7-H4 comprises: a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6. In some embodiments, the antibody or antigen binding fragment thereof comprises a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 45 and SEQ ID NO: 34, respectively, or a functional variant thereof. In some embodiments, the antibody or antigen binding fragment thereof comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 51; and a light chain comprising the amino acid sequence of SEQ ID NO: 44. In some embodiments, the antibody or antigen binding fragment thereof comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 48; and a light chain comprising the amino acid sequence of SEQ ID NO: 44.

[0038] In specific embodiments of the kits and pharmaceutical compositions described herein, the bispecific checkpoint inhibitor comprises: a) a first binding domain that specifically binds to PD-1; and b) a second binding domain that specifically binds to T cell immunoreceptor with Ig and ITIM domains (TIGIT). In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain comprising a HCDR1 having the amino acid sequence of SEQ ID NO: 56, a HCDR2 having the amino acid sequence of SEQ ID NO: 57, and a HCDR3 having the amino acid sequence of SEQ ID NO: 58, and a light chain variable domain comprising a LCDR1 having the amino acid sequence of SEQ ID NO: 59, a LCDR2 having the amino acid sequence of SEQ ID NO: 60 and a LCDR3 having the amino acid sequence of SEQ ID NO: 61.AZ Ref: B7H4-102-PCT01-NP

[0039] In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 62 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 64. In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 62 and a light chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 64. In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain having the amino acid sequence of SEQ ID NO: 63 and a light chain having the amino acid sequence of SEQ ID NO: 65. In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 63 and a light chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 65.

[0040] In some embodiments, the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain comprising a HCDR1 having the amino acid sequence of SEQ ID NO: 66, a HCDR2 having the amino acid sequence of SEQ ID NO: 67, and a HCDR3 having the amino acid sequence of SEQ ID NO: 68, and a light chain variable domain comprising a LCDR1 having the amino acid sequence of SEQ ID NO: 69, a LCDR2 having the amino acid sequence of SEQ ID NO: 70, and a LCDR3 having the amino acid sequence of SEQ ID NO: 71. In some embodiments, the second binding domain that specifically binds TIGIT comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 72 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 74. In some embodiments, the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 72 and a light chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 74. In some embodiments, the second binding domain that specifically binds to TIGIT comprises a heavy chain having the amino sequence of SEQ ID NO: 73 and a light chain having the amino acid sequence of SEQ ID NO: 75. In some embodiments, the second binding domain that specifically binds to TIGIT comprises a heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQAZ Ref: B7H4-102-PCT01-NPID NO: 73 and a light chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 75.

[0041] In specific embodiments of the kits and pharmaceutical compositions described herein, the PARP1 inhibitor is AZD5305.

[0042] The present disclosure is also directed to use of an antibody-drug conjugate in the manufacture of a medicament for use in treating cancer in combination with a bispecific checkpoint inhibitor and a PARP1 inhibitor, wherein the antibody-drug conjugate, the bispecific checkpoint inhibitor and the PARP1 inhibitor are as defined herein. The present disclosure is also directed to use of a bispecific checkpoint inhibitor in the manufacture of a medicament for use in treating cancer in combination with an antibody-drug conjugate and a PARP1 inhibitor, wherein the bispecific checkpoint inhibitor, the antibody-drug conjugate and the PARP1 inhibitor are as defined herein. The present disclosure is also directed to use of a PARP1 inhibitor in the manufacture of a medicament for use in treating cancer in combination with an antibody-drug conjugate and a bispecific checkpoint inhibitor, wherein the PARP1 inhibitor, the antibody-drug conjugate and the bispecific checkpoint inhibitor are as defined herein. The present disclosure is also directed to an antibody-drug conjugate in combination with a bispecific checkpoint inhibitor and a PARP1 inhibitor for use in treating cancer, wherein the antibody-drug conjugate, the bispecific checkpoint inhibitor and the PARP1 inhibitor are as defined herein. In some embodiments, the cancer is selected from endometrial cancer, ovarian cancer, breast cancer, biliary tract cancer, NSCLC (squamous and / or adenocarcinoma), uterine cancer, testicular cancer, bladder cancer, head and neck cancer, melanoma, renal cell carcinoma, pancreatic cancer, prostate cancer, cervical cancer, hematological cancer, cholangiocarcinoma, gastrointestinal cancer such as gastric cancer and colorectal cancer, and lung cancer.

[0043] In specific embodiments according to all aspects of the disclosure, the bispecific checkpoint inhibitor is rilvegostomig.

[0044] In specific embodiments according to all aspects of the disclosure, the ADC is AZD8205, the bispecific checkpoint inhibitor is rilvegostomig and the PARP1 inhibitor is AZD5305.AZ Ref: B7H4-102-PCT01-NPBRIEF DESCRIPTION OF THE DRAWINGS

[0045] The following drawings form part of the present specification and are included to further demonstrate exemplary embodiments of certain aspects of the present disclosure.

[0046] FIG. 1 shows flow cytometry histograms for expression of A) B7-H4, B) PD-L1 and C) PVR on the surface of RL95-2 adenosquamous endometrial carcinoma cells. D) confocal microscopy of RL95-2 GFP expressing 3D spheroid.

[0047] FIGs. 2A and 2B show green fluorescent protein intensity measurements (2A) or the normalized green fluorescence intensity (2B) of RL95-2 GFP spheroids co-cultured with tumor reactive peripheral blood mononuclear cells (PBMCs) and the indicated drugs alone or in combinations. In FIG. 2B, AUC was calculated and the mean of five replicate co-culture wells is shown. Representative data of seven independent experiments with n = 5 unique donors, *p <0.05by Mann- Whitney statistical test.

[0048] FIG. 3 shows the study design and dosing regimen for the study in the MC38_hB7-H4 clone Pl A9 model, as described in Example 2. Each row shown is a different treatment group. AZD8205 isotype is a non-targeting control human IgGl antibody conjugated to the topoisomerase warhead SG3932 at a drug:antibody ratio (DAR) of 8. Dosing with isotype control, AZD8205, mAZD2936 and AZD5305 are depicted via the square, triangle, circle and bar symbols respectively.

[0049] FIGs. 4A-4C show mean tumor volumes ± standard error of the mean (SEM) for each treatment group during the indicated time period. FIG. 4A presents tumor volume measurements in monotherapies, doublets therapies, and triplet therapy in the same graph. FIG.4B compares data from monotherapies and doublet therapies. FIG. 4C shows tumor volume reduction in the triplet therapy. Long dashed line indicates AZD8205 or isotype dosing, while short dashed lines indicate mAZD2936 dosing. AZD5305 was dosed daily throughout the period shown. “AZD2936” in this specific context means the murine AZD2936 surrogate.

[0050] FIG. 5 shows a waterfall plot of the individual responses for each animal in each group and the Overall Response Rate (ORR%) for the group as defined by RECIST criteria.AZ Ref B7H4-102-PCT01-NP“AZD2936” in this specific context means the murine AZD2936 surrogate.

[0051] FIGs. 6A and 6B show addition of AZD5305 further enhanced the anti-tumor activity of the combination of AZD8205 and mAZD2936, as demonstrated by Tumor Growth Inhibition (6A) and Event Free Survival (6B). Event free survival with event defined as a 200% increase in tumor volume from randomization. N = 8-12 / group, *p <0.05, **p <0.01. Statistical significance was determined by ANOVA with post testing or Log-rank test for survival.

[0052] FIG. 7 shows high rate of complete responses by triple combination of AZD8205, mAZD2936, and AZD5305 on two different dose schedules. Solid lines represent untreated controls in each graph. Dashed lines represent the treated samples, as indicated above each graph.

[0053] FIG. 8 shows pharmacodynamic study performed in the MC38 huB7H4 model. Flow cytometry on tumors was carried out 11 days post dosing initiation. N = 8-12 per group, *p <0.05, **p <0.01, ***p <0.001, ****p <0.0001. Statistical significance was determined by ANOVA with post testing.DETAILED DESCRIPTION

[0054] Unless otherwise defined herein, scientific, and technical terms used in the present disclosure shall have the meanings that are commonly understood by one of ordinary skill in the art. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular.

[0055] As used herein, “a” or “an” may mean one or more. As used herein, when used in conjunction with the word “comprising,” the words “a” or “an” may mean one or more than one. As used herein, “another” or “a further” may mean at least a second or more.

[0056] The use of the term “or” in the claims is used to mean “and / or,” unless explicitly indicated to refer only to alternatives or the alternatives are mutually exclusive, although the disclosure supports a definition that refers to only alternatives and “and / or.”AZ Ref: B7H4-102-PCT01-NP

[0057] As used herein, the terms “comprising” (and any variant or form of comprising, such as “comprise” and “comprises”), “having” (and any variant or form of having, such as “have” and “has”), “including” (and any variant or form of including, such as “includes” and “include”) or “containing” (and any variant or form of containing, such as “contains” and “contain”) are inclusive or open-ended and do not exclude additional, unrecited elements or method steps.

[0058] Throughout this application, the term “about” is used to indicate that a value includes the inherent variation of error for the method / device being employed to determine the value, or the variation that exists among the study subjects. Typically, the term “about” is meant to encompass approximately or less than 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20% or higher variability (either “greater than” or “less than” the indicated value), depending on the situation. In some aspects, one of skill in the art will understand the level of variability indicated by the term “about,” due to the context in which it is used herein. It should also be understood that use of the term “about” also includes the specifically recited value.

[0059] The use of the term “for example” and its corresponding abbreviation “e.g.,” (whether italicized or not) means that the specific terms recited are non-limiting, representative examples and aspects of the disclosure.

[0060] Ranges provided herein, of any type, include all values within a particular range described and values about an endpoint for a particular range. As used herein, “between” is a range inclusive of the ends of the range. For example, a number between x and y explicitly includes the numbers x and y, and any numbers that fall within x and y.

[0061] The term “treating” or “treatment” refers to administering a compound or pharmaceutical composition to a subject in order to effect an alteration or improvement of a disease, disorder, or condition in the subject.

[0062] The terms “subject”, “individual” and “patient” are used interchangeably herein to refer to a mammalian subject. In one aspect the “subject” is a human, domestic animals, farm animals, sports animals, and zoo animals, e.g., humans, non-human primates, dogs, cats, guinea pigs, rabbits, rats, mice, horses, cattle, etc. In particular embodiments, the subject is a cynomolgusAZ Ref: B7H4-102-PCT01-NPmonkey Macaca fascicularis). In particular embodiments, the subject is a human. The subject may not have been previously diagnosed as having cancer. Alternatively, the subject may have been previously diagnosed as having cancer. The subject may also be one who exhibits disease risk factors, or one who is asymptomatic for cancer. The subject may also be one who is suffering from or is at risk of developing cancer. The subject may have been previously administered a cancer therapy.

[0063] The term “efficacy" means the ability to produce a desired effect.Treatment of Cancers

[0064] In some aspects, the present disclosure provides a method of treating cancer in a human subject in need thereof, comprising administering to the human subject: a) an antibodydrug conjugate (ADC) comprising: i) an antibody or antigen binding fragment thereof which binds to a B7-H4 polypeptide, ii) a cleavable linker, and iii) a cytotoxic agent; b) a bispecific checkpoint inhibitor, and c) a PARP1 inhibitor. In particular embodiments, the checkpoint inhibitor is a bispecific binding protein having a first binding domain that specifically binds to Programmed cell death protein 1 (PD-1).Antibody-Drug Conjugates Binding B7-H4

[0065] The antibody or antigen binding fragment thereof of the ADC can be used to target a cancer cell. The antibody or antigen binding fragment may have high affinity for a B7-H4 polypeptide both in vitro and in vivo, and thus may advantageously be used in methods for targeting or detecting a B7-H4 epitope.

[0066] B7-H4 (also known as V-set domain-containing T-cell activation inhibitor 1, encoded by the VTCN1 gene) is a transmembrane polypeptide of the B7 family of co-stimulatory proteins. B7-H4 is understood to be expressed on the surface of antigen-presenting cells for interactions with ligands of immune cells (e.g., T-lymphocytes, with CD28 being a potential ligand). B7-H4 has been observed to be highly expressed on cells of various cancer types and is thought to be a tumor-associated antigen. Furthermore, B7-H4 expression is not limited to a particular cancer type, such that it represents a target antigen for treating a broad spectrum ofAZ Ref: B7H4-102-PCT01-NPcancer types.

[0067] In some aspects, a cancer referred to herein is a cancer characterized by the expression (which may be overexpression) of a B7-H4 molecule. In other words, a cancer referred to herein may comprise a cancerous cell that expresses B7-H4 polypeptide. The cancerous cell may be comprised within a tumor. A “B7-H4 polypeptide” may comprise the full length polypeptide sequence of B7-H4 (e.g., SEQ ID NO:55), or may comprise a fragment of B7-H4 of any length of the full length polypeptide sequence of B7-H4 (e.g., comprising a polypeptide sequence of 5%, 15%, 25%, 35%, 45%, 55%, 65%, 75%, 85% or 95% of the full length polypeptide sequence of B7-H4) which comprises an epitope which can bind (e.g. be bound by) an antibody or antigen binding fragment of the disclosure. The B7-H4 polypeptide may comprise a sequence having 75%, 80%, 85%, 90% or 90% sequence identity to the sequence of SEQ ID NO:55. In specific embodiments according to all aspects, the B7-H4 polypeptide comprises the sequence of SEQ ID NO: 55.

[0068] The term “antibody” covers monoclonal antibodies and fragments thereof (e.g., exhibiting the desired biological activity). In specific embodiments according to all aspects, an antibody of the present disclosure is a monoclonal antibody. In specific embodiments according to all aspects, the antibody is a fully human monoclonal antibody. In other embodiments, methods of the disclosure may employ polyclonal antibodies. In particular, an antibody is a protein including at least one or two, heavy (H) chain variable regions (abbreviated herein as VHC, or more simply VH), and at least one or two light (L) chain variable regions (abbreviated herein as VLC, or more simply as VL). The VHC and VLC regions can be further subdivided into regions of hypervariability, termed “complementarity determining regions” (CDR), interspersed with regions that are more conserved, termed “framework regions” (FR). The extent of the framework region and CDRs has been precisely defined. In some embodiments, each VHC and VLC is composed of three CDRs and four FRs, arranged from amino-terminus to carboxyterminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The VHC or VLC chain of the antibody can further include all or part of a heavy or light chain constant region. In specific embodiments according to all aspects, the antibody is a tetramer of two heavy immunoglobulin chains and two light immunoglobulin chains, wherein the heavy and lightAZ Ref: B7H4-102-PCT01-NPimmunoglobulin chains are interconnected by, e.g., disulfide bonds. There are at least two techniques for determining CDRs: (1) an approach based on cross-species sequence variability (i.e., Kabat et al. Sequences of Proteins of Immunological Interest, (5th ed., 1991, National Institutes of Health, Bethesda Md.)); and (2) an approach based on crystallographic studies of antigen-antibody complexes (Al-lazikani et al. (1997) J. Molec. Biol. 273:927-948)). In addition, combinations of these two approaches are sometimes used in the art to determine CDRs.

[0069] The " Kabat numbering system" is generally used when referring to a residue in the variable domain (approximately residues 1-107 of the light chain and residues 1-113 of the heavy chain) (e.g., Kabat et al., Sequences of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)).

[0070] The amino acid position numbering as in Kabat, refers to the numbering system used for heavy chain variable domains or light chain variable domains of the compilation of antibodies in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991). Using this numbering system, the actual linear amino acid sequence can contain fewer or additional amino acids corresponding to a shortening of, or insertion into, a FW or CDR of the variable domain. For example, a heavy chain variable domain can include a single amino acid insert (residue 52a according to Kabat) after residue 52 of H2 and inserted residues (e.g., residues 82a, 82b, and 82c, etc. according to Kabat) after heavy chain FW residue 82.

[0071] The Kabat numbering of residues can be determined for a given antibody by alignment at regions of homology of the sequence of the antibody with a "standard" Kabat numbered sequence. Chothia refers instead to the location of the structural loops (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)). The end of the Chothia CDR-H1 loop, when numbered using the Kabat numbering convention, varies between H32 and H34 depending on the length of the loop (this is because the Kabat numbering scheme places the insertions at H35A and H35B; if neither 35 A nor 35B is present, the loop ends at 32; if only 35 A is present, the loop ends at 33; if both 35 A and 35B are present, the loop ends at 34). The AbM hypervariable regions represent a compromise between the Kabat CDRs and Chothia structural loops, and are used by Oxford Molecular's AbM antibody modeling software. The table below lists the positions of the aminoAZ Ref: B7H4-102-PCT01-NPacids comprising the variable regions of the antibodies in each system.Region Kabat AbM ChothiaLCDR1 L24-L34 L24-L34 L24-L34LCDR2 L50-L56 L50-L56 L50-L56LCDR3 L89-L97 L89-L97 L89-L97HCDR11H31-H35B H26-H35B H26-H32..34HCDR12H31-H35 H26-H35 H26-H32HCDR2 H50-H65 H50-H58 H52-H56HCDR3 H95-H102 H95-H102 H95-H102¹Kabat Numbering2Chothia Numbering

[0072] ImMunoGeneTics (IMGT) also provides a numbering system for the immunoglobulin variable regions, including the CDRs. See, e.g., Lefranc, M. P. et al., Dev. Comp. Immunol. 27: 55-77(2003). The IMGT numbering system is based on an alignment of more than 5,000 sequences, structural data, and characterization of hypervariable loops and allows for easy comparison of the variable and CDR regions for all species. According to the IMGT numbering schema, VH-CDR1 is at positions 26 to 35, VH-CDR2 is at positions 51 to 57, VH-CDR3 is at positions 93 to 102, VL-CDR1 is at positions 27 to 32, VL-CDR2 is at positions 50 to 52, and VL-CDR3 is at positions 89 to 97.

[0073] As used throughout the specification the VH CDRs sequences described correspond to the classical Kabat numbering locations, namely Kabat VH-CDR1 is at positions 31-35, VH-CDR2 is at positions 50-65, and VH-CDR3 is at positions 95-102. VL-CDR1, VL-CDR2 and VL-CDR3 also correspond to classical Kabat numbering locations, namely positions 24-34, SO-56 and 89-97, respectively.

[0074] The heavy chain constant region typically includes three domains, CHI, CH2 and CH3. The light chain constant region is typically comprised of one domain, CL. The variable region of the heavy and light chains contains a binding domain that interacts with an antigen. The term “antibody” includes intact immunoglobulins of types IgA, IgG, IgE, IgD, IgM (as well as subtypes thereof), wherein the light chains of the immunoglobulin may be of types kappa or lambda. The term antibody, as used herein, also refers to a portion of an antibody that retains binding to an antigen, e.g., a molecule in which one or more immunoglobulin chains is not fullAZ Ref: B7H4-102-PCT01-NPlength, but which retains binding to an antigen. Examples of binding portions encompassed within the term antibody include (i) a Fab fragment, a monovalent fragment consisting of the VLC, VHC, CL and CHI domains; (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fc fragment consisting of the VHC and CHI domains; (iv) a Fv fragment consisting of the VLC and VHC domains of a single arm of an antibody, (v) a dAb fragment (Ward et al, Nature 341: 544-546, 1989), which consists of a VHC domain; and (vi) an isolated complementarity determining region (CDR) having sufficient framework to bind, e.g. an antigen binding portion of a variable region. An antigen binding portion of a light chain variable region and an antigen binding portion of a heavy chain variable region, e.g., the two domains of the Fv fragment, VLC and VHC, can be joined, using recombinant methods, by a synthetic linker that enables them to be made as a single protein chain in which the VLC and VHC regions pair to form monovalent molecules (known as single chain Fv (scFv); see e.g., Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85: 5879-5883). Such single chain antibodies are also encompassed within the term antibody. These may be obtained using conventional techniques known to those skilled in the art, and the portions are screened for utility in the same manner as are intact antibodies.

[0075] It is well-known in the art that various minor post-translational modifications may occur to the heavy chain of an antibody during bioproduction. For instance, a lysine residue at the carboxyl terminus of the heavy chain may be removed. It is also known that two amino acid residues (glycine and lysine) at the carboxyl terminus of the heavy chain of an antibody may be removed during bioproduction, and a proline residue newly located at the carboxyl terminus may be amidated. It is also known that an N-terminal glutamine can convert to a pyroglutamate. Disclosures herein to full heavy chains defined by sequence may encompass such equivalent variants, as would be well understood by the person skilled in the art.

[0076] In some embodiments, the antibody or antigen binding fragment is one or more selected from a murine antibody, a humanized antibody, a chimeric antibody, a monoclonal antibody, a polyclonal antibody, a recombinant antibody, a multispecific antibody, or a combination thereof.AZ Ref: B7H4-102-PCT01-NP

[0077] In some embodiments, the antigen-binding fragment is one or more selected from a Fv fragment, an Fab fragment, an F(ab')2 fragment, an Fab' fragment, a dsFv fragment, an scFv fragment, an sc(Fv)2 fragment, or a combination thereof. In some embodiments, the antigen binding fragment thereof of the disclosure is an scFv.

[0078] In some embodiments, the antibody or antigen binding fragment thereof can bind to B7-H4 molecules across species, e.g., the antibody or fragment can bind to mouse B7-H4, rat B7-H4, rabbit, human B7-H4 and / or cynomolgus monkey B7-H4. In some embodiments, the antibody or fragment can bind to human B7-H4 and cynomolgus monkey B7-H4. In some embodiments, the antibody or antigen binding fragment can also bind to mouse B7-H4.

[0079] In some embodiments, the antibody or antigen binding fragment thereof can specifically bind to B7-H4, e.g., human B7-H4 and cynomolgus monkey B7-H4, but does not specifically bind to human B7-H1, B7-H2, and / or B7-H3. The term "does not specifically bind" means that the antibody or antigen binding fragment thereof described herein does not substantially bind to one of more of said molecules (e.g., human B7-H1 molecule, a human B7-H2 molecule, a human B7-H3 molecule, or a combination thereof). The term "substantially no" when used in the context of binding herein may mean less than 5%, 2%, 1%, 0.5% or 0.1% of cells expressing one or more of said molecules in a cell culture become bound by the antibody or antigen binding fragment thereof described herein (upon contact therewith). Suitably, the term "substantially no" when used in the context of binding herein may mean no such cells become bound.

[0080] In specific embodiments according to all aspects, the antibody or antigen binding fragment thereof of the ADC comprises a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequence of: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively, or a functional variant thereof. An antibody or antigen binding fragment thereof comprising said sequences may be referred to as “ZY0EQD-E02” or “EQD-E02” herein.AZ Ref: B7H4-102-PCT01-NP

[0081] In specific embodiments according to all aspects, the antibody or antigen binding fragment thereof of the ADC comprises a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12, respectively, or a functional variant thereof. An antibody or antigen binding fragment thereof comprising said sequences may be referred to as “ZY0EPQ-E02” or “EPQ-E02” herein.

[0082] In specific embodiments according to all aspects, the antibody or antigen binding fragment thereof of the ADC comprises a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, and SEQ ID NO: 18, respectively, or a functional variant thereof. An antibody or antigen binding fragment thereof comprising said sequences may be referred to as “ZY0EOB-F05” or “EOB-F05” herein.

[0083] In specific embodiments according to all aspects, the antibody or antigen binding fragment thereof of the ADC comprises a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 24, respectively, or a functional variant thereof. An antibody or antigen binding fragment thereof comprising said sequences may be referred to as “ZY0EO5-E07” or “EO5-E07” herein.

[0084] In specific embodiments according to all aspects, the antibody or antigen binding fragment thereof of the ADC comprises a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, and SEQ ID NO: 30, respectively, or a functional variant thereof. An antibody or antigen binding fragment thereof comprising said sequences may be referred to as “ZY0EP0-C07” or “EP0-C07” herein.

[0085] As used herein, a “functional variant” of an antibody or antigen binding fragment thereof refers to an amino acid sequence containing one to three amino acid(s) difference from a reference antibody SEQ ID NO, while still retaining the antigen binding capability of the reference antibody.AZ Ref: B7H4-102-PCT01-NP

[0086] In some embodiments, the functional variant comprises an amino acid sequence having one to three amino acid(s) difference from a reference SEQ ID NO. In some embodiments, the functional variant comprises an amino acid sequence having one amino acid difference from a reference SEQ ID NO. In some embodiments, the functional variant comprises an amino acid sequence having two amino acids difference from a reference SEQ ID NO. In some embodiments, the functional variant comprises an amino acid sequence having three amino acids difference from a reference SEQ ID NO. In some embodiments, the difference is a deletion. In some embodiments, the difference is a mutation. In some embodiments, the difference is an addition. In further embodiments, the difference is a combination of any of deletion, mutation, or addition of amino acid(s).

[0087] The antibody or antigen binding fragment thereof having CDRs with the amino acid sequences disclosed above may have beneficial advantages, such as high targeting specificity or binding activity toward B7-H4 peptide on the surface of cancer cells.

[0088] Additionally or alternatively, an antibody or antigen binding fragment thereof described herein may be described by means of a variable heavy (VH) chain and a variable light (VL) chain thereof.

[0089] In specific embodiments according to all aspects, the antibody or antigen binding fragment thereof of the ADC comprises: a variable heavy (VH) chain and a variable light (VL) chain comprising the amino acid sequence of SEQ ID NO: 45 and SEQ ID NO: 34, respectively, or a functional variant thereof; a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 33 and SEQ ID NO: 34, respectively, or a functional variant thereof; a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 43 and SEQ ID NO: 34, respectively, or a functional variant thereof; a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 46 and SEQ ID NO: 34, respectively, or a functional variant thereof; a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 47 and SEQ ID NO: 34, respectively, or a functional variant thereof; a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 31, and SEQ ID NO: 32, respectively, or a functional variant thereof; a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 35 and SEQ ID NO: 36, respectively, or a functional variant thereof; a VH chainAZ Ref: B7H4-102-PCT01-NPand a VL chain comprising the amino acid sequence of SEQ ID NO: 37 and SEQ ID NO: 38, respectively, or a functional variant thereof; or a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 39 and SEQ ID NO: 40, respectively, or a functional variant thereof.

[0090] In some embodiments, the antibody or antigen binding fragment thereof of the ADC comprises: (i) a variable heavy chain comprising an amino acid sequence having at least 70%, 75%, 80%, 90%, 95% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 31, 33, 35, 37, or 39, or a functional variant thereof; and (ii) a variable light chain comprising an amino acid sequence having at least 70%, 75%, 80%, 90%, 95% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 32, 34, 36, 38, or 40, or a functional variant thereof.

[0091] In some embodiments, the antibody or antigen binding fragment thereof of the ADC comprises: (i) a variable heavy chain comprising an amino acid sequence having at least 70%, 75%, 80%, 90%, 95% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 31, 33, 35, 37, 39, 43, 45, 46, or 47, or a functional variant thereof; and (ii) a variable light chain comprising an amino acid sequence having at least 70%, 75%, 80%, 90%, 95% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 32, 34, 36, 38, or 40, or a functional variant thereof.

[0092] In some embodiments, the antibody or antigen binding fragment thereof of the ADC may comprise: (i) a variable heavy chain comprising an amino acid sequence having at least 70%, 75%, 80%, 90%, 95% or 100% sequence identity to the amino acid sequence SEQ ID NO:33, or a functional variant thereof; and (ii) a variable light chain comprising an amino acid sequence having at least 70%, 75%, 80%, 90%, 95% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 34, or a functional variant thereof.

[0093] In some embodiments, the antibody or antigen binding fragment thereof of the ADC may comprise: (i) a variable heavy chain comprising an amino acid sequence having at least 70%, 75%, 80%, 90%, 95% or 100% sequence identity to the amino acid sequence SEQ ID NO: 45, or a functional variant thereof; and (ii) a variable light chain comprising an amino acidAZ Ref: B7H4-102-PCT01-NPsequence having at least 70%, 75%, 80%, 90%, 95% or 100% sequence identity to the amino acid sequence of SEQ ID NO: 34, or a functional variant thereof.

[0094] Advantageously, the antibody or antigen binding fragment of the disclosure may target a broader spectrum of B7-H4 expressing cells when compared with existing (commercially) available antibodies reported to target B7-H4.

[0095] In some embodiments, antibody or antigen binding fragment thereof described herein is capable of binding to B7-H4 as an integral component of a cancer cell (for example, B7-H4 as an integral component of a cell membrane of a cancer cell).

[0096] In some embodiments, the antibody or antigen binding fragment thereof of the ADC binds an OVCAR4 cell line. In some embodiments, an antibody or antigen binding fragment thereof described herein may bind to an OVCAR4 cell line and / or a CHO cell line (e.g. which may lack an exogenous nucleic acid encoding B7-H4). For example, the antibody or antigen binding fragment thereof binds to a B7-H4 (e.g. a B7-H4 epitope) of an OVCAR4 cell line and / or a CHO cell line (e.g. which may lack an exogenous nucleic acid encoding B7-H4).

[0097] In some embodiments, the antibody or antigen binding fragment thereof binds to an OVCAR4 cell line and / or CHO cell line (e.g. which may lack an exogenous nucleic acid encoding B7-H4) with higher affinity when compared to one or more antibody selected from E Biosciences 14-5949 anti-Human B7H4 mouse IgG, US biological B0000-35B anti Human B7H4 mouse IgG, R and D systems AF2514 anti-Mouse B7H4 goat IgGl, Sigma SAB2500141 anti B7H4 Goat IgGl, Isotype 1 CAT004 SP06-003, Isotype 2 R and D Normal goat IgG control (AB-108C), AdD serotec MCA2632, Epitomics 2516-1, eBiosciences, 145972-82, eBioscience 145970-85, or a combination thereof. The affinity (e.g. binding affinity) can be measured by any suitable method of measuring binding affinity described herein.

[0098] The OVCAR4 cell line is a human ovary carcinoma cell line, the CHO cell line is an epithelial cell line derived from the ovary of the Chinese hamster, and are widely obtainable.

[0099] In some embodiments, the antibody or antigen binding fragment thereof of the ADC comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 41.AZ Ref: B7H4-102-PCT01-NPIn some embodiments, the antibody or antigen binding fragment thereof of the ADC comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 52. In some embodiments, the antibody or antigen binding fragment thereof of the ADC comprises a light chain constant region comprising the amino acid sequence of SEQ ID NO: 42.

[0100] In some embodiments, the antibody or antigen binding fragment thereof of the ADC comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 51; and a light chain comprising the amino acid sequence of SEQ ID NO: 44.

[0101] In some embodiments, the antibody or antigen binding fragment thereof of the ADC comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 48; and a light chain comprising the amino acid sequence of SEQ ID NO: 44.

[0102] The “E02-GL” antibody has the CDR sequences (e.g. corresponds to) of ZY0EQD-E02 (“GL” means the antibody has been germlined). E02-GL comprises the VH chain sequence of SEQ ID NO: 45, e.g. a germlined version of SEQ ID NO: 43, and the VL chain sequence of SEQ ID NO: 34. E02-GL has the heavy chain sequence of SEQ ID NO: 51 and the light chain sequence of SEQ ID NO: 44. “E02-GL” conjugated to the topoisomerase I payload SG3932 as described herein is therefore referred to as “E02-GL-SG3932”. In a specific embodiment according to all aspects, the clone E02-GL is conjugated to the topoisomerase I payload SG3932 at an average Drug Antibody Ratio (DAR) of 8: this embodiment of the ADC may also be termed “AZD8205” (also known as “Puxitatug samrotecan”).

[0103] In some embodiments, the antibody or antigen binding fragment thereof of the ADC is a monoclonal antibody.

[0104] In some embodiments, the antibody or antigen binding fragment thereof of the ADC is a humanized monoclonal antibody.

[0105] Methods for the preparation of the anti-B7H4 antibodies and antigen binding fragments thereof disclosed herein are described in WO Publication No. W02022 / 053650, the disclosure of which is hereby incorporated by reference in its entirety.AZ Ref: B7H4-102-PCT01-NPLinkers

[0106] In specific embodiments according to all aspects, the antibody or antigen binding fragment thereof is conjugated to the cytotoxic agent by a linker. As used herein, “conjugated” means linked via a covalent or ionic bond. In some embodiments, the linker is attached (e.g. conjugated) in a cleavable manner to an amino residue, for example, an amino acid of the antibody or antigen binding fragment of the ADC. In some embodiments, the cleavable linker of the ADC is an mp-PEG8-val-ala linker. As indicated in the name, the mp-PEG8-val-ala linker has 8 consecutive polyethylene glycol units followed by a valine-alanine (val-ala) dipeptide that attaches to the cytotoxic agent.

[0107] In some embodiments, the linker is represented by RL, and is selected from:la, whereQ is linked to the cytotoxic agent and has a formula:, where Qxis such that Q is an amino-acid residue, a dipeptide residue, a tripeptide residue or a tetrapeptide residue;X is:where a = 0 to 5, bl = 0 to 16, b2 = 0 to 16, cl = 0 or 1, c2 = 0 or 1, d = 0 to 5, wherein at least bl or b2 = 0 (i.e., only one of bl and b2 may not be 0) and at least cl or c2 = 0 (i.e., only one of cl and c2 may not be 0);AZ Ref: B7H4-102-PCT01-NPGLis a linker for connecting to an antibody or antigen binding fragment thereof described herein; and(lb):, whereRL1and RL2are independently selected from H and methyl, or together with the carbon atom to which they are bound form a cyclopropylene or cyclobutylene group; and e is 0 or 1.

[0108] In some embodiments, Q is an amino acid residue. The amino acid may be a natural amino acid or a non-natural amino acid. For example, Q may be selected from: Phe, Lys, Vai, Ala, Cit, Leu, He, Arg, and Trp, where Cit is citrulline.

[0109] In some embodiments, Q comprises a dipeptide residue. The amino acids in the dipeptide may be any combination of natural amino acids and non-natural amino acids. In some embodiments, the dipeptide comprises natural amino acids. Where the linker is a cathepsin labile linker, the dipeptide is the site of action for cathepsin-mediated cleavage. The dipeptide then is a recognition site for cathepsin.

[0110] In some embodiments, Q is selected from:NH-Phe-Lys-C=o,NH-Vai-Ala-c=o,NH-Val-Lys-c <),NH-Ala-Lys-c NH-Val-Cit-c NH-Phe-Cit-c NH-Leu-Cit-c NH-I1 e-Cit-c NH-Phe-Arg-c=o,NH-Trp-Cit-c=o, andNH-Gly-Val-c=o; where Cit is citrulline. In some embodiments, Q is selected from:NH-Phe-Lys-c=o,NH-Val-Ala-C=o,NH-Val-Lys-C=o,NH-Ala-Lys-c=o, andNH-Val-Cit-c=o. In specific embodiments, Q is selected fromNH-Phe-Lys-c=o,NH-Val-Cit-c=oandNH-Val-Ala-c=o.

[0111] Other suitable dipeptide combinations include:NH-Gly-Gly-C°,NH-Gly-Val-c°,NH-Pro-Pro-c=o, andNH-Val-Glu-c=o. Other dipeptide combinations may be used, including those described by Dubowchik et al., Bioconjugate Chemistry, 2002, 13,855-869, which is hereby incorporated herein by reference.AZ Ref: B7H4-102-PCT01-NP

[0112] In some embodiments, Q is a tripeptide residue. The amino acids in the tripeptide may be any combination of natural amino acids and non-natural amino acids. In some embodiments, the tripeptide comprises natural amino acids. Where the linker is a cathepsin labile linker, the tripeptide is the site of action for cathepsin-mediated cleavage. The tripeptide then is a recognition site for cathepsin. Tripeptide linkers of particular interest are:NH-Glu-Val-Ala-C=o,NH-Glu-Val-Cit-c=o,NH-aGlu-Val-Ala-c=o, andNH-aGlu-Val-Cit-c=o.

[0113] In some embodiments, Q is a tetrapeptide residue. The amino acids in the tetrapeptide may be any combination of natural amino acids and non-natural amino acids. In some embodiments, the tetrapeptide comprises natural amino acids. Where the linker is a cathepsin labile linker, the tetrapeptide is the site of action for cathepsin-mediated cleavage. The tetrapeptide then is a recognition site for cathepsin. Tetrapeptide linkers of particular interest are:NH-Gly-Gly-Phe-Glyc=o; andNH-Gly-Phe-Gly-Glyc=o. In some embodiments, the tetrapeptide is:NH-Gly-Gly-Phe-Glyc=o.

[0114] In the above representations of peptide residues,NH- represents the N-terminus, and -c=orepresents the C-terminus of the residue. The C-terminus binds to, for example, the NH of the cytotoxic agent.

[0115] Glu represents the residue of glutamic acid, i.e.:oaGlu represents the residue of glutamic acid when bound via the a-chain, i.e.:O^OHOAZ Ref: B7H4-102-PCT01-NP

[0116] In some embodiments, the amino acid side chain is chemically protected, where appropriate. The side chain protecting group may be a group as discussed above. Protected amino acid sequences are cleavable by enzymes. For example, a dipeptide sequence comprising a Boc side chain-protected Lys residue is cleavable by cathepsin. In some embodiments, protecting groups for the side chains of amino acids are well known and are described in the Novabiochem Catalog, and as described above.

[0117] In some embodiments of X, a may be 0, 1, 2, 3, 4 or 5. In particular embodiments, a is 0 to 3. More particularly, a may be 0 or 1. In particular embodiments, a is 0. In some embodiments, bl may be 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16. Particularly, bl may be 0 to 12. More particularly, bl may be 0 to 8, and may be 0, 2, 3, 4, 5 or 8. In some embodiments, b2 may be 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16. Particularly, b2 may be 0 to 12. More particularly, b2 may be 0 to 8, and may be 0, 2, 3, 4, 5 or 8. In some embodiments, cl may be 0 or 1. In some embodiments, c2 may be 0 or 1. In particular embodiments, only one of cl and c2 may not be 0. In some embodiments, d may be 0, 1, 2, 3, 4 or 5. Particularly, d may be 0 to 3. More particularly, d may be 1 or 2. In a particular embodiment, d is 2. In a particular embodiment, d is 5.

[0118] In some embodiments of X, a is 0, bl is 0, cl is 1, c2 is 0 and d is 2, and b2 may be from 0 to 8. For instance, b2 may be 0, 2, 3, 4, 5 or 8. In some embodiments of X, a is 1, b2 is 0, cl is 0, c2 is 0 and d is 0, and bl may be from 0 to 8. For instance, bl may be 0, 2, 3, 4, 5 or 8. In some embodiments of X, a is 0, bl is 0, cl is 0, c2 is 0 and d is 1, and b2 may be from 0 to 8. For instance, b2 may be 0, 2, 3, 4, 5 or 8. In some embodiments of X, bl is 0, b2 is 0, cl is 0, c2 is 0 and one of a and d is 0. The other of a and d is from 1 to 5. For instance, the other of a and d may be 1. Alternatively, the other of a and d may be 5. In some embodiments of X, a is 1, b2 is 0, cl is 0, c2 is 1, d is 2, and bl may be from 0 to 8. For instance, b2 may be 0, 2, 3, 4, 5 or 8.

[0119] As described above RL1and RL2may be independently selected from H and methyl, or together with the carbon atom to which they are bound form a cyclopropylene or cyclobutylene group.

[0120] In some embodiments, both RL1and RL2are H. In some embodiments, RL1is H and RL2is methyl. In some embodiments, both RL1and RL2are methyl. In some embodiments, RL1AZ Ref: B7H4-102-PCT01-NPand RL2together with the carbon atom to which they are bound form a cyclopropylene group. In some embodiments, RL1and RL2together with the carbon atom to which they are bound form a cyclobutylene group.

[0121] In particular embodiments according to the formula lb, e is 0. In other embodiments, e is 1 and the nitro group may be in any available position of the ring. In some embodiments, it is in the ortho position. In some embodiments, it is in the para position.

[0122] In some embodiments, GLmay be selected from:(G“-') 0 (GL6) 0° \xS--Nxo(G11'2) O (G")Vt ^Ar,zBr—xo(tn 0 (G“)AV-k o0(G13-1) (CH Ns— sX30(N02)where the NO2 group is optional(GL3-2) (GL1°)S—H(N02)AZ Ref: B7H4-102-PCT01-NPwhere the NO2 group is optional(G^3-3) (G^1)S—\ _O2N-\^= / where the NO2 group is optional O(GL3-4) (GL12)S—o2Ndwhere the NO2 group is optional(G14) t _ 0 (G113)XVN"N\Hal N — IHiWhere Hal = I, Br, Cl(GL5) (GL14)H2I\L XHal—where Ar represents a C5-6 arylene group, e.g., phenylene, and X represents C1-4 alkyl.

[0123] In some embodiments, GLis selected from GL land GL1'2. In one embodiment, GLis GL l.

[0124] In some embodiments, RLis selected from:AZ Ref: B7H4-102-PCT01-NPAZ Ref: B7H4-102-PCT01-NP

[0125] In some embodiments, the linker is represented by RLL, and in particular embodiments is selected from:AZ Ref B7H4-102-PCT01-NPla', whereQ and X are as defined above and GLLis a linker connected to an antibody or antigen binding fragment thereof described herein (e.g., the Ligand Unit); andlb', whereRL1and RL2are as defined above; and p is an integer of from 1 to 20.

[0126] In some embodiments, GLLmay be selected from:CBA0 £CBAjxo(GLL1-2) 0 (GLL8'2) N UBA^ArCEA) yyxo(GLL2) 0 (GLL9-1)*NCBAj N"”V--X o0 CEA(GLL3-1) (G^-2)CBA< A, N *N-^H CBAAZ Ref: B7H4-102-PCT01-NP(G^3-2) (GLL1°)(GLL-4) CBA< _ (GLL11)1 \ HX— NO 3^A L0(G"s) p (GI LI2)CBA$ / / °H 1 L / (GLL6) 0 (GLL13)CBA|__^^p H(GLL7)CBA^_ (GLL14) H twhere Ar represents a C5-6 arylene group, e.g., phenylene anc X represents C1-4 alkyl.

[0127] In some embodiments, GLLis selected from GLL land GLL1'2. In one embodiment, GLLis GLL1’1.

[0128] In some embodiments, RLLis a group derived from the RLgroups above.

[0129] In some embodiments where compounds described herein are provided in a single enantiomer or in an enantiomerically enriched form, the enantiomerically enriched form has an enantiomeric ratio greater than 60:40, 70:30; 80:20 or 90:10. In some embodiments, the enantiomeric ratio is greater than 95:5, 97:3 or 99:1.AZ Ref: B7H4-102-PCT01-NPDrug loading

[0130] The average number of drugs per antibody in preparations of ADC from conjugation reactions may be characterized by conventional means such as UV, reverse phase HPLC, HIC, mass spectroscopy, ELISA assay, and electrophoresis. The quantitative distribution of ADC in terms of p may also be determined. By ELISA, the averaged value of p in a particular preparation of ADC may be determined (Hamblett et al (2004) Clin. Cancer Res. 10:7063-7070; Sanderson et al (2005) Clin. Cancer Res. 11: 843 -852). However, the distribution of p (drug) values is not discernible by the antibody-antigen binding and detection limitation of ELISA. Also, ELISA assay for detection of antibody-drug conjugates does not determine where the drug moieties are attached to the antibody, such as the heavy chain or light chain fragments, or the particular amino acid residues. In some instances, separation, purification, and characterization of homogeneous ADC where p is a certain value from ADC with other drug loadings may be achieved by means such as reverse phase HPLC or electrophoresis. Such techniques are also applicable to other types of conjugates.

[0131] Typically, fewer than the theoretical maximum of drug moieties are conjugated to an antibody during a conjugation reaction. An antibody may contain, for example, many lysine residues that do not react with the Drug Linker. Only the most reactive lysine groups may react with an amine-reactive linker reagent. Also, only the most reactive cysteine thiol groups may react with a thiol-reactive linker reagent. Generally, antibodies do not contain many, if any, free and reactive cysteine thiol groups which may be linked to a drug moiety. Most cysteine thiol residues in the antibodies of the compounds exist as disulfide bridges and must be reduced with a reducing agent such as dithiothreitol (DTT) or TCEP, under partial or total reducing conditions. The loading (drug / antibody ratio) of an ADC may be controlled in several different manners, including: (i) limiting the molar excess of Drug Linker relative to antibody, (ii) limiting the conjugation reaction time or temperature, and (iii) partial or limiting reductive conditions for cysteine thiol modification.

[0132] Certain antibodies have reducible interchain disulfides, i.e. cysteine bridges.Antibodies may be made reactive for conjugation with linker reagents by treatment with a reducing agent such as DTT (dithiothreitol). Each cysteine bridge will thus form, theoretically,AZ Ref: B7H4-102-PCT01-NPtwo reactive thiol nucleophiles. Additional nucleophilic groups can be introduced into antibodies through the reaction of lysines with 2-iminothiolane (Traut’s reagent) resulting in conversion of an amine into a thiol. Reactive thiol groups may be introduced into the antibody (or fragment thereof) by engineering one, two, three, four, or more cysteine residues (e.g., preparing mutant antibodies comprising one or more non-native cysteine amino acid residues). US 7521541 teaches engineering antibodies by introduction of reactive cysteine amino acids.

[0133] Cysteine amino acids may be engineered at reactive sites in an antibody and which do not form intrachain or intermolecular disulfide linkages (Junutula, et al., 2008b Nature Biotech., 26(8):925-932; Dornan et al (2009) Blood 114(13):2721-2729; US 7521541; US 7723485; W02009 / 052249). The engineered cysteine thiols may react with Drug Linkers of the present disclosure which have thiol-reactive, electrophilic groups such as maleimide or alpha-halo amides to form ADC with cysteine engineered antibodies. The location of the drug unit can thus be designed, controlled, and known. The drug loading can be controlled since the engineered cysteine thiol groups typically react with drug-linker reagents in high yield. Engineering an IgG antibody to introduce a cysteine amino acid by substitution at a single site on the heavy or light chain gives two new cysteines on the symmetrical antibody.Cytotoxic agent

[0134] The cytotoxic agent of the ADC may be referred to herein as an “agent” or “active agent.” In some embodiments, the cytotoxic agent is a drug. In some embodiments, the cytotoxic agent of the ADC is a topoisomerase inhibitor, a tubulysin derivative, a pyrrolobenzodiazepine, or a combination thereof.

[0135] In some embodiments, the topoisomerase inhibitor is a compound of formula A:AZ Ref: B7H4-102-PCT01-NPas a single enantiomer or in an enantiomerically enriched form.

[0136] In some embodiments, the disclosure provides a conjugate comprising the following topoisomerase inhibitor derivative (A*, the Drug Unit):

[0137] The present disclosure provides the cytotoxic agent (Dug Unit) of the ADC, which is connected to a Ligand Unit of the ADC via the linker (linking unit) described above. In some embodiments, the Ligand Unit is an antibody or antigen binding fragment thereof. In some embodiments, the present disclosure also provides A* with the linker attached, and intermediates for their synthesis, as well as the released payload.

[0138] In specific embodiments according to all aspects, the cytotoxic agent is a topoisomerase inhibitor and wherein the cleavable linker and cytotoxic agent of the ADC together comprise a compound of formula I:AZ Ref: B7H4-102-PCT01-NPand salts and solvates thereof. RLis the cleavable linker described above.

[0139] In some embodiments, the ADC is of formula IV:L - (DL)P(IV)or a pharmaceutically acceptable salt or solvate thereof,wherein L is the i) antibody or antigen binding fragment thereof, DLis a drug linker unit comprising the ii) cleavable linker and the iii) cytotoxic agent, and DLis of formula III:wherein the cleavable linker RLLis as described above.

[0140] In some embodiments, the drug loading (p) of the cytotoxic agent to the antibody or antibody binding fragment is an integer from 1 to about 10.

[0141] In some embodiments, the topoisomerase inhibitor is a compound with the formulaAZ Ref: B7H4-102-PCT01-NP VI:where Q as described above in related to the linker RL.

[0142] In specific embodiments according to all aspects, the linker and cytotoxic agent together comprise the following compound:HOH 0(SG3932).

[0143] In other embodiments, the linker and cytotoxic agent are together one of the following compounds:AZ Ref: B7H4-102-PCT01-NP(SG4052); and / or

[0144] The cytotoxic agent is typically linked to, or “loaded onto” the antibody or antigenbinding fragment. The agent loading (p) is the average number of agent(s) per antibody or antigen-binding fragment (e.g., the Ligand Unit).

[0145] The average number of agents per antibody (or antigen-binding fragment) in preparations of ADCs from conjugation reactions may be characterized by conventional means such as UV, reverse phase HPLC, HIC, mass spectroscopy, ELISA assay, and electrophoresis.AZ Ref: B7H4-102-PCT01-NPThe quantitative distribution of ADC in terms of p may also be determined. By ELISA, the averaged value of p in a particular preparation of ADC may be determined (Hamblett et al (2004) Clin. Cancer Res. 10:7063-7070; Sanderson et al (2005) Clin. Cancer Res. 11:843-852). In some instances, separation, purification, and characterization of homogeneous ADC, where p is a certain value from ADC with other drug loadings, may be achieved by means such as reverse phase HPLC or electrophoresis. Such techniques are also applicable to other types of conjugates.

[0146] Cysteine amino acids may be engineered at reactive sites in an antibody (or antigenbinding fragment thereof) and which, in some embodiments, do not form intrachain or intermolecular disulfide linkages (Junutula, et al., 2008b Nature Biotech., 26(8):925-932; Dornan et al (2009) Blood 114(13):2721-2729; US 7521541; US 7723485; W02009 / 052249). The engineered cysteine thiols may react with a linker within an agent (e.g., of formula I below) which may have thiol-reactive, electrophilic groups such as maleimide or alpha-halo amides to form ADC with cysteine engineered antibodies. The location of the drug unit can thus be designed, controlled, and known. The drug loading can be controlled since the engineered cysteine thiol groups typically react with drug-linker reagents in high yield. Engineering an IgG antibody to introduce a cysteine amino acid by substitution at a single site on the heavy or light chain gives two new cysteines on the symmetrical antibody. A drug loading near 2 can be achieved with near homogeneity of the conjugation product ADC.

[0147] Where more than one nucleophilic or electrophilic group of the antibody or antigen binding fragment thereof reacts with an agent, then the resulting product may be a mixture of ADC compounds with a distribution of agent units attached to an antibody, e.g., 1, 2, 3, etc. Liquid chromatography methods such as polymeric reverse phase (PLRP) and hydrophobic interaction (HIC) may separate compounds in the mixture by agent loading value. Preparations of ADC with a single agent loading value (p) may be isolated.

[0148] Thus, the antibody-drug conjugate compositions of the disclosure may include mixtures of antibody-drug conjugates where the antibody or antigen binding fragment thereof has one or more agent moieties and where the agent moieties may be attached to the antibody or antigen binding fragment thereof at various amino acid residues.AZ Ref: B7H4-102-PCT01-NP

[0149] In some embodiments, the average number of agents per antibody (or antigen-binding fragment thereof) is in the range 1 to 20. In some embodiments, the range is selected from 1 to 10, 2 to 10, 2 to 8, 2 to 6, and 4 to 10. In some embodiments, there is one agent per antibody (or antigen-binding fragment thereof). In some embodiments, the number of agents per antibody (or antigen-binding fragment thereof) can be expressed as a ratio of agent (i.e., drug) to antibody. This ratio is referred to as the Drug to Antibody Ratio (DAR). The DAR is the average number of drugs (i.e., agents) linked to each antibody. In some embodiments of the present disclosure, the DAR is in the range 1 to 20. In some embodiments, the range of DAR is selected from 1 to 10, 2 to 10, 2 to 8, 2 to 6, and 4 to 10. In some embodiments, the DAR is between about 1 and about 8. In specific embodiments according to all aspects, the DAR is about 8. In particular embodiments, the DAR is 8.

[0150] In specific embodiments according to all aspects, the ADC is AZD8205 (also known as “Puxitatug samrotecan”). AZD8205 may also be termed “E02-GL-SG3932”, as defined herein. E02-GL-SG3932 and methods for the preparation thereof are described in WO Publication No. W02022 / 053650, the disclosure of which is hereby incorporated by reference in its entirety.Bispecific binding proteins

[0151] The term “bispecific binding protein” means a protein that has binding specificities for at least two independent antigens (or targets) or different epitopes within the same antigen. Exemplary bispecific binding proteins may bind to two different epitopes of a target, or may bind two different targets. Other such binding proteins may combine a first target binding site with a second binding site for another target. The bispecific binding protein may be a bispecific antibody. In particular and according to all aspects, the bispecific binding protein may be a bispecific checkpoint inhibitor.

[0152] In the present disclosure, the term "bispecific checkpoint inhibitor" refers to checkpoint inhibitors that have binding specificities for at least two independent antigens (or targets) or different epitopes within the same antigen. Checkpoint inhibitors (or “immuno-oncology (IO) agents”) are agents that inhibit the immune suppression system and activate antitumor immunityAZ Ref: B7H4-102-PCT01-NP(Menon S. et al., Cancers (2016) 8, 106; Pardoll DM., Nat Rev Cancer (2012) 12, 252-264; Wolchok JD., Cell (2015) 162, 937). Targets for immune mediated therapy in oncology include PD-1 (programmed cell death protein- 1) and TIGIT (T cell immunoreceptor with Ig and ITIM domains), an immune receptor present on some T cells and natural killer cells (NKs). Examples of known immune checkpoint inhibitors include anti -PD-1 antibodies nivolumab (WO 2006 / 121168) and pembrolizumab (WO 2008 / 156712), each of which are hereby incorporated by reference in their entirety. In some embodiments, the bispecific checkpoint inhibitor provides additive and / or synergistic therapeutic effects derived from targeting two antigens simultaneously, with the administration of a single manufactured molecule.

[0153] In some embodiments, the antibodies provided herein are monovalent bispecific antibodies (MBab). The monovalent bispecific antibody scaffolds described herein provide a superior platform for the generation of bispecific antibodies that fulfill all the benefits associated with bispecific antibodies while reducing the potential therapeutic risks mentioned above due to their monovalent nature. Furthermore, the MBabs provided herein are readily expressed, stable, and are likely to have low immunogenicity. As used herein, the term “monovalent bispecific,” which may be abbreviated “MBab,” refers to bispecific antibodies, where each arm can specifically bind to a different target antigen, and for a given pair of different target antigens (A and B), the MBab can bind to one of each. In some embodiments, monovalent bispecific antibodies can specifically bind to two independent antigens (or targets) or two independent epitopes on the same antigen. Typically, monovalent bispecific antibodies comprise two different variable regions. In some embodiments, the binding affinity for the two independent antigens is about the same. In some embodiments, the binding affinities for the two independent antigens are different.

[0154] In specific embodiments according to all aspects, the bispecific binding protein comprises: (a) a first binding domain that specifically binds to PD-1; and (b) a second binding domain that specifically binds to T cell immunoreceptor with Ig and ITIM domains (TIGIT).PD-1 Binding Domains

[0155] The term Programmed cell death protein 1 (PD-1) refers to an approximately 31 kD type I membrane protein that is a member of the extended CD28 / CTLA4 family of T cell regulatorsAZ Ref: B7H4-102-PCT01-NP(see Ishida et al., “Induced Expression of PD-1, A Novel Member of the Immunoglobulin Gene Superfamily, Upon Programmed Cell Death,” EMBO J. 11: 3887-95 (1992)). PD-1 is expressed on activated T cells, B cells, and monocytes and at low levels in natural killer (NK) T cells. Programmed death ligand 1 (PD-L1) is a ligand of PD-1. When PD-L1 binds PD-1 on T cells, it prevents the T cells from killing the cells displaying PD-L1 (see Han et al.," PD-l / PD-Ll pathway: current researches in cancer," Am J. Cancer Res., 10(3):727-742 (2020)). PD-L1 is an approximate 33 kD type 1 transmembrane glycoprotein that is a member of the B7 family (see Sanmamed et al. " Inducible expression of B7-H1 (PD-L1) and its selective role in tumor site immune modulation,". Cancer J. 20:256-261 (2014)). Antagonistic inhibition of the PD-1 / PD-L1 interaction increases T cell activation, enhancing recognition and elimination of tumor cells by the host immune system. Such antagonistic inhibition of the PD-1 / PD-L1 interaction can be achieved using agents that bind to either PD-1 or PD-L1, blocking the interaction. The amino acid sequence of the human PD-1 protein is provided as UNIPROT ID no. Q15116, while the amino acid sequence of the human PD-L1 is provided as UNIPROT ID no. Q9NZQ7.

[0156] In specific embodiments according to all aspects, the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain comprising a HCDR1 having the amino acid sequence of SEQ ID NO: 56, a HCDR2 having the amino acid sequence of SEQ ID NO: 57, and a HCDR3 having the amino acid sequence of SEQ ID NO: 58, and a light chain variable domain comprising a LCDR1 having the amino acid sequence of SEQ ID NO: 59, a LCDR2 having the amino acid sequence of SEQ ID NO: 60 and a LCDR3 having the amino acid sequence of SEQ ID NO: 61.

[0157] In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 62 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 64.

[0158] In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain having an amino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the amino acid sequence of SEQ ID NO: 62. In some embodiments, the first binding domain that specifically binds to PD-1 comprises a light chain variable domain having an amino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98,AZ Ref: B7H4-102-PCT01-NPor 99% identical to the amino acid sequence of SEQ ID NO: 64.

[0159] In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain having the amino acid sequence of SEQ ID NO: 63 and a light chain having the amino acid sequence of SEQ ID NO: 65.

[0160] In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain having an amino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the amino acid sequence of SEQ ID NO: 63. In some embodiments, the first binding domain that specifically binds to PD-1 comprises a light chain having an amino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the amino acid sequence of SEQ ID NO: 65.

[0161] In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain that is encoded by the nucleic acid sequence of SEQ ID NO: 76 and a light chain variable domain is encoded by the nucleic acid sequence of SEQ ID NO: 78.

[0162] In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain that is encoded by a nucleic acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 76 and a light chain variable domain that is encoded by a nucleic acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 78.

[0163] In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain that is encoded by the nucleic acid sequence of SEQ ID NO: 77 and a light chain that is encoded by the nucleic acid sequence of SEQ ID NO: 79.

[0164] In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain that is encoded by a nucleic acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 77 and a light chain that is encoded by a nucleic acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 79.AZ Ref: B7H4-102-PCT01-NPTIGIT Binding Domains

[0165] The term T cell immunoreceptor with Ig and ITIM domains (TIGIT) refers to an immune receptor present on some T cells and Natural Killer Cells (NK). TIGIT is upregulated by immune cells, including activated T cells, natural killer cells, and regulatory T cells.

[0166] In specific embodiments according to all aspects, the second binding domain of the bispecific checkpoint inhibitor specifically binds to TIGIT. In some embodiments, the second binding domain comprises a heavy chain variable domain comprising a HCDR1 having the amino acid sequence of SEQ ID NO: 66, a HCDR2 having the amino acid sequence of SEQ ID NO: 67, and a HCDR3 having the amino acid sequence of SEQ ID NO: 68, and a light chain variable domain comprising a LCDR1 having the amino acid sequence of SEQ ID NO: 69, a LCDR2 having the amino acid sequence of SEQ ID NO: 70, and a LCDR3 having the amino acid sequence of SEQ ID NO: 71.

[0167] In some embodiments, the second binding domain that specifically binds TIGIT comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 72 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 74.

[0168] In some embodiments, the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain having an amino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the amino acid sequence of SEQ ID NO: 72. In some embodiments, the second binding domain that specifically binds to TIGIT comprises a light chain variable domain having an amino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the amino acid sequence of SEQ ID NO: 74.

[0169] In some embodiments, the second binding domain that specifically binds to TIGIT comprises a heavy chain having the amino acid sequence of SEQ ID NO: 73 and a light chain having the amino acid sequence of SEQ ID NO: 75.

[0170] In some embodiments, the second binding domain that specifically binds to TIGIT comprises a heavy chain having an amino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the amino acid sequence of SEQ ID NO: 73. In some embodiments, the second binding domain that specifically binds to TIGIT comprises a light chain having anAZ Ref: B7H4-102-PCT01-NPamino acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to the amino acid sequence of SEQ ID NO: 75.

[0171] In some embodiments, the second binding domain that specifically binds TIGIT comprises a heavy chain variable domain that is encoded by the nucleic acid sequence of SEQ ID NO: 80 and a light chain variable domain that is encoded by the nucleic acid sequence of SEQ ID NO: 82.

[0172] In some embodiments, the second binding domain that specifically binds TIGIT comprises a heavy chain variable domain that is encoded by a nucleic acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 80 and a light chain variable domain that is encoded by a nucleic acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 82.

[0173] In some embodiments, the second binding domain that specifically binds to TIGIT comprises a heavy chain that is encoded by the nucleic acid sequence of SEQ ID NO: 81 and a light chain that is encoded by the nucleic acid sequence of SEQ ID NO: 83.

[0174] In some embodiments, the second binding domain that specifically binds to TIGIT comprises a heavy chain that is encoded by a nucleic acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 26 and a light chain that is encoded by a nucleic acid sequence that is at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to SEQ ID NO: 28.Rilvegostomig

[0175] In specific embodiments according to all aspects, the bispecific checkpoint inhibitor is the anti -PD-1 / TIGIT bispecific checkpoint inhibitor named “rilvegostomig” (also “AZD2936”) that comprises the sequences of monoclonal antibody LO115 (PD-1) and VARI (TIGIT) as described in WO2022 / 229919 which is incorporated herein by reference.Rilvegostomig comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO: 63, a first light chain comprising the amino acid sequence of SEQ ID NO: 65, and a second heavy chain comprising the amino acid sequence of SEQ ID NO: 73, and a second light chainAZ Ref: B7H4-102-PCT01-NPcomprising the amino acid sequence of SEQ ID NO: 75.Aspects of Bispecific Proteins Binding PD-1 and TIGIT

[0176] The bi specific binding protein having the PD-1 binding domain and the TIGIT binding domain can be varied. In some embodiments, the bispecific binding protein comprises an aglycosylated Fc region. In some embodiments, the bispecific binding protein comprises a deglycosylated Fc region. In some embodiments, the bispecific binding protein comprises an Fc region which has reduced fucosylation or is afucosylated. In some embodiments, the bispecific binding protein is a full-length antibody. In some embodiments, the antibody is an IgG antibody. In some embodiments, the antibody is an IgGl antibody. In some embodiments, the antibody is human or humanized. In some embodiments, the bispecific binding protein comprises a kappa light chain constant region. In some embodiments, the bispecific binding protein comprises a lambda light chain constant region.

[0177] In some aspects, the disclosure provides a nucleic acid comprising a nucleotide sequence encoding a bispecific binding protein as described herein. In some aspects, the disclosure provides a host cell comprising a nucleic acid as described herein. In some aspects, the disclosure provides a host cell comprising a vector comprising a nucleic acid as described herein. In some aspects, the disclosure provides a host cell having a nucleic acid as described herein integrated into its genome. In some aspects, the disclosure provides a bispecific binding protein produced by a host cell as described herein.PARP1 inhibitors

[0178] Poly adenosine diphosphate-ribose polymerase (PARP) inhibitors (PARPi) with clinical activity as monotherapy selectively inhibit and trap PARP at sites of single-stranded deoxyribonucleic acid (DNA) damage, preventing single-stranded DNA repair, and causing replication-dependent DNA double strand breaks (Pommier et al, Sci Transl Med.2016;8(362):362psl7). Normal cells have the ability to repair the damage accurately and efficiently via the homologous recombination repair (HRR) pathway. In cancer cells that are homologous recombination deficient (HRD), such as those with deleterious BRCA1 / 2 mutations,AZ Ref: B7H4-102-PCT01-NPtreatment with PARPi leads to an increase in genomic instability. This instability will ultimately lead to cancer cell death while sparing normal cells.

[0179] According to all aspects, an ADC and bispecific checkpoint inhibitor as described herein is administered in combination with an inhibitor of PARPI ((Poly (ADP -Ribose) Polymerase 1). In specific embodiments according to all aspects, the inhibitor of PARPI is AZD5305. The language "inhibit," "inhibition" or "inhibiting" includes a decrease in the baseline activity of a biological activity or process.

[0180] The term " AZD5305" refers to a compound with the chemical name of 5-[4-[(7-ethyl-6-oxo-5H-l,5-naphthyridin-3-yl)methyl]piperazin-l-yl]-N-methyl-pyridine-2-carboxamide and structure shown below:

[0181] AZD5305 may also be referred to as “saruparib”.

[0182] Preparation of AZD5305 is disclosed in U. S. Publication. No. US 2021 / 0040084 Al, the disclosure of which is incorporated by reference in its entirety. In some embodiments, a free base of AZD5305 is administered to a subject. In some embodiments, a pharmaceutically acceptable salt of AZD5305 is administered to a subject. In some embodiments, a crystalline AZD5305 is administered to a subject. In some embodiments, crystalline Form A AZD5305 is administered to a subject.

[0183] A "pharmaceutical composition" comprising AZD5305 includes compositions comprising an active ingredient and a pharmaceutically acceptable excipient, carrier or diluent, wherein the active ingredient is AZD5305 or a pharmaceutically acceptable salt thereof.Types of Cancers

[0184] In some embodiments, the cancer is one or more selected from endometrial cancer, ovarian cancer, breast cancer, biliary tract cancer, NSCLC (squamous and / or adenocarcinoma), uterine cancer, testicular cancer, bladder cancer, head and neck cancer, melanoma, renal cell carcinoma, pancreatic cancer, prostate cancer, cervical cancer, hematological cancer,AZ Ref: B7H4-102-PCT01-NPcholangiocarcinoma, gastrointestinal cancer such as gastric cancer and colorectal cancer, and lung cancer. In some embodiments, the cancer is a breast cancer selected from hormone receptorpositive (HR+) breast cancer, human epidermal growth factor receptor 2 positive (HER2+) breast cancer, and triple negative breast cancer (TNBC). In some embodiments, the cancer is homologous recombination deficient (HRD) cancer. In some embodiments, the cancer comprises one or more cells having a mutation in an HRD gene selected from BRC 1, BRCA2, ATM, BRIP1, BARD1, CDK12, CHEK1, CHEK2, FANCL, BAI. 2, PPP2R2A, RAD51B, RAD51C, RAD51D, and RAD54L. In some embodiments, the mutated HRD gene is selected from BRC 1, BRCA2, an ATM.Kits and pharmaceutical compositions

[0185] In related aspects, there is also provided a kit or pharmaceutical composition comprising an ADC as described herein, a bispecific checkpoint inhibitor as described herein, and a PARP1 inhibitor as described herein. There is further embraced use of said kit or said pharmaceutical composition in the methods of the present disclosure.

[0186] Thus, in specific embodiments, the antibody-drug conjugate comprises a drug-linker represented by the following formula:(SG3932),and wherein the drug-linker is conjugated to the antibody or antigen binding fragment thereof. In some embodiments, the antibody or antigen binding fragment thereof that binds to B7-H4 comprises: a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ IDAZ Ref: B7H4-102-PCT01-NPNO: 5, and SEQ ID NO: 6. In some embodiments, the antibody or antigen binding fragment thereof comprises a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 45 and SEQ ID NO: 34, respectively, or a functional variant thereof. In some embodiments, the antibody or antigen binding fragment thereof comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 51; and a light chain comprising the amino acid sequence of SEQ ID NO: 44. In some embodiments, the antibody or antigen binding fragment thereof comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 48; and a light chain comprising the amino acid sequence of SEQ ID NO: 44.

[0187] In specific embodiments of the kits and pharmaceutical compositions described herein, the bispecific checkpoint inhibitor comprises: a) a first binding domain that specifically binds to PD-1; and b) a second binding domain that specifically binds to T cell immunoreceptor with Ig and ITIM domains (TIGIT). In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain comprising a HCDR1 having the amino acid sequence of SEQ ID NO: 56, a HCDR2 having the amino acid sequence of SEQ ID NO: 57, and a HCDR3 having the amino acid sequence of SEQ ID NO: 58, and a light chain variable domain comprising a LCDR1 having the amino acid sequence of SEQ ID NO: 59, a LCDR2 having the amino acid sequence of SEQ ID NO: 60 and a LCDR3 having the amino acid sequence of SEQ ID NO: 61.

[0188] In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 62 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 64. In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 62 and a light chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 64. In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain having the amino acid sequence of SEQ ID NO: 63 and a light chain having the amino acid sequence of SEQ ID NO: 65. In some embodiments, the first binding domain that specifically binds to PD-1 comprises a heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 63 and a light chain having an amino acid sequence that is at least 90%AZ Ref: B7H4-102-PCT01-NPidentical to the amino acid sequence of SEQ ID NO: 65.

[0189] In some embodiments, the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain comprising a HCDR1 having the amino acid sequence of SEQ ID NO: 66, a HCDR2 having the amino acid sequence of SEQ ID NO: 67, and a HCDR3 having the amino acid sequence of SEQ ID NO: 68, and a light chain variable domain comprising a LCDR1 having the amino acid sequence of SEQ ID NO: 69, a LCDR2 having the amino acid sequence of SEQ ID NO: 70, and a LCDR3 having the amino acid sequence of SEQ ID NO: 71. In some embodiments, the second binding domain that specifically binds TIGIT comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 72 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 74. In some embodiments, the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 72 and a light chain variable domain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 74. In some embodiments, the second binding domain that specifically binds to TIGIT comprises a heavy chain having the amino sequence of SEQ ID NO: 73 and a light chain having the amino acid sequence of SEQ ID NO: 75. In some embodiments, the second binding domain that specifically binds to TIGIT comprises a heavy chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 73 and a light chain having an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 75.

[0190] In specific embodiments of the kits and pharmaceutical compositions described herein, the PARP1 inhibitor is AZD5305.

[0191] In some embodiments, a kit comprises an isolated (e.g., purified) antibody or antibody binding fragment as described herein. In some embodiments, a kit comprises an isolated (e.g., purified) ADC as described herein and / or an isolated (e.g., purified) bispecific checkpoint inhibitor as described herein. In some embodiments, the kit comprises one or more containers. The kit may provide the anti-B7H4 antibody or antibody binding fragment and the linker and / or cytotoxic agent individually (e.g., the agent is not conjugated to the anti-B7H4 antibody or antibody binding fragment, but is in a form suitable for conjugation thereto in orderAZ Ref: B7H4-102-PCT01-NPto form the ADC of the disclosure); optionally wherein the kit is further provided with instructions and / or reagents for conjugating the agent to the anti-B7H4 antibody or antibody binding fragment. In some embodiments, the kit comprises all of the components necessary and / or sufficient to administer the combination of ADC, bispecific checkpoint inhibitor and PARP1 inhibitor to a subject. In some embodiments, the kit comprises all of the instructions necessary and / or sufficient to administer the combination of ADC, bispecific checkpoint inhibitor and PARP1 inhibitor to a subject.

[0192] In specific embodiments according to all aspects of the disclosure, the ADC is AZD8205, the bispecific checkpoint inhibitor is rilvegostomig and the PARP1 inhibitor is AZD5305.

[0193] A variety of further modifications and improvements in and to the methods and kits of the present disclosure will be apparent to those skilled in the art. The following non-limiting embodiments are specifically envisioned:Embodiment 1: A method of treating cancer in a human subj ect in need thereof, comprising administering to the human subject:A) an antibody-drug conjugate (ADC) comprising:i) an antibody or antigen binding fragment thereof which binds to a B7-H4 polypeptide, comprising:a) a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively, or a functional variant thereof;b) a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12, respectively, or a functional variant thereof;c) a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3AZ Ref: B7H4-102-PCT01-NPcomprising the amino acid sequence of SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, and SEQ ID NO: 18, respectively, or a functional variant thereof;d) a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 24, respectively, or a functional variant thereof; ore) a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, and SEQ ID NO: 30, respectively, or a functional variant thereof;ii) a cleavable linker, andiii) a cytotoxic agent;B) a bispecific checkpoint inhibitor; andC) a PARP1 inhibitor.Embodiment 2: The method of embodiment 1, wherein the bispecific checkpoint inhibitor is a binding protein having a first binding domain that specifically binds to Programmed Death-1 (PD-1).Embodiment 3: The method of embodiment 1 or 2, wherein the antibody or antigen binding fragment thereof of the ADC comprises:a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively, or a functional variant thereof.Embodiment 4: The method of any one of embodiments 1 to 3, wherein the antibody or antigen binding fragment thereof of the ADC comprises:a variable heavy (VH) chain and a variable light (VL) chain comprising the amino acid sequence of SEQ ID NO: 45 and SEQ ID NO: 34, respectively, or a functionalAZ Ref: B7H4-102-PCT01-NPvariant thereof;a variable heavy (VH) chain and a variable light (VL) chain comprising the amino acid sequence of SEQ ID NO: 33 and SEQ ID NO: 34, respectively, or a functional variant thereof;a variable heavy (VH) chain and a variable light (VL) chain comprising the amino acid sequence of SEQ ID NO: 43 and SEQ ID NO: 34, respectively, or a functional variant thereof;a variable heavy (VH) chain and a variable light (VL) chain comprising the amino acid sequence of SEQ ID NO: 46 and SEQ ID NO: 34, respectively, or a functional variant thereof;a variable heavy (VH) chain and a variable light (VL) chain comprising the amino acid sequence of SEQ ID NO: 47 and SEQ ID NO: 34, respectively, or a functional variant thereof;a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 31, and SEQ ID NO: 32, respectively, or a functional variant thereof;a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 35 and SEQ ID NO: 36, respectively, or a functional variant thereof;a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 37 and SEQ ID NO: 38, respectively, or a functional variant thereof; ora VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 39 and SEQ ID NO: 40, respectively, or a functional variant thereof.Embodiment 5: The method of any one of embodiments 1 to 4, wherein the antibody or antigen binding fragment thereof of the ADC comprises:a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 45 and SEQ ID NO: 34, respectively, or a functional variant thereof.Embodiment 6: The method of any one of embodiments 1 to 5, wherein the antibody or antigen binding fragment thereof of the ADC comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 41.AZ Ref: B7H4-102-PCT01-NPEmbodiment 7: The method of any one of embodiments 1 to 5, wherein the antibody or antigen binding fragment thereof of the ADC comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 52.Embodiment 8: The method of any one of embodiments 1 to 7, wherein the antibody or antigen binding fragment thereof of the ADC comprises a light chain constant region comprising the amino acid sequence of SEQ ID NO: 42.Embodiment 9: The method of any one of embodiments 1 to 5, wherein the antibody or antigen binding fragment thereof of the ADC comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 51; and a light chain comprising the amino acid sequence of SEQ ID NO: 44.Embodiment 10: The method of any one of embodiments 1 to 5, wherein the antibody or antigen binding fragment thereof of the ADC comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 48; and a light chain comprising the amino acid sequence of SEQ ID NO: 44.Embodiment 11: The method of any one of embodiments 1 to 10, wherein the antibody or antigen binding fragment thereof of the ADC is a monoclonal antibody.Embodiment 12: The method of any one of embodiments 1 to 11, wherein the cleavable linker of the ADC is an mp-PEG8-val-ala linker.Embodiment 13: The method of any one of embodiments 1 to 12, wherein the cytotoxic agent of the ADC is a topoisomerase inhibitor, a tubulysin derivative, a pyrrolobenzodiazepine, or a combination thereof.Embodiment 14: The method of any one of embodiments 1 to 13, wherein the cytotoxicAZ Ref: B7H4-102-PCT01-NPagent of the ADC is a topoisomerase inhibitor.Embodiment 15: The method of any one of embodiments 1 to 14, wherein the cytotoxic agent is a topoisomerase inhibitor and wherein the ii) cleavable linker and iii) cytotoxic agent of the ADC comprise a compound of formula I:and salts and solvates thereof, wherein RLis the cleavable linker, which is selected from:wherein Q is:, where Qxis such that Q is an amino-acid residue, a dipeptide residue, a tripeptide residue or a tetrapeptide residue;wherein X is:C(=O)where a = 0 to 5, bl = 0 to 16, b2 = 0 to 16, cl = 0 or 1, c2 = 0 or 1, d = 0 to 5, at least bl or b2 = 0, at least cl or c2 = 0; andAZ Ref: B7H4-102-PCT01-NPwherein GLis a linker for connecting to the antibody or antigen binding fragment;L2Nwherein RL1and RL2are independently selected from H and methyl, or together with the carbon atom to which they are bound form a cyclopropylene or cyclobutylene group, and e is 0 or 1.Embodiment 16: The method of embodiment 15, wherein RLis of formula la.Embodiment 17: The method of embodiment 16, wherein Q is:(a) an amino acid residue selected from: Phe, Lys, Vai, Ala, Cit, Leu, He, Arg, and Trp; or(b) a dipeptide residue selected from:NH-Phe-Lys-C°,XII-Val-Ala-cn-Val-Lys-C°,xnAla-Lys-cNH-Val-Cit-c=o,XII-Phe-Cit-c C),XII-Leu-Cit-cNH-Ile-Cit-c=o,NH-Phe-Arg-c=o,NH-Trp-Cit-c=o, andNH-Gly-Val-C=o; or(c) a tripeptide residue selected from:NH-Glu-Val-Ala-C=o,NH-Glu-Val-Cit-c=o,AZ Ref: B7H4-102-PCT01-NPNH-aGlu-Val-Ala-c=o, andNH-aGlu-Val-Cit-c=o; or(d) a tetrapeptide residue selected from:NH-Gly-Gly-Phe-Glyc=o; andNH-Gly-Phe-Gly-Glyc=oEmbodiment 18: The method of embodiment 16 or 17, wherein a is:(a) 0 to 3; or(b) 0 or 1; or(c) 0.Embodiment 19: The method of any one of embodiments 16 to 18, wherein bl is:(a) 0 to 8; or(b) 0; or(c) 2; or(d) 3; or(e) 4; or(f) 5; or(g) 8.Embodiment 20: The method of any one of embodiments 16 to 19, wherein b2 is:(a) 0 to 8; or(b) 0; or(c) 2; or(d) 3; or(e) 4; or(f) 5; or(g) 8.Embodiment 21: The method of any one of embodiments 15 to 20, wherein:AZ Ref: B7H4-102-PCT01-NP(a) cl is 0 or 1;(b) c2 is 0 or 1; and(c) at least one of cl and c2 is 0.Embodiment 22: The method of any one of embodiments 15 to 21, wherein d is:(a) 0 to 3; or(b) 1 or 2; or(c) 2; or(d) 5.Embodiment 23: The method of any one of embodiments 15 to 22, wherein:(a) a is 0, bl is 0, cl is 1, c2 is 0 and d is 2, and b2 is 0, 2, 3, 4, 5 or 8; or(b) a is 1, b2 is 0, cl is 0, c2 is 0 and d is 0, and bl is 0, 2, 3, 4, 5 or 8; or(c) a is 0, bl is 0, cl is 0, c2 is 0 and d is 1, and b2 is 0, 2, 3, 4, 5 or 8; or(d) b 1 is 0, b2 is 0, cl is 0, c2 is 0, one of a and d is 0, and the other of a and d is 1 or 5; or(e) a is 1, b2 is 0, cl is 0, c2 is 1, d is 2, and bl is 0, 2, 3, 4, 5 or 8.Embodiment 24: The method of any one of embodiments 15 to 23, wherein GLis selectedAZ Ref: B7H4-102-PCT01-NP(GL2) 0 (GL8)0 V— k°^ o V 0(GL34) (GL9) N3s— s>(N02)where the NO2 group is optional(GL3-2) (GL1°)s—H(NO2)where the NO2 group is optional(GL3-3) (GL11)s—XO2N-\2_ / Z~—0 where the NO2 group is optional(GL3-4) (GL12)S—o2Ndwhere the NO2 group is optional(GL4) _ 0 (GL13) Y ZN-N Hal N — |N-NZ? H *Where Hal = I, Br, ClAZ Ref: B7H4-102-PCT01-NP(GL5) 0 (GL14) H2N. \Hal—where Ar represents a C5-6 arylene group, and X represents C1-4 alkyl.Embodiment 25: The method of embodiment 24, wherein GLis selected from GL land GL1‘ 2Embodiment 26: The method of embodiment 15, wherein RLis of formula lb, and:(a) both RL1and RL2are H; or(b) RL1is H and RL2is methyl; or(c) both RL1and RL2are methyl; or(d) wherein RL1and RL2together with the carbon atom to which they are bound form a cyclopropylene group; or(e) wherein RL1and RL2together with the carbon atom to which they are bound form a cyclobutylene group.Embodiment 27: The method of embodiment 1, wherein the ADC is of formula IV:L - (DL)P(IV)or a pharmaceutically acceptable salt or solvate thereof,wherein L is the i) antibody or antigen binding fragment thereof, DLis a drug linker unit comprising the ii) cleavable linker and the iii) cytotoxic agent, and DLis of formula III:AZ Ref: B7H4-102-PCT01-NPwherein RLis the cleavable linker selected from:(ia’):0la', where Q and X are as defined in embodiment 15 and GLLis a linker connected to the antibody or antigen binding fragment; and, where RL1and RL2are as defined in embodiment 15; andp is an integer of from 1 to 20.AZ Ref: B7H4-102-PCT01-NP (GLL1-2) o (GLL8-2)NzCBAVf ^ArCEA< yYxo(GLL2) 0 (GLL94)'C f / xo(GLL34) CBA|_SX (GLL9-2)N-^ CEA(GLL3-2) (GLL1°)T*. --x (GLL-4) CBA| _ (GLL11)1 \ HX— No Ji \ (GLL5) (GLL12)<^1 0°d HN i^^A. Z L7f (GLL6)CBAI__^^ 0 (GLL13)YOHAZ Ref: B7H4-102-PCT01-NP(GLL7)where Ar represents a C5-6 arylene group and X represents C1-4 alkyl.Embodiment 29: The method of embodiment 28, wherein GLLis selected from GLL1and GLL1-2Embodiment 30: The method of any one of embodiments 27 to 29, wherein the drug loading (p) of the cytotoxic agent to the antibody or antibody binding fragment is an integer from 1 to about 10.Embodiment 31: The method of any one of embodiments 14 to 30, wherein the topoisomerase inhibitor is a compound of formula A:H2NOH Oas a single enantiomer or in an enantiomerically enriched form.Embodiment 32: The method of any one of embodiments 14 to 30, wherein the topoisomerase inhibitor is a compound with the formula VI:AZ Ref: B7H4-102-PCT01-NPVIwhere Q is as defined in embodiment 15 or 17.Embodiment 33: The method of any one of embodiments 1 to 32, wherein the ii) cleavable linker and iii) cytotoxic agent together comprise the following compound:OH 0(SG3932).Embodiment 34: The method of any one of embodiments 1 to 33, wherein the bispecific checkpoint inhibitor comprises:a) a first binding domain that specifically binds to PD-1; and b) a second binding domain that specifically binds to T cell immunoreceptor with Ig and ITIM domains (TIGIT).Embodiment 35: The method of embodiment 34, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain comprising a HCDR1 having the amino acid sequence of SEQ ID NO: 56, a HCDR2 having the aminoAZ Ref: B7H4-102-PCT01-NPacid sequence of SEQ ID NO: 57, and a HCDR3 having the amino acid sequence of SEQ ID NO: 58, and a light chain variable domain comprising a LCDR1 having the amino acid sequence of SEQ ID NO: 59, a LCDR2 having the amino acid sequence of SEQ ID NO: 60 and a LCDR3 having the amino acid sequence of SEQ ID NO: 61.Embodiment 36: The method of embodiment 34 or 35, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 62 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 64.Embodiment 37: The method of any one of embodiments 34 to 36, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain having the amino acid sequence of SEQ ID NO: 63 and a light chain having the amino acid sequence of SEQ ID NO: 65.Embodiment 38: The method of any one of embodiments 34 to 37, wherein the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain comprising a HCDR1 having the amino acid sequence of SEQ ID NO: 66, a HCDR2 having the amino acid sequence of SEQ ID NO: 67, and a HCDR3 having the amino acid sequence of SEQ ID NO: 68, and a light chain variable domain comprising a LCDR1 having the amino acid sequence of SEQ ID NO: 69, a LCDR2 having the amino acid sequence of SEQ ID NO: 70, and a LCDR3 having the amino acid sequence of SEQ ID NO: 71.Embodiment 39: The method of any one of embodiments 34 to 38, wherein the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 72 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 74.Embodiment 40: The method of any one of embodiments 34 to 39, wherein the secondAZ Ref: B7H4-102-PCT01-NPbinding domain that specifically binds to TIGIT comprises a heavy chain having the amino sequence of SEQ ID NO: 73 and a light chain having the amino acid sequence of SEQ ID NO: 75.Embodiment 41: The method of any one of embodiments 1 to 40, wherein the bispecific checkpoint inhibitor is a human or humanized bispecific antibody or antigen-binding fragment thereof.Embodiment 42: The method of any one of embodiments 1 to 41, wherein the bispecific checkpoint inhibitor comprises an IgG heavy chain constant region.Embodiment 43: The method of embodiment 42, wherein the IgG heavy chain constant region comprises L234F, L235E and P331S.Embodiment 44: The method of embodiment 42 or 43, wherein the IgG heavy chain constant region is an IgGl heavy chain constant region.Embodiment 45: The method of any one of embodiments 1 to 44, wherein the bispecific checkpoint inhibitor comprises an aglycosylated Fc region.Embodiment 46: The method of any one of embodiments 1 to 44, wherein the bispecific checkpoint inhibitor comprises a deglycosylated Fc region.Embodiment 47: The method of any one of embodiments 1 to 44, wherein the bispecific checkpoint inhibitor comprises an Fc region which has reduced fucosylation or is afucosylated.Embodiment 48: The method of any one of embodiments 1 to 47, wherein the bispecific checkpoint inhibitor comprises a kappa light chain constant region.AZ Ref: B7H4-102-PCT01-NPEmbodiment 49: The method of any one of embodiments 1 to 48, wherein the bispecific checkpoint inhibitor comprises a lambda light chain constant region.Embodiment 50: The method of any one of embodiments 1 to 49, wherein the bispecific checkpoint inhibitor is an antibody.Embodiment 51: The method of embodiment 50, wherein the antibody is an IgG antibody.Embodiment 52: The method of embodiment 51, wherein the antibody is an IgGl antibody.Embodiment 53: The method of any one of embodiments 50 to 52, wherein the antibody is humanized.Embodiment 54: The method of any one of embodiments 1 to 53, wherein the PARP1 inhibitor is AZD5305 or a pharmaceutically acceptable salt thereof.Embodiment 55: The method of any one of embodiments 1 to 54, wherein the cancer comprises a cancer cell which expresses B7-H4.Embodiment 56: The method of any one of embodiments 1 to 55, wherein the cancer is selected from endometrial cancer, ovarian cancer, breast cancer, biliary tract cancer, NSCLC (squamous and / or adenocarcinoma), uterine cancer, testicular cancer, bladder cancer, head and neck cancer, melanoma, renal cell carcinoma, pancreatic cancer, prostate cancer, cervical cancer, hematological cancer, cholangiocarcinoma, gastrointestinal cancer such as gastric cancer and colorectal cancer, and lung cancer.Embodiment 57: The method of any one of embodiments 1 to 56, wherein the cancer is a breast cancer selected from hormone receptor-positive (HR+) breast cancer, human epidermal growth factor receptor 2 positive (HER2+) breast cancer, and triple negative breast cancer (TNBC).AZ Ref: B7H4-102-PCT01-NPEmbodiment 58: The method of any one of embodiments 1 to 57, wherein the cancer is homologous recombination deficient (HRD) cancer.Embodiment 59: The method of embodiment 58, wherein the cancer comprises one or more cells having a mutation in an HRD gene selected from BRCA1, BRCA2, ATM, BRIP 1, BARD1, CDK12, CHEK1, CHEK2, FANCL, PALB2, PPP2R2A, RAD51B, RAD51C, RAD51D, and RAD 54L.Embodiment 60: The method of embodiment 59, wherein the mutated HRD gene is selected from BRCA1, BRCA2, and ATM.Embodiment 61: A kit comprising: 1) an antibody-drug conjugate comprising an antibody or antigen binding fragment thereof which binds to a B7-H4 polypeptide, 2) a bispecific checkpoint inhibitor, and 3) a PARP1 inhibitor, wherein the antibody-drug conjugate, bi specific checkpoint inhibitor and PARP1 inhibitor are each as defined in any one of embodiments 1 to 54.Embodiment 62: The kit of embodiment 61, wherein the antibody-drug conjugate comprises a drug-linker represented by the following formula:(SG3932),and wherein the drug-linker is conjugated to the antibody or antigen binding fragment thereof.AZ Ref: B7H4-102-PCT01-NPEmbodiment 63: The kit of embodiment 61 or 62, wherein the antibody or antigen binding fragment thereof that binds to B7-H4 comprises: a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6.Embodiment 64: The kit of embodiment 63, wherein the antibody or antigen binding fragment thereof comprises a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 45 and SEQ ID NO: 34, respectively, or a functional variant thereof.Embodiment 65: The kit of embodiment 64, wherein the antibody or antigen binding fragment thereof comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 51; and a light chain comprising the amino acid sequence of SEQ ID NO: 44.Embodiment 66: The kit of embodiment 64, wherein the antibody or antigen binding fragment thereof comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 48; and a light chain comprising the amino acid sequence of SEQ ID NO: 44.Embodiment 67: The kit of any one of embodiments 61 to 66, wherein the bispecific checkpoint inhibitor comprises:a) a first binding domain that specifically binds to PD-1; and b) a second binding domain that specifically binds to T cell immunoreceptor with Ig and ITIM domains (TIGIT).Embodiment 68: The kit of embodiment 67, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain comprising a HCDR1 having the amino acid sequence of SEQ ID NO: 56, a HCDR2 having the amino acid sequence of SEQ ID NO: 57, and a HCDR3 having the amino acid sequence of SEQ ID NO: 58, and a light chain variable domain comprising a LCDR1 having the aminoAZ Ref: B7H4-102-PCT01-NPacid sequence of SEQ ID NO: 59, a LCDR2 having the amino acid sequence of SEQ ID NO: 60 and a LCDR3 having the amino acid sequence of SEQ ID NO: 61.Embodiment 69: The kit of embodiment 68, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 62 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 64.Embodiment 70: The kit of embodiment 68 or 69, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain having the amino acid sequence of SEQ ID NO: 63 and a light chain having the amino acid sequence of SEQ ID NO: 65.Embodiment 71: The kit of any one of embodiments 67 to 70, wherein the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain comprising a HCDR1 having the amino acid sequence of SEQ ID NO: 66, a HCDR2 having the amino acid sequence of SEQ ID NO: 67, and a HCDR3 having the amino acid sequence of SEQ ID NO: 68, and a light chain variable domain comprising a LCDR1 having the amino acid sequence of SEQ ID NO: 69, a LCDR2 having the amino acid sequence of SEQ ID NO: 70, and a LCDR3 having the amino acid sequence of SEQ ID NO: 71.Embodiment 72: The kit of any one of embodiments 67 to 71, wherein the second binding domain that specifically binds TIGIT comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 72 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 74.Embodiment 73: The kit of any one of embodiments 67 to 72, wherein the second binding domain that specifically binds to TIGIT comprises a heavy chain having the amino sequence of SEQ ID NO: 73 and a light chain having the amino acid sequence of SEQ ID NO: 75.AZ Ref: B7H4-102-PCT01-NPEmbodiment 74: A pharmaceutical composition comprising: 1) an antibody-drug conjugate comprising an antibody or antigen binding fragment thereof which binds to a B7-H4 polypeptide, 2) a bispecific checkpoint inhibitor, and 3) a PARP1 inhibitor, wherein the antibody-drug conjugate, bispecific checkpoint inhibitor and PARP1 inhibitor are each as defined in any one of embodiments 61 to 73.Embodiment 75: Use of an antibody-drug conjugate in the manufacture of a medicament for use in treating cancer in combination with a bispecific checkpoint inhibitor and a PARP1 inhibitor, wherein the antibody-drug conjugate, the bispecific checkpoint inhibitor and the PARP1 inhibitor are as defined in any one of embodiments 61 to 73.Embodiment 76: Use of a bispecific checkpoint inhibitor in the manufacture of a medicament for use in treating cancer in combination with an antibody-drug conjugate and a PARP1 inhibitor, wherein the bispecific checkpoint inhibitor, the antibody-drug conjugate and the PARP1 inhibitor are as defined in any one of embodiments 61 to 73.Embodiment 77: Use of a PARP1 inhibitor in the manufacture of a medicament for use in treating cancer in combination with an antibody-drug conjugate and a bispecific checkpoint inhibitor, wherein the PARP1 inhibitor, the antibody-drug conjugate and the bispecific checkpoint inhibitor are as defined in any one of embodiments 61 to 73.Embodiment 78: An antibody-drug conjugate in combination with a bispecific checkpoint inhibitor and a PARP1 inhibitor for use in treating cancer, wherein the antibody-drug conjugate, the bispecific checkpoint inhibitor and the PARP1 inhibitor are as defined in any one of embodiments 61 to 73.Embodiment 79: The use of any one of embodiments 75 to 77, or the antibody-drug conjugate in combination with a bispecific checkpoint inhibitor and a PARP1 inhibitor for use of embodiment 78, wherein the cancer is as defined in any one of embodimentsAZ Ref: B7H4-102-PCT01-NP55 to 60.Embodiment 80: The use of embodiment 79, or the antibody-drug conjugate in combination with a bispecific checkpoint inhibitor and a PARP1 inhibitor for use of embodiment 79, wherein the cancer is selected from endometrial cancer, ovarian cancer, breast cancer, biliary tract cancer, NSCLC (squamous and / or adenocarcinoma), uterine cancer, testicular cancer, bladder cancer, head and neck cancer, melanoma, renal cell carcinoma, pancreatic cancer, prostate cancer, cervical cancer, hematological cancer, cholangiocarcinoma, gastrointestinal cancer such as gastric cancer and colorectal cancer, and lung cancer.Embodiment 81: The method, kit, pharmaceutical composition, use or ADC for use of any preceding embodiment, wherein the bispecific checkpoint inhibitor is rilvegostomig.Embodiment 82: The method, kit, pharmaceutical composition, use or ADC for use of any preceding embodiment, wherein:a) the ADC is AZD8205;b) the bispecific checkpoint inhibitor is rilvegostomig; andc) the PARP1 inhibitor is AZD5305.SequencesTable 1SEQ Sequences DescriptionID NO.1 GYYWN ZY0EQD-E02; HCDR1 2 EINHSGSTSYNPSLKS ZY0EQD-E02; HCDR2 3 VLYNWNVDS ZY0EQD-E02; HCDR3 4 RASQDIRNDVG ZY0EQD-E02; LCDR1 5 AASRLQS ZY0EQD-E02; LCDR2 6 LQHNSYPRT ZY0EQD-E02; LCDR3 7 GYYWN ZY0EPQ-E02; HCDR18 EINHSGSTNYNPSLKS ZY0EPQ-E02; HCDR2AZ Ref: B7H4-102-PCT01-NP9 NLYNWNLDS ZY0EPQ-E02; HCDR3 10 RASQGIRNDLG ZY0EPQ-E02; LCDR1 11 VASSLQS ZY0EPQ-E02; LCDR2 12 LQHNSYPRT ZY0EPQ-E02; LCDR3 13 SGGYYWS ZY0EOB-F05; HCDR1 14 NIYYSGSTYYNPSLKS ZY0EOB-F05; HCDR2 15 EKALATVTPSGYENYYTVDV ZY0EOB-F05; HCDR3 16 WASQGISSYLA ZY0EOB-F05; LCDR1 17 AASTLQS ZY0EOB-F05; LCDR2 18 QHLNSYPLT ZY0EOB-F05; LCDR3 19 SGGYYWS ZY0EO5-E07; HCDR1 20 NIYYSGSTYYNPSLKS ZY0EO5-E07; HCDR2 21 EKALASVIPSGYENYYVVDV ZY0EO5-E07; HCDR3 22 WASQGIAGYLA ZY0EO5-E07; LCDR1 23 AASTLQS ZY0EO5-E07; LCDR2 24 QHLNSYPLT ZY0EO5-E07; LCDR3 25 DYYMS ZY0EP0-C07; HCDR1 26 YISSSGSTIYYTDSVKG ZY0EP0-C07; HCDR2 27 DGVGFDY ZY0EP0-C07; HCDR3 28 RASQSVSSSYLA ZY0EP0-C07; LCDR1 29 AASSRAT ZY0EP0-C07; LCDR230 QQYGSSPLYT ZY0EP0-C07; LCDR3SEQ ID NO: 31 (ZY0EPQ-E02, variable heavy chain) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWNWIRQPPGKGLEWIGEINHSGSTNYNPSLK SRVTILVDTSKNQFSLKLSSVTAADTAVYYCARNLYNWNLDSWGQGTLVTVSSSEQ ID NO: 32 (ZY0EPQ-E02, variable light chain) DIQMTQSPSSLSASVGDRVTITCRASQGIRNDLGWYQQKPGRAPKRLIYVASSLQSGVPSRFSGSG SGTEFTLTISSLQPEDFATYYCLQHNSYPRTFGQGTKVEIKSEQ ID NO: 33 (ZY0EQD-E02, variable heavy chain, e.g. pre-germlining) QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWNWIRQPPGKGLEWIGEINHSGSTSYNPSLK SRVTISVDTSKNQFSLKLSSVTAADTAVYYCARVLYNWNVDSWGQGTLVTVSSSEQ ID NO: 34 (ZY0EQD-E02, variable light chain) DIQMTQSPSSLSASVGDRVTITCRASQDIRNDVGWYQQKPGKAPKRLIYAASRLQSGVPSRFSGS GSGTEFTLTISSLQPEDFATYYCLQHNSYPRTFGQGTKVEIKSEQ ID NO: 35 (ZY0EOB-F05, variable heavy chain)AZ Ref: B7H4-102-PCT01-NPQVQLQESGPGLVKPSQTLSLTCTVSDGSISSGGYYWSWIRQHPGKGLEWIGNIYYSGSTYYNPSL KSRVTISVDTSKNQFSLKLNSVTAADTAVYYCATEKALATVTPSGYENYYTVDVWGQGTTVTVssSEQ ID NO: 36 (ZY0EOB-F05, variable light chain) DIQLTQSPSFLSASVGDRVTITCWASQGISSYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGSG SGTEFTLTISSLQPEDFATYYCQHLNSYPLTFGGGTKVEIKSEQ ID NO: 37 (ZY0EO5-E07, variable heavy chain) QVQLQESGPGLVKPSQTLSLTCTVSGGSISSGGYYWSWIRQHPGKGLEWIGNIYYSGSTYYNPSL KSRVTISVDTSKNQFSLKLSSVTAADTAVYYCAREKALASVIPSGYENYYVVDVWGQGTTVTVS SSEQ ID NO: 38 (ZY0EO5-E07, variable light chain) DIQLTQSPSFLSASVGGRVTITCWASQGIAGYLAWYQQKPGKAPKLLIYAASTLQSGVPSRFSGS GSGTEFTLTISSLQPEDFATYYCQHLNSYPLTFGGGTKVEIKSEQ ID NO: 39: (ZY0EP0-C07, variable heavy chain) QVQLVESGGVLVKPGGSLRLSCAASGFTLSDYYMSWIRQAPGMGLEWVSYISSSGSTIYYTDSV KGRFTISRDSAKNSLYLQMNSLRAEDTAVYYCARDGVGFDYWGQGTLVTVSSSEQ ID NO: 40 (ZY0EP0-C07, variable light chain) EIVLTQSPGTLSLFPGERATLSCRASQSVSSSYLAWYQQKPGQSPRLLIYAASSRATGIPDRFSGSG SGTDFTLTISRLEPEDFAVYYCQQYGSSPLYTFGQGTKLEIKSEQ ID NO: 41 (Maia heavy chain constant region, cysteine insertion underlined):ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSCVFLFPPKP KDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT QKSLSLSPGKSEQ ID NO: 42 (light chain constant region) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDS TYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECAZ Ref: B7H4-102-PCT01-NPSEQ ID NO: 43 (ZY0EQD-E02, variable heavy chain, e.g. pre-germlining, e.g. variant of SEQ ID NO: 33 / SEQ ID NO: 45) QVQLQQWGAGLLKPSETLSLTCTVYGGSFSGYYWNWIRQPPGRGLEWIGEINHSGSTSYNPSLK SRITISIDTSKNQFSLKLSSVTAADTAVYYCARVLYNWNVDSWGQGTLVTVSSSEQ ID NO: 44 (ZY0EQD-E02, light chain):DIQMTQSPSSLSASVGDRVTITCRASQDIRNDVGWYQQKPGKAPKRLIYAASRLQSGVPSRFSGS GSGTEFTLTISSLQPEDFATYYCLQHNSYPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASV VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGECSEQ ID NO: 45 (EQD-E02 GL, variable heavy chain, GL = germlined) QVQLQQWGAGLLKPSETLSLACTVYGGSFSGYYWNWIRQPPGKGLEWIGEINHSGSTSYNPSLK SRVTISVDTSKNQFSLKLSSVTAADTAVYYCARVLYNWNVDSWGQGTLVTVSSSEQ ID NO: 46 (EQD-E02-GLY, variable heavy chain, GLY = germlined with a Y substitution) QVQLQQWGAGLLKPSETLSLACTVYGGSFSGYYWNWIRQPPGKGLEWIGEIYHSGSTSYNPSLK SRVTISVDTSKNQFSLKLSSVTAADTAVYYCARVLYNWNVDSWGQGTLVTVSSSEQ ID NO: 47 (EQD-E02-GLQ, variable heavy chain, GLQ = germlined with a Q substitution) QVQLQQWGAGLLKPSETLSLACTVYGGSFSGYYWNWIRQPPGKGLEWIGEIQHSGSTSYNPSLK SRVTISVDTSKNQFSLKLSSVTAADTAVYYCARVLYNWNVDSWGQGTLVTVSSSEQ ID NO: 48 (E02-GL-Maia-heavy chain, cysteine insertion underlined) QVQLQQWGAGLLKPSETLSLACTVYGGSFSGYYWNWIRQPPGKGLEWIGEINHSGSTSYNPSLK SRVTISVDTSKNQFSLKLSSVTAADTAVYYCARVLYNWNVDSWGQGTLVTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSCVFLFPPKPKDTLMISRTPE VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSEQ ID NO: 49 (E02-GLY-Maia-heavy chain, GLY = germlined with a Y substitution)QVQLQQWGAGLLKPSETLSLACTVYGGSFSGYYWNWIRQPPGKGLEWIGEIYHSGSTSYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARVLYNWNVDSWGQGTLVTVSSASTKGPSVFPLAZ Ref: B7H4-102-PCT01-NPAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSCVFLFPPKPKDTLMISRTPE VTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSEQ ID NO: 50 (E02-GLQ-Maia-heavy chain, GLQ = germlined with a Q substitution) QVQLQQWGAGLLI< PSETLSLACTVYGGSFSGYYWNWIRQPPGI< GLEWIGEIQHSGSTSYNPSLI< SRVTISVDTSKNQFSLKLSSVTAADTAVYYCARVLYNWNVDSWGQGTLVTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSCVFLFPPKPKDTLMISRTPE VTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSEQ ID NO: 51 (E02-GL-WT-heavy chain) QVQLQQWGAGLLKPSETLSLACTVYGGSFSGYYWNWIRQPPGKGLEWIGEINHSGSTSYNPSLK SRVTISVDTSKNQFSLKLSSVTAADTAVYYCARVLYNWNVDSWGQGTLVTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK CKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNG QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSEQ ID NO: 52 (heavy chain constant region) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSCVFLFPPKP KDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT QKSLSLSPGKSEQ ID NO: 53 (Human B7H4 nucleic acid sequence, including 5’ and 3’ UTR) GCCACCatggcttccctggggcagatcctcttctggagcataattagcatcatcattattctggctggagcaattgcactcatcattggctttggtatttca gggagacactccatcacagtcactactgtcgcctcagctgggaacattggggaggatggaatcctgagctgcacttttgaacctgacatcaaactttctgat atcgtgatacaatggctgaaggaaggtgttttaggcttggtccatgagttcaaagaaggcaaagatgagctgtcggagcaggatgaaatgttcagaggcc ggacagcagtgtttgctgatcaagtgatagttggcaatgcctctttgcggctgaaaaacgtgcaactcacagatgctggcacctacaaatgttatatcatca cttctaaaggcaaggggaatgctaaccttgagtataaaactggagccttcagcatgccggaagtgaatgtggactataatgccagctcagagaccttgcg gtgtgaggctccccgatggttcccccagcccacagtggtctgggcatcccaagttgaccagggagccaacttctcggaagtctccaataccagctttgagAZ Ref: B7H4-102-PCT01-NPctgaactctgagaatgtgaccatgaaggttgtgtctgtgctctacaatgttacgatcaacaacacatactcctgtatgatgaaaatgacattgccaaagcaa caggggatatcaaagtgacagaatcggagatcaaaaggcggagtcacctacagctgctaaactcaaaggcttctctgtgtgtctcttctttctttgccatca gctgggcacttctgcctctcagcccttacctgatgctaaaaTAATAASEQ ID NO: 54 (Human B7H4 nucleic acid sequence, coding sequence) atggctccctggggcagatcctctctggagcataattagcatcatcatatctggctggagcaattgcactcatcattggctttggtatttcagggagacac tccatcacagtcactactgtcgcctcagctgggaacatggggaggatggaatcctgagctgcacttttgaacctgacatcaaacttctgatatcgtgatac aatggctgaaggaaggtgttttaggcttggtccatgagttcaaagaaggcaaagatgagctgtcggagcaggatgaaatgttcagaggccggacagca gtgtttgctgatcaagtgatagtggcaatgcctctttgcggctgaaaaacgtgcaactcacagatgctggcacctacaaatgttatatcatcacttctaaagg caaggggaatgctaaccttgagtataaaactggagccttcagcatgccggaagtgaatgtggactataatgccagctcagagaccttgcggtgtgaggct ccccgatggttcccccagcccacagtggtctgggcatcccaagttgaccagggagccaacttctcggaagtctccaataccagctttgagctgaactctg agaatgtgaccatgaaggtgtgtctgtgctctacaatgtacgatcaacaacacatactcctgtatgatgaaaatgacatgccaaagcaacaggggatat caaagtgacagaatcggagatcaaaaggcggagtcacctacagctgctaaactcaaaggctctctgtgtgtctcttctttctttgccatcagctgggcact ctgcctctcagcccttacctgatgctaaaaSEQ ID NO: 55 (Human B7H4 polypeptide sequence; UniProt Accession No.: Q7Z7D3) MASLGQILFWSIISIIIILAGAIALIIGFGISGRHSITVTTVASAGNIGEDGILSCTFEPDIKLSDIVIQWL KEGVLGLVHEFKEGKDELSEQDEMFRGRTAVFADQVIVGNASLRLKNVQLTDAGTYKCYIITSK GKGNANLEYKTGAFSMPEVNVDYNASSETLRCEAPRWFPQPTVVWASQVDQGANFSEVSNTSF ELNSENVTMKVVSVLYNVTINNTYSCMIENDIAKATGDIKVTESEIKRRSHLQLLNSKASLCVSSF FAISWALLPLSPYLMLKTable 2: Sequences of PD-l / TIGIT BispecificSEQ SequenceID NO56 PD1 LO115 DYGMHHCDR157 PD1 LO115 YISSGSYTIYSADSVKGHCDR258 PD1 LO115 RAPNSFYEYYFDYHCDR359 PD1 LO115 SASSKHTNLYWSRHMYLCDR160 PD1 LO115 LTSNRATLCDR261 PD1 LO115 QQWSSNPFTLCDR3AZ Ref: B7H4-102-PCT01-NPPD1 LO115 EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYGMHWVRQAPGKG variable heavy LEWVAYISSGSYTIYSADSVKGRFTISRDNAKNSLYLQMNSLRAED chain TAVYYCARRAPNSFYEYYFDYWGQGTTVTVSSPD1 LO115 EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYGMHWVRQAPGKG Hole heavy LEWVAYISSGSYTIYSADSVKGRFTISRDNAKNSLYLQMNSLRAED chain TAVYYCARRAPNSFYEYYFDYWGQGTTVTVSSASTKGPSVFPLAP SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCD KTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSH EDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVCTLPPSREEM TKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS FFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK PD1 LO115 QIVLTQSPATLSLSPGERATLSCSASSKHTNLYWSRHMYWYQQKP variable light GQAPRLLIYLTSNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYC chain QQWSSNPFTFGQGTKLEIKPD1 LO115 QIVLTQSPATLSLSPGERATLSCSASSKHTNLYWSRHMYWYQQKP WT Kappa light GQAPRLLIYLTSNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYC chain QQWSSNPFTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCL LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC TIGIT VARI SYAMH HCDR1TIGIT VARI VIS YAGEVKYYAD S VKG HCDR2TIGIT VARI DPLPLHYYGMDV HCDR3TIGIT VARI SGSSSNIGRRPVN LCDR1TIGIT VARI SQNQRPS LCDR2TIGIT VARI AVWDDIGRVLQ LCDR3TIGIT VARI QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYAMHWVRQAPGKGvariable heavy LEWVAVISYAGEVKYYADSVKGRFTISRDNSKNTLYLQMNSLRAE chain DTAVYYCARDPLPLHYYGMDVWGQGTTVTVSSAZ Ref: B7H4-102-PCT01-NPTIGIT VARI QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYAMHWVRQAPGKGKnob heavy LEWVAVISYAGEVKYYADSVKGRFTISRDNSKNTLYLQMNSLRAE chain DTAVYYCARDPLPLHYYGMDVWGQGTTVTVSSASTKGPSVCPLA PSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSV DKTHTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPCREE MTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K TIGIT VARI QSVLTQPPSASGTPGQRVTISCSGSSSNIGRRPVNWYQQLPGTAPKvariable light LLIYSQNQRPSGVPDRFSGSQSGTSASLAISGLQSEDEADYFCAVW chain DDIGRVLQLGGGTQLTVLTIGIT VARI QSVLTQPPSASGTPGQRVTISCSGSSSNIGRRPVNWYQQLPGTAPKengineered LLIYSQNQRPSGVPDRFSGSQSGTSASLAISGLQSEDEADYFCAVW Lambda light DDIGRVLQLGGGTQLTVLGQPKAAPSVTLFPPCSEELQANKATLV chain CLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYL SLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTEVS PD1 LO115 GAGGTGCAGCTGGTGGAATCCGGCGGAGGACTGGTGCAGCCTG variable heavy GCGGCTCCCTGAGACTGTCTTGCGCCGCCTCCGGCTTCACATTC chain TCCGACTACGGCATGCACTGGGTCCGACAGGCCCCTGGAAAGG GCCTGGAATGGGTGGCCTACATCTCCTCCGGCTCCTACACCATC TACTCCGCCGACTCCGTGAAGGGCCGGTTCACCATCTCCCGGGA CAACGCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGCGG GCCGAGGACACAGCCGTGTACTACTGTGCCAGACGGGCCCCTA ACTCCTTCTACGAGTACTACTTCGACTACTGGGGCCAGGGCACC ACCGTGACCGTGTCCTCT PD1 LO115 GAGGTGCAGCTGGTGGAATCCGGCGGAGGACTGGTGCAGCCTG Hole heavy GCGGCTCCCTGAGACTGTCTTGCGCCGCCTCCGGCTTCACATTC chain TCCGACTACGGCATGCACTGGGTCCGACAGGCCCCTGGAAAGG GCCTGGAATGGGTGGCCTACATCTCCTCCGGCTCCTACACCATC TACTCCGCCGACTCCGTGAAGGGCCGGTTCACCATCTCCCGGGA CAACGCCAAGAACTCCCTGTACCTGCAGATGAACTCCCTGCGG GCCGAGGACACAGCCGTGTACTACTGTGCCAGACGGGCCCCTA ACTCCTTCTACGAGTACTACTTCGACTACTGGGGCCAGGGCACC ACCGTGACCGTGTCCTCTGCTAGCACCAAAGGTCCGAGCGTTTT TCCGCTGGCACCGAGCAGCAAAAGCACCTCTGGTGGCACCGCA GCACTGGGTTGTCTGGTGAAAGATTATTTTCCGGAACCGGTTAC CGTTTCTTGGAATAGCGGTGCACTGACCAGCGGTGTTCATACCT TTCCGGCAGTTCTGCAGAGCAGCGGTCTGTATAGCCTGTCTAGC GTTGTTACCGTTCCGAGCAGCAGCCTGGGCACCCAGACCTATAT TTGCAATGTGAATCATAAACCGAGCAATACAAAAGTTGATAAA CGCGTTGAACCGAAAAGCTGTGACAAAACTCACACGTGCCCAC CGTGCCCAGCACCTGAGTTCGAGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCAZ Ref: B7H4-102-PCT01-NPTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCT GAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATA ATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGT ACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTG AATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCC CAGCCAGCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCC CCGAGAACCACAGGTCTGCACCCTGCCCCCATCCCGGGAGGAG ATGACCAAGAACCAGGTCAGCCTGAGCTGCGCGGTCAAAGGCT TCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCA GCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCC GACGGCTCCTTCTTCCTCGTTAGCAAGCTCACCGTGGACAAGAG CAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCAT GAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGT CTCCGGGTAAATGA PD1 LO115 CAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCCCTGAGCCC variable light TGGCGAGAGAGCCACCCTGAGCTGCTCCGCCTCCTCCAAGCAC chain ACCAACCTGTACTGGTCCCGGCACATGTACTGGTATCAGCAGA AGCCCGGCCAGGCCCCTCGGCTGCTGATCTACCTGACCTCTAAC CGGGCCACCGGCATCCCTGCCAGATTCTCCGGCTCTGGCTCCGG CACCGACTTCACCCTGACCATCTCCAGCCTGGAACCCGAGGACT TCGCCGTGTACTACTGCCAGCAGTGGTCCTCCAACCCCTTCACC TTCGGCCAGGGCACCAAGCTGGAAATCAAG PD1 LO115 CAGATCGTGCTGACCCAGTCCCCTGCCACCCTGTCCCTGAGCCC WT Kappa light TGGCGAGAGAGCCACCCTGAGCTGCTCCGCCTCCTCCAAGCAC chain ACCAACCTGTACTGGTCCCGGCACATGTACTGGTATCAGCAGA AGCCCGGCCAGGCCCCTCGGCTGCTGATCTACCTGACCTCTAAC CGGGCCACCGGCATCCCTGCCAGATTCTCCGGCTCTGGCTCCGG CACCGACTTCACCCTGACCATCTCCAGCCTGGAACCCGAGGACT TCGCCGTGTACTACTGCCAGCAGTGGTCCTCCAACCCCTTCACC TTCGGCCAGGGCACCAAGCTGGAAATCAAGCGTACGGTGGCTG CACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAA TCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCC AGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAAT CGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGG ACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGC AGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCAT CAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAG AGTGTTAG TIGIT VARI CAGGTGCAGCTGGTGGAGTCTGGAGGAGGCGTGGTCCAGCCTGvariable heavy GGAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTC chain AGTAGCTATGCTATGCACTGGGTCCGCCAGGCTCCAGGCAAGG GGCTGGAGTGGGTGGCAGTTATATCATATGCGGGGGAGGTGAA ATACTACGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGA GACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGA GAGCTGAGGACACGGCTGTGTATTACTGTGCGAGAGACCCGCT ACCGCTACATTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCAAZ Ref: B7H4-102-PCT01-NPTIGIT VARI CAGGTGCAGCTGGTGGAGTCTGGAGGAGGCGTGGTCCAGCCTGKnob heavy GGAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTC chain AGTAGCTATGCTATGCACTGGGTCCGCCAGGCTCCAGGCAAGG GGCTGGAGTGGGTGGCAGTTATATCATATGCGGGGGAGGTGAA ATACTACGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGA GACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGA GAGCTGAGGACACGGCTGTGTATTACTGTGCGAGAGACCCGCT ACCGCTACATTACTACGGTATGGACGTCTGGGGCCAAGGGACC ACGGTCACCGTCTCCTCAGCGTCGACCAAAGGTCCGAGCGTGT GCCCGCTGGCACCGAGCAGCAAAAGCACCTCTGGTGGCACCGC AGCACTGGGTTGTCTGGTGAAAGATTATTTTCCGGAACCGGTTA CCGTTTCTTGGAATAGCGGTGCACTGACCAGCGGTGTTCATACC TTTCCGGCAGTCCTGCAGAGCAGCGGTCTGTATAGCCTGTCTAG CGTTGTTACCGTTCCGAGCAGCAGCCTGGGCACCCAGACCTATA TTTGCAATGTGAATCATAAACCGAGCAATACCAAAGTTGATAA ACGCGTTGAACCGAAAAGCGTGGACAAAACTCACACGTGCCCA CCGTGCCCAGCACCTGAGTTCGAGGGGGGACCGTCAGTCTTCCT CTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCC CTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCC TGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCAT AATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACG TACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCT GAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTC CCAGCCAGCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGC CCCGAGAACCACAGGTCTACACCCTGCCCCCATGCCGGGAGGA GATGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGC TTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGC AGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTC CGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGA GCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCA TGAGGCTCTGCACAACCACTACACGCAGAAGAGCTTAAGCCTG TCTCCGGGTAAATGA TIGIT VARI CAGTCTGTGCTGACTCAGCCTCCCTCAGCGTCTGGGACCCCCGGvariable light GCAGAGGGTCACCATCTCTTGTTCTGGAAGCAGCTCCAATATCG chain GAAGGAGGCCTGTAAACTGGTACCAGCAGCTCCCAGGAACGGC CCCCAAACTCCTCATCTATAGTCAGAATCAGCGGCCCTCAGGGG TCCCTGACCGATTCTCTGGCTCCCAGTCTGGCACCTCAGCCTCC CTGGCCATCAGTGGGCTCCAGTCTGAGGATGAGGCTGATTATTT CTGTGCAGTTTGGGATGACATCGGCCGCGTCCTGCAGTTAGGCG GAGGCACCCAGCTGACCGTCCTA TIGIT VARI CAGTCTGTGCTGACTCAGCCTCCCTCAGCGTCTGGGACCCCCGGengineered GCAGAGGGTCACCATCTCTTGTTCTGGAAGCAGCTCCAATATCG Lambda light GAAGGAGGCCTGTAAACTGGTACCAGCAGCTCCCAGGAACGGC chain CCCCAAACTCCTCATCTATAGTCAGAATCAGCGGCCCTCAGGGG TCCCTGACCGATTCTCTGGCTCCCAGTCTGGCACCTCAGCCTCC CTGGCCATCAGTGGGCTCCAGTCTGAGGATGAGGCTGATTATTT CTGTGCAGTTTGGGATGACATCGGCCGCGTCCTGCAGTTAGGCG GAGGCACCCAGCTGACCGTCCTAGGTCAGCCCAAGGCGGCCCCCTCGGTCACTCTGTTCCCGCCCTGCTCTGAGGAGCTTCAAGCCAAZ Ref: B7H4-102-PCT01-NPACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGA GCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGG CGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAA GTACGCGGCCAGCAGCTACCTGAGCCTGACGCCTGAGCAGTGG AAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGA GCACCGTGGAGAAGACAGTGGCCCCTACAGAAGTGTCATGA84 Murine QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYAMHWVRQAPGKG AZD2936 LEWVAVISYAGEVKYYADSVKGRFTISRDNSKNTLYLQMNSLRAE surrogate DTAVYYCARDPLPLHYYGMDVWGQGTTVTVSSASTTAPSVYPLA TIGIT heavy PVCGDTTGS SVTLGCLVKGYFPEPVTLTWNSGSLS SGVHTFPAVLQ chain SDLYTLSSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGPTIKP CPPCKCPAPNAAGGPSVFIFPPKIKDVLMISLSPIVTCVVVDVSEDD PDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQDWM SGKEFKCKVNNKDLGAPIERTISKPKGSVRAPQVCVLPPPEEEMTK KQVTLSCAVTDFMPEDIYVEWTNNGKTELNYKNTEPVLDSDGSYF MVSKLRVEKKNWVERNSYSCSVVHEGLHNHHTTKSFSRTPGK85 Murine QSVLTQPPSASGTPGQRVTISCSGSSSNIGRRPVNWYQQLPGTAPK AZD2936 LLIYSQNQRPSGVPDRFSGSQSGTSASLAISGLQSEDEADYFCAVW surrogate DDIGRVLQLGGGTQLTVLGQPKLSPSVTLFPPSSEELETNKATLVC TIGIT light TITDFYPGVVTVDWKVDGTPVTQGMETTQPSKQSNNKYMASSYL chain TLTARAWERHSSYSCQVTHEGHTVEKSLSRADCS86 Murine QVQLQQSGAELVKPGSSVKISCKASGYTFTSHFIHWIKQQPGNGLE AZD2936 WIGGIYPGDGDTEYNQQFNGKATLTADKSSSTAYMRLSSLTSEDS surrogate PD 1 AVYFCATRVPSYWFFDFWGPGTMVTVSSASTTAPSVCPLAPVSGD heavy chain TTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPAVLQSDLYT LSSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGPTIKPCPPCK CPAPNAAGGPSVFIFPPKIKDVLMISLSPIVTCVVVDVSEDDPDVQIS WFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQDWMSGKEFK CKVNNKDLGAPIERTISKPKGSVRAPQVYVLPPCEEEMTKKQVTL WCMVTDFMPEDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSK LRVEKKNWVERNSYSCSVVHEGLHNHHTTKSFSRTPGK87 Murine DVALTQTPVAQPVTLGDQASISCRSSQSLVHSNGRTYLEWYLQKP AZD2936 GQSPQLLIYKVSNRFSGVPDRFIGSGSGSDFTLTISRVEPEDLGVYY surrogate PD 1 CFQATHDPNTFGAGTKLELKRADAAPTVSIFPPCSEQLTSGGASVV light chain CFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNESEXAMPLES

[0194] The combination therapy disclosed herein will now be further explained by reference to the following non-limiting examples.AZ Ref: B7H4-102-PCT01-NPExample 1: AZD5305 synergizes with AZD8205 and AZD2936 to inhibit endometrial tumor spheroid growth in a 3D endometrial tumor spheroid-peripheral blood mononuclear cell co-culture model

[0195] An in vitro 3D RL95-2 endometrial tumor spheroid-peripheral blood mononuclear cell co-culture model was used to compare the anti-tumor efficacy of various doublet and triplet combinations of AZD8205, AZD5305, and AZD2936 (rilvegostomig).

[0196] RL95-2 human endometrial adenosquamous carcinoma cells were stained with fluorescently conjugated monoclonal antibodies with specificity for B7 Homolog 4 (B7-H4), Programmed Death Ligand 1 (PD-L1) and Poliovirus Receptor (PVR) and evaluated for surface expression of the target proteins by flow cytometry. RL95-2 cells were shown to express B7-H4 (FIG. 1, panel A), which is the target of AZD8205, as well as PD-L1 (FIG. 1, panel B) and PVR (FIG. 1, panel C), the ligands for PD-1 and TIGIT, respectively, which are the targets of rilvegostomig. RL95-2 cells were engineered to express Green Fluorescent Protein (GFP) by lentiviral transduction with a vector encoding the GFP DNA sequence. When plated in ultra-low attachment u-bottom multiwell plates, RL95-2-GFP cells spontaneously form tumor spheroids which display linear growth for up to 20 days in culture and develop secondary structure which can be observed by spinning disk confocal microscopy (Nikon) (FIG. 1, panel D).

[0197] For co-culture experiments with RL95-2-GFP spheroids, peripheral blood mononuclear cells (PBMCs) were isolated from the blood of healthy donors and educated to react to RL95-2 spheroids by incubation with chemical free cell lysates of the parental RL95-2 cell line in the presence of AIM-V media containing human serum and a cocktail of human cytokines. RL95-2 endometrial tumor spheroids were formed for 72 hours prior to co-culture with tumor reactive PBMCs and doublet or triplet treatments with 10 pg / ml AZD8205, 1 nM rilvegostomig, and 100 nM AZD5305 alone or in combination, and the fluorescent images of each well were captured every 4 hours in an Incucyte SX5 high content imager (Sartorius) for up to 14 days. As shown in FIG. 2A, the triplet combination of AZD8205 + AZD5305 + AZD2936 controlled the tumor and prevented its growth more effectively than any other treatment or combination of treatments.AZ Ref: B7H4-102-PCT01-NP

[0198] The fluorescence intensity of the tumor spheroids was evaluated at the end of the assay as a relative measure of viable spheroid volume. As shown in FIG.2B, the triplet combination of AZD8205 + AZD5305 + AZD2936 significantly enhanced the anti-tumor efficacy of any of the tested doublet or single treatments.Example 2: AZD8205 combines with AZD5305 and a murine surrogate of AZD2936 for enhanced anti-tumor efficacy in a human B7-H4 overexpressing colon carcinoma mouse model

[0199] AZD8205 (an anti-B7H4 topoisomerase inhibitor ADC), AZD5305 (a PARP1 inhibitor) and a murine surrogate of AZD2936 (an anti-PD-l / TIGIT bispecific checkpoint inhibitor) were tested in a colon cancer mouse model for efficacy.

[0200] MC38 mouse colon carcinoma cells were engineered by lentiviral transduction with a vector encoding the human B7-H4 gene to express human B7-H4 protein (SEQ ID NO: 55). After single cell cloning and in vivo evaluation, MC-38_huB7-H4 clone Pl A9 was selected for in vivo combination experiments. MC38_huB7-H4 clone Pl A9 cells were implanted subcutaneously in the flank of female C57BL / 6 at an inoculum of 2xl06cells per mouse. Mice were kept in groups of no more than six mice per cage with continuous access to food and water. Ten mice per group were tested with each treatment. Drug treatment was administered as shown in the schematic in FIG.3. Control mice were either untreated, administered vehicle (sterile phosphate buffered saline) or a non-targeting control human IgGl antibody conjugated to the topoisomerase inhibitor payload SG3932 at a drug:antibody ratio (DAR) of 8. Per FIG.3, individual agents were administered as follows: AZD8205 7 mg / kg single dose intravenously, murine AZD2936 10 mg / kg twice weekly intraperitoneally for two weeks, AZD5305 0.1 mg / kg orally once daily for 21 days. Murine AZD2936 (comprising sequences of SEQ ID NOs: 84-87) is a surrogate rilvegostomig molecule, since rilvegostomig is not mouse cross-reactive. Mean tumor volume was measured using digital calipers and the average volume across each treatment group is shown in FIGs. 4A-4C. Whilst the respective monotherapies and doublet therapies demonstrated a reduction in mean tumour volume versus control, with the doublet therapies outperforming the monotherapies, the triple combination of AZD8205 + AZD5305 + mAZD2936 near-ablated tumour growth during the same timeframe.AZ Ref: B7H4-102-PCT01-NP

[0201] Complete Responses (CRs: i.e. the absence of a measurable tumor at the end of the study) were calculated for each treatment group, per the RECIST criteria. No CRs were observed in the ADC isotype control group or monotherapy treatment groups, whilst 1 CR was observed in each of the doublet therapy treatment arms. However, of the 10 mice treated with the triplet therapy (AZD8205 + AZD5305 + mAZD2936), 6 CRs were observed.

[0202] Overall Response Rate (ORR) was further evaluated by RECIST criteria using the best percent change from baseline in tumor volume for each animal. As seen in FIG. 5, the triplet combination treatment delivered a 70% overall response rate which was more than the additive response rates of the monotherapies with each doublet therapy, suggesting therapeutic synergy with the triplet combination.

[0203] Tumor Growth Inhibition (TGI) was calculated from the tumor volume (TV) data at day 29 post tumor implantation using the isotype ADC treated animals as the control group. The equation for TGI is: TGI = 100 - ((Average TVcontrol / TVtreated) * 100). As shown in FIG.6A, the triple combination of AZD8205 + AZD5305 + mAZD2936 inhibited tumor growth significantly more effectively than any of the doublet treatments. Statistical significance between groups was determined by one-way ANOVA with Holm-Sidak multiple comparisons testing. N = 8-12 per group, *p <0.05, **p <0.01.

[0204] Event free survival was determined with the tumor progression event was reached when tumor volumes reached 400% or more compared to their baseline measure. FIG. 6B further confirms the triple combination of AZD8205 + AZD5305 + mAZD2936 was significantly more effective than any of the doublet treatments. Statistical differences in median survival between groups were determined by the Logrank test. N = 8-12 per group, *p <0.05, **p <0.01.Example 3: Combination of AZD8205, mAZD2936 and AZD5305 modulates the tumor immune microenvironment

[0205] A repeat study of identical design to the study described in Example 2 was carried out, with an additional group assessing a gap scheduled treatment regimen where AZD5305 was administered on days 8-14 post AZD8205 and mAZD2936 treatment initiation (see FIG. 7). AnAZ Ref: B7H4-102-PCT01-NPadditional cohort of animals was also included for pharmacodynamic assessment of the post treatment tumor immune composition (FIG. 8).

[0206] At Day 11 post AZD8205 and mAZD2936 treatment initiation tumors were harvested and dissociated to single cells by enzymatic digestion prior to immunostaining with fluorescent conjugated anti-mouse antibodies specific for markers of immune cell identity and activation status. Stained samples were evaluated by multi-color flow cytometry on a BD FACS Symphony A5 cytometer and data analyzed by FlowJo software. In the efficacy cohort, the combination of AZD8205 and mAZD2936 was highly effective with 4 out of 6 mice achieving a complete response (CR) (FIG. 7). The day 8-14 AZD5305 gap schedule dosing in combination AZD8205 and mAZD2936 resulted in 3 of 6 CRs, while continuous daily treatment with AZD5305 in combination with AZD8205 and mAZD2936 resulted in 6 of 6 animals achieving CRs (FIG. 7).

[0207] In the pharmacodynamic cohort, triple combination therapy with AZD8205, mAZD2936 and AZD5305 resulted in significant increases in the proportion of T cells among tumor infiltrating immune cells regardless of dose schedule (FIG. 8). The proportion of immune cells consisting of CD3 positive T cells was significantly increased by the triple combination therapy, regardless of dose schedule. The proportion of immune cells consisting of cytotoxic CD8 positive T cells was significantly increased for the doublet combination of AZD8205 and mAZD2936 as well as the triple combination, again regardless of dose schedule. Among myeloid populations, the proportion of immune cells that consisted of co-stimulatory molecule (CD86) expressing macrophages was significantly increased while the proportion of cells consisting of immunosuppressive granulocytic myeloid derived suppressor cells (gMDSCs) was significantly decreased by triple combination therapy with AZD8205, mAZD2936 and AZD5305, regardless of dose schedule (FIG. 8). The results suggest that the triple combination treatment results in early infiltration of effector T cells and change in myeloid composition toward an antitumor phenotype.Example 4: A Phase I / IIa Multi-center, Open-label Dose-Escalation and Expansion Study of AZD8205 in combination with Saruparib (AZD5305) and Rilvegostomig (AZD2936) -BLUESTARAZ Ref: B7H4-102-PCT01-NP

[0208] The study will evaluate the safety, tolerability, pharmacokinetics, pharmacodynamics, and antitumor activity of AZD8205 when given in combination with saruparib (AZD5305) with or without rilvegostomig (AZD2936) in participants with B7-H4-expressing advanced tumors.

[0209] The study will comprise a Part A and a Part B. Part A will be a dose-escalation part evaluating AZD8205 in combination with saruparib with or without rilvegostomig. Doseescalation cohorts may be expanded with additional participants to evaluate further the safety, tolerability, PK, PD, and biological activity of AZD8205 in combination with saruparib with or without rilvegostomig. Part B will be a dose-expansion part to further evaluate anti-tumor activity.

[0210] Primary outcomes include: observing the number of patients with adverse events, observing the number of patients with serious adverse events, observing the number of patients experiencing dose-limiting toxicity and observing the number of patients with changes from baseline laboratory findings, ECGs and vital signs.

[0211] Secondary outcomes include:Table 3: Secondary Outcome MeasuresOutcome Measure DescriptionObjective Response Rate (ORR) The percentage of patients with a confirmed investigator assessed complete or partial response according to response criteria in solid tumours (RECIST 1.1).Duration of response (DoR) The time from the date of first response until date of disease progression (RECIST 1.1) or death in the absence of disease progression.Progression free Survival (PFS) The time from first dose until RECIST 1.1defined disease progression or cessation of study treatment.Disease Control Rate at 12 weeks (DCR- Percentage of patients with confirmed CR or PR 12) or having SD maintained for >= 11 weeks from first dose (RECIST 1.1).Overall Survival (OS) The time from the date of the first dose of study treatment until death due to any cause.Pharmacokinetics of saruparib in To characterize the PK plasma concentration and combination with AZD8205 with or PK parameters of saruparib, including but notwithout rilvegostomig limited to AUC, Cmax, tmax, clearance, and half-AZ Ref: B7H4-102-PCT01-NPlife, as data allow, of saruparib when given in combination with AZD8205 with or without rilvegostomig.Pharmacokinetics of rilvegostomig in To characterize the PK serum concentrations and combination with AZD8205 and saruparib PK parameters (where applicable) of rilvegostomig when given in combination with AZD8205 and saruparib.Immunogenicity of rilvegostomig in The number and percentage of participants whocombination with AZD8205 and saruparib develop AD As.

[0212] Inclusion criteria include:• Age > 18 years• Relapsed / metastatic solid tumors treated with prior adequate standard of care therapy for tumor type and stage of disease or where in the opinion of the Investigator, a clinical trial is the best option for the next treatment based on response and / or tolerability to prior therapy.• Measurable disease per RECIST vl.1• Eastern Cooperative Oncology Group (ECOG) Performance Status: 0-1• Life expectancy > 12 weeks• Adequate bone marrow, hepatic, and renal function• Minimum body weight > 30 kg (for participants enrolled in cohorts including rilvegostomig only).• Histologically or cytologically confirmed metastatic or locally advanced / recurrent breast cancer, ovarian cancer, biliary tract cancer, endometrial cancer or squamous non-small cell lung cancer.

[0213] Exclusion criteria include:Treatment with any of the following:AZ Ref: B7H4-102-PCT01-NPo Nitrosourea or mitomycin C within 6 weeks prior to the first dose of study treatmento Any investigational agents or study drugs from a previous clinical study within 5 half-lives or 28 days (whichever is shorter) prior to the first dose of study treatmento Any other anticancer treatment within the following time periods prior to the first dose of study intervention:■ Cytotoxic treatment: 21 days■ Non-cytotoxic drugs: 21 days or 5 half-lives (whichever is shorter)■ Biological products including immuno-oncology agents: 28 days• Spinal cord compression or a history of leptomeningeal carcinomatosis.• Brain metastases unless treated, asymptomatic, stable, and not requiring continuous corticosteroids at a dose of > 10 mg prednisone / day or equivalent for at least 4 weeks prior to start of study.• Active infection including tuberculosis and HBV, HCV or HIV• History of (non-infectious) ILD / pneumonitis that required steroids, has current ILD / pneumonitis, or where suspected ILD / pneumonitis cannot be ruled out by imaging at screening.• Clinically severe pulmonary compromise resulting from intercurrent pulmonary illnesses• Participants with any of the following cardiac criteria:o History of arrhythmia which is symptomatic or requires treatment (NCI CTCAE v5.0 Grade 3); symptomatic or uncontrolled atrial fibrillation, or asymptomatic sustained ventricular tachycardia.AZ Ref: B7H4-102-PCT01-NPo Uncontrolled hypertension.o Acute coronary syndrome / acute myocardial infarction, unstable angina pectoris, coronary intervention procedure with percutaneous coronary intervention, or coronary artery bypass grafting within 6 months.o History of brain perfusion problems (eg, carotid stenosis) or stroke, or transient ischemic attack in the last 6 months prior to screening.o Symptomatic heart failure (NYHA class > 2).o Prior or current cardiomyopathy.o Severe valvular heart disease.o Mean resting QTcF > 470 msec.o Risk factors for QTc prolongation or risk of arrhythmic events such as heart failure, congenital long QT syndrome, family history of long QT syndrome or unexplained sudden death under 40 years of age.• Concomitant use of medications or herbal supplements known to be strong cytochrome P (CYP) 3A4 inducers / inhibitors.• Any history of persisting (> 2 weeks) severe cytopenia due to any cause• Patients with any known predisposition to bleeding• Patients with history of myelodysplastic syndrome (MDS) / acute myeloid leukemia(AML) or with features suggestive of MDS / AML (as determined by prior diagnosticinvestigation).AZ Ref: B7H4-102-PCT01-NP• Refractory nausea and vomiting, chronic gastrointestinal diseases, inability to swallow the formulated product or previous significant bowel resection that would preclude adequate absorption of saruparib.

Claims

AZ Ref: B7H4-102-PCT01-NPCLAIMS1. A method of treating cancer in a human subject in need thereof, comprising administering to the human subject:A) an antibody-drug conjugate (ADC) comprising:i) an antibody or antigen binding fragment thereof which binds to a B7-H4 polypeptide, comprising:f) a heavy chain CDR1 (HCDR1), a heavy chain CDR2 (HCDR2), a heavy chain CDR3 (HCDR3), a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3) comprising the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively, or a functional variant thereof;g) a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12, respectively, or a functional variant thereof;h) a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, and SEQ ID NO: 18, respectively, or a functional variant thereof;i) a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 24, respectively, or a functional variant thereof; orj) a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, and SEQ ID NO: 30, respectively, or a functional variant thereof;ii) a cleavable linker, andiii) a cytotoxic agent;AZ Ref: B7H4-102-PCT01-NPB) a bispecific checkpoint inhibitor; andC) a PARP1 inhibitor.

2. The method of claim 1, wherein the bispecific checkpoint inhibitor is a binding protein having a first binding domain that specifically binds to Programmed Death-1 (PD-1).

3. The method of claim 1 or 2, wherein the antibody or antigen binding fragment thereof of the ADC comprises:a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively, or a functional variant thereof.

4. The method of any one of claims 1 to 3, wherein the antibody or antigen binding fragment thereof of the ADC comprises:a variable heavy (VH) chain and a variable light (VL) chain comprising the amino acid sequence of SEQ ID NO: 45 and SEQ ID NO: 34, respectively, or a functional variant thereof;a variable heavy (VH) chain and a variable light (VL) chain comprising the amino acid sequence of SEQ ID NO: 33 and SEQ ID NO: 34, respectively, or a functional variant thereof;a variable heavy (VH) chain and a variable light (VL) chain comprising the amino acid sequence of SEQ ID NO: 43 and SEQ ID NO: 34, respectively, or a functional variant thereof;a variable heavy (VH) chain and a variable light (VL) chain comprising the amino acid sequence of SEQ ID NO: 46 and SEQ ID NO: 34, respectively, or a functional variant thereof;a variable heavy (VH) chain and a variable light (VL) chain comprising the amino acid sequence of SEQ ID NO: 47 and SEQ ID NO: 34, respectively, or a functional variant thereof;a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO:AZ Ref: B7H4-102-PCT01-NP31, and SEQ ID NO: 32, respectively, or a functional variant thereof;a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 35 and SEQ ID NO: 36, respectively, or a functional variant thereof;a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 37 and SEQ ID NO: 38, respectively, or a functional variant thereof; ora VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 39 and SEQ ID NO: 40, respectively, or a functional variant thereof.

5. The method of any one of claims 1 to 4, wherein the antibody or antigen binding fragment thereof of the ADC comprises:a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 45 and SEQ ID NO: 34, respectively, or a functional variant thereof.

6. The method of any one of claims 1 to 5, wherein the antibody or antigen binding fragment thereof of the ADC comprises a heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 41 or 52, or wherein the antibody or antigen binding fragment thereof of the ADC comprises a light chain constant region comprising the amino acid sequence of SEQ ID NO: 42.

7. The method of any one of claims 1 to 5, wherein the antibody or antigen binding fragment thereof of the ADC comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 48 or 51; and a light chain comprising the amino acid sequence of SEQ ID NO: 44.

8. The method of any one of claims 1 to 7, wherein the antibody or antigen binding fragment thereof of the ADC is a monoclonal antibody.

9. The method of any one of claims 1 to 8, wherein the cleavable linker of the ADC is an mp-PEG8-val-ala linker.AZ Ref: B7H4-102-PCT01-NP10. The method of any one of claims 1 to 9, wherein the cytotoxic agent of the ADC is a topoisomerase inhibitor, a tubulysin derivative, a pyrrolobenzodiazepine, or a combination thereof.

11. The method of any one of claims 1 to 10, wherein the cytotoxic agent of the ADC is a topoisomerase inhibitor.

12. The method of any one of claims 1 to 11, wherein the ii) cleavable linker and iii) cytotoxic agent together comprise the following compound:

13. The method of any one of claims 1 to 12, wherein the bispecific checkpoint inhibitor comprises:a) a first binding domain that specifically binds to PD-1; and b) a second binding domain that specifically binds to T cell immunoreceptor with Ig and ITIM domains (TIGIT).

14. The method of claim 13, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain comprising a HCDR1 having the amino acid sequence of SEQ ID NO: 56, a HCDR2 having the amino acid sequence of SEQ ID NO: 57, and a HCDR3 having the amino acid sequence of SEQ ID NO: 58, and a light chain variable domain comprising a LCDR1 having the amino acid sequence of SEQ ID NO: 59, a LCDR2 having the amino acid sequence of SEQ ID NO: 60 and a LCDR3 havingAZ Ref: B7H4-102-PCT01-NPthe amino acid sequence of SEQ ID NO: 61.

15. The method of claim 13 or 14, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 62 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 64.

16. The method of any one of claims 13 to 15, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain having the amino acid sequence of SEQ ID NO: 63 and a light chain having the amino acid sequence of SEQ ID NO: 65.

17. The method of any one of claims 13 to 16, wherein the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain comprising a HCDR1 having the amino acid sequence of SEQ ID NO: 66, a HCDR2 having the amino acid sequence of SEQ ID NO: 67, and a HCDR3 having the amino acid sequence of SEQ ID NO: 68, and a light chain variable domain comprising a LCDR1 having the amino acid sequence of SEQ ID NO: 69, a LCDR2 having the amino acid sequence of SEQ ID NO: 70, and a LCDR3 having the amino acid sequence of SEQ ID NO: 71.

18. The method of any one of claims 13 to 17, wherein the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 72 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 74.

19. The method of any one of claims 13 to 18, wherein the second binding domain that specifically binds to TIGIT comprises a heavy chain having the amino sequence of SEQ ID NO: 73 and a light chain having the amino acid sequence of SEQ ID NO: 75.

20. The method of any one of claims 1 to 19, wherein the bispecific checkpoint inhibitor is a human or humanized bispecific antibody or antigen-binding fragment thereof.AZ Ref: B7H4-102-PCT01-NP21. The method of any one of claims 1 to 20, wherein the PARP1 inhibitor is AZD5305 or a pharmaceutically acceptable salt thereof.

22. The method of any one of claims 1 to 21, wherein the cancer comprises a cancer cell which expresses B7-H4.

23. The method of any one of claims 1 to 22, wherein the cancer is selected from endometrial cancer, ovarian cancer, breast cancer, biliary tract cancer, NSCLC (squamous and / or adenocarcinoma), uterine cancer, testicular cancer, bladder cancer, head and neck cancer, melanoma, renal cell carcinoma, pancreatic cancer, prostate cancer, cervical cancer, hematological cancer, cholangiocarcinoma, gastrointestinal cancer such as gastric cancer and colorectal cancer, and lung cancer.

24. The method of any one of claims 1 to 23, wherein the cancer is a breast cancer selected from hormone receptor-positive (HR+) breast cancer, human epidermal growth factor receptor 2 positive (HER2+) breast cancer, and triple negative breast cancer (TNBC).

25. The method of any one of claims 1 to 24, wherein the cancer is homologous recombination deficient (HRD) cancer.

26. The method of claim 25, wherein the cancer comprises one or more cells having a mutation in an HRD gene selected from BRCA1, BRCA2, ATM, BRIP1, BARD1, CDK12, CHEK1, CHEK2, FANCL, PALB2, PPP2R2A, RAD51B, RAD51C, RAD51D, and RAD54L.

27. The method of claim 26, wherein the mutated HRD gene is selected from BRCA1, BRCA2, an ATM.AZ Ref: B7H4-102-PCT01-NP28. A kit comprising: 1) an antibody-drug conjugate comprising an antibody or antigen binding fragment thereof which binds to a B7-H4 polypeptide, 2) a bispecific checkpoint inhibitor, and 3) a PARP1 inhibitor, wherein the antibody-drug conjugate, bispecific checkpoint inhibitor and PARP1 inhibitor are each as defined in any one of claims 1 to 54.

29. The kit of claim 28, wherein the antibody-drug conjugate comprises a drug-linker represented by the following formula:(SG3932),and wherein the drug-linker is conjugated to the antibody or antigen binding fragment thereof.

30. The kit of claim 28 or 29, wherein the antibody or antigen binding fragment thereof that binds to B7-H4 comprises: a HCDR1, a HCDR2, a HCDR3, a LCDR1, a LCDR2, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6.

31. The kit of claim 30, wherein the antibody or antigen binding fragment thereof comprises a VH chain and a VL chain comprising the amino acid sequence of SEQ ID NO: 45 and SEQ ID NO: 34, respectively, or a functional variant thereof.

32. The kit of claim 31, wherein the antibody or antigen binding fragment thereof comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 48 or 51; and a lightAZ Ref: B7H4-102-PCT01-NPchain comprising the amino acid sequence of SEQ ID NO: 44.

33. The kit of any one of claims 28 to 32, wherein the bispecific checkpoint inhibitor comprises:a) a first binding domain that specifically binds to PD-1; and b) a second binding domain that specifically binds to T cell immunoreceptor with Ig and ITIM domains (TIGIT).

34. The kit of claim 33, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain comprising a HCDR1 having the amino acid sequence of SEQ ID NO: 56, a HCDR2 having the amino acid sequence of SEQ ID NO: 57, and a HCDR3 having the amino acid sequence of SEQ ID NO: 58, and a light chain variable domain comprising a LCDR1 having the amino acid sequence of SEQ ID NO: 59, a LCDR2 having the amino acid sequence of SEQ ID NO: 60 and a LCDR3 having the amino acid sequence of SEQ ID NO: 61.

35. The kit of claim 34, wherein the first binding domain that specifically binds to PD-1 comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 62 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 64.

36. The kit of claim 34 or 35, wherein the first binding domain that specifically binds to PD- 1 comprises a heavy chain having the amino acid sequence of SEQ ID NO: 63 and a light chain having the amino acid sequence of SEQ ID NO: 65.

37. The kit of any one of claims 33 to 36, wherein the second binding domain that specifically binds to TIGIT comprises a heavy chain variable domain comprising a HCDR1 having the amino acid sequence of SEQ ID NO: 66, a HCDR2 having the amino acid sequence of SEQ ID NO: 67, and a HCDR3 having the amino acid sequence of SEQ ID NO: 68, and a light chain variable domain comprising a LCDR1 having the aminoAZ Ref: B7H4-102-PCT01-NPacid sequence of SEQ ID NO: 69, a LCDR2 having the amino acid sequence of SEQ ID NO: 70, and a LCDR3 having the amino acid sequence of SEQ ID NO: 71.

38. The kit of any one of claims 33 to 37, wherein the second binding domain that specifically binds TIGIT comprises a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 72 and a light chain variable domain having the amino acid sequence of SEQ ID NO: 74.

39. The kit of any one of claims 33 to 38, wherein the second binding domain that specifically binds to TIGIT comprises a heavy chain having the amino sequence of SEQ ID NO: 73 and a light chain having the amino acid sequence of SEQ ID NO: 75.

40. The method or kit of any one of claims 1-39, wherein:a) the ADC is AZD8205;b) the bispecific checkpoint inhibitor is rilvegostomig; andc) the PARP1 inhibitor is AZD5305.

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