ENTPD2 antibodies, combination therapies, and methods of using the antibodies and combination therapies

1. An antibody or antigen binding fragment thereof that specifically binds to human ENTPD2 protein, wherein the antibody or antigen binding fragment thereof comprises a heavy chain complementary determining region 1 (HCDR1), a heavy chain complementary determining region 2 (HCDR2), a heavy chain complementary determining region 3 (HCDR3), a light chain complementary determining region 1 (LCDR1), a light chain complementary determining region 2 (LCDR2), and a light chain complementary determining region 3 (LCDR3), wherein the antibody or antigen binding fragment thereof is selected from any one of the following:

1) an antibody or antigen binding fragment thereof comprising:

an HCDR1 sequence comprising SEQ ID NO: 1,

an HCDR2 sequence comprising SEQ ID NO: 2,

an HCDR3 sequence comprising SEQ ID NO: 3,

an LCDR1 sequence comprising SEQ ID NO: 14,

an LCDR2 sequence comprising SEQ ID NO: 15, and

an LCDR3 sequence comprising SEQ ID NO: 16;

2) an antibody or antigen binding fragment thereof comprising:

an HCDR1 sequence comprising SEQ ID NO: 4,

an HCDR2 sequence comprising SEQ ID NO: 5,

an HCDR3 sequence comprising SEQ ID NO: 3,

an LCDR1 sequence comprising SEQ ID NO: 17,

an LCDR2 sequence comprising SEQ ID NO: 18, and

an LCDR3 sequence comprising SEQ ID NO: 19; or,

3) an antibody or antigen binding fragment thereof comprising:

an HCDR1 sequence comprising SEQ ID NO: 7,

an HCDR2 sequence comprising SEQ ID NO: 8,

an HCDR3 sequence comprising SEQ ID NO: 9,

an LCDR1 sequence comprising SEQ ID NO: 20,

an LCDR2 sequence comprising SEQ ID NO: 18, and

an LCDR3 sequence comprising SEQ ID NO: 16.

2. The antibody or antigen binding fragment thereof of claim 1 , wherein the antibody or antigen binding fragment thereof is selected from any one of the following:

1) an antibody or antigen binding fragment thereof comprising a heavy chain variable region (VH) comprising SEQ ID NO: 10 or a sequence at least about 95% or more identical thereto, and a light chain variable region (VL) comprising SEQ ID NO: 21 or a sequence at least about 95% or more identical thereto;

2) an antibody or antigen binding fragment thereof comprising a VH comprising SEQ ID NO: 25 or a sequence at least about 95% or more identical thereto, and a VL comprising SEQ ID NO: 29 or a sequence at least about 95% or more identical thereto;

3) an antibody or antigen binding fragment thereof comprising a VH comprising SEQ ID NO: 33 or a sequence at least about 95% or more identical thereto, and a VL comprising SEQ ID NO: 29 or a sequence at least about 95% or more identical thereto;

4) an antibody or antigen binding fragment thereof comprising a VH comprising SEQ ID NO: 25 or a sequence at least about 95% or more identical thereto, and a VL comprising SEQ ID NO: 78 or a sequence at least about 95% or more identical thereto; or,

5) an antibody or antigen binding fragment thereof comprising a VH comprising SEQ ID NO: 233 or a sequence at least about 95% or more identical thereto, and a VL comprising SEQ ID NO: 237 or a sequence at least about 95% or more identical thereto.

3. The antibody or antigen binding fragment thereof of claim 1 , wherein the antibody or antigen binding fragment thereof is selected from any one of the following:

1) an antibody comprising a heavy chain comprising SEQ ID NO: 12 or a sequence at least about 95% or more identical thereto, and a light chain comprising SEQ ID NO: 23 or a sequence at least about 95% or more identical thereto;

2) an antibody comprising a heavy chain comprising SEQ ID NO: 27 or a sequence at least about 95% or more identical thereto, and a light chain comprising SEQ ID NO: 31 or a sequence at least about 95% or more identical thereto;

3) an antibody comprising a heavy chain comprising SEQ ID NO: 35 or a sequence at least about 95% or more identical thereto, and a light chain comprising SEQ ID NO: 31 or a sequence at least about 95% or more identical thereto;

4) an antibody comprising a heavy chain comprising SEQ ID NO: 27 or a sequence at least about 95% or more identical thereto, and a light chain comprising SEQ ID NO: 80 or a sequence at least about 95% or more identical thereto; Q.L,

5) an antibody comprising a heavy chain comprising SEQ ID NO: 235 or a sequence at least about 95% or more identical thereto, and a light chain comprising SEQ ID NO: 239 or a sequence at least about 95% or more identical thereto.

4. The antibody or antigen binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment thereof binds to human ENTPD2 protein with a dissociation constant (K D ) of less than 10 nM as measured by surface plasmon resonance (SPR).

5. The antibody or antigen binding fragment thereof of claim 1 , wherein the antibody or antigen binding fragment thereof inhibits human ENTPD2 enzymatic activity by at least 40%.

6. The antibody or antigen binding fragment thereof of claim 1 , wherein the antibody or antigen binding fragment thereof comprises a modified Fc region that has reduced antibody-dependent cellular cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC) activity compared to the parent antibody.

7. A pharmaceutical composition comprising the antibody or antigen binding fragment thereof of claim 1 and a pharmaceutically acceptable carrier.

8. A method of treating an ENTPD2+ cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the pharmaceutical composition of claim 7 .

9. The method of claim 8 , wherein the ENTPD2+ cancer is colorectal cancer (CRC), gastric cancer, esophageal cancer, lung cancer, breast cancer, or ovarian cancer.

10. The method of claim 8 , wherein the pharmaceutical composition, is administered to the subject through an intravenous, intratumoral or subcutaneous route.

11. A method of stimulating an immune response in a subject, the method comprising administering to the subject the pharmaceutical composition of claim 7 , in an amount effective to stimulate the immune response.

12. The method of claim 8 further comprising administering to the subject at least one additional therapeutic agent or procedure.

13. The method of claim 12 , wherein the at least one additional therapeutic agent is a PD-1 inhibitor, wherein the PD-1 inhibitor is selected from PDR001, Nivolumab, Pembrolizumab, Pidilizumab, MEDI0680, REGN2810, TSR-042, PF-06801591, BGB-A317, or AMP-224.

14. The method of claim 12 , wherein the at least one additional therapeutic agent is a PD-L1 inhibitor wherein the PD-L1 inhibitor is selected from FAZ053, Atezolizumab, Avelumab, Durvalumab, or BMS-936559.

15. The method of claim 12 , wherein the at least one additional therapeutic agent is an A2AR antagonist, wherein the A2AR antagonist is selected from:

i. an anti-CD73 antibody molecule, or antigen-binding fragment thereof, optionally wherein the anti-CD73 antibody is selected from:

a. an anti-CD73 antibody molecule comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 295 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 296, or an amino acid sequence at least 85%, 90%, 95% identical or higher to SEQ ID NO: 295 or 296;

b. an anti-CD73 antibody molecule comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 299 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 300, or an amino acid sequence at least 85%, 90%, 95% identical or higher to SEQ ID NO: 299 or 300;

c. an anti-CD73 antibody molecule comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 302 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 303, or an amino acid sequence at least 85%, 90%, 95% identical or higher to SEQ ID NO: 302 or 303;

d. an anti-CD73 antibody molecule comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 304 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 305, or an amino acid sequence at least 85%, 90%, 95% identical or higher to SEQ ID NO: 304 or 305;

e. an anti-CD73 antibody molecule comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 306 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 307, or an amino acid sequence at least 85%, 90%, 95% identical or higher to SEQ ID NO: 306 or 307; or

f. an anti-CD73 antibody molecule comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 308 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 309, or an amino acid sequence at least 85%, 90%, 95% identical or higher to SEQ ID NO: 308 or 309; or

ii. PBF509/NIR178, CPI444/V81444, AZD4635/HTL-1071, Vipadenant, GBV-2034, AB928, Theophylline, Istradefylline, Tozadenant/SYN-115, KW-6356, ST-4206, and Preladenant/SCH 420814; or

iii. 5-bromo-2,6-di-(1H-pyrazol-1-yl)pyrimidine-4-amine, or a pharmaceutically acceptable salt thereof; (S)-7-(5-methylfuran-2-yl)-34(6-(((tetrahydrofuran-3-yl)oxy)methyl)pyridin-2-yl)methyl)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-5-amine, or a pharmaceutically acceptable salt thereof; (R)-7-(5-methylfuran-2-yl)-3-((6-(((tetrahydrofuran-3-yl)oxy)methyl)pyridin-2-yl)methyl)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-5-amine, or racemate thereof, or a pharmaceutically acceptable salt thereof; 7-(5-methylfuran-2-yl)-3-((6-(((tetrahydrofuran-3-yl)oxy)methyl)pyridin-2-yl)methyl)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-5-amine, or a pharmaceutically acceptable salt thereof; and 6-(2-chloro-6-methylpyridin-4-yl)-5-(4-fluorophenyl)-1,2,4-triazin-3-amine, or a pharmaceutically acceptable salt thereof.

16. The method of claim 12 , wherein the at least one additional therapeutic agent is selected from:

i. a CTLA-4 inhibitor, optionally wherein the CTLA-4 inhibitor is selected from Ipilimumab or Tremelimumab;

ii. a TIM-3 inhibitor, optionally wherein the TIM-3 inhibitor is selected from MBG453, TSR-022, or LY3321367;

iii. a LAG-3 inhibitor, optionally wherein the LAG-3 inhibitor is selected from LAG525, BMS-986016, TSR-033, MK-4280 or REGN3767;

iv. a GITR agonist, optionally wherein the GITR agonist is selected from GWN323, BMS-986156, MK-4166, MK-1248, TRX518, INCAGN1876, AMG 228, or INBRX-110;

v. an anti-CD3 multispecific antibody molecule, optionally wherein the anti-CD3 multispecific antibody molecule is an anti-CD3×anti-CD123 bispecific antibody molecule (e.g., XENP14045), or an anti-CD3×anti-CD20 bispecific antibody molecule (e.g., XENP13676);

vi. a cytokine molecule, optionally wherein the cytokine molecule is IL-15 complexed with a soluble form of IL-15 receptor alpha (IL-15Ra);

vii. a macrophage colony-stimulating factor (M-CSF) inhibitor, optionally wherein the M-CSF inhibitor is MCS110;

viii. a CSF-1R inhibitor, optionally wherein the CSF-1R inhibitor is BLZ945;

ix. an inhibitor of indoleamine 2,3-dioxygenase (IDO) and/or tryptophan 2,3-dioxygenase (TDO);

x. a TGF-β inhibitor;

xi. an oncolytic virus; or,

xii. a chimeric antigen receptor (CAR) T-cell therapy.

17. The method of claim 12 , wherein the at least one additional therapeutic agent is selected from: 1) a protein kinase C (PKC) inhibitor; 2) a heat shock protein 90 (HSP90) inhibitor; 3) an inhibitor of a phosphoinositide 3-kinase (PI3K) and/or target of rapamycin (mTOR); 4) an inhibitor of cytochrome P450; 5) an iron chelating agent; 6) an aromatase inhibitor; 7) an inhibitor of p53 8) an apoptosis inducer; 9) an angiogenesis inhibitor; 10) an aldosterone synthase inhibitor; 11) a smoothened (SMO) receptor inhibitor; 12) a prolactin receptor (PRLR) inhibitor; 13) a Wnt signaling inhibitor; 14) a CDK4/6 inhibitor; 15) a fibroblast growth factor receptor 2 (FGFR2)/fibroblast growth factor receptor 4 (FGFR4) inhibitor; 16) an inhibitor of macrophage colony-stimulating factor (M-CSF); 17) an inhibitor of one or more of c-KIT, histamine release, Flt3 or PKC; 18) an inhibitor of one or more of VEGFR-2, PDGFRbeta, c-KIT or Raf kinase C; 19) a somatostatin agonist and/or a growth hormone release inhibitor; 20) an anaplastic lymphoma kinase (ALK) inhibitor; 21) an insulin-like growth factor 1 receptor (IGF-1R) inhibitor; 22) a P-Glycoprotein 1 inhibitor; 23) a vascular endothelial growth factor receptor (VEGFR) inhibitor; 24) a BCR-ABL kinase inhibitor; 25) an FGFR inhibitor; 26) an inhibitor of CYP11B2; 27) a HDM2 inhibitor; 28) an inhibitor of a tyrosine kinase; 29) an inhibitor of c-MET; 30) an inhibitor of JAK; 31) an inhibitor of DAC; 32) an inhibitor of 11β-hydroxylase; 33) an inhibitor of IAP; 34) an inhibitor of PIM kinase; 35) an inhibitor of Porcupine; 36) an inhibitor of BRAF; 37) an inhibitor of HER3; 38) an inhibitor of MEK; or 39) an inhibitor of a lipid kinase.

18. The method of claim 8 , wherein administering the pharmaceutical composition has one or more of the following effects:

(a) increased number of CD45+ CD4− CD8+ CD69+ CD25+ cells in a tumor or lesion site in the subject;

(b) increased number of CD45+ CD8− CD4+ FOXP3− CD69+ CD25+ cells in a tumor or lesion site in the subject;

(c) decreased plasma MCP1 or IL-1β level in the subject; or

(d) increased MCP1 level in a tumor or lesion site in the subject.