Compositions and methods for neoadjuvant treatment in cancer

Abstract

The present invention relates to methods of using these compounds for the treatment of cancer, including in combination with antibodies that inhibit the enzymatic activity of human CD39 and anti-PD(L)1 antibodies. The anti-CD39 antibody and anti-PD(L)1 antibody can be administered as neoadjuvant therapy for the treatment of lung cancer. The present invention also relates to dosing regimens for the combined treatment of anti-CD39 antibody and anti-PD(L)1 antibody.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 63 / 385,628, filed December 1, 2022, any drawings of which are incorporated herein by reference in their entirety.

[0002] Sequence Listing Reference This application has been submitted with an electronic Sequence Listing. The Sequence Listing is provided under the name "INN.276XPCT.xml," created on November 13, 2023, and is 28,938 bytes in size. The information in the electronic version of this Sequence Listing is incorporated herein by reference in its entirety.

[0003] The present invention relates to antibodies that inhibit the enzymatic activity of human CD39 and methods of using these compounds to treat cancer. [Background technology]

[0004] Eight distinct ENTPD genes encode members of the NTPDase protein family. Individual NTPDase subtypes differ in their subcellular location and functional properties. Cell membrane-bound nucleoside triphosphate diphosphohydrolases regulate nucleotide levels on the cell surface by hydrolyzing the c- and b-phosphates of nucleotides. NTPDase 1 (ectonucleoside triphosphate diphosphohydrolase 1) (also known as CD39 / ENTPD1 or vascular CD39) works with another enzyme, CD73 (ecto-5'-nucleotidase), to hydrolyze extracellular adenosine triphosphate (ATP) and extracellular adenosine diphosphate (ADP) to generate adenosine, which binds to adenosine receptors and inhibits T cell and natural killer (NK) cell responses, thereby suppressing the immune system. Adenosine generation via the CD73 / CD39 pathway is recognized as a key mechanism for the immunosuppressive function of regulatory T cells (Tregs). In some human cancers, CD39 +Treg numbers are increasing and several in vivo models have been used to demonstrate the role of CD39 in promoting tumor growth and metastasis. + The importance of Tregs has been demonstrated. However, CD39 is also expressed by tumor cells, and CD39+ tumor cells may mediate immunosuppression via the adenosine pathway. CD39 in cancer cells exhibits ATPase activity and generates adenosine together with CD73. CD73 + CD39 + Cancer cells inhibited the proliferation of CD4 and CD8 T cells and the generation of cytotoxic effector CD8 T cells (CTLs) in a CD39- and adenosine-dependent manner.

[0005] It has been reported that CD39 expression is increased in some solid cancers (colorectal cancer, head and neck cancer, pancreatic cancer) and chronic lymphocytic leukemia. Antibodies that bind to and inhibit the enzymatic activity of CD39 are disclosed in, for example, WO2018 / 167267, WO2019 / 243252, WO2019 / 178269, WO2019 / 127935, and WO2021 / 037037.

[0006] In many cases of cancer, the disease may be deemed surgically resectable at the time of diagnosis or may become surgically resectable after initial radiation or chemotherapy. For many tumors, surgery often offers the best chance of cure.

[0007] Globally, approximately 1.5 million new cases of lung cancer are diagnosed annually, with approximately 85% of these cases being non-small cell lung cancer (NSCLC). Although surgery is considered the best treatment option, approximately 20–30% of NSCLC patients have surgically resectable disease (Molina et al., Mayo Clin. Proc. 83(5): 584–94 (2008); Burdett S. Lancet. 2014;383:1561–1571). Adjuvant chemotherapy after resection of NSCLC is standard practice to reduce the risk of disease recurrence. For patients with limited-stage disease (stages I, II, and IIIA), the 5-year survival rate for patients who undergo surgical resection and chemotherapy is 51%, with an absolute benefit of 5.4% in 5-year survival, especially for patients with good performance status (PS) (Provencio et al. 2011 J Thorac Dis. 3(3): 197–204). There is evidence that identifying a patient's minimal residual disease (MRD) status through detection of postoperative circulating tumor DNA (ctDNA) can accurately predict disease recurrence. It has been proposed that durvalumab may be particularly beneficial in high-risk NSCLC patients in whom MRD is detected via ctDNA isolation after complete resection. Furthermore, neoadjuvant chemotherapy is used in some cases. Burdett S. Lancet. 2014;383:1561-1571 concluded that preoperative chemotherapy significantly improves overall survival, time to distant recurrence, and recurrence-free survival in patients with stage IB-IIIA resectable NSCLC. Such neoadjuvant chemotherapy has been proposed as having the potential to reduce tumor size, increase operability, and eradicate micrometastases. Furthermore, neoadjuvant chemotherapy may be more effective if the blood supply to the tumor is still intact before surgical resection, and chemotherapy may be better tolerated if patients have not yet recovered from major surgery.Although several clinical trials have evaluated neoadjuvant chemotherapy alone and five have evaluated both neoadjuvant and postoperative chemotherapy, usually in responders (see Burnett 2014 above), it has been concluded that the potential benefits of neoadjuvant chemotherapy must be balanced against possible toxic effects.

[0008] There remains a need in the art for improved cancer treatment. Many patients with locally advanced disease (stage III) have disease deemed unresectable at diagnosis, and the desire is to make the disease more operable or resectable. For patients with stage II-IIIA and select IIIB disease, despite having resectable disease, surgery and postoperative adjuvant chemotherapy (SoC) results in a 5-year disease-free survival (DFS) rate of only approximately 40% (Wakelee et al., Lancet. Oncol. 18(12): 1610-23 (2017)). Summary of the Invention

[0009] The present disclosure provides methods of treating cancer and / or preventing recurrence of cancer in a patient in need thereof, e.g., in stage I-III NSCLC, optionally in stage IB-IIIA NSCLC, optionally in stage II or IIIA NSCLC. The present disclosure also provides dosing regimens of anti-CD39 antibodies and anti-PD(L)1 antibodies indicated for such treatment, e.g., in resectable tumors and cancers.

[0010] The present disclosure provides a method for treating a tumor or cancer and / or preventing the recurrence of a tumor or cancer in a patient in need of such treatment or prevention, comprising administering an anti-CD39 antibody and an anti-PD(L)1 antibody, wherein the anti-CD39 antibody and anti-PD(L)1 antibody are administered as neoadjuvant therapy (preoperative therapy). In one embodiment, the method comprises administering an anti-CD39 antibody, an anti-PD(L)1 antibody, and a chemotherapeutic agent, wherein the anti-CD39 antibody, anti-PD(L)1 antibody, and the chemotherapeutic agent are administered as neoadjuvant therapy. The present disclosure further provides a method for treating a tumor or cancer and / or preventing the recurrence of a tumor or cancer in a patient in need of such treatment or prevention, comprising administering an anti-CD39 antibody and an anti-PD(L)1 antibody, wherein the anti-CD39 antibody and anti-PD(L)1 antibody are administered as neoadjuvant therapy and further as adjuvant therapy. The treatment regimens of the present disclosure may be characterized as comprising: (a) administering to the patient an anti-CD39 antibody and an anti-PD(L)1 antibody (and optionally, further an effective amount of chemotherapy) prior to surgical tumor resection, or if the patient has not undergone surgical tumor resection; and (b) administering to the patient an anti-CD39 antibody and an anti-PD(L)1 antibody after surgical tumor resection. In one embodiment, the tumor or cancer is characterized as one that is considered surgically resectable. In one embodiment, the tumor or cancer is characterized as having the potential to become surgically resectable (e.g., locally advanced and / or stage IIIB NSCLC).

[0011] In one embodiment, the anti-CD39 antibody is an antibody comprising the amino acid sequence of SEQ ID NOs: 2-7, optionally the antibody comprises the amino acid sequence of SEQ ID NOs: 8 and 9, and optionally the antibody comprises the amino acid sequence of SEQ ID NOs: 10 and 11. In one embodiment, the anti-PD(L)1 antibody is durvalumab.

[0012] This method is particularly advantageous for treating lung cancer, particularly non-small cell lung cancer (NSCLC). In either embodiment, the cancer can optionally be characterized as resectable NSCLC. In one embodiment, the cancer is stage II or IIIA NSCLC, for example, resectable stage II or IIIA NSCLC. In one embodiment, the cancer is stage I, II, or IIIA NSCLC. In another aspect, the cancer can optionally be characterized as unresectable and / or locally advanced NSCLC (e.g., stage III or IIIB NSCLC).

[0013] In one embodiment, the treatment regimen herein provides for administration of the anti-CD39 antibody at the same dose (e.g., 2250 mg or 3000 mg) in both the adjuvant and neoadjuvant settings. In one embodiment, the treatment regimen herein allows for the administration of the anti-CD39 antibody and the anti-PD(L)1 antibody at the same respective doses (e.g., a fixed dose of 2250 mg or 3000 mg for the anti-CD39 antibody and a fixed dose of 1500 mg for durvalumab) in both the adjuvant and neoadjuvant settings. Furthermore, the regimen allows for the administration of the anti-CD39 antibody (and the anti-PD(L)1 antibody) every three weeks as a neoadjuvant and every four weeks as an adjuvant (at the same doses).

[0014] In one embodiment, a method is provided for administering an anti-CD39 antibody comprising the amino acid sequence of SEQ ID NO:2-7, 8-9, or 10-11, wherein the antibody is administered Q3w or Q4w at a dose of 3000 mg. In one embodiment, a method is provided for administering an anti-CD39 antibody comprising the amino acid sequence of SEQ ID NO:2-7, 8-9, or 10-11, wherein the antibody is administered Q3w or Q4w at a dose of 2250 mg.

[0015] In one embodiment, a method is provided for administering an anti-CD39 antibody comprising the amino acid sequence of SEQ ID NO:2-7, 8-9, or 10-11, wherein the antibody is administered (a) on day 1 of one or more three-week cycles, and (b) on day 1 of one or more four-week cycles, in each case wherein the antibody is administered at a dose of 2250 mg or 3000 mg. Optionally, the method is a method for treating cancer or preventing cancer recurrence. Optionally, the method is a method of administering the anti-CD39 antibody in combination with an anti-PD(L)1 antibody.

[0016] In one embodiment, a method of administering an anti-CD39 antibody and an anti-PD(L)1 antibody is provided, wherein the anti-CD39 antibody and the anti-PD(L)1 antibody are administered (a) on day 1 of a 3-week cycle for one or more cycles, and (b) on day 1 of a 4-week cycle for one or more cycles, in each case, the anti-CD39 antibody is administered at a fixed dose of 3000 mg (or optionally, 2250 mg), and optionally further, the anti-PD(L)1 antibody is durvalumab and is administered at a fixed dose of 1500 mg.

[0017] In one embodiment, a method of treating cancer or preventing the recurrence of cancer is provided, comprising administering an anti-CD39 antibody and an anti-PD(L)1 antibody, wherein the anti-CD39 antibody and the anti-PD(L)1 antibody are administered as neoadjuvant therapy, and optionally further as adjuvant therapy, wherein the neoadjuvant therapy comprises one or more cycles of administering the anti-CD39 antibody and the anti-PD(L)1 antibody on day 1 of a three-week cycle, and the adjuvant therapy comprises one or more cycles of administering the anti-CD39 antibody and the anti-PD(L)1 antibody on day 1 of a four-week cycle, in each case, the anti-CD39 antibody is administered at a fixed dose of 3000 mg (or optionally 2250 mg), and optionally further wherein the anti-PD(L)1 antibody is durvalumab and is administered at a fixed dose of 1500 mg. In one embodiment, the neoadjuvant therapy comprises four cycles. In one embodiment, the neoadjuvant therapy comprises at least 2, 4, 8, 12, and / or up to 12 cycles.

[0018] These aspects are described in more detail in the description set forth herein, and additional aspects, features and advantages will become apparent from the description set forth herein. [Brief explanation of the drawings]

[0019] [Figure 1] Figure 1 shows CD39 immunohistochemical staining of 50 FFPE samples from squamous NSCLC (sqNSCLC) and 50 adenocarcinoma NSCLC (adNSCLC). CD39 expression was scored based on frequency and intensity. Staining is shown for stromal, immune cell, and total expression. Neither CD39 staining nor tumor cell staining was observed. Panel A shows the CD39 total score, which is the sum of stromal, immune, and tumor expression scores (0–60). Panel B shows the stromal score, which is the sum of vascular (0–12) and connective tissue (0–12) expression scores. Panel C shows the immune score as the sum of small immune cell (0–12) and large immune cell (0–12) scores. For each case, the expression score is indicated by the stage of cancer (stage I, II, or III). [Figure 2] FIG. 2 shows ATP release from squamous NSCLC tumor cells when the tumor cells were separately incubated with different chemotherapies. [Figure 3]Figure 3A shows the extracellular ATP (eATP) release from H1703 (CD39-) cells incubated with recombinant huCD39 to mimic soluble CD39 in the tumor microenvironment, in the presence or absence of anti-CD39 antibody (IPH5201), and then treated with docetaxel. Docetaxel induced a strong release of eATP, which was reduced in the presence of added recombinant human CD39 protein and restored in the presence of added IPH5201 antibody. Figure 3B shows eATP release from OAW42 (CD39+) cells incubated with anti-CD39 antibody (IPH5201) and treated with a range of docetaxel doses. Docetaxel induced only slight eATP release at the highest concentrations in these CD39-expressing cells, but significant eATP accumulation occurred in the presence of 10 μg / mL or 50 μg / mL IPH5201. [Figure 4] The left and right panels of Figure 4 show the percentage of CD39 expression and quantification of adenosine in MCA205 tumor cells harvested for CD39 expression analysis after transplantation of MCA205 tumors into human CD39 knock-in mice and treatment with anti-CD39 antibody (moIPH5201) or isotype control. A significant proportion of cells expressed CD39 (left panel), and animals treated with moIPH5201 had less intratumoral adenosine than controls (left panel). [Figure 5] FIG. 5 shows an experiment evaluating tumor growth of MC38 tumors in a cohort of huCD39 knock-in mice following gemcitabine + / - anti-CD39 antibody (moIPH5201) treatment. [Figure 6] Figures 6, 7, and 8, respectively, show experiments evaluating tumor growth of MC38 tumors in a cohort of huCD39 knock-in mice following treatment with gemcitabine + / - anti-PD-L1 + / - anti-CD39 antibody (moIPH5201). [Figure 7] Same as above [Figure 8] Same as above [Figure 9]Figure 9 shows a pooled analysis of three series of experiments assessing tumor growth of MC38 tumors in huCD39 knock-in mice following treatment with gemcitabine + / - anti-PD-L1 + / - anti-CD39 antibody (moIPH5201). [Figure 10] FIG. 10 shows the indirect response PD model (inhibition against Kin) constructed to describe the relationship between IPH5201 concentration and free mCD39 on monocytes. [Figure 11A] Figure 11A shows that an anti-CD39 antibody (IPH5201) saturated binding of over 300 mg of soluble CD39 in human patients. Each line represents data from one patient. The increase in total soluble CD39 (not shown) is consistent with increased stabilization of soluble CD39 by antibody binding. A similar trend was observed in combination treatment with durvalumab. [Figure 11B] FIG. 11B shows that anti-CD39 antibody (IPH5201) saturated the binding of membrane-bound CD39 on immune cells from samples of human patients treated with 3000 mg of IPH5201. [Figure 12] Figure 12 shows the pharmacokinetics (PK) of IPH5201 in human patients as monotherapy (left panel) or in combination with durvalumab (right panel). The X-axis represents days after first dose, and the Y-axis represents serum IPH5201 concentration. The curves from bottom to top represent fixed doses of IPH5201: 100 mg, 300 mg, 1000 mg, and 3000 mg. The PK of IPH5201 was nonlinear below 300 mg and linear above 1000 mg. [Figure 13] Figure 13 shows a schematic diagram of the treatment regimen in human clinical trials. Neoadjuvant therapy involves a fixed dose of 3000 mg of IPH5201 and a fixed dose of 1500 mg of durvalumab administered Q3W for four cycles in combination with chemotherapy (CT). Adjuvant therapy involves a fixed dose of 3000 mg of IPH5201 and a fixed dose of 1500 mg of durvalumab administered Q4W for up to 12 cycles. DETAILED DESCRIPTION OF THE INVENTION

[0020] definition When "comprising" is used, it may optionally be replaced with "consisting essentially of" or "consisting of."

[0021] Human CD39 (also known as "vascular" CD39, NTPdase1, ENTPD1, ​​ATPDase, and vascular ATP diphosphohydrolase) exhibits ATPase activity. CD39 hydrolyzes extracellular ATP and extracellular ADP to AMP, which is further converted to adenosine by another enzyme, 5-prime nucleotidase. The amino acid sequence of the "vascular" human CD39 mature polypeptide chain is shown in Genbank under accession number P49961 (the entire disclosure of which is incorporated herein by reference) and is as follows: MEDTKESNVK TFCSKNILAI LGFSSIIAVI ALLAVGLTQN KALPENVKYG IVLDAGSSHT SLYIYKWPAE KENDTGVVHQ VEECRVKGPG ISKFVQKVNE IGIYLTDCME RAREVIPRSQ HQETPVYLGA TAGMRLLRME SEELADRVLD VVERSLSNYP FDFQGARIIT GQEEGAYGWI TINYLLGKFS QKTRWFSIVP YETNNQETFG ALDLGGASTQ VTFVPQNQTI ESPDNALQFR LYGKDYNVYT HSFLCYGKDQ ALWQKLAKDI QVASNEILRD PCFHPGYKKV VNVSDLYKTP CTKRFEMTLP FQQFEIQGIG NYQQCHQSIL ELFNTSYCPY SQCAFNGIFL PPLQGDFGAF SAFYFVMKFL NLTSEKVSQE KVTEMMKKFC AQPWEEIKTS YAGVKEKYLS EYCFSGTYIL SLLLQGYHFT ADSWEHIHFI GKIQGSDAGW TLGYMLNLTN MIPAEQPLST PLSHSTYVFL MVLFSLVLFT VAIIGLLIFH KPSYFWKDMV (SEQ ID NO: 1).

[0022] In the context of this specification, "neutralize" or "neutralizing," when referring to a CD39 polypeptide (e.g., "neutralize CD39," "neutralize the activity of CD39," or "neutralize the enzymatic activity of CD39"), refers to a process by which the ATP hydrolysis (ATPase) activity of CD39 is inhibited. This particularly includes inhibition of CD39-mediated production of AMP and / or ADP, i.e., inhibition of CD39-mediated catabolism of ATP to AMP and / or ADP. In the case of membrane-bound CD39, this may be measured directly or indirectly, for example, in a cellular assay that measures the ability of a test compound to inhibit the conversion of ATP to AMP and / or ADP. In the case of soluble CD39, this may be measured directly or indirectly, for example, by incubating recombinant soluble CD39 described herein with a test compound and then measuring the conversion of ATP to AMP and / or ADP. For example, as described herein, the disappearance of ATP and / or the generation of AMP may be assessed, for example, by quantifying luminescence units, which are proportional to the amount of ATP present. In one embodiment, e.g., with reference to the assays described herein (e.g., ATP depletion and / or AMP production), the antibody preparation causes at least a 60% reduction in the conversion of ATP to AMP, or causes at least a 70% reduction in the conversion of ATP to AMP, or causes at least an 80% or 90% reduction in the conversion of ATP to AMP.

[0023] Whenever "cancer treatment" or the like is referred to in reference to an anti-CD39 binding agent (e.g., an antibody), this may include: (a) a method of treating cancer, comprising the step of administering (at least one treatment) to an individual, mammal, particularly a human, in need of such treatment an anti-CD39 binding agent (preferably in a pharmaceutically acceptable carrier substance) in a dose that allows for cancer treatment (a therapeutically effective amount), preferably in a dose (amount) as specified herein; (b) the use of an anti-CD39 binding agent for the treatment of cancer (particularly in a human) or an anti-CD39 binding agent for use in the treatment; (c) the use of an anti-CD39 binding agent for the manufacture of a medicament for the treatment of cancer, optionally comprising mixing the anti-CD39 binding agent with a pharmaceutically acceptable carrier, or a method of using an anti-CD39 binding agent for the manufacture of a medicament for the treatment of cancer or a medicament comprising an effective dose of an anti-CD39 binding agent suitable for the treatment of cancer; or (d) any combination of a), b), and c), according to subject matter that may be patentable in the country in which this application is filed.

[0024] As used herein, the term "antigen-binding domain" refers to a domain comprising a three-dimensional structure capable of immunospecifically binding to an epitope. Thus, in one embodiment, this domain may comprise a hypervariable region, optionally a VH and / or VL domain, of an antibody chain, optionally at least a VH domain. In another embodiment, the binding domain may comprise at least one complementarity-determining region (CDR) of an antibody chain. In another embodiment, the binding domain may comprise a polypeptide domain from a non-immunoglobulin scaffold.

[0025] The term "antibody," as used herein, can include polyclonal and monoclonal antibodies. Depending on the type of constant domain in the heavy chain, antibodies are assigned to one of five major classes: IgA, IgD, IgE, IgG, and IgM. Some of these are further divided into subclasses or isotypes (e.g., IgG1, IgG2, IgG3, IgG4, and the like). A typical immunoglobulin (antibody) structural unit comprises a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one "light" chain (approximately 25 kDa) and one "heavy" chain (approximately 50-70 kDa). The N-terminus of each chain defines a variable region of approximately 100-110 or more amino acids primarily responsible for antigen recognition. The terms variable light chain (VL) and variable heavy chain (VH) refer to these light and heavy chains, respectively. The heavy chain constant domains corresponding to the different classes of immunoglobulins are called "alpha," "delta," "epsilon," "gamma," and "mu," respectively. The subunit structures and three-dimensional configurations of the different classes of immunoglobulins are well known. IgG is the representative class of antibody used herein because it is the most common antibody in the physiological situation and is most easily produced in the laboratory. In some cases, the antibody is a monoclonal antibody. Particular examples of antibodies are humanized, chimeric, human, or otherwise human-suitable antibodies. "Antibody" also includes any fragment or derivative of any of the antibodies described herein.

[0026] The term "specifically binds" means that the antibody can bind to a binding partner, e.g., CD39, preferably in a competitive binding assay, when assessed using a recombinant form of either the protein, an epitope therein, or the native protein present on the surface of an isolated target cell. Competitive binding assays and other methods for determining specific binding are further described below and are well known in the art.

[0027] When an antibody is said to "compete" with a particular monoclonal antibody (e.g., an antibody having the amino acid sequence of SEQ ID NO: 2-7, 8-9, or 10-11), this means that the antibody competes with the monoclonal antibody in a binding assay using either a recombinant CD39 molecule or a surface-expressed CD39 molecule. For example, if a test antibody reduces the binding of a reference antibody to a CD39 polypeptide or a CD39-expressing cell, respectively, in a binding assay, the antibody is said to "compete" with the reference antibody.

[0028] The term "affinity," as used herein, refers to the strength of antibody binding to an epitope. The affinity of an antibody is given by the dissociation constant, Kd, ​​defined as [Ab] x [Ag] / [Ab-Ag], where [Ab-Ag] is the molar concentration of the antibody-antigen complex, [Ab] is the molar concentration of unbound antibody, and [Ag] is the molar concentration of unbound antigen. The affinity constant, Ka, is defined as 1 / Kd. Methods for determining the affinity of mAbs can be found in Harlow, et al., Antibody: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1988), Coligan et al., eds., Current Protocols in Immunology, Greene Publishing Assoc. and Wiley Interscience, NY, (1992, 1993), and Muller, Meth. Enzymol. 92:589-601 (1983), which are incorporated herein by reference in their entireties. One standard method known in the art for determining the affinity of mAbs is the use of surface plasmon resonance (SPR) screening (such as analysis with a BIAcore™ SPR analyzer).

[0029] In the context of this specification, a "determinant" refers to a site of interaction or binding on a polypeptide.

[0030] The term "epitope" refers to an antigenic determinant, an area or region on an antigen to which an antibody binds. A protein epitope can include amino acid residues involved in direct binding as well as those effectively blocked by a specific antigen-binding antibody or peptide, i.e., within the "footprint" of an antibody. This is the simplest form or smallest structural region on a complex antigen molecule that can combine with, for example, an antibody or receptor. Epitopes can be linear or conformational. The term "linear epitope" is defined as an epitope composed of amino acid residues that are adjacent in a linear sequence of amino acids (primary structure). The term "conformational or conformational epitope" is defined as an epitope composed of amino acid residues that are not all adjacent and therefore represent discrete portions of the linear sequence of amino acids that are brought into close proximity with each other by molecular folding (secondary, tertiary, and / or quaternary structure). Conformational epitopes depend on three-dimensional structure. Thus, the term "conformational" is often used interchangeably with "structure."

[0031] The term "drug" is used herein to refer to a chemical compound, a mixture of chemical compounds, a biological macromolecule, or an extract made from biological material. The term "therapeutic agent" refers to a drug that has biological activity.

[0032] For purposes herein, a "humanized" antibody refers to an antibody in which one or more human immunoglobulin constant and variable framework regions are fused with the binding regions, e.g., CDRs, of an animal immunoglobulin. Such antibodies are designed to retain the binding specificity of the non-human antibody from which the binding regions are derived, but to avoid an immune response against the non-human antibody.

[0033] The term "hypervariable region" when used herein refers to the amino acid residues of an antibody which are responsible for antigen-binding. The hypervariable regions generally comprise amino acid residues from the "complementarity determining regions" or "CDRs" (e.g., residues 24-34 (L1), 50-56 (L2), and 89-97 (L3) in the light chain variable domain and residues 31-35 (H1), 50-65 (H2), and 95-102 (H3) in the heavy chain variable domain; Kabat et al. 1991) and / or "hypervariable loops" (e.g., residues 26-32 (L1), 50-52 (L2), and 91-96 (L3) in the light chain variable domain and residues 26-32 (H1), 53-55 (H2), and 96-101 (H3) in the heavy chain variable domain; Chothia and Lesk, J. Mol. Biol. 1987;196:901-917) or similar systems for determining essential amino acids involved in antigen binding. Generally, the numbering of amino acid residues in this region is performed according to the method described by Kabat et al., supra. Phrases such as "Kabat position," "variable domain residue numbering as in Kabat," and "according to Kabat" refer herein to this numbering system for heavy chain variable domains or light chain variable domains. Using the Kabat numbering system, the actual linear amino acid sequence of a peptide may contain fewer or additional amino acids corresponding to shortening of, or insertion into, a FR or CDR of the variable domain. For example, a heavy chain variable domain may contain a single amino acid insertion after residue 52 of CDR H2 (residue 52a according to Kabat) and an insertion of residues after heavy chain FR residue 82 (e.g., residues 82a, 82b, and 82c according to Kabat). The Kabat numbering of residues can be determined for a particular antibody by alignment of the antibody's sequence with the "standard" Kabat numbering sequence in regions of affinity.

[0034] "Framework" or "FR" residues, as used herein, refer to the regions of an antibody variable domain excluding the regions defined as CDRs. Each antibody variable domain framework can be further divided into contiguous regions (FR1, FR2, FR3, and FR4) separated by the CDRs.

[0035] The terms "Fc domain," "Fc portion," and "Fc region" refer to the C-terminal fragment of an antibody heavy chain, e.g., from about amino acid (aa) 230 to about aa 450 of the human gamma (γ) heavy chain, or its corresponding sequences in other types of antibody heavy chains (e.g., α, δ, ε, and μ in the case of human antibodies), or naturally occurring allotypes thereof. Unless otherwise specified, the generally accepted Kabat amino acid numbering for immunoglobulins is used throughout this disclosure (see Kabat et al. (1991) Sequences of Protein of Immunological Interest, 5th ed., United States Public Health Service, National Institutes of Health, Bethesda, MD).

[0036] The terms "isolated," "purified," or "biologically pure" refer to material that is substantially or essentially free from components that normally accompany it as found in its native state. Purity and homogeneity are typically determined using analytical chemistry techniques such as polyacrylamide gel electrophoresis or high performance liquid chromatography. A protein that is the predominant species present in a preparation is substantially purified.

[0037] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. This term applies to amino acid polymers in which one or more amino acid residues are artificial chemical mimetics of the corresponding naturally occurring amino acids, as well as to naturally occurring and non-naturally occurring amino acid polymers.

[0038] The term "recombinant," when used with reference to, for example, a cell or a nucleic acid, protein (e.g., an antibody or antibody fragment), or vector, indicates that the cell, nucleic acid, protein, or vector has been modified by the introduction of a heterologous nucleic acid or protein or the alteration of a native nucleic acid or protein, or that the cell is derived from a cell so modified. Thus, for example, a recombinant cell expresses genes that are not found within the native (non-recombinant) form of the cell, or expresses native genes that are otherwise aberrantly expressed, poorly expressed, or not expressed at all.

[0039] In the context of this specification, the term antibody that "binds" to a polypeptide or epitope refers to an antibody that binds to said determinant with specificity and / or affinity.

[0040] The terms "identity" or "identical," when used in the context of the sequences of two or more polypeptides, refer to the degree of sequence relatedness between the polypeptides as determined by the number of matches between two or more strings of amino acid residues. "Identity" measures the percent of exact matches between the smaller of two or more sequences, using gap alignments (if any) handled by a particular mathematical model or computer program (i.e., "algorithm"). The identity of related polypeptides can be readily calculated by known methods. Such methods include, but are not limited to, those described in Computational Molecular Biology, Lesk, AM, ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, DW, ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part 1, Griffin, AM and Griffin, HG, eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; Sequence Analysis Primer, Gribskov, M. and DevereuX, J., eds., M. Stockton Press, New York, 1991; and Carillo et al., SIAM J. Applied Math. 48, 1073 (1988).

[0041] Methods for determining identity are designed to maximize the match between the sequences tested. Methods for determining identity are described in publicly available computer programs. Computer program methods for determining identity between two sequences include the GCG program package, including GAP (Devereux et al., Nucl. Acid. Res. 12, 387 (1984); Genetics Computer Group, University of Wisconsin, Madison, Wis.), BLASTP, BLASTN, and FASTA (Altschul et al., J. Mol. Biol. 215, 403-410 (1990)). The BLASTX program is available from the National Center for Biotechnology Information (NCBI) and other sources (BLAST Manual, Altschul et al. NCB / NLM / NIH Bethesda, Md. 20894; Altschul et al., supra). The well-known Smith-Waterman algorithm can also be used to determine identity.

[0042] Cancer Treatment The present disclosure generally relates to methods of treating cancer in patients using an anti-CD39 antibody in combination with an anti-PD(L)1 antibody as neoadjuvant therapy, and optionally further as adjuvant therapy. This treatment method is therefore useful as preoperative therapy, and optionally further as postoperative therapy, in patients with cancers or tumors that are resectable, potentially resectable, or potentially becoming resectable. Optionally, the preoperative use is further combined with postoperative chemotherapy. This method is particularly advantageous in the treatment of lung cancer, particularly non-small cell lung cancer (NSCLC). In one embodiment, the cancer is stage II or IIIA NSCLC, e.g., resectable stage II or IIIA NSCLC. This method is therefore particularly advantageous in the treatment of tumors or cancers for which surgery is the primary treatment.

[0043] In one embodiment, a method for reducing or inhibiting tumor growth in a subject in need thereof is disclosed, comprising administering to the subject therapeutically effective amounts of an anti-CD39 antibody, an anti-PD(L)1 antibody, and optionally a chemotherapeutic agent. In one embodiment, a method for treating cancer, particularly stage II or IIIA NSCLC, in a subject in need thereof is disclosed, comprising administering to the subject therapeutically effective amounts of an anti-CD39 antibody, an anti-PD(L)1 antibody, and optionally a chemotherapeutic agent. In one embodiment, the anti-CD39 antibody, the anti-PD(L)1 antibody, and optionally the chemotherapeutic agent are administered as preoperative therapy. In one embodiment, the anti-CD39 antibody and the anti-PD(L)1 antibody are administered as preoperative therapy and as postoperative therapy.

[0044] The methods and regimens herein can be useful, for example, for treating and / or preventing tumor or cancer recurrence in patients, preventing metastasis or metastatic recurrence, preventing tumor or cancer progression or growth, and / or improving or increasing survival, time to recurrence (e.g., time to distant recurrence), and / or recurrence-free survival. Patients can optionally be identified as having surgically resectable cancer or tumors. The methods and regimens herein can also be useful for making tumors more accessible to surgery and / or increasing the likelihood of complete resection. In some cases, the methods herein are characterized as methods that increase the likelihood of no residual disease (e.g., minimal residual disease MRD) and / or ctDNA negativity (no detectable ctDNA), for example, after surgical resection. The methods and regimens herein may also be useful, for example, to increase or enhance anti-tumor immune responses, inhibit the enzymatic activity of CD39 (e.g., in tumors), increase intratumoral concentrations of ATP, decrease intratumoral concentrations of adenosine, increase the activity and / or activity of T cells, NK cells, tumor-infiltrating NK cells, tumor-infiltrating T cells, and / or dendritic cells.

[0045] As used herein, an anti-CD39 antibody is an antibody that neutralizes CD39. "Neutralizing" or "neutralization," when referring to a CD39 polypeptide (e.g., "neutralizing CD39," "neutralizing the activity of CD39," or "neutralizing the enzymatic activity of CD39"), refers to the process by which the ATP hydrolysis (ATPase) activity of CD39 is inhibited. Such antibodies have been reported in several publications mentioned above, the disclosures of whose amino acid sequences are incorporated herein by reference. Some antibodies can neutralize membrane-bound CD39 by inhibiting domain movement in membrane-bound CD39 (memCD39), but do not similarly affect the activity of soluble CD39 protein (sCD39). While sCD39 is a monomer, memCD39 has been reported to be a homomultimer, and further, the transmembrane domain in memCD39 has been reported to undergo dynamic movement that indicates a functional relationship with the active site. Consequently, unlike sCD39, memCD39 may exhibit a configuration that allows antibody-mediated neutralization. One possibility is that functional neutralization requires the use of a bivalent antibody that simultaneously binds two memCD39 molecules (e.g., within a memCD39 homomultimer).

[0046] The anti-CD39 antibodies described herein bind to an epitope present on human CD39 protein expressed on the surface of cells, including tumor cells (e.g., these anti-CD39 antibodies can compete with the anti-CD39 antibodies of SEQ ID NOs: 10 and 11 for binding to an epitope on CD39), potently inhibit the enzymatic (ATPase activity) activity of the CD39 enzyme bound to the cell membrane (cell surface-expressed CD39), and the anti-CD39 antibodies described herein further inhibit the enzymatic (ATPase activity) activity of soluble (extracellular domain) human CD39 protein. The antibodies thereby mediate potent neutralization of CD39 activity in an individual by neutralizing both membrane-bound and soluble CD39 protein, including soluble CD39 released or excreted from tumor cells, thereby reducing immunosuppression, e.g., for the treatment of cancer and / or infectious diseases.

[0047] In one embodiment, the anti-CD39 antibody comprises (a) the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NOs: 2, 3, and 4, respectively, and (b) the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NOs: 5, 6, and 7, respectively.

[0048] In one embodiment, a human CD39 antibody is characterized as comprising: an HCDR1 comprising the amino acid sequence DYNMH (SEQ ID NO: 2), or a sequence of at least three or four consecutive amino acids thereof, wherein optionally, one or more of these amino acids may be substituted with a different amino acid; an HCDR2 comprising the amino acid sequence YIVPLNGGSTFNQKFKG (SEQ ID NO: 3), or a sequence of at least four, five, six, seven, eight, nine, or ten consecutive amino acids thereof, wherein optionally, one or more of these amino acids may be substituted with a different amino acid, optionally with an aspartic acid substitution at Kabat position 61 and optionally with a lysine substitution at Kabat position 65; an HCDR3 comprising the amino acid sequence GGTRFAY (SEQ ID NO: 4), or a sequence of at least four, five, or six consecutive amino acids thereof, wherein optionally, one or more of these amino acids may be substituted with a different amino acid. an LCDR1 comprising the amino acid sequence: RASESVDNFGVSFMY (SEQ ID NO: 5) or a sequence of at least 4, 5, 6, 7, 8, 9, or 10 consecutive amino acids thereof, optionally wherein one or more of these amino acids may be substituted with a different amino acid, and optionally wherein an arginine is substituted at Kabat position 24; an LCDR2 region comprising the amino acid sequence: GASNQGS (SEQ ID NO: 6) or a sequence of at least 4, 5, or 6 consecutive amino acids thereof, optionally wherein one or more of these amino acids may be substituted with a different amino acid; and / or an LCDR3 region comprising the amino acid sequence: QQTKEVPYT (SEQ ID NO: 7) or a sequence of at least 4, 5, 6, 7, or 8 consecutive amino acids thereof, optionally wherein one or more of these amino acids may be deleted or substituted with a different amino acid.

[0049] In any embodiment, the CDR positions may be according to Kabat numbering.

[0050] In one embodiment, the anti-CD39 antibody comprises the hypervariable regions, and optionally the CDRs, of the antibody having the VH and VL of SEQ ID NOs: 8 and 9 as shown below.

[0051] Anti-CD39 VH: QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYNMHWVRQAPGQRLEWIGYIVPLNGGSTFNQKFKGRATITVDTSARTAYMELSSLRSEDTAVYYCARGGTRFAYWGQGTLVTVSS (SEQ ID NO: 8). Anti-CD39 VL: DIVMTQSPDSLAVSLGERATINCRASESVDNFGVSFMYWFQQKPGQPPKLLIYGASNQGSGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQTKEVPYTFGGGTKVEIK (SEQ ID NO: 9).

[0052] In one embodiment, the anti-CD39 antibody comprises CDR1, CDR2, and CDR3 having the amino acid sequences set forth in SEQ ID NOs: 2, 3, and 4, respectively, and a human IGHV1-3 gene, e.g., IGHV1-3 * a heavy chain variable region (VH) comprising the amino acid sequences of frameworks FR1, FR2, and FR3 derived from the human IGHJ1 gene, e.g., IGHJ1 * and CDR1, CDR2, and CDR3 having the respective amino acid sequences set forth in SEQ ID NOs: 5, 6, and 7, and a human IGKV4-1 (e.g., IGK4-1 * 01) gene, and optionally human IGKJ4 (e.g., IGKJ4 *The VH may be characterized by an antigen-binding domain comprising a light chain variable region (VL) further comprising a framework 4 (FR4) amino acid sequence derived from the VH (VL-01) gene. The VH further comprises one or more amino acid substitutions of residues present in the human framework sequence with different residues (e.g., residues present in the non-human framework) at Kabat positions selected from the group consisting of 48, 67, 71, and 76. In one embodiment, the VH comprises one or more amino acid substitutions in heavy chain CDR2, e.g., at Kabat positions 60 and / or 64. Optionally, the residue at position 60 is serine (e.g., CDR2 comprises an N60S substitution). Optionally, the residue present at Kabat position 64 is glutamine (e.g., CDR2 comprises a K64Q substitution). In one embodiment, the residue present at Kabat position 24 in the VL is lysine (e.g., CDR1 comprises an R24K substitution). Optionally, a phenylalanine is present at Kabat position 36 in the VL.

[0053] In one embodiment, the VH comprises an isoleucine residue at Kabat position 48, an alanine residue at Kabat position 67, a valine at Kabat position 71 and an arginine at Kabat position 76.

[0054] In one embodiment, the VL comprises a phenylalanine at Kabat position 36 (FR2). In one embodiment, the VL comprises a lysine at Kabat position 24 (CDR1).

[0055] In any embodiment, the anti-CD39 antibody may be characterized as comprising a heavy chain variable region (VH) comprising an amino acid sequence at least 70%, 80%, 85%, 90%, 92%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:8 and a light chain variable region (VL) comprising an amino acid sequence at least 70%, 80%, 85%, 90%, 92%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO:8 and SEQ ID NO:9.

[0056] In one embodiment, the anti-CD39 antibody comprises a heavy chain variable region (VH) comprising CDR1, CDR2, and CDR3 having the respective amino acid sequences set forth in SEQ ID NOs: 2, 3, and 4 and a human framework (e.g., FR1, FR2, FR3, and FR4 of human origin); and CDR1, CDR2, and CDR3 having the respective amino acid sequences set forth in SEQ ID NOs: 5, 6, and 7 and a human framework (e.g., FR1, FR2, FR3, and FR4 of human origin). and a light chain variable region (VL) comprising an amino acid sequence at least 70%, 80%, 85%, 90%, 92%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 8, and the light chain variable region (VL) comprises an amino acid sequence at least 70%, 80%, 85%, 90%, 92%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 9.

[0057] In any embodiment, the VH can be characterized as containing a substitution at one, two, three, or all of Kabat positions 48, 67, 71, and 76. In one embodiment, the residue at position 48 is an isoleucine (e.g., an M48I substitution). In one embodiment, the residue at position 67 is an alanine (e.g., a V67A substitution). In one embodiment, the residue at position 71 is a valine (e.g., an R71V substitution). In one embodiment, the residue at position 76 is an arginine (e.g., an S76R substitution). In any embodiment, the VL can be characterized as containing a substitution at Kabat position 36. In one embodiment, the residue at position 36 is a phenylalanine (e.g., a Y36F substitution).

[0058] In one embodiment, the anti-CD39 antibody comprises a heavy chain comprising an amino acid sequence at least 70%, 80%, 85%, 90%, 92%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 10, and a light chain comprising an amino acid sequence at least 70%, 80%, 85%, 90%, 92%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 11.

[0059] Anti-CD39 heavy chain QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYNMHWVRQAPGQRLEWIGYIVPLNGGSTFNQKFKGRATITVDTSARTAYMELSSLRSEDTAVYYCARGGTRFAYWGQGTLVT VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPEAEGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 10). Anti-CD39 light chain DIVMTQSPDSLAVSLGERATINCRASESVDNFGVSFMYWFQQKPGQPPKLLIYGASNQGSGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQTKEVPYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 11).

[0060] In one embodiment, the anti-CD39 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 and a light chain comprising the amino acid sequence of SEQ ID NO: 11. In one embodiment, the anti-CD39 antibody is IPH5201.

[0061] In any embodiment, the anti-CD39 antibody can be characterized as binding to, inhibiting, or neutralizing the ATPase activity of soluble CD39 protein (sCD39). In one embodiment, the sCD39 protein lacks the two transmembrane domains (i.e., the transmembrane domains near the N- and C-termini) found in membrane-bound CD39. In one embodiment, the sCD39 protein is a non-membrane-bound sCD39 protein found in the circulation, e.g., in human individuals. In one embodiment, the CD39-derived sequence of the sCD39 protein comprises or consists of the Thr38-Val478 fragment of CD39. A Thr38-Val478 protein with a C-terminal His tag is commercially available from R&D Systems, Inc. (product number 4397-EN). In one embodiment, the protein, antibody, or antibody fragment inhibits the ATPase activity of sCD39 when incubated with sCD39 in solution, e.g., in tumor cell supernatant. In one embodiment, the protein, antibody, or antibody fragment specifically binds to human CD39 protein, both in soluble (extracellular domain protein) and membrane-bound form.

[0062] The anti-CD39 antibodies described herein contain a human Fc domain that has been modified to reduce or substantially eliminate binding to one or more (or all) of human Fcγ receptors, such as human CD16, CD32a, CD32b, and CD64. CD39 inhibitory activity is independent of ADCC, CDC, or toxin-mediated depletion of CD39-expressing cells. These antibodies are therefore used as "pure" CD39 blocking agents with immunomodulatory activity.

[0063] As used herein, the term "PD-1" refers to the programmed death 1 (PD-1) protein (also called "programmed cell death 1"), an inhibitory member of the CD28 receptor family, which also includes CD28, CTLA-4, ICOS, and BTLA. The complete nucleotide sequence of human PD-1 is shown in GenBank Accession No. U64863 as follows: MQIPQAPWPVVWAVLQLGWRPGWFLDSPDRPWNPPTFFPALLVVTEGDNATFTCSFSNTSESFVLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGRDFHMSVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVTERRAEVPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVLAVICSRAARGTIGARRTGQPLKEDPSAVPVFSVDYGELDFQWREKTPEPPVPCVPEQTEYATIVFPSGMGTSSPARRGSADGPRSAQPLRPEDGHCSWPL (SEQ ID NO: 12).

[0064] "PD-1" also includes any mutant, derivative, or isoform of the PD-1 gene or the protein encoded by PD-1. PD-1 is expressed on activated B cells, T cells, and myeloid cells (Okazaki et al. (2002) Curr. Opin. Immunol. 14: 391779-82; Bennett et al. (2003) J Immunol 170:711-8). Two ligands for PD-1, PD-L1 and PD-L2, have been identified and have been shown to downregulate T cell activation upon binding to PD-1 (Freeman et al. (2000) J Exp Med 192:1027-34; Latchman et al. (2001) Nat Immunol 2:261-8; Carter et al. (2002) Eur J Immunol 32:634-43). Both PD-L1 and PD-L2 are B7 homologs that bind to PD-1 but do not bind to other CD28 family members.

[0065] The complete nucleotide sequence of human PD-L1 is shown in UniProtKB / Swiss-Prot, identifier Q9NZQ7-1, as follows: MRIFAVFIFMTYWHLLNAFTVTVPKDLYVVEYGSNMTIECKFPVEKQLDLAALIVYWEMEDKNIIQFVHGEEDLKVQHSSYRQRARLLKDQLSLGNAALQITDVKLQDAGVYRCMISYGGADYKRITVKVNAPYNKINQRILVVDPVTSEHELTCQAEGYPKAEVIWTSSDHQVLSGKTTTTNSKREEKLFNVTSTLRINTTTNEIFYCTFRRLDPEENHTAELVIPELPLAHPPNERTHLVILGAILLCLGVALTFIFRLRKGRMMDVK KCGIQDTNSKKQSDTHLEET (SEQ ID NO: 13).

[0066] PD-L1 is abundant in a variety of human cancers. PD-1 / PD-L1 interaction results in a reduction in tumor-infiltrating lymphocytes, reduced T-cell receptor-mediated proliferation, and immune evasion by cancer cells. Immune suppression can be reversed by inhibiting the local interaction between PD-1 and PD-L1, and the effects are additive when the interaction between PD-1 and PD-L2 is also blocked.

[0067] Anti-PD(L)1 antibodies are antibodies that bind to PD-1 or PD-L1. The antibodies can neutralize PD-1 or reduce the inhibitory activity of human PD-1. "Reducing the inhibitory activity of human PD-1," "neutralizing PD-1," or "neutralizing the inhibitory activity of human PD-1" refers to the process by which PD-1 interacts with one or more of its binding partners, such as PD-L1 or PD-L2, resulting in inhibition of its signaling ability. Antibodies that neutralize the inhibitory activity of PD-1 reduce, block, inhibit, suppress, or interfere with signaling resulting from the interaction of PD-1 with one or more of its binding partners, particularly PD-L1. Such agents can enhance T cell effector functions, such as proliferation, cytokine production, and / or cytotoxicity, by reducing negative costimulatory signals mediated by or through cell surface proteins expressed on T lymphocytes. PD-1 neutralizing agents can interact with PD-1 and / or one or more of its binding partners, such as PD-L1 and PD-L2.

[0068] In some embodiments, the anti-PD(L)1 antibody is an anti-PD-L1 monoclonal antibody that inhibits the binding of PD-L1 to PD-1. In some embodiments, the anti-PD(L)1 antibody is an anti-PD-1 monoclonal antibody that inhibits the binding of PD-1 to PD-L1.

[0069] In some embodiments, the anti-PD(L)1 antibody is YW243.55.S70, MPDL3280A (atezolizumab, Tecentriq®), MDX-1105, or durvalumab (MEDI4736, Imfinzi®). MDX-1105, also known as BMS-936559, is an anti-PD-L1 antibody described in WO2007 / 005874. Antibody YW243.55.S70 is an anti-PD-L1 antibody described in WO2010 / 077634. ​​Examples of anti-PD-L1 antibodies useful in the methods disclosed herein, and methods for their production, are also described in WO2010 / 077634A1 and U.S. Patent No. 8,217,149, which are incorporated herein by reference.

[0070] In some embodiments, the anti-PD(L)1 antibody is a PD-L1 antibody that is durvalumab. Durvalumab (MEDI4736, Imfinzi™) is a human monoclonal antibody against human PD-L1 that can block the binding of PD-L1 to both the PD-1 and CD80 receptors. Disclosures regarding durvalumab are described in U.S. Patent Nos. 8,779,108 and 9,493,565, which are incorporated herein by reference. Durvalumab has heavy and light chains with the amino acid sequences of SEQ ID NO: 16 and SEQ ID NO: 17, respectively. The heavy chain variable region of durvalumab is set forth in SEQ ID NO: 14, and the light chain variable region of durvalumab is set forth in SEQ ID NO: 15.

[0071] In another embodiment, the anti-PD(L)1 antibody is an anti-PD-L1 antibody (or antigen-binding portion thereof) that competes with durvalumab for binding to PD-L1. In some embodiments, the anti-PD-L1 antibody binds to the same epitope as durvalumab. In a particular embodiment, the anti-PD-L1 antibody has the same heavy and light chain CDRs as durvalumab.

[0072] In some embodiments, the anti-PD(L)1 antibody (e.g., a durvalumab-derived drug) comprises (i) a heavy chain variable region of SEQ ID NO: 14, or an amino acid sequence at least 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 99% identical thereto, and (ii) a light chain variable region of SEQ ID NO: 15, or an amino acid sequence at least 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 99% identical thereto. In some embodiments, the PD-1 neutralizing agent (e.g., an agent derived from durvalumab) comprises (i) a heavy chain of SEQ ID NO: 16, or an amino acid sequence at least 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 99% identical thereto, and (ii) a light chain of SEQ ID NO: 17, or an amino acid sequence at least 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 99% identical thereto. In some embodiments, the PD-1 neutralizing agent comprises H-CDR1, H-CDR2, and / or H-CDR3 sequences from a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 14. In some embodiments, the PD-1 neutralizing agent comprises L-CDR1, L-CDR2, and / or L-CDR3 sequences from a light chain variable region comprising the amino acid sequence of SEQ ID NO: 15.

[0073] In some embodiments, the anti-PD(L)1 antibody comprises heavy chain H-CDR1, H-CDR2, and H-CDR3 domains having the amino acid sequences of SEQ ID NOs: 18-20, respectively, and light chain L-CDR1, L-CDR2, and L-CDR3 domains having the amino acid sequences of SEQ ID NOs: 21-23, respectively.

[0074] Durvalumab heavy chain variable region: EVQLVESGGGLVQPGGSLRLSCAASGFTFSRYWMSWVRQAPGKGLEWVANIKQDGSEKYYVDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAREGGWFGELAFDYWGQGTLVTVSS (SEQ ID NO: 14).

[0075] Durvalumab light chain variable region: EIVLTQSPGTLSLSPGERATLSCRASQRVSSSYLAWYQQKPGQAPRLLIYDASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSLPWTFGQGTKVEIK (SEQ ID NO: 15).

[0076] Durvalumab heavy chain: EVQLVESGGGLVQPGGSLRLSCAASGFTFSRYWMSWVRQAPGKGLEWVANIKQDGSEKYYVDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAREGGWFGELAFDYWGQGT LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTC PPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPASIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 16).

[0077] Durvalumab light chain: EIVLTQSPGTLSLSPGERATLSCRASQRVSSSYLAWYQQKPGQAPRLLIYDASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSLPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 17).

[0078] Durvalumab, heavy chain CDR: H-CDR1: GFTFSRYWMS (SEQ ID NO: 18) H-CDR2: NIKQDGSEKYYVDSVKG (SEQ ID NO: 19) H-CDR3: EGGWFGELAFDY (SEQ ID NO: 20) Durvalumab, light chain CDR: L-CDR1: RASQRVSSSYLA (SEQ ID NO: 21) L-CDR2: DASSRAT (SEQ ID NO: 22) L-CDR3: QQYGSLPWT (SEQ ID NO: 23)

[0079] In another embodiment, the anti-PD(L)1 antibody is atezolizumab (MPDL3280A, Tecentriq®, CAS Registry Number: 1422185-06-5). In some embodiments, the anti-PD-L1 antibody has a heavy chain variable region comprising the amino acid sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSS (SEQ ID NO: 24) or EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSASTK (SEQ ID NO: 25) and a light chain variable region comprising the following amino acid sequence: DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKR (SEQ ID NO: 26) Includes.

[0080] In some embodiments, the anti-PD(L)1 antibody comprises (i) a heavy chain or heavy chain variable region of SEQ ID NO: 27, or an amino acid sequence at least 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 99% identical thereto, and (ii) a light chain or light chain variable region of SEQ ID NO: 28, or an amino acid sequence at least 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 99% identical thereto.

[0081] EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTL VTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKQVSLTCLVKGFYPSDIAVEWESNGQPENYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 27). DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH KVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 28).

[0082] In some embodiments, the anti-PD(L)1 antibody is an anti-PD-1 antibody that inhibits the binding of PD-1 to PD-L1. In some embodiments, the anti-PD-1 antibody is nivolumab. Nivolumab (also known as Opdivo®, formerly known as 5C4, BMS-936558, MDX-1106, or ONO-4538) is a fully human IgG4(S228P) PD-1 immune checkpoint inhibitor antibody that selectively interferes with the interaction of PD-1 with PD-L1 and PD-L2 ligands, thereby blocking the downregulation of anti-tumor T cell function (U.S. Patent No. 8,008,449; Wang et al., (2014) Cancer Immunol Res. 2(9):846-56). In another embodiment, the anti-PD-1 antibody or a fragment thereof competes with nivolumab for binding to PD-1. In some embodiments, the anti-PD-1 antibody binds to the same epitope as nivolumab. In certain embodiments, the anti-PD-1 antibody has the same heavy and light chain CDRs as nivolumab.

[0083] In another embodiment, the anti-PD-1 antibody is pembrolizumab. Pembrolizumab (also known as "KEYTRUDA®," lambrolizumab, and MK-3475) is a humanized monoclonal IgG4 antibody directed against the human cell surface receptor PD-1. Pembrolizumab is described, for example, in U.S. Patent No. 8,900,587. Pembrolizumab is FDA-approved for the treatment of relapsed or refractory melanoma and advanced NSCLC. In another embodiment, the anti-PD-1 antibody (or antigen-binding portion thereof) competes with pembrolizumab for binding to PD-1. In some embodiments, the anti-PD-1 antibody binds to the same epitope as pembrolizumab. In certain embodiments, the anti-PD-1 antibody has the same heavy and light chain CDRs as pembrolizumab.

[0084] In another embodiment, the anti-PD-1 antibody is cemiplimab.

[0085] A chemotherapeutic agent refers to a compound useful in the treatment of cancer, e.g., lung cancer, NSCLC, optionally stage II NSCLC and / or stage IIIA NSCLC. For example, the chemotherapeutic agent can be a compound known to be useful in the neoadjuvant treatment of NSCLC, optionally resectable or II or IIIA NSCLC. When used, e.g., as neoadjuvant therapy, chemotherapy can be used at known or standard doses and frequencies for such chemotherapy. For examples of neoadjuvant chemotherapy and regimens, see, e.g., Burdett S., 2014, supra. In some embodiments, the chemotherapy used in the neoadjuvant treatment herein comprises a platinum-based chemotherapy agent. A "platinum-based" chemotherapy agent includes an organic compound containing platinum as an integral part of the molecule. Typically, platinum-based chemotherapy agents are platinum coordination complexes. Platinum-based chemotherapy agents are sometimes referred to in the art as "platins." Examples of platinum-based chemotherapeutic agents include, but are not limited to, cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin tetranitrate, phenanthriplatin, picoplatin, lipoplatin, and satraplatin. In some embodiments, a platinum-based chemotherapeutic agent (e.g., cisplatin or carboplatin) can be administered in combination with one or more additional chemotherapeutic agents, such as a nucleoside analog (e.g., gemcitabine).

[0086] For example, platinum-based chemotherapy can include a platinum-based chemotherapy agent (e.g., cisplatin or carboplatin) and, optionally, one or more additional chemotherapy agents, such as a nucleoside analog (e.g., gemcitabine), a taxane (e.g., paclitaxel), or an antifolate (e.g., pemetrexed).

[0087] In some embodiments, the chemotherapy used in the neoadjuvant treatment herein comprises gemcitabine.

[0088] In some embodiments, the chemotherapy used in the neoadjuvant treatment herein comprises the administration of two or more chemotherapy drugs. In some embodiments, the chemotherapy comprises the administration of at least one or two drugs selected from paclitaxel, carboplatin, pemetrexed, or cisplatin. In some embodiments, the chemotherapy comprises or consists of the administration of carboplatin and paclitaxel. In some embodiments, the chemotherapy comprises or consists of the administration of cisplatin and gemcitabine. In some embodiments, the chemotherapy comprises or consists of the administration of cisplatin and pemetrexed. In some embodiments, the chemotherapy comprises or consists of the administration of carboplatin and pemetrexed.

[0089] As disclosed herein, the present disclosure provides a method of treating a tumor or cancer and / or preventing recurrence of a tumor or cancer in a patient in need thereof, e.g., a patient with a resectable cancer or tumor, comprising administering an anti-CD39 antibody and an anti-PD(L)1 antibody, wherein the anti-CD39 antibody and the anti-PD(L)1 antibody are administered as neoadjuvant therapy. In one embodiment, the method comprises administering an anti-CD39 antibody, an anti-PD(L)1 antibody, and a chemotherapeutic agent, wherein the anti-CD39 antibody, the anti-PD(L)1 antibody, and the chemotherapeutic agent are administered as neoadjuvant therapy.

[0090] The present disclosure further provides a method of treating a tumor or cancer and / or preventing recurrence of a tumor or cancer in a patient in need thereof, comprising administering an anti-CD39 antibody and an anti-PD(L)1 antibody, wherein the anti-CD39 antibody and the anti-PD(L)1 antibody are administered as neoadjuvant therapy and then as adjuvant therapy.

[0091] The term "patient" is intended to include humans and non-human animals, particularly mammals. In some embodiments, the present disclosure relates to providing treatments (methods of treatment or pharmaceutical formulations for use as described herein) for patients with neoplastic and / or cancerous disorders, optionally with surgically resectable tumors and / or cancers. In some embodiments, the tumor is a lung tumor (e.g., non-small cell lung cancer (NSCLC)). In some embodiments, the non-small cell lung tumor is squamous cell carcinoma, adenocarcinoma, or large cell carcinoma. In some embodiments, the patient has stage I, stage II, or stage III non-small cell lung tumor. The method can be used as a first-line treatment (1L treatment, 1-L treatment, 1L, or 1-L), i.e., the first treatment administered for a disease, particularly a cancer, described herein. A first-line treatment is specific for a particular cancer type or subtype or stage of cancer. A first-line treatment can also be part of a standard treatment set. A first-line treatment is generally accepted as the best treatment for a disease, particularly a cancer, as described herein. If the first-line treatment does not cure the disease or causes severe side effects, subsequent alternative treatment lines may be used. In some embodiments, the present disclosure provides first-line treatment methods for cancer (methods of treatment, or pharmaceutical formulations for use as described herein).

[0092] In some embodiments, the patient has not previously undergone resection of the non-small cell lung tumor prior to receiving the neoadjuvant treatment of the present disclosure, hi some embodiments, the patient has undergone resection, optionally complete resection, of the non-small cell lung tumor prior to receiving the adjuvant therapy of the present disclosure.

[0093] Additionally, treatment regimens adapted for administering an anti-CD39 antibody and an anti-PD(L)1 antibody as neoadjuvant therapy and / or adjuvant therapy are provided. Accordingly, in the administration, treatment, or prevention methods herein, the method can be identified as comprising administering to a patient an effective amount of a treatment regimen comprising an anti-CD39 antibody and an anti-PD(L)1 antibody, e.g., a treatment regimen described herein comprising an anti-CD39 antibody and an anti-PD(L)1 antibody. Accordingly, provided herein is a method for treating lung cancer in a patient, comprising administering to the patient an effective amount of a treatment regimen comprising an anti-CD39 antibody and an anti-PD(L)1 antibody, and optionally further chemotherapy, wherein the treatment regimen is neoadjuvant therapy. Optionally, the method further comprises administering to the patient an effective amount of a treatment regimen comprising an anti-CD39 antibody and an anti-PD(L)1 antibody, wherein the treatment regimen is adjuvant therapy. Doses and dosing intervals used in treatment regimens can be further specified herein, including advantageous regimens in which the same doses of anti-CD39 antibody and anti-PD(L)1 antibody can be used in both neoadjuvant and adjuvant settings.

[0094] Thus, in one embodiment, the treatment regimen of the present disclosure is characterized by comprising administering to the patient an anti-CD39 antibody and an anti-PD(L)1 antibody (and optionally further chemotherapy) prior to surgical tumor resection. The treatment can further comprise administering to the patient an anti-CD39 antibody and an anti-PD(L)1 antibody after surgical tumor resection. In one embodiment, the anti-PD(L)1 antibody is durvalumab. The tumor or cancer can optionally be identified as a tumor or cancer that is considered surgically resectable.

[0095] In any embodiment herein, a method and / or effective amount may be specified to include an amount of a combination of an anti-PD(L)1 antibody and an anti-CD39 antibody (and optionally further chemotherapy) that achieves a therapeutic outcome. In some examples, an effective amount of a therapeutic agent or combination of therapeutic agents is an amount of a therapeutic agent or combination of therapeutic agents that achieves the clinical endpoint of improved overall response rate (ORR), complete response (CR), pathological complete response (pCR), partial response (PR), improved survival (e.g., disease-free survival (DFS), disease-specific survival (DSS), distant metastasis-free survival, progression-free survival (PFS), and / or overall survival (OS)), improved duration of response (DOR), improved time to decline in function and quality of life (QoL), and / or ctDNA clearance. The improvement (e.g., response rate (e.g., ORR, CR, and / or PR), survival rate (e.g., DFS, DSS, distant metastasis-free survival, PFS, and / or OS), DOR, improvement in time to deterioration of function and QoL, undetectable ctDNA and / or ctDNA clearance) can be compared to an appropriate standard, such as observation or standard treatment (e.g., treatment not including an anti-CD39 antibody (e.g., treatment with a placebo, treatment with an anti-PD(L)1 antibody)). In some cases, the improvement (e.g., response rate (e.g., ORR, CR, and / or PR), survival rate (e.g., EFS, DFS, DSS, distant metastasis-free survival, PFS, and / or OS), DOR, improvement in time to deterioration of function and QoL, and / or ctDNA clearance or undetectable ctDNA status) can be compared to observation.

[0096] As used herein, "complete response" and "CR" refer to the disappearance of all target lesions.

[0097] As used herein, "partial response" and "PR" refer to a reduction in the sum of the longest diameters (SLD) of target lesions by at least a specified value (e.g., 30%), referenced to the pre-treatment baseline SLD.

[0098] As used herein, "overall response rate," "objective response rate," and "ORR" refer interchangeably to the sum of the CR rate and the PR rate.

[0099] As used herein, "disease-free survival" and "DFS" refer to the length of time a patient lives without cancer recurrence after primary treatment (e.g., surgical resection).

[0100] As used herein, "disease-specific survival" and "DSS" refer to the length of time that a patient has not died from a particular disease (e.g., NSCLC). In some cases, DSS may be defined as the time from randomization to death from NSCLC (e.g., by investigator's assessment of the cause of death). As used herein, "distant metastasis-free survival" refers to the length of time from either the date of diagnosis or the date of treatment initiation that a patient is still alive and the cancer has not spread to other parts of the body. In some cases, distant metastasis-free survival is defined as the time from randomization to the diagnosis of distant metastasis (i.e., non-regional metastasis) or death from any cause.

[0101] As used herein, "progression-free survival" and "PFS" refer to the length of time during and after treatment during which the cancer does not worsen. PFS includes periods during which the patient experiences CR or PR as well as periods of stable disease.

[0102] As used herein, "overall survival" and "OS" refer to the length of time a patient is still alive, either from the date of diagnosis of a disease (e.g., cancer) or from the date of treatment initiation. For example, OS can be defined as the time from randomization to death from any cause.

[0103] As used herein, the terms "duration of response" and "DOR" refer to the length of time from evidence of tumor response to disease progression or death from any cause, whichever occurs first.

[0104] In one embodiment, the treatment regimen herein allows for administration of the anti-CD39 antibody at the same dose (e.g., a fixed dose of 2250 mg or 3000 mg of anti-CD39 antibody) in both the adjuvant and neoadjuvant settings. Furthermore, the regimen allows for administration of the anti-CD39 antibody every three weeks as a neoadjuvant and every four weeks as an adjuvant (the same dosage in both neoadjuvant and adjuvant therapy).

[0105] In one embodiment, a method of administering an anti-CD39 antibody (e.g., an anti-CD39 antibody having the features described herein; comprising the amino acid sequence of SEQ ID NOs:2-7, SEQ ID NOs:8 and 9, or SEQ ID NOs:10 and 11) is provided, wherein the antibody is administered Q3w and / or Q4w at a fixed dose of 3000 mg. In one embodiment, the anti-CD39 antibody is administered Q3w at a fixed dose of 3000 mg as neoadjuvant therapy and Q4w at a dose of 3000 mg as adjuvant therapy. In another embodiment, a method of administering an anti-CD39 antibody (e.g., an anti-CD39 antibody having the features described herein; comprising the amino acid sequence of SEQ ID NOs:2-7, SEQ ID NOs:8 and 9, or SEQ ID NOs:10 and 11) is provided, wherein the antibody is (a) administered Q3w for one or more administrations and (b) administered Q4w for one or more administrations, in each case the antibody is administered at a fixed dose of 3000 mg.

[0106] In one embodiment, a method of administering an anti-CD39 antibody (e.g., an anti-CD39 antibody having the features described herein; comprising the amino acid sequence of SEQ ID NOs:2-7, SEQ ID NOs:8 and 9, or SEQ ID NOs:10 and 11) is provided, wherein the antibody is administered Q3w and / or Q4w at a fixed dose of 2250 mg. In one embodiment, the anti-CD39 antibody is administered Q3w at a fixed dose of 2250 mg as neoadjuvant therapy and Q4w at a dose of 2250 mg as adjuvant therapy. In another embodiment, a method of administering an anti-CD39 antibody (e.g., an anti-CD39 antibody having the features described herein; comprising the amino acid sequence of SEQ ID NOs:2-7, SEQ ID NOs:8 and 9, or SEQ ID NOs:10 and 11) is provided, wherein the antibody is administered (a) Q3w for one or more administrations and (b) Q4w for one or more administrations, in each case the antibody is administered at a fixed dose of 2250 mg.

[0107] Optionally, in any embodiment herein, the method is characterized as a method of treating cancer and / or preventing recurrence of cancer. Optionally, the method is characterized as a method of administering an anti-CD39 antibody in combination with an anti-PD(L)1 antibody (and / or chemotherapy). Optionally, in any embodiment, the method is characterized as a method of administering an anti-CD39 antibody (and optionally further an anti-PD(L)1 antibody, optionally further chemotherapy) in combination with surgery, and optionally further radiation therapy.

[0108] Preferably, the anti-CD39 antibody is administered intravenously (iv). Preferably, the anti-PD(L)1 antibody is administered intravenously.

[0109] In some embodiments, the doses of anti-PD(L)1 antibody and chemotherapy administered to a patient may vary depending, in part, on the size (weight, body surface, or organ size) and condition (age and general health) of the patient.

[0110] In some embodiments, the anti-PD(L)1 antibody is durvalumab and is administered at a fixed dose of 1500 mg. Durvalumab is marketed by AstraZeneca as Imfinzi. (商標) It is sold as.

[0111] A neoadjuvant treatment regimen may therefore include an anti-CD39 antibody administered at a fixed dose of 2250 mg or 3000 mg every three weeks and durvalumab administered at a fixed dose of 1500 mg every three weeks.

[0112] An adjuvant therapy treatment regimen may therefore include an anti-CD39 antibody administered at a fixed dose of 2250 mg or 3000 mg every four weeks and durvalumab administered at a fixed dose of 1500 mg every four weeks.

[0113] In certain embodiments, the anti-CD39 antibody and anti-PD(L)1 antibody, and optionally further chemotherapy, are administered simultaneously, concurrently, separately, or sequentially. In some embodiments, the anti-CD39 antibody and anti-PD(L)1 antibody are administered before chemotherapy. In further embodiments, the anti-CD39 antibody and anti-PD(L)1 antibody are administered concurrently with chemotherapy. Advantageously, the anti-CD39 antibody and anti-PD(L)1 antibody are administered on the same day, e.g., on day 1 of a 3-week cycle or day 1 of a 4-week cycle for neoadjuvant treatment and adjuvant treatment, respectively. Optionally, for neoadjuvant treatment, the anti-CD39 antibody and anti-PD(L)1 antibody are administered on the same day as chemotherapy (e.g., on day 1 of a 3-week cycle).

[0114] Thus, neoadjuvant therapy can include one or more cycles of treatment with an anti-CD39 antibody having the heavy and light chain amino acid sequences of SEQ ID NOs: 10 and 11, respectively, administered at a fixed dose of 2250 mg or 3000 mg, and durvalumab administered at a fixed dose of 1500 mg, where the anti-CD39 antibody and durvalumab are administered once on day 1 of a three-week cycle. Neoadjuvant therapy can include, for example, two, three, four or more such cycles of treatment.

[0115] The adjuvant therapy can include one or more cycles of treatment with an anti-CD39 antibody having the heavy and light chain amino acid sequences of SEQ ID NOs: 10 and 11, respectively, administered at a fixed dose of 2250 mg or 3000 mg, and durvalumab administered at a fixed dose of 1500 mg, where the anti-CD39 antibody and durvalumab are administered once on day 1 of a 4-week cycle. The adjuvant therapy can include, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or more such cycles, or at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 cycles. In one embodiment, up to 12 cycles of the anti-CD39 antibody and durvalumab are administered as adjuvant therapy.

[0116] Thus, when anti-CD39 and anti-PD(L)1 antibodies are used as neoadjuvant and adjuvant therapy, treatment may therefore include: (a) administering one or more doses of an anti-CD39 antibody having heavy and light chain amino acid sequences of SEQ ID NOs: 10 and 11, respectively, at a fixed dose of 2250 mg or 3000 mg, and durvalumab at a fixed dose of 1500 mg, as neoadjuvant therapy, wherein the anti-CD39 antibody and durvalumab are administered on day 1 of a 3-week cycle; and (b) administering one or more doses of an anti-CD39 antibody having heavy and light chain amino acid sequences of SEQ ID NOs: 10 and 11, respectively, at a fixed dose of 2250 mg or 3000 mg, and durvalumab at a fixed dose of 1500 mg, as adjuvant therapy, wherein the anti-CD39 antibody and durvalumab are administered on day 1 of a 4-week cycle.

[0117] Adjuvant therapy: Anti-CD39 antibody and anti-PD(L)1 antibody can be initiated soon after surgical resection of the tumor or cancer, preferably within 10 weeks of surgical resection. Adjuvant therapy can be continued for up to 12 cycles or more, for example, until disease progression, or until a desired response (e.g., CR) is achieved, or until ctDNA clearance.

[0118] In some embodiments, anti-CD39 antibodies and anti-PD(L)1 antibodies are used independently or regardless of the mutation status of the epidermal growth factor receptor (EGFR) gene and / or the anaplastic lymphoma kinase (ALK) gene (e.g., as neoadjuvant and optionally adjuvant therapy). In some embodiments, anti-CD39 antibodies and anti-PD(L)1 antibodies are used to treat patients who lack EGFR mutations (EGFR wild-type) and / or patients who lack ALK gene rearrangements (ALK wild-type).

[0119] In some embodiments, anti-CD39 antibodies and anti-PD(L)1 antibodies are used independently or regardless of the patient's PD-L1 expression status (e.g., as neoadjuvant and optionally adjuvant therapy). In some embodiments, anti-CD39 antibodies and anti-PD(L)1 antibodies are used as neoadjuvant and optionally adjuvant therapy for the treatment of patients with detectable PD-L1 expression, e.g., patients with a CPS score of at least 1 (CPS≧1).

[0120] PD-L1 expression can include any detectable level of PD-L1 protein on the cell surface or PD-L1 mRNA expression within cells or tissues. PD-L1 protein expression can be detected using a diagnostic PD-L1 antibody by IHC assay or flow cytometry of tumor tissue sections. Alternatively, PD-L1 protein expression by tumor cells can be detected by PET imaging using a binding agent.

[0121] One approach employs a simple binary endpoint of positive or negative PD-L1 expression, with a positive result defined as the percentage of tumor cells showing histological evidence of cell surface membrane staining. Tumor tissue sections are counted as PD-L1 positive if PD-L1 expression is present on ≥1% of total tumor cells.

[0122] In another approach, PD-L1 expression in tumor tissue sections is quantified on tumor cells and infiltrating immune cells, primarily lymphocytes. The percentage of tumor cells and infiltrating immune cells showing membrane staining is separately quantified at <5%, 5–9%, and then in 10% increments up to 100%. PD-L1 expression in immune infiltrates is reported as a semiquantitative measure called the adjusted inflammation score (AIS), which is determined by multiplying the percentage of membrane-staining cells by the intensity of the infiltration and graded as absent (0), mild (score 1, rare lymphocytes), moderate (score 2, focal infiltration of the tumor with lymphohistiocytic aggregates), or severe (score 3, diffuse infiltration). Tumor tissue sections are counted as positive for PD-L1 expression in immune infiltrates if their AIS score is 5 or higher.

[0123] Several scoring systems for the PD-L1 protein are commonly used. The "tumor proportion score (TPS)" refers to the proportion of tumor cells expressing PD-L1 on the cell membrane at any intensity (weak, moderate, or strong). Linear partial or complete staining of the cell membrane is interpreted as PD-L1 positivity. The "mononuclear inflammatory density score (MIDS)" refers to the ratio of the number of PD-L1-expressing mononuclear inflammatory cells (MIC) (large and small lymphocytes, monocytes, and macrophages within tumor nests and adjacent supporting stroma) infiltrating or adjacent to the tumor to the total number of tumor cells. MIDS is scored on a scale of 0 to 4, with 0 = absent; 1 = present but less than 1 MIC per 100 tumor cells (<1%); 2 = at least 1 MIC per 100 tumor cells but less than 1 MIC per 10 tumor cells (1–9%); 3 = at least 1 MIC per 10 tumor cells but fewer MICs than tumor cells (10–99%); and 4 = at least more MICs than tumor cells (≥100%). The "combined positive score (CPS)" refers to the ratio of the number of PD-L1-positive tumor cells and PD-L1-positive mononuclear inflammatory cells (MICs) (numerator) within tumor nests and adjacent supporting stroma compared to the total number of tumor cells (denominator; i.e., the number of PD-L1-positive tumor cells and PD-L1-negative tumor cells). PD-L1 expression of any intensity is considered positive, i.e., weak (1+), moderate (2+), or strong (3+).

[0124] When anti-CD39 antibodies and anti-PD(L)1 antibodies are used as adjuvant therapy (e.g., in addition to neoadjuvant therapy), the treatment of the present disclosure may be advantageous for patients who are minimal residual disease positive (MRD+) after surgical resection. For example, after neoadjuvant therapy and surgery, optionally, (a) determine whether the patient is minimal residual disease positive (MRD+); and (b) if the patient is identified as MRD+, the anti-CD39 antibody and anti-PD(L)1 antibody can be administered to the patient as adjuvant therapy. In one embodiment, the anti-CD39 antibody and anti-PD(L)1 antibody are administered within 10 weeks of tumor resection.

[0125] A patient's MRD status can be determined using methods known in the art (see, e.g., Abbosh et al. (2017); Chaudhuri et al. (2017)). In some embodiments, a patient's MRD status can be determined using a multi-step assay. First, whole exome sequencing (WES) is performed on DNA extracted from the patient's tumor tissue, and germline mutations are controlled for by WES of the patient's whole blood. A personalized panel of frequently expressed tumor variants is then developed for the patient. This panel is then used to identify the presence of these variants in circulating tumor DNA (ctDNA) extracted from the patient's plasma. If the panel detects the tumor variants, the patient is considered MRD+. This personalized approach allows for highly sensitive detection of a patient's tumor variants from DNA extracted from plasma.

[0126] In some embodiments, determining whether a patient is minimal residual disease positive (MRD+) is determined by: (a) sequencing all or part of the patient's tumor genome or exome to define clonal and / or subclonal mutations in the tumor; (b) defining a set of reagents that detect the presence of DNA from the tumor through the presence of clonal and / or subclonal mutations; and (c) analyzing a sample containing tumor-derived DNA obtained from the patient after tumor removal and the defined set of reagents to determine whether the tumor has recurred by detecting clonal and / or subclonal mutations in the sample. The presence and / or increase of clonal and / or subclonal mutations characteristic of the tumor in a sample from the patient indicates whether the tumor has recurred. The clonal and / or subclonal mutations characteristic of a patient's tumor are defined by sequencing all or part of the whole genome and / or exome of DNA obtained from the tumor, in some cases after the tumor has been removed from the patient. The presence and / or elevation of clonal and / or subclonal mutations in a sample obtained from a patient is analyzed using a set of reagents designed or defined to detect the presence of DNA from the tumor via the presence of specific clonal and / or subclonal mutations identified for the subject of interest.

[0127] In some embodiments, sequencing is performed on a tumor biopsy, all or part of a tumor, or one or more subsections of a tumor, cell-free DNA (cfDNA), ctDNA, exosome-derived tumor DNA, or circulating tumor cells from a subject. In some embodiments, sequencing is performed on a tumor or subsections thereof after tumor removal. In some embodiments, all or part of the genome or exome of at least two subsections of a tumor are sequenced, and clonal and / or subclonal mutations are defined based on which mutations occur in which tumor subsections. In some embodiments, the defined reagent set includes multiplex PCR primers, and the analysis is multiplex PCR. In some embodiments, sequencing is performed on plasma obtained from a patient, or the sample analyzed is a plasma sample from a patient.

[0128] MRD, as demonstrated by ctDNA detection, can reveal the presence of clinically indiscernible residual tumor after curative-intent treatment (surgery ± chemotherapy / radiotherapy). Detecting MRD in the absence of radiological evidence of disease offers an opportunity for early therapeutic intervention. Anti-CD39 and anti-PD(L)1 antibodies, for example, can be administered as soon as possible after tumor resection, preferably within 10 weeks. MRD+ patients have inferior recurrence-free survival rates compared with MRD- patients. Therefore, MRD+ patients may benefit from earlier therapeutic intervention and treatment expansion, including immunotherapy alone or in combination with chemotherapy. Furthermore, MRD- patients (the majority of whom are cured by surgery alone) can be spared more intensive treatment and the resulting unnecessary toxicity.

[0129] It will be understood that the anti-CD39 antibody and anti-PD(L)1 antibody can be incorporated into the pharmaceutical formulation at any suitable concentration (e.g., 1 mg / ml to 500 mg / ml, wherein the formulation has a pH of 2.0 to 10.0). The anti-CD39 antibody and anti-PD(L1) antibody can be included in the same or separate pharmaceutical formulations. The formulations may further comprise a buffer system, preservatives, tonicity agents, chelating agents, stabilizers, and surfactants. In one embodiment, the pharmaceutical formulation is an aqueous formulation, i.e., a formulation containing water. Such formulations are generally solutions or suspensions. In a further embodiment, the pharmaceutical formulation is an aqueous solution. The term "aqueous formulation" is defined as a formulation containing at least 50% w / w water. Similarly, the term "aqueous solution" is defined as a solution containing at least 50% w / w water, and the term "aqueous suspension" is defined as a suspension containing at least 50% w / w water.

[0130] In some cases, the pharmaceutical formulation is a lyophilized formulation, to which the physician or patient adds solvents and / or diluents before use. In some cases, the pharmaceutical formulation is a ready-to-use dried formulation (e.g., lyophilized or spray-dried) that does not require prior dissolution.

[0131] In a further aspect, the pharmaceutical formulation comprises an aqueous solution of such an antibody and a buffer, wherein the antibody is present at a concentration of 1 mg / mL or greater, and the pH of the formulation is about 2.0 to about 10.0. Optionally, the pH of the formulation is in a range selected from the list consisting of about 2.0 to about 10.0, about 3.0 to about 9.0, about 4.0 to about 8.5, about 5.0 to about 8.0, and about 5.5 to about 7.5. In a further embodiment, the buffer is selected from the group consisting of sodium acetate, sodium carbonate, citrate, glycylglycine, histidine, glycine, lysine, arginine, sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium phosphate, and tris(hydroxymethyl)aminomethane, bicine, tricine, malic acid, succinate, maleic acid, fumaric acid, tartaric acid, aspartic acid, or a mixture thereof. Each of these specific buffers constitutes an alternative embodiment of the invention.

[0132] In a further embodiment, the formulation further comprises a pharmaceutically acceptable preservative. In a further embodiment, the formulation further comprises an isotonicity agent. In a further embodiment of the invention, the formulation also comprises a chelating agent. In a further embodiment, the formulation further comprises a stabilizer. In a further embodiment, the formulation further comprises a surfactant. For convenience, reference is made to Remington: The Science and Practice of Pharmacy, 19th edition, 1995.

[0133] Other ingredients may be present in the pharmaceutical formulation of the present invention. Such additional ingredients may include wetting agents, emulsifiers, antioxidants, bulking agents, isotonicity adjusting agents, chelating agents, metal ions, oily vehicles, proteins (e.g., human serum albumin, gelatin, or proteins), and zwitterions (e.g., amino acids such as betaine, taurine, arginine, glycine, lysine, and histidine). Such additional ingredients, of course, should not adversely affect the overall stability of the pharmaceutical formulation of the present invention.

[0134] Administration of pharmaceutical compositions according to the invention can be via any of several routes of administration, for example, intravenously. Appropriate antibody formulations can also be determined by examining experience with other therapeutic monoclonal antibodies already in development.

[0135] In one aspect, provided herein is a kit, e.g., a kit comprising: (i) a pharmaceutical composition comprising an anti-CD39 antibody, such as an anti-CD39 antibody comprising the VH and VL amino acid sequences of SEQ ID NOs: 8 and 9, respectively, and an anti-PD(L)1 antibody, such as durvalumab; or (ii) a first pharmaceutical composition comprising an anti-PD(L)1 antibody, such as durvalumab, and a second pharmaceutical composition comprising an anti-CD39 antibody, such as an anti-CD39 antibody comprising the VH and VL amino acid sequences of SEQ ID NOs: 8 and 9, respectively; or (iii) a pharmaceutical composition comprising an anti-CD39 antibody, such as an anti-CD39 antibody comprising the VH and VL amino acid sequences of SEQ ID NOs: 8 and 9, respectively, and instructions for administering the anti-CD39 antibody together with an anti-PD(L)1 antibody (e.g., durvalumab), e.g., as neoadjuvant and / or adjuvant therapy, optionally to NSCLC; or (iv) A pharmaceutical composition comprising an anti-PD(L)1 antibody (e.g., durvalumab), and instructions for administering the anti-PD(L)1 antibody together with an anti-CD39 antibody (e.g., an anti-CD39 antibody comprising the VH and VL amino acid sequences of SEQ ID NOs: 8 and 9, respectively), for example as neoadjuvant and / or adjuvant therapy, optionally to NSCLC.

[0136] In any embodiment, the kit can optionally further comprise a chemotherapy, e.g., a chemotherapy comprising a platinum agent, gemcitabine, e.g., a chemotherapy comprising a combination of chemotherapy agents such as carboplatin and paclitaxel, cisplatin and gemcitabine, cisplatin and pemetrexed, or carboplatin and pemetrexed.

[0137] The pharmaceutical composition may optionally be identified as comprising a pharmaceutically acceptable carrier. The anti-CD39 antibody or anti-PD(L)1 antibody may optionally be identified as being present in a therapeutically effective amount compatible with use in any of the methods herein. The anti-CD39 antibody may optionally be identified as comprising the CDR amino acid sequences of SEQ ID NOs: 2-7. The anti-CD39 antibody may optionally be identified as comprising the heavy and light chain amino acid sequences of SEQ ID NOs: 10 and 11. The kit may also optionally include instructions, including, for example, a dosing schedule (e.g., a Q3w and / or Q4w schedule disclosed herein), to enable a healthcare professional (e.g., a doctor, nurse, or patient) to administer the composition contained therein to a patient with cancer (e.g., a solid tumor, particularly NSCLC, stage II or III NSCLC, resectable or not yet surgically resected NSCLC). In any embodiment, the kit may optionally include instructions for administering the anti-CD39 antibody simultaneously, separately, or sequentially with the anti-PD(L)1 antibody. The instructions may optionally further specify that the anti-CD39 antibody and the anti-PD(L)1 antibody are administered simultaneously, separately, or sequentially with chemotherapy (e.g., neoadjuvant therapy). The kit may also include a syringe.

[0138] The kit may be specified as including one or more containers (eg, single-use vials or pre-filled syringes) containing the specified pharmaceutical compositions or antibodies.

[0139] A specified amount or dose (e.g., 3000 mg, 2250 mg, or 1500 mg) can be specified to be provided in multiple vials. For example, a vial of anti-CD39 antibody can contain 375 mg of anti-CD39 antibody. A vial of durvalumab can contain 120 mg or 500 mg of durvalumab (e.g., 120 mg in 2.4 mL or 500 mg in 10 mL).

[0140] Optionally, the kit includes multiple packages of single-dose pharmaceutical compositions, each containing an effective amount of an anti-CD39 antibody and / or an anti-PD(L)1 antibody, for single administration according to the methods provided above. Instruments or devices necessary for administering the pharmaceutical compositions may also be included in the kit. For example, the kit may provide one or more pre-filled syringes containing an amount of an anti-CD39 antibody or anti-PD(L)1 antibody.

[0141] In one embodiment, the invention provides a kit for treating cancer or tumor in a human patient, the kit comprising one or more single-use vials containing an anti-CD39 antibody comprising the H-CDR1, H-CDR2 and H-CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 8 and the L-CDR1, L-CDR2 and L-CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 9, and optionally instructions for using the anti-CD39 antibody in any of the methods described herein (e.g., at the dosages and frequencies described herein).

[0142] In one embodiment, the invention provides a kit for treating cancer or tumor in a human patient, optionally wherein the cancer or tumor is NSCLC (e.g., stage II or III NSCLC, resectable NSCLC), comprising: (a) one or more vials containing an anti-CD39 antibody comprising the H-CDR1, H-CDR2, and H-CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:8, and the L-CDR1, L-CDR2, and L-CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:9; and / or (b) one or more vials containing a dose of an anti-PD-L1 antibody, optionally durvalumab; and (c) optionally, instructions for using the anti-CD39 antibody and / or the anti-PD(L)1 antibody in any of the methods described herein.

[0143] In one embodiment, each vial of anti-CD39 antibody contains 375 mg of anti-CD39 antibody. In one embodiment, the kit contains one or more sets of six vials of anti-CD39 antibody. In one embodiment, the kit contains one or more sets of eight vials of anti-CD39 antibody. In one embodiment, the kit contains four sets of six or eight vials of anti-CD39 antibody for use in neoadjuvant therapy. In one embodiment, the kit contains at least four sets (e.g., four, six, eight, ten, or twelve sets) of six or eight vials of anti-CD39 antibody for use in adjuvant therapy.

[0144] In one embodiment, the vial of durvalumab contains 500 mg of durvalumab. In one embodiment, the kit contains one or more sets of three vials of durvalumab. In one embodiment, the kit contains four sets of three vials of durvalumab for use in neoadjuvant therapy. In one embodiment, the kit contains at least four sets (e.g., 4, 6, 8, 10, or 12 sets) of three vials of durvalumab for use in adjuvant therapy.

[0145] In one embodiment, the invention provides a kit for treating cancer or tumor in a human patient, optionally wherein the cancer or tumor is NSCLC (e.g., stage II or III NSCLC, resectable NSCLC), comprising: (a) a dose of an anti-CD39 antibody comprising the H-CDR1, H-CDR2, and H-CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO: 8, and the L-CDR1, L-CDR2, and L-CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO: 9; and / or (b) the dose of the anti-PD-L1 antibody, and optionally, the dose of durvalumab; and (c) optionally, instructions for using the anti-CD39 antibody and / or the anti-PD(L)1 antibody in any of the methods described herein.

[0146] In one embodiment, the dose of the anti-CD39 antibody may be a fixed dose of 2250 mg or 3000 mg. In one embodiment, the dose of durvalumab may be a fixed dose of 1500 mg.

[0147] In one embodiment, the invention provides a kit for treating cancer or tumor, e.g., in a human patient, optionally wherein the cancer or tumor is NSCLC (e.g., stage II or III NSCLC, resectable NSCLC), comprising: (a) one or more containers (e.g., vials) containing 3000 mg or 2250 mg of an anti-CD39 antibody comprising the H-CDR1, H-CDR2, and H-CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:8, and the L-CDR1, L-CDR2, and L-CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:9; and / or (b) one or more containers (e.g., vials) containing 1500 mg of durvalumab; and (c) optionally, instructions for using the anti-CD39 antibody and / or the anti-PD(L)1 antibody in any of the methods described herein.

[0148] In any embodiment, the instructions may specify administering the anti-CD39 antibody Q3w or Q4w. In one embodiment, the instructions specify administering the anti-CD39 antibody Q3w as neoadjuvant therapy and Q4w as adjuvant therapy. In one embodiment, the instructions specify administering the anti-PD(L)1 antibody Q3 or Q4. In one embodiment, the instructions specify administering the anti-PD(L)1 antibody Q3w as neoadjuvant therapy and Q4w as adjuvant therapy. In one embodiment, the instructions specify administering the anti-CD39 antibody and the anti-PD(L)1 antibody on the same day (e.g., on day 1 of a 3-week cycle in the neoadjuvant setting, or on day 1 of a 4-week cycle in the adjuvant setting). In one embodiment, the instructions specify administering the anti-CD39 antibody and the anti-PD(L)1 antibody for 4 cycles as neoadjuvant therapy. In one embodiment, the instructions specify that the anti-CD39 antibody and the anti-PD(L)1 antibody can be administered in combination with chemotherapy in a neoadjuvant setting. The anti-CD39 antibody can optionally be identified as comprising the heavy and light chain variable region amino acid sequences of SEQ ID NOs: 8 and 9, or the heavy and light chain amino acid sequences of SEQ ID NOs: 10 and 11.

[0149] Optionally, the kit further comprises a dose of chemotherapy, for example, chemotherapy comprising a platinum drug, gemcitabine, or chemotherapy comprising the following chemotherapy drug combinations: carboplatin and paclitaxel, cisplatin and gemcitabine, cisplatin and pemetrexed, or carboplatin and pemetrexed. [Example]

[0150] Example 1: CD39 expression in early and late stage NSCLC biopsies CD39 staining was performed on 50 FFPE specimens of squamous NSCLC (sqNSCLC) and 50 adenocarcinoma NSCLC (adNSCLC) using Abcam's antibody EPR20627 clone. CD39 expression scoring (0-12) is a combination of staining frequency (0-4) and intensity (1-3).

[0151] The results are shown in Figure 1: Panel A shows that the CD39 total score is the sum of the stromal, immune, and tumor expression scores (0-60). Panel B shows that the stromal score is the sum of the vascular (0-12) and connective tissue (0-12) expression scores. Panel C shows the immune score as the sum of the small immune cell score (0-12) and large immune cell score (0-12). In each case, the expression score is indicated by the stage of cancer (stage I, II, or III).

[0152] Both sqNSCLC and adNSCLC, including early-stage biopsies, showed staining in stromal and immune cells. The overall score was higher in sqNSCLC. CD39 staining was absent or poor in tumor cells.

[0153] Example 2: ATP release from squamous NSCLC tumor cells after chemotherapy treatment This experiment aimed to investigate ATP release from squamous NSCLC tumor cells after chemotherapy treatment. The squamous NSCLC H1703 tumor cell line was separately incubated with different chemotherapies. ATP release was measured 24 hours after cisplatin treatment, 72 hours after carboplatin treatment, 60 hours after oxaliplatin treatment, 48 hours after pemetrexed treatment, 48 hours after gemcitabine treatment, 72 hours after 5-FU treatment, 24 hours after paclitaxel treatment, and 40 hours after docetaxel treatment. Extracellular ATP release in the culture supernatant was measured using a luminescence-based assay (CellTiter-Glo®) and expressed as Luminescence Arbitrary Units (AU). Mean ± SD.

[0154] The results are shown in Figure 2, which shows ATP release after treatment of the H1703 tumor cell line (sqNSCLC) with different chemotherapeutics: cisplatin, carboplatin, oxaliplatin, pemetrexed, gemcitabine, 5-Fu, paclitaxel, and docetaxel. Each chemotherapeutic drug induced significant ATP release in the NSCLC tumor cell line.

[0155] Example 3: IPH5201 was able to accumulate ATP released after chemotherapy treatment. H1703 (CD39-) cells were incubated with recombinant human CD39 protein (huCD39) (400 ng / mL) to mimic soluble CD39 in the tumor microenvironment, with or without 10 μg / mL IPH5201 (a humanized anti-CD39 antibody with heavy and light chain amino acid sequences of SEQ ID NOs: 10 and 11, respectively), and then treated with docetaxel (0.1 μM). Extracellular ATP (eATP) release was measured 40 hours after docetaxel treatment. The results are shown in Figure 3A. In H1703 cells that do not express CD39, docetaxel induced a strong release of eATP, which was reduced in the presence of added recombinant human CD39 protein and restored by the addition of IPH5201 antibody.

[0156] OAW42 (CD39+) cells were incubated with 10 μg / mL or 50 μg / mL IPH5201 and treated with a dose range of docetaxel. eATP release was measured 30 hours after docetaxel treatment. eATP was measured in cell culture supernatants using a luminescence-based assay (CellTiter-Glo®) and expressed as Luminescence Arbitrary Units (AU). Mean ± SD. Results are shown in Figure 3B. In CD39-expressing OAW42 cells, docetaxel induced at best a modest release of eATP at the highest concentration, whereas significant eATP accumulation occurred in the presence of 10 μg / mL or 50 μg / mL IPH5201.

[0157] Example 4: CD39 is expressed in the tumor microenvironment of a mouse model, and moIPH5201 significantly reduces tumor adenosine levels in vivo After assessing the expression of human CD39 in MCA205 tumors implanted in human CD39 knock-in (huCD39KI) mice, we measured intratumoral adenosine levels after in vivo treatment with the antibody moIPH5201, which has a mouse constant region containing VH and VL or SEQ ID NOs: 8 and 9 and an Fc domain with amino acid substitutions to eliminate binding to mouse Fcγ receptors, thereby assessing whether moIPH5201 can reduce intratumoral adenosine levels in vivo.

[0158] Human CD39 expression in MCA205 tumors harvested 16 days after implantation was assessed by IHC using the EPR20627 clone from Abcam B&C. MCA205 tumors were implanted into huCD39KI mice. Mice were treated with moIPH5201 (20 mg / kg) or isotype control on days 7 and 14 after tumor implantation. Tumors were excised on day 16. The results are shown in the left panel of Figure 4 and demonstrate that a significant proportion of cells express CD39.

[0159] Quantification of adenosine in harvested MCA205 tumors was performed using precolumn derivatization and liquid chromatography-mass spectrometry (LC-MS / MS) as described in Goodwin et al. 2019 Anal Biochem 568:78-88. The results are shown in the right panel of Figure 4 and demonstrate that moIPH5201-treated animals had less intratumoral adenosine compared to controls.

[0160] Blockade of CD39 enzyme activity was measured using the Wachstein-Meisel CD39 enzyme assay (brown color indicates enzyme activity). Tissues from moIPH5201-treated animals were less brown, indicating decreased CD39 enzyme activity.

[0161] Example 5: Anti-CD39 improves the antitumor efficacy of chemotherapy and anti-PD-L1 in vivo In vivo experiments were performed using human CD39 knock-in (huCD39KI) mice, which were genetically engineered to express human CD39 instead of mouse CD39. MC38 cancer cell lines were subcutaneously implanted into these mice. Mice bearing established tumors were treated with an isotype control (IC) antibody, an anti-human CD39 antibody (moIPH5201, described above), gemcitabine chemotherapy, and an anti-mouse PD-L1 antibody (an antibody that binds to mouse PD-L1 with an amino acid substitution in the Fc domain to eliminate binding to mouse Fcγ receptors).

[0162] In vivo antitumor effects of moIPH5201 and gemcitabine 1x10 huCD39KI mice of either sex 6 MC38 cancer cells were implanted subcutaneously. Treatment began after randomization. Mice were treated with gemcitabine (25 mg / kg) or PBS twice weekly for 2 weeks (days 8, 11, 15, and 18 after cell implantation) and with anti-human CD39 antibody (moIPH5201) or the corresponding isotype control (IC) antibody (400 μg / mouse) once weekly for 4 weeks.

[0163] On day 7 after tumor cell implantation, mice were randomized into four groups, and the mean tumor volume was 72 mm 3 ±27mm 3 The individual tumor volumes ranged from 37 to 130 mm 3 It was between.

[0164] The results are shown in Figure 5. Both gemcitabine alone and the combination of gemcitabine and moIPH5201 significantly and dramatically delayed tumor growth compared to the control group (p<0.001). moIPH5201 did not affect tumor growth compared to the control and did not improve the antitumor effect of gemcitabine. No complete responses were observed in this experiment. No excess toxicity was observed in either group.

[0165] In vivo antitumor effects of chemotherapy, moIPH5201, and anti-PD-L1 antibody The antitumor effects of different combinations of gemcitabine, moIPH5201, and anti-PD-L1 antibody treatment were evaluated as combined treatments. Three independent experiments were performed to evaluate the three combinations.

[0166] Experiment 1 1x10 huCD39KI mice of either sex 6 MC38 tumor cells were implanted sc. Treatment began after randomization of mice. Mice were treated with gemcitabine (25 mg / kg) or PBS twice weekly for 2 weeks, anti-mouse PD-L1 antibody or corresponding IC antibody (200 μg / mouse) twice weekly for 3 weeks, and anti-human CD39 antibody (moIPH5201) or corresponding IC antibody (400 μg / mouse) once weekly for 4 or 5 weeks depending on the experiment. Seven days after tumor cell implantation, the mean tumor volume was 62 mm 3 ±23mm 3 and individual tumor volumes of 31–117 mm 3 At some time during the study, mice were randomized into five groups.

[0167] No evidence of increased toxicity was observed in either group. One mouse in the gemcitabine + moIPH5201 group lost more than 10% of its body weight but gradually regained its initial weight after the last gemcitabine treatment.

[0168] Gemcitabine, as a single agent or in all three combinations tested, significantly delayed tumor growth compared to control (p<0.001). Neither anti-PD-L1 nor moIPH5201 improved the antitumor efficacy of gemcitabine as single agents, but the addition of both anti-PD-L1 and moIPH5201 to gemcitabine significantly improved efficacy over gemcitabine alone (p<0.05 excluding control group). Within the groups, moIPH5201 did not statistically significantly improve the efficacy of gemcitabine plus anti-PD-L1, and anti-PD-L1 did not statistically significantly improve the antitumor efficacy of gemcitabine plus moIPH5201.

[0169] The results are shown in Figure 6. Both anti-PD-L1 and moIPH5201 drugs, when combined with gemcitabine, increased the number of complete responses compared to gemcitabine alone. Furthermore, the benefit of moIPH5201 over the antitumor effect of gemcitabine + anti-PD-L1 was supported by the complete response (CR) rates: 6 / 10 (60%) in the gemcitabine + anti-PD-L1 + moIPH5201 group and 3 / 10 (30%) in the gemcitabine + anti-PD-L1 treated group. Arrows indicate the administration of different drugs.

[0170] Experiment 2 As in Experiment 1, 1x10 6 MC38 tumor cells were implanted sc. Treatment began after randomization of mice. Mice were treated with gemcitabine (25 mg / kg) or PBS twice weekly for 2 weeks, anti-mouse PD-L1 antibody or corresponding IC antibody (200 μg / mouse) twice weekly for 3 weeks, and anti-human CD39 antibody (moIPH5201) or corresponding IC antibody (400 μg / mouse) once weekly for 4 or 5 weeks depending on the experiment. Seven days after tumor cell implantation, the mean tumor volume was 64 mm 3 ±23mm 3 and individual tumor volumes of 31–115 mm 3 At some time during the study, mice were randomized into five groups.

[0171] As in previous studies, gemcitabine monotherapy and combination therapy significantly slowed tumor growth compared with controls (p<0.001), with no excess toxicity observed in either group.

[0172] Interestingly, when the untreated control group was excluded from statistical analysis, moIPH5201 tended to improve the antitumor efficacy of the gemcitabine + anti-PD-L1 combination (difference marginally significant, p<0.1). Similar to the previous experiment, the addition of both anti-PD-L1 + moIPH5201 to gemcitabine induced a superior effect than gemcitabine alone (difference marginally significant, p<0.1 when the control group was excluded).

[0173] The results are shown in Figure 7. The CR rate was 4 / 10 in the triplet treatment group compared with 3 / 10 in the gemcitabine + anti-PD-L1 treatment group. Given these results, this experiment was replicated a third time in the same setting, and the results are presented in the next section.

[0174] Experiment 3 As in experiments 1 and 2, 1x10 6 MC38 tumor cells were implanted sc. Treatment began after randomization of mice. Mice were treated with gemcitabine (25 mg / kg) or PBS twice weekly for 2 weeks, anti-mouse PD-L1 antibody or corresponding IC antibody (200 μg / mouse) twice weekly for 3 weeks, and anti-human CD39 antibody (moIPH5201) or corresponding IC antibody (400 μg / mouse) once weekly for 4 or 5 weeks depending on the experiment. Seven days after tumor cell implantation, the mean tumor volume was 74 mm 3 ±21mm 3 and individual tumor volumes of 40–121 mm 3 At some point during the study, the mice were randomized into five groups. No signs of excess toxicity were observed in any of the groups.

[0175] The results are shown in Figure 8. As in both previous experiments, gemcitabine significantly inhibited tumor growth compared to the control, both as a single agent and in all combinations tested (p<0.001). Neither anti-PD-L1 nor moIPH5201 improved the antitumor efficacy of gemcitabine as single agents; however, the addition of both anti-PD-L1 and moIPH5201 to gemcitabine induced a superior response compared to gemcitabine alone (the difference was marginally significant, p<0.1, excluding the control group). moIPH5201 did not statistically significantly improve the efficacy of gemcitabine plus anti-PD-L1. Thus, in this experiment, the combination of anti-PD-L1 with gemcitabine increased the number of complete responses compared to gemcitabine alone, whereas the combination of moIPH5201 with gemcitabine alone or gemcitabine plus anti-PD-L1 did not increase the number of CRs.

[0176] Pooled analysis Three experiments were pooled and the results are shown in Figure 9, which shows tumor growth of MC38 tumors in huCD39KI mice after treatment with gemcitabine + / - anti-PD-L1 + / - moIPH5201. MC38 tumor-bearing huCD39KI mice were randomized on day 7 and then treated with 25 mg / kg of gemcitabine or PBS ip, 200 μg of anti-mouse PD-L1 antibody or corresponding isotype control antibody ip, and 400 μg of moIPH5201 or corresponding IC antibody iv. The graph shows tumor growth for each individual (n=31 / group). CR: complete response.

[0177] A pooled experiment conducted in the MC38 tumor model (n=3), which responds to gemcitabine chemotherapy, showed that neither anti-PD-L1 nor moIPH5201 alone improved the anti-tumor efficacy of gemcitabine (not statistically significant), but the triple combination of anti-PD-L1, moIPH5201, and gemcitabine improved the anti-tumor efficacy compared to gemcitabine alone (p<0.05). Furthermore, moIPH5201 improved the anti-tumor efficacy of the gemcitabine and anti-PD-L1 combination (p<0.05; linear mixed-effects model analysis). This therapeutic efficacy was also supported by the complete response rate, which was observed in only 42% (13 / 31) of mice treated with gemcitabine and anti-PD-L1, compared to 55% (17 / 31) of mice treated with the triple combination.

[0178] In summary, IPH5201 blocks the enzymatic activity of CD39, reduces intratumoral adenosine levels, increases extracellular ATP release by tumor cells following chemotherapy treatment, and ultimately improves antitumor efficacy in preclinical models in combination with chemotherapy and a blocking anti-PDL1 antibody. Taken together, the CD39 expression profile in early-stage NSCLC and preclinical combination data support the clinical evaluation of IPH5201 in combination with durvalumab and chemotherapy in patients with early-stage NSCLC.

[0179] Example 6: Results of Phase 1 Human Clinical Trials of Anti-CD39 Antibodies and Design of New Treatment Regimen of Anti-CD39 Antibodies for Human Therapy A first-in-human, multicenter, non-randomized, open-label Phase 1 study was conducted to evaluate the safety, efficacy, pharmacokinetics (PK), and pharmacodynamics (PD) of IPH5201 ± durvalumab (Imfinzi™) in patients with advanced solid tumors. The study consisted of two consecutive dose-escalation parts: Part 1: Escalating doses of IPH5201 (100, 300, 1000, and 3000 mg) every 3 weeks (Q3W). Part 2: IPH5201 (300, 1000, and 3000 mg) and durvalumab 1500 mg Q3W escalating doses.

[0180] The PD cohort enrolled patients with advanced squamous cell lung carcinoma or advanced pancreatic ductal adenocarcinoma at the top two dose levels in Parts 1 and 2. The primary endpoints were safety and tolerability. Key secondary endpoints were preliminary antitumor activity as measured by objective response and disease control per RECIST v1.1, PK, and immunogenicity. Exploratory endpoints included efficacy as measured by duration of response and progression-free survival (PFS) per RECIST v1.1, overall survival, and biomarker assessments.

[0181] The main inclusion criteria were: · Adults aged 18 or over. Histologically or cytologically confirmed advanced solid tumors. At least one measurable lesion per RECIST v1.1 -Diseases that are refractory to standard treatment or for which no standard treatment exists. Eastern Cooperative Oncology Group Performance Status of 0 or 1. Archived or fresh tumor samples. The main exclusion criteria were: Previous treatment with any drug that targets CD73, CD39, or adenosine receptors. - Have received any conventional or investigational anti-cancer drug treatment within 21 days of the first scheduled dose. · History of autoimmune or inflammatory disorders within the past 5 years. Cardiac and vascular criteria, including the presence of acute coronary syndrome or thromboembolic event within 6 months prior to enrollment, congestive heart failure, serious arrhythmias requiring medication, or uncontrolled hypertension. ·Untreated metastasis to the central nervous system.

[0182] PK and PD modeling PK / PD modeling and simulation were used for dose selection. PK data for IPH5201 from 45 patients and PD data for free membrane CD39 (mCD39) occupancy on monocytes from 43 patients were available. A population PK model was developed, and the PK of IPH5201 was best described by a two-compartment model with parallel linear and saturable kinetics.

[0183] Next, to explain the relationship between IPH5201 concentration and free mCD39 on monocytes, we used an indirect response PD model (K in A model for the inhibition of CD39 free+ on monocytes was constructed (Figure 10). Increasing the concentration of the drug IPH5201 decreased the CD39 free+ (%) on monocytes, and the model parameters are shown in the table below. The model accurately predicted the CD39 free+ (%) on monocytes. The maximum inhibition of CD39 free+, Imax = 0.926; IC50 = 2.44 μg / mL, defines that IPH5201 produces 50% of the maximum inhibition rate. The final indirect response model was used for parametric simulations. [Table 1]

[0184] A total of 499 simulations were performed using this model to predict mCD39 occupancy on monocytes and suggest Q3W or Q4W dosing that would achieve at least 85% occupancy in the majority of patients and maintain sufficient exposure time at steady state. [Table 2]

[0185] At 3000 mg q3w, 95% of patients will achieve 85% mCD39 occupancy on monocytes for the entire dosing interval (100% of the time), and at 3000 mg q4w, 91.8% of patients will achieve 85% mCD39 occupancy at 100% of the exposure time. At 2000 mg q3w, patient coverage will reach >90%, but not at Q4W (84% patient coverage). However, at 2250 mg q4w, patient coverage achieving 85% mCD39 occupancy at 100% of the exposure time is expected to be approximately >90%.

[0186] safety Overall, 57 patients received treatment (IPH5201, n=38; IPH5201 + durvalumab, n=19). No DLTs were observed during dose escalation. Treatment-emergent adverse events (TEAEs) occurred in 96.5% of patients, with 40.4% experiencing grade 3 or higher TEAEs. The most common TEAEs were fatigue (28.1%), decreased appetite (26.3%), infusion-related reactions (21.1%), anemia (19.3%), and tumor pain (19.3%). Treatment-related adverse events (TRAEs) occurred in 66.7% of patients (37 / 57 (64.9%) related to IPH5201; 11 / 19 (57.9%) related to durvalumab). Grade 3 TRAEs occurred in 10.5% of patients, but no grade 4 TRAEs were observed. The most common TRAEs were infusion-related reactions (21.1%), fatigue (17.5%), nausea, arthralgia, tumor pain, and pruritus (8.8% each). There were no treatment-related deaths. The maximum tolerated dose (MTD) was not reached. No significant correlation was observed between dose and the incidence of adverse events of at least grade 3.

[0187] Pharmacokinetics and Pharmacodynamics CD39 occupancy by IPH5201 in samples from patients was determined by assessing free soluble CD39 and free membrane-bound CD39.

[0188] To assess free soluble CD39, a monoclonal anti-CD39 antibody that competes with IPH5201 is coated onto a microtiter plate, thereby allowing free CD39 to be captured. A second, non-competitive anti-CD39 monoclonal antibody carrying a biotin label is used to bind to a site on CD39 distinct from the capture antibody. Bound biotin molecules are detected by the addition of a streptavidin-horseradish peroxidase (SA-HRP) conjugate. Tetramethylbenzidine enzyme substrate is added to produce a colorimetric reaction, which is measured at a wavelength of 450 nm. The free CD39 concentration in the sample is calculated as 1 / Y 2 Determine by relating color intensity and CD39 concentration by interpolating from the standard curve using a 4-parameter curve fit with weight values ​​of .

[0189] To assess free CD39 on monocytes and B cells, blood samples were spiked with an antibody containing the VH and VL domains of IPH5201 and a mouse Fc domain, labeled with a fluorescent dye. A second, non-competitive anti-CD39 monoclonal antibody labeled with a different fluorescent dye was used to bind to a different site on CD39 than the capture antibody. CD14, CD19, and CD45 were detected by incubation with anti-CD14, CD19, and CD45 monoclonal antibodies labeled with different fluorescent dyes, respectively. Binding was detected by flow cytometry.

[0190] The results are shown in Figures 11A and 11B. C1D1 indicates before the first cycle of treatment, C1D2 indicates after the first cycle of treatment, C1D8 indicates day 8 of the first cycle, C2D1 indicates day 1 of the second cycle, and C3D1 indicates day 1 of the third cycle. EOT indicates end of treatment, LLOQ indicates lower limit of quantification, and V1 indicates visit 1 (screening). Figure 11A shows that IPH5201 saturated soluble CD39 binding at doses above 300 mg. Each line represents data from one patient. The increase in total soluble CD39 (not shown) is consistent with increased stabilization of soluble CD39 by antibody binding. A similar trend was observed with combination treatment with durvalumab. Figure 11B shows that IPH5201 saturated membrane-bound CD39 binding on immune cells at 3000 mg. In monocytes (right panel), administration of 100 mg IPH5201 resulted in a complete rebound of free CD39 on C2D1, and administration of 300 mg IPH5201 resulted in a partial rebound of free CD39 on C2D1. In B cells, only administration of 100 mg IPH5201 resulted in a rebound of free CD39 on C2D1.

[0191] Figure 12 shows the pharmacokinetics (PK) of IPH5201 monotherapy (left panel) or in combination with durvalumab (right panel). The X-axis represents days after first dose, and the Y-axis represents serum IPH5201 concentration. In the left panel, the curves from bottom to top represent doses of 100 mg, 300 mg, 1000 mg, and 3000 mg of IPH5201. In the right panel, the curves from bottom to top represent doses of 300 mg, 1000 mg (up to day 21), and 3000 mg of IPH5201. The PK of IPH5201 was nonlinear below 300 mg and linear above 1000 mg. In 6 / 40 patients (n=2, 100 mg; n=1, 1000 mg; n=3, 3000 mg), free soluble CD39 exceeded the lower limit of quantification on or after day 2 of the first cycle.

[0192] Tumor samples were collected from patients with pancreatic cancer and CD39 enzyme activity was assessed. CD39 enzyme activity was assessed using the Wachstein-Meisel assay to detect the presence of phosphate hydrolyzed from ATP due to CD39 enzyme activity. Enzyme activity was assessed in the tumor, stroma, and vasculature using a semiquantitative scoring method. A pathologist comprehensively assessed and determined whether there was a significant decrease at day 15 compared to the screening biopsy. The table below shows that tumor CD39 enzyme activity was decreased in five of eight patients with available samples. Administration of 3000 mg of IPH5201 demonstrated a decrease in enzyme activity. [Table 3]

[0193] In summary, PK / PD modeling and simulations predicted that dosing 3000 mg Q3W or Q4W with or without durvalumab would achieve monocyte mCD39 occupancy of 85% or greater in more than 90% of patients and maintain sufficient exposure time (i.e., 100% of the dosing interval) at steady state. Dosing of 3000 mg IPH5201 Q3W or Q4W therefore offers the advantage of reducing the risk of dosing errors and allowing same-day administration of durvalumab, both as monotherapy and in combination with durvalumab.

[0194] Example 7: Design of a Phase 2 Single-Arm Study of IPH5201 in Resectable NSCLC Based on these results, a phase 2 single-arm clinical trial of IPH5201 in resectable NSCLC was planned.

[0195] Key eligibility criteria were resectable (stage II / IIIA) NSCLC, no prior treatment, Eastern Cooperative Oncology Group (ECOG) performance status (PS) of 0 or 1, any CPS status with PDL1 expression, and EGFR / ALK wild-type.

[0196] The primary endpoints are pathological complete response (pCR) and safety. Secondary endpoints include feasibility of surgery, major pathological response (mPR), objective response rate (ORR), event-free survival (EFS), disease-free survival (DFS), median and landmark overall survival (OS) (12 months), as well as evaluation of PK and anti-drug antibodies (ADA) of IPH5201. In this study, a safety study will be conducted on the first six patients.

[0197] Figure 13 shows a schematic diagram of the treatment regimen. For neoadjuvant therapy, IPH5201 and durvalumab are combined with chemotherapy (CT), with a fixed dose of 3000 mg of IPH5201 (alternatively, 2250 mg) and 1500 mg of durvalumab administered Q3W for four cycles. CT includes carboplatin / paclitaxel, cisplatin / gemcitabine, cisplatin / pemetrexed, or carboplatin / pemetrexed. Within 40 days of the final dose of study drug in neoadjuvant therapy, patients undergo surgical tumor resection with or without additional radiation therapy. Within 10 weeks after surgery, patients will receive adjuvant therapy with IPH5201 and durvalumab Q4w for up to 12 cycles or until progressive disease, at a fixed dose of 3000 mg IPH5201 (2250 mg if used alternatively) and 1500 mg durvalumab.

[0198] All references cited herein, including publications, patent applications, and patents, are herein entirely incorporated by reference to the same extent as if each reference was individually and specifically indicated to be incorporated by reference and were set forth in its entirety herein (to the maximum extent permitted by law), notwithstanding any individual provided incorporation of any specific reference elsewhere herein.

[0199] The use of the words "a," "an," and "the" and similar referents should be construed to encompass both the singular and the plural unless otherwise indicated herein or clearly contradicted by context.

[0200] Unless otherwise specified, all exact values ​​provided herein represent the corresponding approximate values ​​(e.g., all representative exact values ​​or measurements provided with respect to a particular factor can also be considered to provide the corresponding approximate measurements, modified by "about" as appropriate).

[0201] Any description herein of any aspect or embodiment using words such as "comprising," "having," "including," or "containing" in reference to one or more elements is intended to provide support for similar aspects or embodiments herein that "consist," "consist essentially of," or "substantially comprise" that particular element or elements, unless otherwise specified or clearly contradicted by the context (e.g., a composition described herein as comprising a particular element should also be understood to describe a composition consisting of that element, unless otherwise specified or clearly contradicted by the context).

[0202] The use of any examples or representative language (e.g., "etc.") provided herein is intended merely to better clarify the invention and does not pose a limitation on the scope of the invention unless otherwise stated. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

Claims

1. 1. A method of treating cancer in a patient in need thereof, comprising: administering to the patient an effective amount of a treatment regimen comprising an anti-CD39 antibody and an anti-PD(L)1 antibody; optionally, the anti-CD39 antibody comprises (a) the HCDR1, HCDR2, and HCDR3 sequences of SEQ ID NO:2, SEQ ID NO:3, and SEQ ID NO:4, respectively, and (b) the LCDR1, LCDR2, and LCDR3 sequences of SEQ ID NO:5, SEQ ID NO:6, and SEQ ID NO:7, respectively; The method wherein the treatment regimen is neoadjuvant therapy.

2. 10. The method of any of the above claims, wherein the neoadjuvant therapy treatment regimen further comprises a chemotherapeutic agent.

3. 3. The method of claim 1 or 2, wherein the treatment regimen is neoadjuvant therapy and adjuvant therapy.

4. 4. The method of claim 1, 2, or 3, wherein the anti-CD39 antibody is administered at a fixed dose of 3000 mg.

5. The method of any one of claims 1 to 4, wherein the anti-CD39 antibody is administered as neoadjuvant therapy once every three weeks.

6. The method according to any one of claims 1 to 5, wherein the anti-CD39 antibody and the anti-PD(L)1 antibody are each administered once every three weeks as neoadjuvant therapy.

7. 10. The method of any of the preceding claims, wherein the anti-CD39 antibody and the anti-PD(L)1 antibody are administered as neoadjuvant therapy on day 1 of a 3-week cycle for 4 cycles.

8. The method of any one of claims 3 to 7, wherein the anti-CD39 antibody is administered as an adjuvant therapy once every four weeks.

9. The method according to any one of claims 3 to 7, wherein the anti-CD39 antibody and the anti-PD(L)1 antibody are each administered once every four weeks as adjuvant therapy.

10. The method of any one of claims 3 to 9, wherein the anti-CD39 antibody is administered as an adjuvant therapy in a fixed dose of 3000 mg.

11. 10. The method of any of the preceding claims, wherein the anti-CD39 antibody and the anti-PD(L)1 antibody are administered as neoadjuvant therapy on day 1 of a 3-week cycle and as adjuvant therapy on day 1 of a 4-week cycle.

12. The method according to claims 3 to 11, wherein the amount of anti-CD39 antibody per dose in the neoadjuvant therapy and the adjuvant therapy is the same.

13. The method according to claims 3 to 12, wherein the amount of anti-PD(L)1 antibody per dose in the neoadjuvant therapy and the adjuvant therapy is the same.

14. The treatment regimen comprises: (a) administering to the patient effective amounts of an anti-CD39 antibody, an anti-PD(L)1 antibody, and optionally a chemotherapeutic agent prior to surgical tumor resection; (b) administering to the patient effective amounts of an anti-CD39 antibody and an anti-PD(L)1 antibody following surgical tumor resection; 10. The method of any preceding claim, comprising:

15. 10. The method of any of the above claims, wherein the anti-PD(L)1 antibody is durvalumab.

16. 16. The method of claim 15, wherein durvalumab is administered at a fixed dose of 1500 mg.

17. A method of administering to a human patient an anti-CD39 antibody comprising heavy and light chains comprising the amino acid sequences of SEQ ID NOs: 8 and 9, respectively, wherein the anti-CD39 antibody is administered at a fixed dose of 3000 mg once every three weeks or once every four weeks.

18. 18. The method of claim 17, wherein the anti-CD39 antibody is administered on day 1 of a three-week cycle for one or more cycles and on day 1 of a four-week cycle for one or more cycles.

19. anti-CD39 is administered in combination with an anti-PD(L)1 antibody; optionally, the anti-PD(L)1 antibody is administered on day 1 of one or more 3-weekly cycles and on day 1 of one or more 4-weekly cycles; optionally, the anti-PD(L)1 antibody is durvalumab; In some cases, durvalumab is administered at a fixed dose of 1500 mg; 19. The method of claim 17 or 18.

20. 10. The method of any of the above claims, wherein the anti-CD39 antibody comprises a heavy chain comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO: 10, and a light chain comprising an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO:

11.

21. 10. The method of any of the above claims, wherein the anti-CD39 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 and a light chain comprising the amino acid sequence of SEQ ID NO:

11.

22. 10. The method of any of the above claims, wherein the anti-PD(L)1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 14 and a light chain comprising the amino acid sequence of SEQ ID NO:

15.

23. 10. The method of any of the above claims, wherein the cancer is surgically resectable.

24. 10. The method of any of the above claims, wherein the cancer is lung cancer.

25. 10. The method of any of the above claims, wherein the cancer is non-small cell lung cancer.

26. 10. The method of any of the above claims, wherein the cancer is stage I-III NSCLC.

27. 10. The method of any of the preceding claims, wherein the patient has not received prior treatment for cancer.

28. The method of claims 24 to 27, wherein the cancer is EGFR / ALK wild-type.

29. 1. A kit for treating cancer or tumors in a human patient, comprising: (a) one or more vials containing an anti-CD39 antibody comprising the H-CDR1, H-CDR2, and H-CDR3 domains of a heavy chain variable region having the sequence set forth in SEQ ID NO:8, and the L-CDR1, L-CDR2, and L-CDR3 domains of a light chain variable region having the sequence set forth in SEQ ID NO:9; and / or (b) one or more vials containing durvalumab, and (c) optionally instructions for using said anti-CD39 antibody and / or durvalumab.

30. 30. The kit of claim 29, wherein each vial of anti-CD39 antibody contains 375 mg of anti-CD39 antibody.

31. 31. The kit of claim 29 or 30, wherein each vial of durvalumab contains 500 mg of durvalumab.

Images