Compositions and methods comprising anti-CD39 antibodies for neoadjuvant treatment of cancer

Abstract

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

Description

[0001] Cross - Reference to Related Applications

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 385,628, filed on December 1, 2022; the entire disclosure of the U.S. Provisional Application is incorporated herein by reference; including any drawings.

[0003] Reference to Sequence Listing

[0004] This application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file named "INN.276XPCT.xml", which was created on November 13, 2023 and is 28,938 bytes. The information in the electronic format of the Sequence Listing is incorporated herein by reference in its entirety. Technical Field

[0005] The present invention relates to antibodies that inhibit the activity of human CD39 enzyme and methods of treating cancer using compounds. Background Art

[0006] Eight different ENTPD genes encode members of the NTPDase protein family. The cellular location and functional properties of each NTPDase subtype are different. Plasma membrane - bound nucleoside triphosphate diphosphohydrolases control the nucleotide levels at the cell surface by hydrolyzing the c and b phosphates of nucleotides. NTPDase1 (ecto - nucleoside triphosphate diphosphohydrolase 1), also known as CD39 / ENTPD1 or vascular CD39, acts together with another enzyme, CD73 (ecto - 5′ - nucleotidase), to hydrolyze extracellular adenosine triphosphate (ATP) and 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. Generation of adenosine through the CD73 / CD39 pathway is considered to be the major mechanism for the immunosuppressive function of regulatory T cells (Tregs). CD39 + The number of Tregs is increased in some human cancers, and CD39 + The importance of Tregs in promoting tumor growth and metastasis has been demonstrated using several in - vivo models. However, CD39 is also expressed by tumor cells, and CD39 + Tumor cells can mediate immunosuppression through the adenosine pathway. CD39 in cancer cells exhibits ATPase activity and, together with CD73, generates adenosine. CD73 + CD39 + Cancer cells inhibit the proliferation of CD4 and CD8 T cells and the generation of cytotoxic effector CD8 T cells (CTLs) in a CD39 - dependent and adenosine - dependent manner.

[0007] Increased CD39 expression has been reported in several solid tumors (colorectal cancer, head and neck cancer, pancreatic cancer) as well as chronic lymphocytic leukemia. Antibodies that bind to and inhibit CD39 enzymatic activity are disclosed, for example, in WO2018 / 167267, WO2019 / 243252, WO2019 / 178269, WO2019 / 127935, and WO2021 / 037037.

[0008] In many cancer cases, the disease may be determined to be surgically resectable at diagnosis or may become surgically resectable after initial radiotherapy or chemotherapy. Surgery often provides the best chance of cure for many tumors.

[0009] Approximately 1.5 million new cases of lung cancer are diagnosed worldwide each year, of which approximately 85% are non-small cell lung cancer (NSCLC). Surgery is considered the best treatment option, but only about 20%-30% of NSCLC patients have 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 the standard practice to reduce the risk of disease recurrence. In the case of limited disease (stage I, II, IIIA), patients who undergo surgical resection and receive chemotherapy achieve a 5-year survival rate of 51%, with an absolute benefit in 5-year survival of 5.4%, especially in patients with good performance status (PS) (Provencio et al. 2011 J Thorac Dis. 3(3): 197-204). There is evidence that identification of the minimal residual disease (MRD) status of patients by detecting postoperative circulating tumor DNA (ctDNA) can accurately predict disease recurrence. Durvalumab has been proposed to be particularly beneficial in patients in the high-risk NSCLC patient population, in whom MRD is detected via ctDNA isolation after complete resection. In addition, in some cases, neoadjuvant chemotherapy is used. Burdett S. Lancet. 2014; 383: 1561-1571 concluded that in stages IB-IIIA, preoperative chemotherapy significantly improves the overall survival, distant recurrence time, and recurrence-free survival of resectable NSCLC. Such preoperative neoadjuvant chemotherapy has been proposed to have the potential to reduce tumor size, increase resectability, and eradicate micrometastases. Neoadjuvant chemotherapy may also be more effective when the tumor blood supply remains intact before surgical resection, and chemotherapy may be better tolerated if the patient does not recover from major surgery. Some clinical trials have evaluated chemotherapy administered only preoperatively, and five trials have evaluated chemotherapy preoperatively and then postoperatively, usually in responders (see Burnett 2014, ibid.), however, the conclusion is that the potential benefits of neoadjuvant chemotherapy will need to be balanced against possible toxic effects.

[0010] There is still a need in the art to improve the treatment of cancer. Many patients with locally advanced disease (stage III) have disease that is considered unresectable at diagnosis and would like to make the disease more operable or resectable. For patients with stage II-IIIA and selected stage IIIB disease, although the disease is resectable, surgery and adjuvant standard of care (SoC) chemotherapy result in a 5-year disease-free survival (DFS) rate of only -40% (Wakelee et al., Lancet. Oncol. 18(12): 1610-23 (2017). SUMMARY OF THE INVENTION

[0011] The present disclosure provides methods for treating cancer and / or preventing cancer recurrence in a patient in need thereof, such as in stage I-III NSCLC, optionally stage 1B-IIIA NSCLC, optionally stage II or IIIA NSCLC. The present disclosure also provides dosing regimens of anti-CD39 antibody and anti-PD(L)1 antibody that are suitable for such treatment, such as in resectable tumors or cancers.

[0012] The present disclosure provides a method for treating a tumor or cancer and / or preventing recurrence of a tumor or cancer in a patient in need thereof, the method 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 (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, the anti-PD(L)1 antibody, and the chemotherapeutic agent are administered as neoadjuvant therapy. The present disclosure also provides a method for treating a tumor or cancer and / or preventing recurrence of a tumor or cancer in a patient in need thereof, the method 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 further as adjuvant therapy. The treatment regimen of the present disclosure can be characterized as comprising: (a) administering an anti-CD39 antibody and an anti-PD(L)1 antibody (and optionally further administering an effective amount of chemotherapy) to a patient before tumor surgical resection or where the patient has not undergone surgical resection; and (b) administering an anti-CD39 antibody and an anti-PD(L)1 antibody to the patient after tumor surgical resection. In one embodiment, the tumor or cancer is characterized as a tumor or cancer that is considered resectable. In one embodiment, the tumor or cancer is characterized as having the potential to become resectable (such as locally advanced and / or stage IIIB NSCLC).

[0013] In one embodiment, the anti-CD39 antibody is an antibody comprising the amino acid sequences of SEQ ID NOS: 2-7, optionally the antibody comprises the amino acid sequences of SEQ ID NOs: 8 and 9, optionally the antibody comprises the amino acid sequences of SEQ ID NOs: 10 and 11. In one embodiment, the anti-PD(L)1 antibody is durvalumab.

[0014] These methods are particularly advantageous in the treatment of lung cancer, particularly non-small cell lung cancer (NSCLC). In any embodiment, the cancer may optionally be characterized as resectable NSCLC. In one embodiment, the cancer is stage II or IIIA NSCLC, such as resectable stage II or IIIA NSCLC. In one embodiment, the cancer is stage I, II, or IIIA NSCLC. In another aspect, optionally the cancer may be characterized as unresectable and / or locally advanced NSCLC (e.g., stage III or IIIB NSCLC).

[0015] In one embodiment, the treatment regimens herein provide for administration of the anti-CD39 antibody at the same dose (e.g., 2250 mg or 3000 mg) in the adjuvant and neoadjuvant settings. In one embodiment, the treatment regimens herein allow for administration of the anti-CD39 antibody and the anti-PD(L)1 antibody at their respective same 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 the adjuvant and neoadjuvant settings. Additionally, these regimens allow for administration of the anti-CD39 antibody (and further administration of the anti-PD(L)1 antibody) every three weeks as a neoadjuvant and every four weeks as an adjuvant (at the same dose).

[0016] In one embodiment, a method of administering an anti-CD39 antibody comprising the amino acid sequence of SEQ ID NO: 2-7, 8-9, or 10-11 is provided, wherein the antibody is administered at a dose of 3000 mg Q3w or Q4w. In one embodiment, a method of administering an anti-CD39 antibody comprising the amino acid sequence of SEQ ID NO: 2-7, 8-9, or 10-11 is provided, wherein the antibody is administered at a dose of 2250 mg Q3w or Q4w.

[0017] In one embodiment, a method for administering an anti-CD39 antibody comprising the amino acid sequence of SEQ ID NO: 2-7, 8-9, or 10-11 is provided, wherein the antibody is (a) administered on day 1 of a 3-week cycle for one or more cycles and (b) administered on day 1 of a 4-week cycle for one or more cycles, wherein in each case the antibody is administered at a dose of 2250 mg or 3000 mg. Optionally, the method is a method of 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.

[0018] In one embodiment, a method for 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 (a) administered on day 1 of a 3-week cycle for one or more cycles and (b) administered on day 1 of a 4-week cycle for one or more cycles, wherein 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.

[0019] In one embodiment, a method of treating cancer or preventing cancer recurrence is provided, the method 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 administering the anti-CD39 antibody and the anti-PD(L)1 antibody on day 1 of a 3-week cycle for one or more cycles, and wherein the adjuvant therapy comprises administering the anti-CD39 antibody and the anti-PD(L)1 antibody on day 1 of a 4-week cycle, wherein 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 4 cycles. In one embodiment, the neoadjuvant therapy comprises at least 2, 4, 8 or 12 cycles, and / or up to 12 cycles.

[0020] These aspects are more fully described in the specification provided herein, and additional aspects, features and advantages will be apparent from the specification provided herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 CD39 staining by immunohistochemistry is shown for 50 squamous cell NSCLC (sqNSCLC) and 50 adenocarcinoma NSCLC (adNSCLC) FFPE samples. CD39 expression scores are based on staining frequency and intensity. Staining is shown for stromal expression, immune cell expression, and total expression. No CD39 staining or poor CD39 staining was observed on tumor cells. Figure A shows the CD39 total score as the sum of stromal, immune, and tumor expression scores (0 - 60). Figure B shows the stromal score as the sum of vascular (0 - 12) and connective tissue (0 - 12) expression scores. Figure C shows the immune score as the sum of small immune cell (0 - 12) and large immune cell (0 - 12) scores. In each case, the expression scores are shown by cancer stage (stage I, II, or III).

[0022] Figure 2 Shown is the amount of ATP released from squamous NSCLC tumor cells when the tumor cells were incubated with different chemotherapies alone.

[0023] Figure 3 A shows the amount of extracellular ATP (eATP) released by H1703 (CD39-) cells incubated with recombinant huCD39 that mimics soluble CD39 in the tumor microenvironment with or without anti-CD39 antibody (IPH5201) and then treated with docetaxel. Docetaxel induces a strong release of eATP, which is reduced in the presence of added recombinant human CD39 protein and then restored in the presence of added IPH5201 antibody.

[0024] Figure 3 B shows the amount of eATP released in OAW42 (CD39+) cells incubated with anti-CD39 antibody (IPH5201) and treated with a range of doses of docetaxel. In these CD39-expressing cells, docetaxel induced at most modest eATP release at the highest concentration, whereas eATP accumulated significantly in the presence of 10 μg / mL or 50 μg / mL of IPH5201.

[0025] Figure 4 The left and right panels show the percentage of MCA205 tumor cells expressing CD39 and quantification of adenosine, respectively, after MCA205 tumors were transplanted into human CD39 knock-in mice, treated with anti-CD39 antibody (moIPH5201) or isotype control, and then harvested for CD39 expression analysis. A significant percentage of cells expressed CD39 (left panel), and moIPH5201-treated animals showed less intratumoral adenosine compared to controls (left panel).

[0026] Figure 5 Shown is an experiment evaluating MC38 tumor growth in a cohort of huCD39 knock-in mice following treatment with gemcitabine + / - anti-CD39 antibody (moIPH5201).

[0027] Figure 6 , Figure 7 and Figure 8 Each shows an experiment evaluating MC38 tumor growth in a cohort of huCD39 knock-in mice after treatment with gemcitabine + / - anti-PD-L1 antibody + / - anti-CD39 antibody (moIPH5201).

[0028] Figure 9 Shown is a pooled analysis of three series of experiments evaluating MC38 tumor growth in huCD39 knock-in mice following treatment with gemcitabine + / - anti-PD-L1 antibody + / - anti-CD39 antibody (moIPH5201).

[0029] Figure 10 shows the establishment of an indirect response PD model for describing the relationship between IPH5201 concentration and free mCD39 on monocytes (inhibition of K in ).

[0030] Figure 11A Shows that the anti-CD39 antibody (IPH5201) saturates the binding of soluble CD39 at ≥ 300 mg in human patients. Each line represents data from one patient. The increase in total soluble CD39 (not shown) is consistent with the increase in soluble CD39 bound by antibody. A similar trend was observed in combination therapy with durvalumab.

[0031] Figure 11B Shows that IPH5201 saturates the binding of membrane-bound CD39 on immune cells from samples of human patients treated with 3000 mg anti-CD39 antibody (IPH5201).

[0032] Figure 12 Shows the pharmacokinetics (PK) of IPH5201 as monotherapy (left panel) or as combination therapy with durvalumab (right panel) in human patients, where the time in days after the first dose is on the x-axis and the serum IPH5201 concentration is on the y-axis. The curves from bottom to top represent fixed doses of 100 mg, 300 mg, 1000 mg, and 3000 mg of IPH5201. The PK of IPH5201 is non-linear at ≤ 300 mg and linear at ≥ 1000 mg.

[0033] Figure 13 Shows a schematic of the treatment regimen for a human clinical trial. As neoadjuvant therapy, IPH5201 and durvalumab are administered in combination with chemotherapy (CT) for 4 cycles at a fixed dose of 3000 mg IPH5201 and a fixed dose of 1500 mg durvalumab Q3W. As adjuvant therapy, IPH5201 and durvalumab are administered at a fixed dose of 3000 mg IPH5201 and a fixed dose of 1500 mg durvalumab Q4W for up to 12 cycles. Detailed Description

[0034] Definition

[0035] When using "comprising", this may optionally be replaced by "consisting essentially of" or "consisting of".

[0036] Human CD39, also known as "vascular" CD39, NTPdasel, ENTPD1, ATPDase, and vascular ATP diphosphohydrolase, exhibits ATPase activity. CD39 hydrolyzes extracellular ATP and ADP to AMP, which is further converted to adenosine by another enzyme, 5'-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:

[0037]

[0038] In the context of this disclosure, when referring to a CD39 polypeptide, "neutralize" or "neutralizing" (e.g., "neutralize CD39", "neutralize the activity of CD39", or "neutralize the enzymatic activity of CD39") refers to a process in which the ATP hydrolysis (ATPase) activity of CD39 is inhibited. This particularly includes inhibiting the generation of AMP and / or ADP mediated by CD39, i.e., inhibiting the catabolism of ATP to AMP and / or ADP mediated by CD39. For membrane-bound CD39, this can be measured, for example, in a cell assay that directly or indirectly measures the ability of a test compound to inhibit the conversion of ATP to AMP and / or ADP. For soluble CD39, this can be measured by incubating recombinant soluble CD39 as described herein with a test compound and directly or indirectly measuring the conversion of ATP to AMP and / or ADP. For example, the disappearance of ATP and / or the generation of AMP can be evaluated as described herein, e.g., by quantifying luminescence units proportional to the amount of ATP present. In one embodiment, an antibody preparation results in at least a 60% reduction, at least a 70% reduction, or at least an 80% or 90% reduction in the conversion of ATP to AMP, e.g., with reference to the assays described herein (e.g., the disappearance of ATP and / or the generation of AMP).

[0039] Whenever reference is made to "treatment of cancer", etc., in connection with an anti-CD39 binder (e.g., an antibody), this can include: (a) a method of treating cancer, the method comprising the step of administering (for at least one treatment) an anti-CD39 binder (preferably in a pharmaceutically acceptable carrier material) in a dose (therapeutically effective amount) that permits treatment of cancer, preferably in a dose (amount) as specified herein, to an individual, a mammal, especially a human, in need of such treatment; (b) the use of an anti-CD39 binder for the treatment of cancer, or an anti-CD39 binder for such treatment (especially in humans); (c) the use of an anti-CD39 binder for the manufacture of a pharmaceutical preparation for the treatment of cancer, a method of manufacturing a pharmaceutical preparation for the treatment of cancer using an anti-CD39 binder (optionally including mixing the anti-CD39 binder with a pharmaceutically acceptable carrier), or a pharmaceutical preparation comprising an effective dose of an anti-CD39 binder suitable for the treatment of cancer; or (d) any combination of a), b), and c) according to the subject matter that permits patent authorization in the country where this application is filed.

[0040] As used herein, the term "antigen-binding domain" refers to a domain that includes a three-dimensional structure capable of immunospecifically binding to an epitope. Thus, in one embodiment, the domain can include hypervariable regions, optionally the VH and / or VL domains of an antibody chain, optionally at least the VH domain. In another embodiment, the binding domain can include at least one complementarity-determining region (CDR) of an antibody chain. In another embodiment, the binding domain can include a polypeptide domain from a non-immunoglobulin scaffold.

[0041] As used herein, the term "antibody" can include polyclonal and monoclonal antibodies. Antibodies are assigned to one of five main classes based on the type of constant domain in the heavy chain: IgA, IgD, IgE, IgG, and IgM. Some of these are further divided into subclasses or isotypes, such as IgG1, IgG2, IgG3, IgG4, etc. Exemplary immunoglobulin (antibody) structural units include tetramers. Each tetramer is composed of two pairs of identical polypeptide chains, each pair having one "light" chain (about 25 kDa) and one "heavy" chain (about 50 - 70 kDa). The N-terminus of each chain defines a variable region of about 100 to 110 or more amino acids that is primarily responsible for antigen recognition. The term variable light chain (V L ) and variable heavy chain (V HThese refer to the light and heavy chains, respectively. The constant domains of the heavy chains corresponding to the different classes of immunoglobulins are designated "α", "δ", "ε", "γ", and "μ", respectively. The subunit structures and three-dimensional configurations of the different classes of immunoglobulins are well known. IgG is an exemplary class of antibodies used herein because they are the most commonly used antibodies in physiological situations and because they are the most easily prepared in a laboratory setting. Optionally, the antibody is a monoclonal antibody. Specific examples of antibodies are humanized, chimeric, human, or otherwise human-compatible antibodies. "Antibody" also includes any fragment or derivative of any of the antibodies described herein.

[0042] The term "specifically binds" means that an antibody can preferably bind to a binding partner, such as CD39, in a competitive binding assay, as evaluated using a protein, a recombinant form of the epitope, or the native protein present on the surface of isolated target cells. Competitive binding assays and other methods for determining specific binding are further described below and are well known in the art.

[0043] When an antibody "competes" 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), it means that the antibody competes with the monoclonal antibody in a binding assay using a recombinant CD39 molecule or a CD39 molecule expressed on the surface. For example, if a test antibody reduces the binding of a reference antibody to a CD39 polypeptide or a CD39-expressing cell in a binding assay, the antibody is said to "compete" with the reference antibody accordingly.

[0044] As used herein, the term "affinity" refers to the strength of binding of an antibody to an epitope. The affinity of an antibody is given by the dissociation constant Kd, defined as [Ab] × [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 K aDefined by 1 / Kd. Methods for determining mAb affinity are described in Harlow et al., Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1988), Coligan et al., eds., Current Protocols in Immunology, Greene Publishing Assoc. and Wiley Interscience, N.Y., (1992, 1993), and Muller, Meth. Enzymol. 92:589-601 (1983), the entire disclosures of which are incorporated herein by reference. A standard method well known in the art for determining the affinity of mAbs is surface plasmon resonance (SPR) screening (such as by analyzing with a BIAcore TM SPR analysis device).

[0045] In the context of this disclosure, a "determinant" refers to a site on a polypeptide that interacts or binds.

[0046] The term "epitope" refers to an antigenic determinant and is the area or region on an antigen to which an antibody binds. A protein epitope can include amino acid residues directly involved in binding as well as amino acid residues that are effectively blocked by a specific antigen-binding antibody or peptide, i.e., amino acid residues within the "footprint" of the antibody. It is the simplest form or smallest structural region on a complex antigen molecule that can combine with, for example, an antibody or a receptor. Epitopes can be linear or conformational / structural. The term "linear epitope" is defined as an epitope composed of contiguous amino acid residues on a linear amino acid sequence (primary structure). The term "conformational or structural epitope" is defined as an epitope composed of amino acid residues that are not all contiguous and thus represent separate portions of a linear amino acid sequence that are brought into proximity to one another by molecular folding (secondary, tertiary, and / or quaternary structure). Conformational epitopes depend on three-dimensional structure. Thus, the term "conformational" is generally used interchangeably with "structural".

[0047] The term "agent" is used herein to denote a chemical compound, a mixture of chemical compounds, a biological macromolecule, or an extract prepared from a biological material. The term "therapeutic agent" refers to an agent having biological activity.

[0048] For the purposes of this disclosure, a "humanized" antibody is an antibody in which one or more constant and variable framework regions of human immunoglobulins are fused to the binding regions, e.g., CDRs, of an animal immunoglobulin. Such antibodies are designed to maintain the binding specificity of the non-human antibody from which the binding regions are derived while avoiding an immune response against the non-human antibody.

[0049] As used herein, the term "hypervariable region" refers to the amino acid residues of an antibody that are responsible for antigen binding. Hypervariable regions generally contain amino acid residues from "complementary determining regions" or "CDRs" (e.g., residues 24-34 (L1), 50-56 (L2), and 89-97 (L3) in the light chain variable domain and 31-35 (H1), 50-65 (H2), and 95-102 (H3) in the heavy chain variable domain; Kabat et al., 1991) and / or those residues from "hypervariable loops" (e.g., residues 26-32 (L1), 50-52 (L2), and 91-96 (L3) in the light chain variable domain and 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 a similar system for determining the essential amino acids responsible for antigen binding. Generally, the numbering of amino acid residues in this region is carried out by the method described by Kabat et al., supra. Phrases such as "Kabat position", "residue numbering in the variable domain as in Kabat", and "according to Kabat" herein refer to this numbering system for the heavy chain variable domain or the light chain variable domain. Using the Kabat numbering system, the actual linear amino acid sequence of a peptide may contain fewer or additional amino acids corresponding to deletions or insertions into the FR or CDR of the variable domain. For example, the heavy chain variable domain may contain a single amino acid insertion after residue 52 of CDR H2 (residue 52a according to Kabat) and residues inserted after heavy chain FR residue 82 (e.g., residues 82a, 82b, and 82c, etc. according to Kabat). The Kabat numbering of the residues of a given antibody can be determined by aligning the sequence of the antibody with the "standard" Kabat numbering sequence at the homologous regions.

[0050] As used herein, the term "framework" or "FR" residues means the regions of an antibody variable domain other than those regions defined as CDRs. Each antibody variable domain framework can be further subdivided into contiguous regions (FR1, FR2, FR3, and FR4) separated by these CDRs.

[0051] The terms "Fc domain", "Fc portion", and "Fc region" refer to the C-terminal fragment of an antibody heavy chain, e.g., approximately amino acids (aa) 230 to approximately aa 450 from a human gamma (γ) heavy chain, or the corresponding sequence in other types of antibody heavy chains (e.g., alpha, delta, epsilon, and mu of human antibodies), or naturally occurring allotypes thereof. Unless otherwise specified, the well-known 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).

[0052] The terms "isolated", "purified", or "biologically pure" refer to a substance that is substantially or essentially free from components that are normally associated with it 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 major species present in a preparation is substantially purified.

[0053] The terms "polypeptide", "peptide", and "protein" are used interchangeably herein and refer to a polymer of amino acid residues. The term applies to amino acid polymers in which one or more amino acid residues are artificial chemical mimics of the corresponding naturally occurring amino acids, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers.

[0054] When used in reference to, for example, a cell or a nucleic acid, protein (e.g., an antibody or antibody fragment), or vector, the term "recombinant" indicates that the cell, nucleic acid, protein, or vector has been modified by the introduction of heterologous nucleic acid or protein or by a change to a native nucleic acid or protein, or the cell is derived from a cell so modified. Thus, for example, a recombinant cell expresses a gene not found in a cell in its native (non-recombinant) form or expresses a native gene that is otherwise abnormally expressed, under-expressed, or not expressed at all.

[0055] In the context of this disclosure, the term antibody that "binds" a polypeptide or epitope designates an antibody that binds the determinant specifically and / or with affinity.

[0056] The term "identity" or "identical," when used in reference to the relationship between the sequences of two or more polypeptides, refers to the degree of sequence relatedness between the polypeptides, as determined by the number of matches between the chains of two or more amino acid residues. "Identity" measures the percentage of identical matches between the smaller sequences of two or more sequences with gap alignments (if any) resolved by a specific 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 the following references: Computational Molecular Biology, Lesk, A.M. ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D.W. ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part 1, Griffin, A.M. and Griffin, H.G. 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).

[0057] The method for determining identity is designed to give the maximum match between the sequences being tested. Methods for determining identity are described in publicly available computer programs. Computer program methods for determining the 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 publicly 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., ibid.). The well-known Smith Waterman algorithm can also be used to determine identity.

[0058] Treatment of cancer

[0059] The present disclosure generally relates to methods of treating cancer in a patient using a combination of an anti-CD39 antibody and an anti-PD(L)1 antibody as neoadjuvant therapy and optionally further as adjuvant therapy. Thus, these treatments can be used as preoperative therapy in patients with resectable or potentially resectable or likely to become resectable cancer or tumors, and optionally further as postoperative therapy. Optionally, the use as preoperative therapy is further combined with postoperative chemotherapy. These methods are 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, such as resectable stage II or IIIA NSCLC. Thus, these methods are particularly advantageous in the treatment of tumors or cancers where surgery is the primary treatment.

[0060] In one embodiment, a method of reducing or inhibiting tumor growth in a subject in need thereof is disclosed, the method comprising administering to the subject a therapeutically effective amount of each of an anti-CD39 antibody, an anti-PD(L)1 antibody, and an optional chemotherapeutic agent. In one embodiment, a method of treating cancer, particularly stage II or IIIA NSCLC, in a subject in need thereof is disclosed, the method comprising administering to the subject a therapeutically effective amount of each of an anti-CD39 antibody, an anti-PD(L)1 antibody, and an optional chemotherapeutic agent. In one embodiment, the anti-CD39 antibody, the anti-PD(L)1 antibody, and the optional chemotherapeutic agent are administered as neoadjuvant therapy. In one embodiment, the anti-CD39 antibody and the anti-PD(L)1 antibody are administered as neoadjuvant and adjuvant therapy.

[0061] The methods and regimens herein can be used, for example, for treating and / or preventing tumor or cancer recurrence in a patient, for preventing metastasis or recurrence of metastasis, for preventing tumor or cancer progression or growth, for preventing tumor or cancer progression or growth, and / or for improving or increasing survival, time to recurrence (e.g., distant recurrence), and / or recurrence-free survival. The patient can optionally be designated as having resectable cancer or tumor. The methods and regimens herein can also be used to make a tumor more surgically viable and / or to increase the likelihood of complete resection. Optionally, the methods herein are characterized as methods of improving, for example, the likelihood of having no residual disease (e.g., minimal residual disease MRD) and / or being ctDNA negative (no detectable ctDNA) after surgical resection. The methods and regimens herein can also be used, for example, to increase or enhance an anti-tumor immune response, inhibit the enzymatic activity of CD39 (e.g., within a tumor), increase the intratumoral concentration of ATP, decrease the intratumoral concentration of adenosine, increase the activity and / or activation of T cells, NK cells, tumor-infiltrating NK cells, tumor-infiltrating T cells, and / or dendritic cells.

[0062] The anti-CD39 antibodies used herein are antibodies that neutralize CD39. When referring to a CD39 polypeptide, "neutralize" (e.g., "neutralize CD39", "neutralize the activity of CD39", or "neutralize the enzymatic activity of CD39") refers to the process in which the ATP hydrolysis (ATPase) activity of CD39 is inhibited. Such antibodies have been reported in several publications (cited supra), and the disclosures of the amino acid sequences of such antibodies are incorporated herein by reference. Some antibodies can neutralize membrane-bound CD39 (memCD39) by inhibiting the domain movement of membrane-bound CD39, yet do not similarly affect the activity of soluble CD39 protein (sCD39). It has been reported that memCD39 occurs as a homomultimer, while sCD39 is a monomer, and furthermore, the transmembrane domain in memCD39 undergoes dynamic movement, which is the basis for the functional relationship with the active site. Thus, unlike sCD39, memCD39 can present a configuration that enables antibody-mediated neutralization. One possibility is that functional neutralization requires the use of a bivalent antibody that binds two memCD39 molecules simultaneously (e.g., within a memCD39 homomultimer).

[0063] The anti-CD39 antibodies described herein bind to epitopes present on human CD39 protein expressed on the surface of cells (including tumor cells) (e.g., they can compete with the anti-CD39 antibodies of SEQ ID NOs: 10 and 11 for binding to epitopes on CD39), and effectively inhibit the enzymatic activity (ATPase activity) of membrane-bound CD39 enzyme (CD39 as expressed on the cell surface), and the anti-CD39 antibodies described herein further inhibit the enzymatic activity (ATPase activity) of soluble (extracellular domain) human CD39 protein. The antibodies thus mediate strong neutralization of CD39 activity in an individual by neutralizing both membrane-bound and soluble CD39 proteins (including soluble CD39 released or shed from tumor cells), thereby reducing immunosuppression, e.g., for the treatment of cancer and / or infectious diseases.

[0064] In one aspect, the anti-CD39 antibody comprises (a) HCDR1, HCDR2, and HCDR3 that are SEQ ID NOs: 2, 3, and 4, respectively, and (b) LCDR1, LCDR2, and LCDR3 sequences that are SEQ ID NOs: 5, 6, and 7, respectively.

[0065] In one aspect, an anti-CD39 antibody can be characterized as comprising: an HCDR1 comprising the amino acid sequence: DYNMH (SEQ ID NO: 2) or a sequence of at least 3 or 4 contiguous amino acids thereof, optionally wherein one or more of these amino acids can be replaced by a different amino acid; an HCDR2 comprising the amino acid sequence: YIVPLNGGSTFNQKFKG (SEQ ID NO: 3) or a sequence of at least 4, 5, 6, 7, 8, 9 or 10 contiguous amino acids thereof, optionally wherein one or more of these amino acids can be replaced by a different amino acid, optionally wherein the asparagine at Kabat position 61 is replaced, optionally wherein the lysine at Kabat position 65 is replaced; an HCDR3 comprising the amino acid sequence: GGTRFAY (SEQ ID NO: 4) or a sequence of at least 4, 5 or 6 contiguous amino acids thereof, optionally wherein one or more of these amino acids can be replaced by 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 contiguous amino acids thereof, optionally wherein one or more of these amino acids can be replaced by a different amino acid, optionally wherein the arginine at Kabat position 24 is replaced; an LCDR2 region comprising the amino acid sequence: GASNQGS (SEQ ID NO: 6) or a sequence of at least 4, 5 or 6 contiguous amino acids thereof, optionally wherein one or more of these amino acids can be replaced by a different amino acid; and / or an LCDR3 region having the amino acid sequence: QQTKEVPYT (SEQ ID NO: 7) or a sequence of at least 4, 5, 6, 7 or 8 contiguous amino acids thereof, optionally wherein one or more of these amino acids are absent or replaced by a different amino acid.

[0066] In any embodiment, CDR positions can be according to Kabat numbering.

[0067] In one aspect, an anti-CD39 antibody comprises the hypervariable regions, optionally the CDRs, of an antibody having VH and VL of SEQ ID NOs: 8 and 9, as follows.

[0068] Anti-CD39 VH:

[0069]

[0070] Anti-CD39 VL:

[0071]

[0072] In one embodiment, the anti-CD39 antibody can be characterized by an antigen-binding domain that comprises: a heavy chain variable region (VH) that comprises CDR1, CDR2, and CDR3 having the respective amino acid sequences shown in SEQ ID NOs: 2, 3, and 4 and framework FR1, FR2, and FR3 amino acid sequences derived from a human IGHV1-3 gene such as IGHV1-3*01 (and optionally an additional framework 4 (FR4) amino acid sequence derived from a human IGHJ1 gene such as IGHJ1*01); and light chain variable region (VL) CDR1, CDR2, and CDR3 that have the respective amino acid sequences shown in SEQ ID NOs: 5, 6, and 7 and framework FR1, FR2, and FR3 amino acid sequences derived from a human IGKV4-1 (such as IGK4-1*01) gene and optionally an additional framework 4 (FR4) amino acid sequence derived from a human IGKJ4 (such as IGKJ4*01) gene. The VH further comprises one or more amino acid substitutions of residues present in the human framework sequence by different residues (e.g., residues present in a 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, such as 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 in the VL at Kabat position 24 is lysine (e.g., CDR1 includes an R24K substitution). Optionally, phenylalanine is present in the VL at Kabat position 36.

[0073] 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.

[0074] In one embodiment, the VL comprises phenylalanine (FR2) at Kabat position 36. In one embodiment, the VL comprises lysine (CDR1) at Kabat position 24.

[0075] In any embodiment, the anti-CD39 antibody can be characterized as comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the heavy chain variable region (VH) comprises an amino acid sequence that is 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 that is 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.

[0076] In one embodiment, the anti-CD39 antibody comprises: a heavy chain variable region (VH) that comprises CDR1, CDR2, and CDR3 having the respective amino acid sequences shown in SEQ ID NOs: 2, 3, and 4 and a human framework (e.g., FR1, FR2, FR3, and FR4 of human origin); and light chain variable region (VL) CDR1, CDR2, and CDR3 that comprise the respective amino acid sequences shown in SEQ ID NOs: 5, 6, and 7 and a human framework (e.g., FR1, FR2, FR3, and FR4 of human origin), wherein (VH) comprises an amino acid sequence that is 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 that is 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.

[0077] In any embodiment, VH can be characterized as comprising 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 isoleucine (e.g., M48I substitution). In one embodiment, the residue at position 67 is alanine (e.g., V67A substitution). In one embodiment, the residue at position 71 is valine (e.g., R71V substitution). In one embodiment, the residue at position 76 is arginine (e.g., S76R substitution). In any embodiment, VL can be characterized as comprising a substitution at Kabat position 36. In one embodiment, the residue at position 36 is phenylalanine (e.g., Y36F substitution).

[0078] In one embodiment, the anti-CD39 antibody comprises a heavy chain and a light chain, the heavy chain comprising an amino acid sequence that is 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 the light chain comprising an amino acid sequence that is 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.

[0079] Anti-CD39 heavy chain

[0080]

[0081]

[0082] Anti-CD39 light chain

[0083]

[0084] In one embodiment, the anti-CD39 antibody comprises a heavy chain and a light chain, the heavy chain comprising the amino acid sequence of SEQ ID NO: 10 and the light chain comprising the amino acid sequence of SEQ ID NO: 11. In one embodiment, the anti-CD39 antibody is IPH5201.

[0085] In any embodiment, the anti-CD39 antibody can be characterized as binding and inhibiting or neutralizing the ATPase activity of soluble CD39 protein (sCD39). In one embodiment, the sCD39 protein lacks the two transmembrane domains found in membrane-bound CD39 (i.e., the transmembrane domains near the N- and C-termini). In one embodiment, sCD39 is a non-membrane-bound sCD39 protein found in the circulation, such as in a human individual. In one embodiment, the CD39-derived sequence of the sCD39 protein comprises or consists of the Thr38-Val478 fragment of CD39. The Thr38-Val478 protein with a C-terminal His tag is commercially available from R&D Systems, Inc. (product number 4397-EN). In one embodiment, when incubated with sCD39 in solution (e.g., in tumor cell supernatant), the protein, antibody or antibody fragment inhibits the ATPase activity of sCD39. In one embodiment, the protein, antibody or antibody fragment specifically binds to human CD39 protein in both soluble (extracellular domain protein) and membrane-bound forms.

[0086] Advantageously, the anti-CD39 antibodies described herein comprise a human Fc domain that is modified to have reduced or substantially lacking binding to human Fcγ receptors, such as one or more (or all) of human CD16, CD32a, CD32b, and CD64. The antibodies exert CD39 inhibitory activity independent of ADCC, CDC, or toxin-mediated depletion of CD39-expressing cells. Thus, these antibodies act as "pure" CD39 blockers and have immunomodulatory activity.

[0087] As used herein, the term "PD-1" refers to the protein programmed death 1 (PD-1) (also known as "programmed cell death 1"), which is an inhibitory member of the CD28 receptor family, which also includes CD28, CTLA-4, ICOS, and BTLA. The complete human PD-1 sequence can be found under GenBank accession number U64863, as follows:

[0088]

[0089] "PD-1" also includes any variants, derivatives, or isotypes of the PD-1 gene or encoded protein. PD-1 is expressed on activated B cells, T cells, and myeloid cells (Okazaki et al. (2002) Curr. Opin. Immunol. 14: 391-779-82; Bennett et al. (2003) J Immunol 170: 711-8). Two ligands for PD-1, PD-L1 and PD-L2, have been identified, and it has been demonstrated that they 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 PD-1 but not other CD28 family members.

[0090] The complete human PD-L1 sequence can be found under UniProtKB / Swiss-Prot, identifier Q9NZQ7-1, as follows:

[0091]

[0092] PD-L1 is abundant in a variety of human cancers. The interaction between PD-1 and PD-L1 results in reduced tumor-infiltrating lymphocytes, reduced T cell receptor-mediated proliferation, and immune evasion of cancer cells. Immunosuppression can be reversed by inhibiting the local interaction between PD-1 and PD-L1, and this effect is additive when the interaction between PD-1 and PD-L2 is also blocked.

[0093] An anti-PD(L)1 antibody is an antibody that binds to PD-1 or PD-L1. The antibody 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 the signal transduction ability of PD-1 is inhibited due to the interaction of PD-1 with one or more of its binding partners, such as PD-L1 or PD-L2. An antibody that neutralizes the inhibitory activity of PD-1 reduces, blocks, inhibits, eliminates, or interferes with signal transduction caused by the interaction of PD-1 with one or more of its binding partners, particularly such as PD-L1. Thus, such agents can reduce negative co-stimulatory signals mediated by cell surface proteins expressed on T lymphocytes, thereby enhancing T cell effector functions such as proliferation, cytokine production, and / or cytotoxicity. A PD-1 neutralizing agent can interact with PD-1 and / or one or more of its binding partners (e.g., PD-L1 and PD-L2).

[0094] 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.

[0095] In some embodiments, the anti-PD(L)1 antibody is YW243.55.S70, MPDL3280A (atezolizumab, ), MDX-1105 or durvalumab (MEDI4736, ). MDX-1105, also known as BMS-936559, is an anti-PD-L1 antibody described in WO2007 / 005874. The antibody YW243.55.S70 is an anti-PD-L1 described in WO2010 / 077634. Examples of anti-PD-L1 antibodies that can be used in the methods disclosed herein and methods for their preparation are also described in WO2010 / 077634A1 and U.S. Patent No. 8,217,149, both of which are incorporated herein by reference.

[0096] In some embodiments, the anti-PD(L)1 is the PD-L1 antibody durvalumab. Durvalumab (MEDI4736, Imfinzi TM ) is a human monoclonal antibody against human PD-L1 that is capable of blocking the binding of PD-L1 to both the PD-1 and CD80 receptors. The disclosures regarding durvalumab can be found in U.S. Patent Nos. 8,779,108 and 9,493,565, both of which are incorporated herein by reference. Durvalumab has a heavy chain and a light chain having amino acid sequences SEQ ID NO: 16 and SEQ ID NO: 17, respectively. The heavy chain variable region of durvalumab is shown as SEQ ID NO: 14, and the light chain variable region of durvalumab is shown as SEQ ID NO: 15.

[0097] 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 certain embodiments, the anti-PD-L1 antibody has the same heavy and light chain CDRs as durvalumab.

[0098] In some embodiments, the anti-PD(L)1 antibody (e.g., an agent derived from durvalumab) comprises (i) the heavy chain variable region of SEQ ID NO: 14, or an amino acid sequence that is at least 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 99% identical thereto, and (ii) the light chain variable region of SEQ ID NO: 15, or an amino acid sequence that is at least 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 99% identical thereto. In some embodiments, the anti-PD-1 neutralizing agent (e.g., an agent derived from durvalumab) comprises (i) the heavy chain of SEQ ID NO: 16, or an amino acid sequence that is at least 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 99% identical thereto, and (ii) the light chain of SEQ ID NO: 17, or an amino acid sequence that is 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 derived from the 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 derived from the light chain variable region comprising the amino acid sequence of SEQ ID NO: 15.

[0099] 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.

[0100] Heavy chain variable region of durvalumab:

[0101]

[0102] Light chain variable region of durvalumab:

[0103] EIVLTQSPGTLSLSPGERATLSCRASQRVSSSYLAWYQQKPGQAPRLLIYDASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSLPWTFGQGTKVEIK (SEQ ID NO: 15).

[0104] Heavy chain of durvalumab:

[0105]

[0106]

[0107] Light chain of durvalumab:

[0108]

[0109] Durvalumab, heavy chain CDR:

[0110] H-CDR1: GFTFSRYWMS (SEQ ID NO: 18)

[0111] H-CDR2: NIKQDGSEKYYVDSVKG (SEQ ID NO: 19)

[0112] H-CDR3: EGGWFGELAFDY SEQ ID NO: 20)

[0113] Durvalumab, light chain CDR:

[0114] L-CDR1: RASQRVSSSYLA (SEQ ID NO: 21)

[0115] L-CDR2: DASSRAT (SEQ ID NO: 22)

[0116] L-CDR3: QQYGSLPWT (SEQ ID NO: 23)

[0117] In another embodiment, the anti-PD(L)1 is atezolizumab (MPDL3280A, CAS Registry Number: 1422185-06-5). In some embodiments, the anti-PD-L1 antibody comprises: a heavy chain variable region comprising the following amino acid sequence:

[0118]

[0119] or

[0120]

[0121] and a light chain variable region comprising the following amino acid sequence:

[0122]

[0123] In some embodiments, the anti-PD(L)1 antibody comprises (i) the heavy chain or heavy chain variable region of SEQ ID NO: 27, or an amino acid sequence that is at least 50%, 60%, 70%, 80%, 90%, 95%, 98% or 99% identical thereto, and (ii) the light chain or light chain variable region of SEQ ID NO: 28, or an amino acid sequence that is at least 50%, 60%, 70%, 80%, 90%, 95%, 98% or 99% identical thereto.

[0124]

[0125] 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 previously named 5C4, BMS-936558, MDX-1106 or ONO-4538) is a fully human IgG4 (S228P) PD-1 immune checkpoint inhibitor antibody that selectively blocks the interaction with the PD-1 ligands (PD-L1 and PD-L2), 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 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 chain and light chain CDRs as nivolumab.

[0126] In another embodiment, the anti-PD-1 antibody is pembrolizumab. Pembrolizumab (also known as 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 has been approved by the FDA for the treatment of recurrent 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.

[0127] In another embodiment, the anti-PD-1 antibody is cemiplimab.

[0128] A chemotherapeutic agent refers to a compound that can be used to treat cancer (e.g., lung cancer, NSCLC, optionally stage II NSCLC and / or stage IIIA NSCLC). For example, a chemotherapeutic agent can be a compound known to be useful for neoadjuvant treatment of NSCLC, optionally resectable or stage II or IIIA NSCLC. When used as neoadjuvant therapy, chemotherapy can be used, for example, at the doses and administration frequencies known or standard 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 includes platinum-based chemotherapeutic agents. A "platinum-based" chemotherapeutic agent includes an organic compound containing platinum as a component of the molecule. Generally, a platinum-based chemotherapeutic agent is a coordination complex of platinum. Platinum-based chemotherapeutic 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 cases, a platinum-based chemotherapeutic agent (e.g., cisplatin or carboplatin) can be administered in combination with one or more additional chemotherapeutic agents, such as, for example, nucleoside analogs (e.g., gemcitabine).

[0129] For example, platinum-based chemotherapy can include a platinum-based chemotherapeutic agent (e.g., cisplatin or carboplatin), and optionally one or more additional chemotherapeutic agents, such as nucleoside analogs (e.g., gemcitabine), taxanes (e.g., paclitaxel), or folic acid antimetabolites (e.g., pemetrexed).

[0130] In some embodiments, the chemotherapy used in the neoadjuvant treatment herein includes gemcitabine.

[0131] In some embodiments, the chemotherapy used in the neoadjuvant treatment herein includes administering two or more chemotherapeutic agents. In some embodiments, the chemotherapy includes administering at least one or two agents selected from the following: paclitaxel, carboplatin, pemetrexed, or cisplatin. In some embodiments, the chemotherapy includes or consists of the administration of carboplatin and paclitaxel. In some embodiments, the chemotherapy includes or consists of the administration of cisplatin and gemcitabine. In some embodiments, the chemotherapy includes or consists of the administration of cisplatin and pemetrexed. In some embodiments, the chemotherapy includes or consists of the administration of carboplatin and pemetrexed.

[0132] 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, such as a patient having a resectable cancer or tumor, the method 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.

[0133] The present disclosure also provides a method of treating a tumor or cancer and / or preventing recurrence of a tumor or cancer in a patient in need thereof, the method 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 further as adjuvant therapy.

[0134] The term "patient" is intended to include humans and non-human animals, particularly mammals. In some embodiments, the present disclosure relates to providing treatment to a patient having a tumor disorder and / or a cancer disorder (such as a treatment method or pharmaceutical formulation used as described herein), optionally a surgically resectable tumor and / or cancer. 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 a stage I, II, or III non-small cell lung tumor. These methods can be used as first-line treatment (1L treatment, 1-L treatment, 1L or 1-L), i.e., the first treatment given for a disease (particularly cancer as described herein). First-line treatment can be specific for a given type or subtype of cancer or a particular cancer stage. First-line treatment can be part of a standard treatment regimen. First-line treatment is generally recognized as the best treatment for a disease, particularly cancer as described herein. If first-line treatment does not cure the disease or it causes severe side effects, then a subsequent treatment route can be used instead. In some embodiments, the present disclosure relates to providing first-line treatment for cancer (such as a treatment method or pharmaceutical formulation used as described herein).

[0135] In some embodiments, prior to receiving neoadjuvant treatment of the present disclosure, the patient has not previously undergone resection of a non-small cell lung tumor. In some embodiments, prior to receiving adjuvant treatment of the present disclosure, the patient has undergone resection of a non-small cell lung tumor, optionally a complete resection.

[0136] Also provided herein are treatment regimens suitable for administering anti-CD39 antibody and anti-PD(L)1 antibody as neoadjuvant therapy and / or adjuvant therapy. Thus, in the methods of administration, treatment, or prevention herein, the method is designated as comprising administering to a patient an effective amount of a treatment regimen comprising anti-CD39 antibody and anti-PD(L)1 antibody, such as the treatment regimen comprising anti-CD39 antibody and anti-PD(L)1 antibody described herein. Accordingly, provided herein is a method of treating a patient with lung cancer, the method comprising administering to the patient an effective amount of a treatment regimen comprising anti-CD39 antibody and anti-PD(L)1 antibody, and optionally further administering 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 anti-CD39 antibody and anti-PD(L)1 antibody, wherein the treatment regimen is adjuvant therapy. The doses and dosing intervals used in the treatment regimen can be specified as further provided herein, including advantageous regimens wherein the same doses of anti-CD39 antibody and anti-PD(L)1 antibody can be used in both neoadjuvant and adjuvant settings.

[0137] Thus, in one embodiment, the treatment regimen of the present disclosure can be characterized as including administering an anti-CD39 antibody and an anti-PD(L)1 antibody (and optionally further administering chemotherapy) to a patient before surgical resection of a tumor. The treatment can also include administering an anti-CD39 antibody and an anti-PD(L)1 antibody to the patient after surgical resection of the tumor. In one embodiment, the anti-PD(L)1 antibody is durvalumab. The tumor or cancer can optionally be designated as a tumor or cancer that is considered surgically resectable.

[0138] In any embodiment herein, the method and / or the effective amount can be designated as including a combination of an anti-PD(L)1 antibody and an anti-CD39 antibody (and optionally further chemotherapy) in an amount that achieves a therapeutic outcome. In some instances, the effective amount of a therapeutic agent or combination of therapeutic agents is the amount of the agent or combination of agents that achieves one or more of the following clinical endpoints: improved overall response rate (ORR), complete response (CR), pathologic 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 functional and quality of life (QoL) time to deterioration, and / or clearance of ctDNA. Improvement (e.g., in response rate (e.g., ORR, CR, and / or PR), survival (e.g., DFS, DSS, distant metastasis-free survival, PFS, and / or OS), DOR, improved functional and QoL time to deterioration, undetectable ctDNA, and / or clearance of ctDNA) can be relative to a suitable reference, e.g., an observation or a reference treatment (e.g., a treatment that does not include an anti-CD39 antibody (e.g., treatment with a placebo, treatment with an anti-PD(L)1 antibody)). In some cases, improvement (e.g., in response rate (e.g., ORR, CR, and / or PR), survival (e.g., EFS, DFS, DSS, distant metastasis-free survival, PFS, and / or OS), DOR, improved functional and QoL time to deterioration, and / or clearance of ctDNA or undetectable ctDNA status) can be relative to an observation.

[0139] As used herein, "complete response" and "CR" refer to the disappearance of all target lesions.

[0140] As used herein, the terms "partial response" and "PR" refer to at least a defined (e.g., 30%) reduction in the sum of the longest diameters (SLD) of the target lesions, with reference to the sum of the longest diameters at baseline before treatment.

[0141] As used herein, "overall response rate," "objective response rate," and "ORR" can be used interchangeably to refer to the sum of the CR rate and the PR rate.

[0142] As used herein, "disease-free survival" and "DFS" refer to the length of time that a patient survives without cancer recurrence after initial treatment (e.g., surgical resection).

[0143] As used herein, "disease-specific survival" and "DSS" refer to the length of time that a patient does not die from a specific disease (e.g., NSCLC). In some cases, DSS can be defined as the time from randomization to death due to NSCLC (e.g., according to the investigator's assessment of the cause of death). As used herein, "distant metastasis-free survival" refers to the length of time from the date of diagnosis or start of treatment, i.e., the time that the patient remains 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 (i.e., non-local) metastasis or death from any cause.

[0144] As used herein, "progression-free survival" and "PFS" refer to the length of time during and after treatment that cancer does not get worse. PFS can include the amount of time that a patient experiences CR or PR, as well as the amount of time that a patient experiences stable disease.

[0145] As used herein, "overall survival" and "OS" refer to the length of time that a patient remains alive from the date of diagnosis of the disease (e.g., cancer) or start of treatment. For example, OS can be defined as the time from randomization to death from any cause.

[0146] As used herein, the terms "duration of response" and "DOR" refer to the length of time from the recording of a tumor response until disease progression or death from any cause (whichever occurs first).

[0147] In one embodiment, the treatment regimens herein allow for the administration of the same dose of anti-CD39 antibody (e.g., a fixed dose of 2250 mg or 3000 mg of anti-CD39 antibody) in the adjuvant and neoadjuvant settings. Additionally, these regimens allow for the administration of the anti-CD39 antibody every three weeks as a neoadjuvant and every four weeks as an adjuvant (at the same dose in both neoadjuvant and adjuvant therapies).

[0148] In one embodiment, a method for administering an anti-CD39 antibody (e.g., an anti-CD39 antibody having the characteristics described herein; an antibody comprising the amino acid sequences of SEQ ID NO: 2-7, SEQ ID NO: 8 and 9 or SEQ ID NO: 10 and 11) is provided, wherein the antibody is administered at a fixed dose of 3000 mg Q3w and / or Q4w. In one embodiment, the anti-CD39 antibody is administered at a fixed dose of 3000 mg as neoadjuvant therapy Q3w and at a fixed dose of 3000 mg as adjuvant therapy Q4w. In another embodiment, a method for administering an anti-CD39 antibody (e.g., an anti-CD39 antibody having the characteristics described herein; an antibody comprising the amino acid sequences of SEQ ID NO: 2-7, SEQ ID NO: 8 and 9 or SEQ ID NO: 10 and 11) is provided, wherein the antibody is (a) administered Q3w, one or more times and (b) administered Q4w, one or more times, wherein in each case the antibody is administered at a fixed dose of 3000 mg.

[0149] In one embodiment, a method for administering an anti-CD39 antibody (e.g., an anti-CD39 antibody having the characteristics described herein; an antibody comprising the amino acid sequences of SEQ ID NO: 2-7, SEQ ID NO: 8 and 9 or SEQ ID NO: 10 and 11) is provided, wherein the antibody is administered at a fixed dose of 2250 mg Q3w and / or Q4w. In one embodiment, the anti-CD39 antibody is administered at a fixed dose of 2250 mg as neoadjuvant therapy Q3w and at a fixed dose of 2250 mg as adjuvant therapy Q4w. In another embodiment, a method for administering an anti-CD39 antibody (e.g., an anti-CD39 antibody having the characteristics described herein; an antibody comprising the amino acid sequences of SEQ ID NO: 2-7, SEQ ID NO: 8 and 9 or SEQ ID NO: 10 and 11) is provided, wherein the antibody is (a) administered Q3w, one or more times and (b) administered Q4w, one or more times, wherein in each case the antibody is administered at a fixed dose of 2250 mg.

[0150] Optionally, in any embodiment herein, the method can be characterized as a method for treating cancer and / or preventing cancer recurrence. 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 administering an anti-PD(L)1 antibody, and optionally further administering chemotherapy) in combination with surgery and optionally additional radiotherapy.

[0151] Preferably, the anti-CD39 antibody is administered intravenously (i.v.). Preferably, the anti-PD(L)1 antibody is administered intravenously.

[0152] In some embodiments, the doses of the anti-PD(L)1 antibody and chemotherapy to be administered to a patient can vary in part according to the patient's size (body weight, body surface, or organ size) and condition (age and general health).

[0153] 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 TM and is supplied to the market.

[0154] Thus, the neoadjuvant therapy treatment regimen can include administering the anti-CD39 antibody at a fixed dose of 2250 mg or 3000 mg every three weeks, and administering durvalumab at a fixed dose of 1500 mg every three weeks.

[0155] Thus, the adjuvant therapy treatment regimen can include administering the anti-CD39 antibody at a fixed dose of 2250 mg or 3000 mg every four weeks, and administering durvalumab at a fixed dose of 1500 mg every four weeks.

[0156] In certain embodiments, the anti-CD39 antibody and the anti-PD(L)1 antibody and optionally additional chemotherapy are administered simultaneously, in parallel, separately, or sequentially. In some embodiments, the anti-CD39 antibody and the anti-PD(L)1 antibody are administered before chemotherapy. In additional embodiments, the anti-CD39 antibody and the anti-PD(L)1 antibody are administered in parallel with chemotherapy. Advantageously, the anti-CD39 antibody and the anti-PD(L)1 antibody are administered on the same day, for example, on the first day of a three-week cycle or on the first day of a four-week cycle for neoadjuvant treatment and adjuvant treatment, respectively. Optionally, for neoadjuvant treatment, the anti-CD39 antibody and the anti-PD(L)1 antibody are administered on the same day as the administration of chemotherapy (e.g., the first day of a three-week cycle).

[0157] Thus, the neoadjuvant therapy can include one or more cycles of treatment in the following manner: administering the anti-CD39 antibody having the respective heavy and light chain amino acid sequences of SEQ ID NO: 10 and 11 at a fixed dose of 2250 mg or 3000 mg and administering durvalumab at a fixed dose of 1500 mg, wherein the anti-CD39 antibody and durvalumab are administered once on the first day of a three-week cycle. The neoadjuvant therapy can include, for example, 2, 3, 4, or more such treatment cycles.

[0158] Adjuvant therapy may include one or more cycles of treatment in the following manner: administering an anti-CD39 antibody having the respective heavy and light chain amino acid sequences of SEQ ID NOs: 10 and 11 at a fixed dose of 2250 mg or 3000 mg and durvalumab at a fixed dose of 1500 mg, wherein the anti-CD39 antibody and durvalumab are administered once on the first day of a four-week cycle. Adjuvant therapy may include, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or more such treatment cycles, or at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 cycles. In one embodiment, the anti-CD39 antibody and durvalumab are administered as adjuvant therapy for up to 12 cycles.

[0159] Thus, when an anti-CD39 antibody and an anti-PD(L)1 antibody are used as neoadjuvant and adjuvant therapy, the treatment may thus include:

[0160] (a) Administering one or more times as neoadjuvant therapy a fixed dose of 2250 mg or 3000 mg of an anti-CD39 antibody having the respective heavy and light chain amino acid sequences of SEQ ID NOs: 10 and 11 and a fixed dose of 1500 mg of durvalumab, wherein the anti-CD39 antibody and durvalumab are administered on the first day of a three-week cycle; and

[0161] (b) Administering one or more times as adjuvant therapy a fixed dose of 2250 mg or 3000 mg of an anti-CD39 antibody having the respective heavy and light chain amino acid sequences of SEQ ID NOs: 10 and 11 and a fixed dose of 1500 mg of durvalumab, wherein the anti-CD39 antibody and durvalumab are administered on the first day of a four-week cycle.

[0162] The adjuvant therapy anti-CD39 antibody and anti-PD(L)1 antibody may be initiated immediately after surgical resection of the tumor or cancer, and preferably, the adjuvant therapy will be conducted within 10 weeks of surgical resection. The adjuvant therapy may continue for up to 12 cycles, or more than 12 cycles, such as until disease progression or until a desired response (e.g., CR) is obtained, or until ctDNA clearance.

[0163] In some embodiments, the anti-CD39 antibody and the anti-PD(L)1 antibody are used independently or irrespective of the mutation status of the epidermal growth factor receptor (EGFR) and / or anaplastic lymphoma kinase (ALK) gene (e.g., as neoadjuvant therapy and optionally as adjuvant therapy). In some embodiments, the anti-CD39 antibody and the anti-PD(L)1 antibody are used to treat patients lacking an EGFR mutation (EGFR wild-type) and / or lacking an ALK gene rearrangement (ALK wild-type).

[0164] In some embodiments, the anti-CD39 antibody and the anti-PD(L)1 antibody are used independently or regardless of the patient's PD-L1 expression status (e.g., as neoadjuvant therapy and optionally as adjuvant therapy). In some embodiments, the anti-CD39 antibody and the anti-PD(L)1 antibody are used as neoadjuvant therapy and optionally as adjuvant therapy for treating patients with detectable PD-L1 expression, such as patients with a CPS score of at least 1 (CPS≥1).

[0165] PD-L1 expression can include any detectable level of expression of PD-L1 protein on the cell surface or PD-L1 mRNA within cells or tissues. PD-L1 protein expression can be detected using a diagnostic PD-L1 antibody in an IHC assay of tumor tissue sections or can be detected by flow cytometry. Alternatively, PD-L1 protein expression in tumor cells can be detected by PET imaging using a binding agent.

[0166] One method employs a simple binary endpoint of PD-L1 expression being positive or negative, with a positive result defined according to the percentage of tumor cells showing histological evidence of cell surface membrane staining. If a tumor tissue section is at least 1% of total tumor cells, it is counted as PD-L1 expression positive.

[0167] In another method, PD-L1 expression in tumor tissue sections is quantified in tumor cells as well as in infiltrating immune cells that mainly comprise lymphocytes. The percentage of tumor cells and infiltrating immune cells showing membrane staining is quantified separately as <5%, 5% to 9%, and then in 10% increments up to 100%. PD-L1 expression in the immune infiltrate is reported as a semi-quantitative measurement called the adjusted inflammation score (AIS), which is determined by multiplying the percentage of membrane-staining cells by the intensity of the infiltrate and graded as none (0), mild (score of 1, sparse lymphocytes), moderate (score of 2, focal infiltration of lymphohistiocytic aggregates into the tumor), or severe (score of 3, diffuse infiltration). If MS≥5, the tumor tissue section is counted as PD-L1 expression positive in the immune infiltrate.

[0168] Several PD-L1 protein scoring systems are commonly used. The "Tumor Proportion Score (TPS)" refers to the percentage of tumor cells expressing PD-L1 at any intensity (weak, moderate, or strong) on the cell membrane. Linear or complete cell membrane staining is interpreted as PD-L1 positive. The "Mononuclear Inflammatory Cell Density Score (MIDS)" refers to the ratio of the number of mononuclear inflammatory cells (MICs) expressing PD-L1 (small and large lymphocytes, monocytes, and macrophages within the tumor nests and adjacent supporting stroma) infiltrating the tumor or adjacent to the tumor to the total number of tumor cells. MIDS is recorded on a scale of 0 to 4, where 0 = none; 1 = present, but less than one MIC per 100 tumor cells (<1%); 2 = at least one MIC per 100 tumor cells, but less than one MIC per 10 tumor cells (1%-9%); 3 = at least one MIC per 10 tumor cells, but fewer MICs than tumor cells (10%-99%); 4 = at least as many MICs as 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 the tumor nests and adjacent supporting stroma to the total number of tumor cells (denominator; i.e., the number of PD-L1-positive and PD-L1-negative tumor cells). PD-L1 expression at any intensity is considered positive, i.e., weak (1+), moderate (2+), or strong (3+).

[0169] When anti-CD39 antibody and anti-PD(L)1 antibody are used as adjuvant therapy (e.g., in addition to neoadjuvant therapy), the treatment of the present disclosure can optionally be beneficial for patients who are positive for minimal residual disease (MRD+) after surgical resection. For example, after neoadjuvant therapy and surgery, it can optionally be determined (a) whether the patient is positive for minimal residual disease (MRD+); and (b) if the patient is identified as MRD+, the patient is administered an anti-CD39 antibody and an anti-PD(L)1 antibody as adjuvant therapy. In one embodiment, the anti-CD39 antibody and the anti-PD(L)1 antibody are administered within 10 weeks of tumor resection.

[0170] The MRD status of a patient can be determined using methods known in the art (see, e.g., Abbosh et al. (2017); Chaudhuri et al. (2017)). In some embodiments, the MRD status of a patient 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 by WES of the patient's whole blood. Then, a personalized panel consisting of highly expressed patient tumor variants is developed. Then, this panel is used to identify the presence of these variants on circulating tumor DNA (ctDNA) extracted from the patient's plasma, and if the panel detects tumor variants, the patient is considered MRD+. This personalized approach allows for the detection of a patient's tumor variants in DNA extracted from the patient's plasma with high sensitivity.

[0171] In some embodiments, determining whether a patient is minimal residual disease positive (MRD+) is determined by the following steps: (a) sequencing all or a portion of the genome or exome of the patient's tumor to define clonal and / or subclonal mutations in the tumor; (b) defining a set of reagents that will detect the presence of DNA from the tumor via the presence of clonal and / or subclonal mutations; and (c) analyzing a sample comprising DNA from the tumor 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 elevation of clonal and / or subclonal mutations in a sample from a patient with a tumor signature indicates whether the tumor has recurred. Clonal and / or subclonal mutations specific to the patient's tumor are defined by sequencing all or a portion of the whole genome and / or exome of DNA from the tumor, in some cases, after the tumor has been resected from the patient. 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 a particular subject of interest, the presence and / or elevation of clonal and / or subclonal mutations in a sample obtained from the patient is analyzed.

[0172] In some embodiments, tumor biopsies, all or part of a tumor, or one or more sub - parts of a tumor, cell - free DNA (cfDNA), ctDNA, tumor DNA from exosomes, or circulating tumor cells from a subject are sequenced. In some embodiments, the tumor or a sub - part thereof is sequenced after tumor removal. In some embodiments, all or part of the genome or exome of at least two sub - parts of a tumor is sequenced, and clonal and / or sub - clonal mutations are defined based on which mutations occur in which tumor sub - parts. In some embodiments, the defined set of reagents includes multiplex PCR primers, and the analysis is multiplex PCR. In some embodiments, plasma obtained from a patient is sequenced, or the sample to be analyzed is a plasma sample from a patient.

[0173] As indicated by the detection of ctDNA, MRD can reveal the presence of residual tumor that is clinically indistinguishable after effective intended therapy (surgery ± chemotherapy / radiotherapy). Detecting MRD in the absence of radiological evidence of disease provides an opportunity for early therapeutic intervention. Anti - CD39 antibody and anti - PD(L)1 antibody can be administered, for example, as soon as possible, preferably within 10 weeks of tumor resection. Compared to MRD - patients, MRD+ patients experience a poorer recurrence - free survival. Thus, MRD+ patients may benefit from earlier intervention and treatment escalation, including immunotherapy alone or in combination with chemotherapy; in addition, MRD - patients (mostly cured only by surgery) can be spared stronger therapies and the resulting unnecessary toxicities.

[0174] It should be understood that anti - CD39 antibody and anti - PD(L)1 antibody can be incorporated into a pharmaceutical formulation at a suitable concentration (e.g., 1 mg / ml to 500 mg / ml, where the pH of the formulation is 2.0 to 10.0). Anti - CD39 antibody and anti - PD(L)1 antibody can be included in the same or separate pharmaceutical formulations. The formulation can further include a buffer system, one or more preservatives, one or more tonicity agents, one or more chelating agents, stabilizers, and surfactants. In one embodiment, the pharmaceutical formulation is an aqueous formulation, i.e., a formulation that includes water. Such formulations are typically solutions or suspensions. In another embodiment, the pharmaceutical formulation is an aqueous solution. The term "aqueous formulation" is defined as a formulation that includes at least 50% w / w water. Similarly, the term "aqueous solution" is defined as a solution that includes at least 50% w / w water, and the term "aqueous suspension" is defined as a suspension that includes at least 50% w / w water.

[0175] Optionally, the pharmaceutical formulation is a lyophilized formulation, and a solvent and / or diluent are added by a physician or patient before use. Optionally, the pharmaceutical formulation is a dry formulation (e.g., freeze - dried or spray - dried) that can be used without any prior dissolution.

[0176] In another 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 higher, and wherein the pH of the formulation is from about 2.0 to about 10.0. Optionally, the pH of the formulation is within the 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 another 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, trimethylglycine, malic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, aspartic acid or mixtures thereof. Each of these specific buffers constitutes an alternative embodiment of the present invention.

[0177] In another embodiment, the formulation further comprises a pharmaceutically acceptable preservative. In another embodiment, the formulation further comprises an isotonic agent. In another embodiment, the formulation further comprises a chelating agent. In another embodiment of the present invention, the formulation further comprises a stabilizer. In another embodiment, the formulation further comprises a surfactant. For convenience, reference is made to Remington: The Science and Practice of Pharmacy, 19th Edition, 1995.

[0178] Other components may be present in the pharmaceutical formulations of the present invention. Such additional components may include wetting agents, emulsifying agents, antioxidants, fillers, tonicity modifiers, 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). Of course, such additional components should not have an adverse effect on the overall stability of the pharmaceutical formulations of the present invention.

[0179] Administration of the pharmaceutical composition according to the present invention can be by any of several routes of administration, such as intravenous administration. A suitable antibody formulation can also be determined by examining the experience of other developed therapeutic monoclonal antibodies.

[0180] In one aspect, provided herein are kits, such as kits comprising:

[0181] (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

[0182] (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

[0183] (iii) a pharmaceutical composition comprising an anti-CD39 antibody (such as an anti-CD39 antibody comprising the corresponding VH and VL amino acid sequences of SEQ ID NOs: 8 and 9), and instructions for administering the anti-CD39 antibody in combination with an anti-PD(L)1 antibody (such as durvalumab), for example as neoadjuvant therapy and / or adjuvant therapy, optionally for NSCLC, or

[0184] (iv) a pharmaceutical composition comprising an anti-PD(L)1 antibody (such as durvalumab), and instructions for administering the anti-PD(L)1 antibody in combination with an anti-CD39 antibody (such as an anti-CD39 antibody comprising the corresponding VH and VL amino acid sequences of SEQ ID NOs: 8 and 9), for example as neoadjuvant therapy and / or adjuvant therapy, optionally for NSCLC.

[0185] In any embodiment, the kit may optionally further comprise a chemotherapy, such as a chemotherapy comprising a platinum agent, gemcitabine, including combinations of chemotherapeutic agents, such as carboplatin and paclitaxel, cisplatin and gemcitabine, cisplatin and pemetrexed, or carboplatin and pemetrexed.

[0186] The pharmaceutical composition may optionally be designated as comprising a pharmaceutically acceptable carrier. The anti-CD39 antibody or the anti-PD(L)1 antibody may optionally be designated as being present in a therapeutically effective amount for any method suitable for the methods herein. The anti-CD39 antibody may optionally be designated as comprising the CDR amino acid sequences of SEQ ID NOs: 2-7. The anti-CD39 antibody may optionally be designated as comprising the heavy and light chain amino acid sequences of SEQ ID NOs: 10 and 11. The kit may optionally further include instructions, for example, including a dosing schedule (such as the Q3w and / or Q4w schedules disclosed herein), to allow a practitioner (such as a physician, nurse, or patient) to administer the compositions contained therein to a patient suffering from cancer (such as a solid tumor, especially NSCLC, stage II or III NSCLC, resectable or non-resected NSCLC). In any embodiment, the kit may optionally include instructions for administering the anti-CD39 antibody and the anti-PD(L)1 antibody simultaneously, separately, or sequentially. The instructions may also optionally specify that the anti-CD39 antibody and the anti-PD(L)1 antibody are administered simultaneously, separately, or sequentially with chemotherapy (such as for neoadjuvant therapy). The kit may also include a syringe.

[0187] The kit can be designated to include one or more containers (such as disposable vials or pre-filled syringes), which contain a designated pharmaceutical composition or antibody.

[0188] A designated amount or dose (such as 3000 mg, 2250 mg or 1500 mg) can be designated to be provided in multiple vials. For example, one vial of anti-CD39 antibody can contain 375 mg of anti-CD39 antibody. One vial of durvalumab can contain 120 mg or 500 mg of durvalumab (such as 120 mg in 2.4 mL or 500 mg in 10 mL).

[0189] Optionally, the kit includes multiple packaged single-dose pharmaceutical compositions, each of the multiple packaged single-dose pharmaceutical compositions containing an effective amount of anti-CD39 antibody and / or anti-PD(L)1 antibody for single administration according to the methods provided above. Instruments or devices necessary for administering the pharmaceutical composition can also be included in the kit. For example, the kit can provide one or more pre-filled syringes containing a certain amount of anti-CD39 antibody or anti-PD(L)1 antibody.

[0190] In one embodiment, the present invention provides a kit for treating cancer or tumor in a human patient, the kit comprising: one or more disposable vials containing an anti-CD39 antibody, the anti-CD39 antibody comprising the H-CDR1, H-CDR2 and H-CDR3 domains of the heavy chain variable region having the sequence shown in SEQ ID NO: 8 and the L-CDR1, L-CDR2 and L-CDR3 domains of the light chain variable region having the sequence shown in SEQ ID NO: 9; and optionally, instructions for using the anti-CD39 antibody (such as at the doses, frequencies described herein) in any of the methods described herein.

[0191] In one embodiment, the present invention provides a kit for treating cancer or tumor in a human patient, optionally wherein the cancer or tumor is NSCLC (such as stage II or III NSCLC, resectable NSCLC, etc.), the kit comprising:

[0192] (a) one or more vials containing an anti-CD39 antibody, the anti-CD39 antibody comprising the H-CDR1, H-CDR2 and H-CDR3 domains of the heavy chain variable region having the sequence shown in SEQ ID NO: 8, and the L-CDR1, L-CDR2 and L-CDR3 domains of the light chain variable region having the sequence shown in SEQ ID NO: 9; and / or

[0193] (b) one or more vials containing an anti-PD-L1 antibody, optionally one dose of durvalumab; and

[0194] (c) Optionally, instructions for using the anti-CD39 antibody and / or the anti-PD(L)1 antibody in any of the methods described herein.

[0195] In one embodiment, a vial of anti-CD39 antibody contains 375 mg of anti-CD39 antibody. In one embodiment, the kit contains one or more sets of 6 vials each of anti-CD39 antibody. In one embodiment, the kit contains one or more sets of 8 vials each of anti-CD39 antibody. In one embodiment, the kit contains 4 sets of 6 vials or 8 vials each of anti-CD39 antibody for neoadjuvant therapy. In one embodiment, the kit contains at least 4 sets (e.g., 4 sets, 6 sets, 8 sets, 10 sets, or 12 sets) of 6 vials or 8 vials each of anti-CD39 antibody for adjuvant therapy.

[0196] In one embodiment, a vial of durvalumab contains 500 mg of durvalumab. In one embodiment, the kit contains one or more sets of 3 vials each of durvalumab. In one embodiment, the kit contains 4 sets of 3 vials each of durvalumab for neoadjuvant therapy. In one embodiment, the kit contains at least 4 sets (e.g., 4 sets, 6 sets, 8 sets, 10 sets, or 12 sets) of 3 vials each of durvalumab for adjuvant therapy.

[0197] In one embodiment, the present 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), the kit comprising:

[0198] (a) A dose of anti-CD39 antibody, the anti-CD39 antibody comprising the H-CDR1, H-CDR2, and H-CDR3 domains of the heavy chain variable region having the sequence shown in SEQ ID NO: 8, and the L-CDR1, L-CDR2, and L-CDR3 domains of the light chain variable region having the sequence shown in SEQ ID NO: 9; and / or

[0199] (b) A dose of anti-PD-L1 antibody, optionally a dose of durvalumab; and

[0200] (c) Optionally, instructions for using the anti-CD39 antibody and / or the anti-PD(L)1 antibody in any of the methods described herein.

[0201] In one embodiment, the dose of anti-CD39 antibody can be a fixed dose of 2250 mg or 3000 mg. In one embodiment, the dose of durvalumab can be a fixed dose of 1500 mg.

[0202] In one embodiment, the present invention provides a kit for treating cancer or a tumor in a human patient, optionally wherein the cancer or tumor is NSCLC (e.g., stage II or III NSCLC, resectable NSCLC), the kit comprising:

[0203] (a) one or more containers (e.g., one or more vials) together containing 3000 mg or 2250 mg of an anti-CD39 antibody, the anti-CD39 antibody comprising H-CDR1, H-CDR2, and H-CDR3 domains of a heavy chain variable region having the sequence shown in SEQ ID NO: 8, and L-CDR1, L-CDR2, and L-CDR3 domains of a light chain variable region having the sequence shown in SEQ ID NO: 9; and / or

[0204] (b) one or more containers (e.g., one or more vials) together containing 1500 mg of durvalumab; and

[0205] (c) optionally, instructions for using the anti-CD39 antibody and / or the anti-PD(L)1 antibody in any of the methods described herein.

[0206] In any embodiment, the instructions may specify anti-CD39 antibody administration Q3w or Q4w. In one embodiment, the instructions specify anti-CD39 antibody Q3w for administration as neoadjuvant therapy and Q4w for administration as adjuvant therapy. In one embodiment, the instructions specify anti-PD(L)1 antibody for administration as Q3 or Q4. In one embodiment, the instructions specify anti-PD(L)1 antibody Q3w for administration as neoadjuvant therapy and Q4w for administration as adjuvant therapy. In one embodiment, the instructions specify that the anti-CD39 antibody and the anti-PD(L)1 antibody are administered on the same day (e.g., on day 1 of a 3-week cycle in the neoadjuvant setting and on day 1 of a 4-week cycle in the adjuvant setting). In one embodiment, the instructions specify that the anti-CD39 antibody and the anti-PD(L)1 antibody are administered for 4 cycles as neoadjuvant therapy. In one embodiment, the instructions specify that the anti-CD39 antibody and the anti-PD(L)1 antibody may be administered in combination with chemotherapy in the neoadjuvant setting. The anti-CD39 antibody may optionally be specified 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.

[0207] Optionally, the kit further comprises a dose of chemotherapy, such as chemotherapy comprising a platinum agent, gemcitabine, including combinations of chemotherapeutic agents, such as carboplatin and paclitaxel, cisplatin and gemcitabine, cisplatin and pemetrexed, or carboplatin and pemetrexed.

[0208] Example

[0209] Example 1: CD39 expression in early and late NSCLC biopsies

[0210] Fifty squamous cell NSCLC (sqNSCLC) and fifty adenocarcinoma NSCLC (adNSCLC) FFPE samples were stained for CD39 using clone EPR20627 from Abcam. The CD39 expression score (0 - 12) is a combination of staining frequency (0 - 4) and intensity (1 - 3).

[0211] The results are shown in Figure 1 : Figure A shows that the total CD39 score is the sum of stromal, immune, and tumor expression scores (0 - 60). Figure B shows that the stromal score is the sum of vascular (0 - 12) and connective tissue (0 - 12) expression scores. Figure C shows the immune score as the sum of small immune cell (0 - 12) and large immune cell (0 - 12) scores. In each case, the expression scores are shown by cancer stage (stage I, II, or III).

[0212] Both sqNSCLC and adNSCLC showed staining of stromal cells and immune cells (including early biopsies). The total score was higher in sqNSCLC. No CD39 staining or poor CD39 staining was observed on tumor cells.

[0213] Example 2: ATP release from squamous NSCLC tumor cells after chemotherapy

[0214] This experiment aimed to study the ATP release of squamous NSCLC tumor cells after chemotherapy treatment. The squamous NSCLCH1703 tumor cell line was incubated alone with different chemotherapies. Measurements of ATP release were performed at 24 hours after cisplatin treatment, 72 hours after carboplatin, 60 hours after oxaliplatin, 48 hours after pemetrexed, 48 hours after gemcitabine, 72 hours after 5 - FU, 24 hours after paclitaxel, and 40 hours after docetaxel. Using a luminescence - based assay to measure extracellular ATP release in the culture supernatant and expressed as luminescence arbitrary units (AU). Mean ± SD.

[0215] The results are shown in Figure 2 which shows the ATP release of the H1703 tumor cell line (sqNSCLC) after treatment with different chemotherapies, cisplatin, carboplatin, oxaliplatin, pemetrexed, gemcitabine, 5 - Fu, paclitaxel, and docetaxel. Each chemotherapeutic agent induced significant ATP release in the NSCLC tumor cell line.

[0216] Example 3: IPH5201 can accumulate released ATP after chemotherapy .

[0217] H1703 (CD39-) cells were incubated with recombinant human CD39 protein (huCD39) (400 ng / mL), which mimics soluble CD39 in the tumor microenvironment, in the presence or absence of 10 μg / mL of IPH5201, a humanized anti-CD39 antibody with the corresponding heavy and light chain amino acid sequences of SEQ ID NO: 10 and 11, and then treated with docetaxel (0.1 μM). Measurement of extracellular ATP (eATP) release was performed 40 hours after docetaxel treatment. The results are shown in Figure 3 Panel A. In H1703 cells that do not express CD39, docetaxel induced strong eATP release, which was reduced in the presence of added recombinant human CD39 protein and then restored in the presence of added IPH5201 antibody.

[0218] OAW42 (CD39+) cells were incubated with 10 μg / mL or 50 μg / mL of IPH5201 and treated with a range of doses of docetaxel. Measurement of eATP release was performed 30 hours after docetaxel treatment. eATP in the cell culture supernatant was measured using a luminescence-based assay and expressed as luminescence arbitrary units (AU). Mean ± SD. The results are shown in Figure 3 Panel B. In OAW42 cells that express CD39, docetaxel induced at most moderate eATP release at the highest concentration, whereas in the presence of 10 μg / mL or 50 μg / mL of IPH5201, eATP accumulated significantly.

[0219] Example 4: CD39 is expressed in the tumor microenvironment of a mouse model and moIPH5201 significantly reduces tumor adenosine levels in vivo

[0220] The expression of human CD39 in MCA205 tumors implanted into human CD39 knock-in (huCD39KI) mice was evaluated, and then the intratumoral level of adenosine was measured after in vivo treatment with moIPH5201, an antibody with VH and VL of SEQ ID NO: 8 and 9 and a murine constant domain including an amino acid substitution to eliminate binding to murine Fcγ receptors in the Fc domain, in order to evaluate the reduction of adenosine levels in tumors by moIPH5201, which could be reduced in vivo.

[0221] Human CD39 expression on MCA205 tumors harvested on day 16 after implantation was evaluated by IHC using clone EPR20627 from Abcam B&C. MCA205 tumors were implanted into huCD39KI mice. On days 7 and 14 after tumor implantation, they were treated with moIPH5201 (20 mg / kg) or isotype control. On day 16, the tumors were harvested. The results are shown in Figure 4In the left panel, a significant percentage of cells express CD39.

[0222] As described by Goodwin et al., adenosine quantification in harvested MCA205 tumors was performed using pre-column derivatization and liquid chromatography-tandem mass spectrometry (LC-MS / MS). 2019 Anal Biochem 568:78-88. The results are shown in Figure 4 In the right panel, animals treated with moIPH5201 showed less intratumoral adenosine compared to the control.

[0223] Blocking of CD39 enzyme activity was determined using the Wachstein-Meisel CD39 enzyme assay (brown indicates enzyme activity). Tissues from animals treated with moIPH5201 showed less brown indicating reduced CD39 enzyme activity.

[0224] Example 5: Anti-CD39 improves the in vivo anti-tumor efficacy of chemotherapy and anti-PD-L1

[0225] In vivo experiments were conducted in human CD39 knock-in (huCD39KI) mice, which were genetically modified to express human CD39 instead of its murine counterpart, murine CD39. These mice were subcutaneously (s.c.) implanted with the MC38 cancer cell line. Mice bearing established tumors were dosed with an isotype control (IC) antibody, an anti-human CD39 antibody (moIPH5201, as described above), gemcitabine chemotherapy, and an anti-mouse PD-L1 antibody (an antibody with an Fc domain containing amino acid substitutions to eliminate binding to murine Fcγ receptors that binds murine PD-L1).

[0226] In vitro anti-tumor efficacy of moIPH5201 and gemcitabine

[0227] Male and female huCD39KI mice were subcutaneously (s.c.) implanted with 1x10 6 MC38 cancer cells. Treatment was initiated after randomization of the mice. Mice were treated twice a week with gemcitabine (25 mg / kg) or PBS (on days 8, 11, 15, and 18 after implantation) for two weeks, and once a week with an anti-human CD39 antibody (moIPH5201) or the corresponding isotype control (IC) antibody (400 μg / mouse) for four weeks.

[0228] On day 7 after tumor cell implantation, the mice were randomly divided into four groups with an average tumor volume of 72 mm 3 ±27 mm 3 and individual tumor volumes ranging from 37 mm 3 to 130 mm 3 .

[0229] The results are shown in Figure 5In the middle. Compared with the control group, both gemcitabine alone and the combination of gemcitabine and moIPH5201 sharply and significantly slowed tumor growth (p<0.001). Compared with the control, moIPH5201 did not alter tumor growth and did not improve the anti-tumor efficacy of gemcitabine. No complete response was observed in this experiment. No evidence of overt toxicity was observed in any group.

[0230] In vitro anti-tumor efficacy of chemotherapy, moIPH5201 and anti-PD-L1 antibody

[0231] The anti-tumor effects of different treatment combinations of gemcitabine, moIPH5201, and anti-PD-L1 antibody were evaluated as combination therapies. Three independent experiments were conducted to evaluate this triple combination.

[0232] Experiment 1

[0233] Male and female huCD39KI mice were s.c. implanted with 1x10 6 MC38s. Treatment was initiated after randomization of the mice. The mice were treated twice a week with gemcitabine (25 mg / kg) or PBS for two weeks, twice a week with anti-mouse PD-L1 antibody or the corresponding isotype control antibody (200 μg / mouse) for three weeks, and once a week with anti-human CD39 antibody (moIPH5201) or the corresponding isotype control antibody (400 μg / mouse) according to the experiment for four or five weeks. The mice were randomly divided into five groups on day 7 after tumor cell implantation, with an average tumor volume of 62 mm 3 ±23 mm 3 and individual tumor volumes ranging from 31 mm 3 to 117 mm 3 in between.

[0234] No evidence of increased toxicity was observed in any group. One mouse in the gemcitabine + moIPH5201 group lost more than 10% of its body weight but gradually regained its initial body weight after the last gemcitabine treatment.

[0235] Compared with the control, gemcitabine as a single agent or in all three tested combinations significantly slowed tumor growth (p<0.001). Neither anti-PD-L1 nor moIPH5201 improved the anti-tumor effect of gemcitabine as a single agent, while adding both anti-PD-L1 + moIPH5201 to gemcitabine induced significantly better efficacy than gemcitabine alone (p < 0.05 when the control group was excluded). In these groups, moIPH5201 did not statistically significantly improve the gemcitabine + anti-PD-L1 efficacy, and anti-PD-L1 did not statistically significantly improve the anti-tumor effect of gemcitabine + moIPH5201.

[0236] The results are shown inFigure 6 Among them, compared with gemcitabine alone, both anti-PD-L1 and moIPH5201 agents increase the number of complete responses when combined with gemcitabine. In addition, the benefit of moIPH5201 on the anti-tumor effect of gemcitabine + anti-PD-L1 is supported by the proportion of complete responses (CR): 6 / 10 (60%) was observed in the gemcitabine + anti-PD-L1 + moIPH5201 group, while 3 / 10 (30%) was observed in the group treated with gemcitabine + anti-PD-L1. Arrows indicate the administration of different agents.

[0237] Experiment 2

[0238] As in Experiment 1, male and female huCD39KI mice were s.c. implanted with 1x10 6 MC38s. Treatment was initiated after randomization of the mice. The mice were treated twice a week with gemcitabine (25 mg / kg) or PBS for two weeks, twice a week with anti-mouse PD-L1 antibody or corresponding isotype control antibody (200 μg / mouse) for three weeks, and once a week with anti-human CD39 antibody (moIPH5201) or corresponding isotype control antibody (400 μg / mouse) according to the experiment for four or five weeks. The mice were randomly divided into five groups on day 7 after tumor cell implantation, with an average tumor volume of 64 mm 3 ±23 mm 3 and individual tumor volumes ranging from 31 mm 3 to 115 mm 3 .

[0239] As in the previous experiment, gemcitabine, as a single agent and in combination therapy, significantly slowed tumor growth compared to the control (p < 0.001). No evidence of overt toxicity was observed in any group.

[0240] Interestingly, when the untreated control group was excluded from the statistical analysis, moIPH5201 tended to improve the anti-tumor efficacy of the combination of gemcitabine + anti-PD-L1 (margin of difference significant, p < 0.1). Similar to the previous experiment, adding both anti-PD-L1 + moIPH5201 to gemcitabine induced better efficacy than gemcitabine alone (margin of difference significant p < 0.1 when the control group was excluded).

[0241] The results are shown in Figure 7 . The proportion of CR was 4 / 10 in the group treated with the triple combination and 3 / 10 in the group treated with gemcitabine + anti-PD-L1. Given these results, the experiment was repeated a third time under the same settings and the results are shown in the next section.

[0242] Experiment 3

[0243] As in Experiments 1 and 2, male and female huCD39KI mice were s.c. implanted with 1x10 6 MC38s. Treatment was initiated after randomization of the mice. The mice were treated twice weekly with gemcitabine (25 mg / kg) or PBS for two weeks, twice weekly with anti-mouse PD-L1 antibody or corresponding isotype control antibody (200 μg / mouse) for three weeks, and once weekly with anti-human CD39 antibody (moIPH5201) or corresponding isotype control antibody (400 μg / mouse) for four or five weeks according to the experiment. Mice were randomized into five groups on day 7 after tumor cell implantation, with a mean tumor volume of 74 mm 3 ±21 mm 3 and individual tumor volumes ranging from 40 mm 3 to 121 mm 3 . No signs of overt toxicity were observed in any group.

[0244] Results are shown in Figure 8 . As in the two previous experiments, gemcitabine, as a single agent and in all combinations tested, significantly inhibited tumor growth compared to control (p < 0.001). Neither anti-PD-L1 nor moIPH5201 improved the anti-tumor effect of gemcitabine as a single agent. However, adding both anti-PD-L1 + moIPH5201 to gemcitabine induced a better efficacy than gemcitabine alone (marginally significant difference p < 0.1 when excluding the control group). moIPH5201 did not statistically significantly improve the efficacy of gemcitabine + anti-PD-L1. Thus, in this experiment, compared to gemcitabine alone, anti-PD-L1 increased the number of complete responses when combined with gemcitabine, but moIPH5201 did not increase the number of CRs when combined with gemcitabine alone or with gemcitabine + anti-PD-L1.

[0245] Pooled analysis

[0246] The three experiments were combined and the results are shown in Figure 9 , showing 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 gemcitabine at 25 mg / kg ip or PBS, anti-mouse PD-L1 at 200 μg ip or corresponding isotype control antibody, and moIPH5201 at 400 μg iv or corresponding isotype control antibody as indicated. The figure shows tumor growth in each individual (n = 31 mice / group). CR: complete response.

[0247] Combined experiments conducted in the MC38 tumor model (n = 3) in response to gemcitabine chemotherapy showed that while neither anti-PD-L1 nor moIPH5201 improved the anti-tumor effect of gemcitabine as a single agent (no statistically significant difference), the anti-PD-L1 + moIPH5201 + gemcitabine triple combination improved anti-tumor efficacy compared to gemcitabine alone (p < 0.05). In addition, moIPH5201 improved the anti-tumor efficacy of the gemcitabine + anti-PD-L1 combination (p < 0.05, linear mixed effects model analysis). This therapeutic benefit was also supported by the proportion of complete responses: 55% (17 / 31) was observed in the group of mice treated with the triple combination, while only 42% (13 / 31) was observed in the group of mice treated with gemcitabine + anti-PD-L1.

[0248] In summary, IPH5201 blocks CD39 enzyme activity, reduces intratumoral adenosine levels and increases extracellular ATP release from tumor cells after chemotherapy treatment, and ultimately improves anti-tumor efficacy in preclinical models in combination with chemotherapy and blocking anti-PDL1 antibodies. Overall, the expression profile of CD39 in early NSCLC and preclinical combination data support the clinical evaluation of IPH5201 in combination with durvalumab and chemotherapy in patients with early NSCLC.

[0249] Example 6: Results from a Phase 1 human trial of an anti-CD39 antibody and a new treatment regimen for anti-CD39 in human therapy Protocol design

[0250] A first-in-human, multi-center, non-randomized, open-label phase 1 study was conducted to evaluate the safety, efficacy, pharmacokinetics (PK) and pharmacodynamics (PD) of IPH5201 ± durvalumab (Imfinzi TM ) in patients with advanced solid tumors. The study consisted of two consecutive dose-escalation parts:

[0251] Part 1: Escalating doses of IPH5201 (100 mg, 300 mg, 1000 mg and 3000 mg), every 3 weeks (Q3W).

[0252] Part 2: Escalating doses of IPH5201 (300 mg, 1000 mg and 3000 mg) and 1500 mg durvalumab, Q3W.

[0253] The PD cohort enrolled patients with advanced squamous cell lung cancer or advanced pancreatic ductal adenocarcinoma at the first two dose levels of Part 1 and Part 2. The primary endpoint was safety and tolerability. Key secondary endpoints included initial anti-tumor activity, PK, and immunogenicity measured by objective response and disease control according to RECIST v1.1. Exploratory endpoints included efficacy measured by duration of response and progression-free survival (PFS), overall survival, and assessment of biomarkers according to RECIST v1.1.

[0254] Key inclusion criteria were:

[0255] · Adults aged ≥ 18 years.

[0256] · Histologically or cytologically confirmed advanced solid tumor.

[0257] · At least one measurable lesion according to RECIST v1.1.

[0258] · Disease refractory to standard therapy or for which no standard therapy exists.

[0259] · Eastern Cooperative Oncology Group performance status of 0 or 1.

[0260] · Archived or fresh tumor samples available.

[0261] Key exclusion criteria were:

[0262] · Prior treatment with any agent targeting CD73, CD39, or adenosine receptor.

[0263] · Treatment with any conventional or investigational anti-cancer therapy within 21 days of the first scheduled dose.

[0264] · Active or prior autoimmune or inflammatory disorder within the past 5 years.

[0265] · Cardiac and vascular criteria, including the presence of acute coronary syndrome or thromboembolic event, congestive heart failure, severe arrhythmia requiring medical treatment, or uncontrolled hypertension within 6 months prior to enrollment.

[0266] · Untreated metastases to the central nervous system.

[0267] PK and PD modeling

[0268] PK / PD modeling and simulation were used to guide dose selection. IPH5201 PK data from 45 patients and PD data on cell surface CD39 (mCD39) occupancy on monocytes from 43 patients were available. A population PK model for IPH5201 was developed using a two-compartment model best described with parallel linear and saturable kinetics.

[0269] Then an indirect response PD model (inhibitory effect on K in ) was established to describe the relationship between the concentration of IPH5201 and the free mCD39 on monocytes ( Figure 10 ). Increasing the concentration of the drug IPH5201 would decrease the free CD39+ (%) on monocytes, and the model parameters are listed in the following table. This model accurately predicts the free CD39+ (%) on monocytes. The maximum inhibition of free CD39+, Imax = 0.926; IC50 = 2.44 μg / mL defines the concentration of IPH5201 that produces 50% of the maximum inhibition. Then the final indirect response model was used for parameter simulation.

[0270] Parameter Value %RSE Production rate constant (1 / h) 20.5 9％ IC50 (ug / mL) 2.44 8％ Maximum inhibition (IMAX) 0.92 1％ Baseline response (%) 92 FIX Proportional error 0.57 0.3％ IIV on KIN 70.6％ 9％ IIV on IMAX 5.9％ 5％ IIV on RS0 15.2％ 34％

[0271] A total of 499 simulations were performed using this model to predict the mCD39 occupancy on monocytes, and the recommended doses of Q3W or Q4W that achieve at least 85% occupancy and maintain sufficient exposure time at steady state in most patients were selected.

[0272]

[0273]

[0274] At 3000 mg q3w, 95% of patients will achieve 85% mCD39 occupancy in monocytes throughout the dosing interval (100% of the duration); at 3000 mg q4w, 91.8% of patients achieve 85% mCD39 occupancy with 100% exposure duration. 2000 mg q3w can achieve >90% patient coverage, but not for Q4W (84% patient coverage). However, it is expected that 2250 mg q4w will achieve approximately >90% patient coverage with 100% exposure duration and 85% mCD39 occupancy.

[0275] Safety

[0276] Overall, 57 patients were treated (IPH5201, n = 38; IPH5201 + durvalumab, n = 19). No DLT was observed during dose escalation. Treatment-emergent adverse events (TEAEs) occurred in 96.5% of patients; 40.4% had ≥ grade 3 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 (TRAE) occurred in 66.7% of patients (37 / 57 (64.9%) related to IPH5201; 11 / 19 (57.9%) related to durvalumab). 10.5% of patients had grade 3 TRAEs; no grade 4 TRAEs. The most common TRAEs were infusion-related reactions (21.1%), fatigue (17.5%), nausea, arthralgia, tumor pain, and pruritus (each 8.8%). 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 at least grade 3 treatment-emergent adverse events.

[0277] Pharmacokinetics and pharmacodynamics

[0278] Occupancy of CD39 by IPH5201 in patient samples was determined by assessing free soluble CD39 and free membrane-bound CD39.

[0279] To assess free soluble CD39, a monoclonal anti-CD39 antibody that competes with IPH5201 was coated onto microtiter plates, allowing capture of free CD39. A second non-competing anti-CD39 monoclonal antibody carrying a biotin tag was used to bind to a site on CD39 different from the capture antibody. Bound biotin molecules were detected by addition of streptavidin horseradish peroxidase (SA-HRP) conjugate. Tetramethylbenzidine enzyme substrate was added to generate a colorimetric reaction, which was measured at a wavelength of 450 nm. The concentration of free CD39 in the sample was determined by interpolation from a standard curve using a four-parameter curve fit of the 1 / Y 2 weight values that relate color intensity to CD39 concentration.

[0280] To assess free CD39 on monocytes and B cells, antibodies with the VH and VL of IPH5201 and the murine Fc domain were incorporated into blood samples and labeled with a fluorochrome. A second non-competing anti-CD39 monoclonal antibody labeled with a different fluorochrome was used to bind to a site on CD39 different from the capture antibody. CD14, CD19, and CD45 were detected by incubation with anti-CD14, CD19, and CD45 monoclonal antibodies labeled with different fluorochromes, respectively. Binding was detected by flow cytometry.

[0281] The results are shown in Figure 11A and Figure 11B . C1D1 indicates before dosing in cycle 1; C1D2 indicates after dosing in cycle 1; C1D8 indicates day 8 of cycle 1; C2D1 indicates day 1 of cycle 2; C3D1 indicates day 1 of cycle 3; EOT indicates end of treatment; LLOQ, lower limit of quantification; V1, visit 1 (at screening). Figure 11A Shows that IPH5201 saturates the binding of soluble CD39 at ≥300 mg. Each line represents data from one patient. The increase in total soluble CD39 (not shown) is consistent with the stabilization of soluble CD39 increased by antibody binding. A similar trend was observed in combination therapy with durvalumab. Figure 11B Shows that IPH5201 saturates the binding of membrane-bound CD39 on immune cells at 3000 mg. In monocytes (right panel), a 100 mg IPH5201 dose resulted in complete rebound of free CD39 at C2D1, and a 300 mg IPH5201 dose resulted in partial rebound of free CD39 at C2D1. In B cells, only a 100 mg IPH5201 dose resulted in a rebound of free CD39 at C2D1.

[0282] Figure 12 Shows the pharmacokinetics (PK) of IPH5201 as monotherapy (left panel) or as combination therapy with durvalumab (right panel), where the x-axis is time in days after the first dose and the y-axis is the serum IPH5201 concentration. In the left panel, the curves from bottom to top represent 100 mg, 300 mg, 1000 mg, and 3000 mg doses of IPH5201; in the right panel, the curves from bottom to top represent 300 mg, 1000 mg (up to the 21-day time point), and 3000 mg doses of IPH5201. The PK of IPH5201 is non-linear at ≤300 mg and linear at ≥1000 mg. Free soluble CD39 was above the lower limit of quantification in 6 / 40 patients on day 2 or later in cycle 1 (n = 2, 100 mg; n = 1, 1000 mg; n = 3, 3000 mg).

[0283] Tumor samples were obtained from patients with pancreatic cancer and CD39 enzyme activity was evaluated. The Wachstein-Meisel assay was used to evaluate CD39 enzyme activity to detect the presence of phosphate esters hydrolyzed from ATP due to the enzyme activity of CD39. Enzyme activity was evaluated by a semi-quantitative scoring method of tumor, stroma, and vasculature. Pathologists performed a comprehensive evaluation to determine whether there was a significant decrease on day 15 compared to screening biopsies. The following table shows a decrease in tumor CD39 enzyme activity in 5 / 8 patients with available samples. A dose of 3000 mg of IPH5201 showed a decrease in enzyme activity.

[0284]

[0285] In summary, PK / PD modeling and simulation predict that the recommended doses of 3000 mg Q3W or Q4W will achieve ≥85% mCD39 occupancy on monocytes in >90% of patients, with or without durvalumab, and maintain sufficient exposure time at steady state (i.e., 100% of the dose interval duration). Therefore, dosing 3000 mg of IPH5201 Q3W or Q4W offers advantages for use in both monotherapy and combination therapy with durvalumab, including reducing the risk of dosing errors and the likelihood of co-administering with durvalumab on the same day.

[0286] Example 7: Phase 2 single-arm study design of IPH5201 in resectable NSCLC

[0287] Based on the results among the foregoing results, a Phase 2 single-arm clinical trial study of IPH5201 in resectable NSCLC was designed.

[0288] The key eligibility criteria were resectable (Stage II / IIIA) NSCLC, no prior treatment, and an Eastern Cooperative Oncology Group (ECOG) performance status (PS) of 0 or 1, all CPS statuses of PDL1 expression, and EGFR / ALK wild-type.

[0289] The primary endpoints were pathologic complete response (pCR) and safety. Secondary endpoints: surgical feasibility; major pathologic response (mPR); objective response rate (ORR); median and landmark (12 months) event-free survival (EFS), disease-free survival (DFS), and overall survival (OS); and the evaluation of PK and anti-drug antibodies (ADA) of IPH5201. The study will include a safety run-in period for the first six patients.

[0290] Figure 13A schematic diagram of a treatment regimen for treatment is shown. As neoadjuvant therapy, IPH5201 and durvalumab are administered in combination with chemotherapy (CT) for 4 cycles at Q3W with a fixed dose of 3000 mg of IPH5201 (2250 mg can be used as an alternative) and a fixed dose of 1500 mg of durvalumab. CT includes a combination of carboplatin / paclitaxel, cisplatin / gemcitabine, cisplatin / pemetrexed, or carboplatin / pemetrexed. Within 40 days of the last dose of the investigational drug in the neoadjuvant therapy, the patient undergoes tumor surgical resection with or without additional radiotherapy. Within 10 weeks of surgery, the patient receives adjuvant therapy with IPH5201 and durvalumab at Q4w at a fixed dose of 3000 mg of IPH5201 (or 2250 mg if used as an alternative) and a fixed dose of 1500 mg of durvalumab for 12 cycles or until progressive disease occurs.

[0291] All references cited herein (including publications, patent applications, and patents) are hereby incorporated by reference in their entirety as if each reference were individually and specifically indicated to be incorporated by reference and fully set forth herein (to the maximum extent permitted by law), regardless of any separate incorporation of a particular document provided elsewhere in this text.

[0292] Unless otherwise indicated herein or clearly contradicted by the context, the terms “a / an” and “the” and similar pronouns used herein shall be construed to cover both the singular and the plural.

[0293] Unless otherwise specified, all exact values provided herein represent corresponding approximate values (e.g., all exact exemplary values provided relative to a particular factor or measurement may be considered to also provide corresponding approximate measurements, modified by “about” where appropriate).

[0294] The description herein of any aspect or embodiment using terms such as “comprising,” “having,” “including,” or “containing” is intended to provide support for a similar aspect or embodiment “consisting of the one or more specific elements,” “consisting essentially of the one or more specific elements,” or “essentially including the one or more specific elements” herein, unless otherwise stated or clearly contradicted by the context (e.g., a composition described herein as including a specific element should be understood to also describe a composition consisting of that element, unless otherwise stated or clearly contradicted by the context).

[0295] Unless otherwise protected by the claims, the use of any and all examples or exemplary language (e.g., "such as") provided herein is only intended to better illustrate the invention and does not limit the scope of the invention. No language in the specification should be construed as indicating that any element not protected by the claims is necessary for the practice of the invention.

Claims

1. A method of treating cancer in a patient in need thereof, the method 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 wherein the anti-CD39 antibody comprises (a) HCDR1, HCDR2 and HCDR3 that are SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4 respectively, and (b) LCDR1, LCDR2 and LCDR3 sequences that are SEQ ID NO: 5, SEQ ID NO: 6 and SEQ ID NO: 7 respectively, wherein the treatment regimen is neoadjuvant therapy.

2. The method according to any one of the preceding claims, wherein the neoadjuvant therapy treatment regimen further comprises a chemotherapeutic agent.

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

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

5. The method according to any one of claims 1 to 4, wherein the anti-CD39 antibody is administered once every 3 weeks as neoadjuvant therapy.

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 3 weeks as neoadjuvant therapy.

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

8. The method according to any one of claims 3 to 7, wherein the anti-CD39 antibody is administered once every 4 weeks as adjuvant therapy.

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 4 weeks as adjuvant therapy.

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

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

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

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

14. The method according to any one of the preceding claims, wherein the treatment regimen comprises: (a) Administering to the patient an effective amount of an anti-CD39 antibody, an anti-PD(L)1 antibody and optionally a chemotherapeutic agent before tumor surgical resection, (b) Administering to the patient an effective amount of an anti-CD39 antibody and an anti-PD(L)1 antibody after tumor surgical resection.

15. The method according to any one of the preceding claims, wherein the anti-PD(L)1 antibody is durvalumab.

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

17. A method of administering an anti-CD39 antibody to a human patient, the anti-CD39 antibody comprising a heavy chain and a light chain comprising the amino acid sequences of SEQ ID NO: 8 and 9, respectively, wherein the anti-CD39 antibody is administered at a fixed dose of 3000 mg once every 3 weeks or once every 4 weeks.

18. The method according to claim 17, wherein the anti-CD39 is administered on day 1 of a 3-week cycle for one or more cycles and on day 1 of a 4-week cycle for one or more cycles.

19. The method according to claim 17 or 18, wherein the anti-CD39 is administered in combination with an anti-PD(L)1 antibody, optionally wherein the anti-PD(L)1 antibody is administered on day 1 of the 3-week cycle for one or more cycles and on day 1 of the 4-week cycle for one or more cycles, optionally wherein the anti-PD(L)1 antibody is durvalumab, optionally wherein durvalumab is administered at a fixed dose of 1500 mg.

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

11.

21. The method according to any one of the preceding 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. The method according to any one of the preceding 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. The method according to any one of the preceding claims, wherein the cancer is surgically resectable.

24. The method according to any one of the preceding claims, wherein the cancer is lung cancer.

25. The method according to any one of the preceding claims, wherein the cancer is non-small cell lung cancer.

26. The method according to any one of the preceding claims, wherein the cancer is stage I-III NSCLC.

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

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

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

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

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

Images