Combining checkpoint inhibitors and oncolytic viruses to treat cancer

Abstract

The present disclosure relates to a method for inhibiting tumor growth in a patient in need thereof. The method comprises administering an oncolytic virus to a patient in combination with a therapeutically effective amount of a PD-1 inhibitor (e.g., cemiplimab or its biological equivalent) and optionally a therapeutically effective amount of a CTLA-4 inhibitor (e.g., ipilimumab or its biological equivalent). In some embodiments, administering the combination enhances tumor inhibition compared to administering either the antibody or the virus alone. In some embodiments, the oncolytic virus is a recombinant vesicular stomatitis virus (VSV), e.g., VV1.

Description

[Technical field]

[0001] The present disclosure relates generally to combination therapies for the treatment of cancer using oncolytic viruses and checkpoint inhibitors, such as programmed death 1 (PD-1) pathway inhibitors and / or cytotoxic T-lymphocyte antigen-4 (CTLA-4) inhibitors. [Background technology]

[0002] VV1 (also known as VSV-IFNβ-NIS) is derived from the vesicular stomatitis virus (VSV), a bullet-shaped, negative-stranded ribonucleic acid (RNA) virus with low human seroprevalence; it has been engineered to selectively replicate in and kill human cancer cells. The virus encodes human interferon-β (IFNβ), which boosts antitumor immune responses and enhances tumor specificity, and the thyroid sodium-iodide symporter (NIS), which allows imaging of viral spread. VSV-IFNβ-NIS is synergistic with different anti-PD-(L)1 antibodies in several tumor models.

[0003] VV1 is a living RNA virus that has been engineered to express both human IFNβ and NIS genes. The virus was constructed by inserting the IFNβ gene downstream of the M gene and the NIS gene (cDNA) downstream of the G protein gene into a full-length infectious molecular clone of Indiana lineage VSV [Non-Patent Document 1]. This virus is not a vaccine. VSV-IFNβ-NIS reproduces in BHK cells with kinetics similar to the parental strain of the virus and can grow to high titers. It selectively reproduces in human cancer cells, many of which are unable to mount an effective antiviral response mediated through the IFNβ pathway. However, IFNβ production from infected cells serves to protect non-cancer cells from the effects of the virus. As a result, the virus becomes directly cytopathic to tumor cells, resulting in rapid lysis accompanied by viral amplification [Non-Patent Document 2; Non-Patent Document 3]. Preclinical studies indicate that IV treatment with oncolytic VSV selectively targets tumor cells and induces acute inflammatory responses and IT immune infiltration [4; 5; 6; 1; 7; 8].

[0004] VSV-IFNβ-NIS-infected tumor cells also express NIS, a membrane ion channel that actively transports iodide into cells. Radioactive iodine uptake by cells expressing NIS provides the basis for in vivo imaging with 99mTc pertechnetate or radioactive iodine I-12, which can reveal the time-dependent profile of VSV-IFNβ-NIS gene expression and the location of VSV-IFNβ-NIS-infected cells during viral propagation and clearance [Non-Patent Document 1]. F18-tetrafluoroborate (F18-TFB) PET imaging technology can also be used to track viral replication and propagation [Non-Patent Document 9]. Very recently, the safety and feasibility of using F18-TFB PET imaging to localize tissues that physiologically express NIS has been shown in healthy human subjects [Non-Patent Document 10].

[0005] Programmed death 1 (PD-1) is an ideal target for immunotherapy because it plays a key role in autoimmunity, tumor immunity and infectious immunity. Blockade of PD-1 with antagonists, including monoclonal antibodies, has been investigated in the treatment of cancer and chronic viral infections. Blockade of PD-1 is also an effective and well-tolerated approach to stimulate immune responses, and therapeutic benefits have been achieved against a variety of human cancers, including melanoma, renal cell carcinoma and non-small cell lung cancer (NSCLC) (Non-Patent Document 11; Non-Patent Document 12; Non-Patent Document 13; Non-Patent Document 14; Non-Patent Document 15; Non-Patent Document 16; Non-Patent Document 17; Non-Patent Document 18; Non-Patent Document 19; Non-Patent Document 20; Non-Patent Document 21; Non-Patent Document 22; Non-Patent Document 23). Monoclonal antibodies against PD-1 are known in the art and are described, for example, in U.S. Pat. No. 6,233,633; ... For example, cemiplimab (REGN2810; also known as LIBTAYO®) is a high-affinity, fully human, hinge-stabilized IgG4P antibody against the PD-1 receptor that potently blocks the interaction of PD-1 with its ligands, PD-L1 and PD-L2.

[0006] Blockade of cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) has been shown to increase the number of activated T cells and allow the proliferation of tumor-specific cytotoxic T cells. CTLA-4 is a type I transmembrane T cell inhibitory checkpoint receptor expressed on normal and regulatory T cells. CTLA-4 negatively regulates T cell activation by outcompeting the stimulatory receptor CD28 from binding to its common natural ligands, B7-1 (CD80) and B7-2 (CD86). Ipilimumab is a recombinant humanized monoclonal antibody that binds to CTLA-4.

[0007] Lung cancer is the leading cause of cancer deaths worldwide

[24] . In the United States, approximately 228,000 patients will be diagnosed with lung cancer in 2019, with an estimated 80-85% of these cases being of NSCLC histology

[25] . The etiologic association of NSCLC with exposure to tobacco carcinogens is particularly relevant for cancer immunotherapy, as current approaches targeting immune checkpoints show increased response in tumors with multiple somatic mutations, as is the case in smoking-induced NSCLC. Conversely, immune checkpoint blockade has been significantly less successful in never-smokers, including the majority of anaplastic lymphoma kinase (ALK)-rearranged or epidermal growth factor receptor (EGFR)-mutated NSCLC

[26] . Most NSCLC patients present at an advanced stage of the disease, are symptomatic at diagnosis, and have an associated poor prognosis with no curative options. In stage IV, systemic palliative treatment is recommended, including cytotoxic chemotherapy, immune checkpoint inhibitors (CPIs) and a range of targeted agents in selected molecularly defined subsets of NSCLC patients. Despite considerable progress in the treatment of NSCLC with anti-PD-(L)1 therapy, advanced disease remains largely incurable.

[0008] Melanoma remains highly prevalent, with over 96,000 new cases diagnosed in the United States in 2019, although overall mortality remains low

[27] . However, despite recent advances with CPI therapy, metastatic melanoma continues to retain a relatively high incidence. Overall survival of patients with advanced melanoma has improved from approximately 9 months prior to 2011 to at least 2 years and possibly more for certain mutations, such as BRAF V600 mutant melanoma

[28] , with some patients experiencing ongoing long-term tumor control. Melanoma is amenable to immunotherapy for a variety of reasons, including extensive tumor infiltration by T cells, high mutational load, and crosstalk between oncogenic signaling pathways and immunobiology. The use of ipilimumab, nivolumab, and pembrolizumab has become the standard of care for patients with advanced metastatic melanoma. Furthermore, despite long-term benefit for some patients, sustained responses remain elusive. Resistance mechanisms to immunotherapy, such as loss of tumor antigen presentation and T cell trafficking, have been observed.

[0009] Head and neck squamous cell carcinoma (HNSCC) is diagnosed in approximately 65,000 patients annually in the United States, with approximately 14,000 deaths secondary to HNSCC each year

[27] . Globally, over 600,000 patients are diagnosed with the disease annually

[24] . Head and neck cancers, referred to as HNSCC, constitute a collection of tumors arising in the squamous epithelial cells that line the mucosal surfaces of the head and neck, and are categorized according to the region of the head or neck in which they occur: oral cavity, pharynx (including the nasopharynx, oropharynx, and hypopharynx), paranasal sinuses, nasal cavity, and salivary glands. Infection with human papillomavirus (HPV), particularly HPV-16 and 18, is a risk factor for developing HNSCC, especially oropharyngeal cancer involving the tonsils or base of the tongue

[29] . In the United States, the incidence of oropharyngeal cancer caused by HPV infection is increasing, while the incidence of oropharyngeal cancer associated with other causes is decreasing

[29] .

[0010] When HNSCC is detected and treated at the earliest stage, it is curable with excellent long-term OS. However, once a patient develops locally recurrent or metastatic disease, HNSCC remains essentially incurable, with a 5-year survival rate of less than 50% [Non-Patent Document 30]. Monoclonal antibodies targeting the T cell immune CPI PD-(L)1 have recently shown substantial antitumor activity in patients with advanced HNSCC. Pembrolizumab received accelerated approval based on response rates and durability of response in patients with recurrent or metastatic HNSCC whose disease progressed after platinum-containing chemotherapy (second-line), regardless of HPV status. The ORR after pembrolizumab in this population is approximately 16% [Non-Patent Document 31]. Furthermore, although recent advances in therapy for advanced HNSCC have prolonged the OS of patients, the disease remains incurable, and pembrolizumab did not increase response rates, only prolonging the duration of response, compared with EXTREME.

[0011] Colorectal cancer (CRC)CRC is one of the most diagnosed cancers worldwide, ranking second in incidence in women and third in men globally

[32] . In the United States, approximately 149,500 new cases of colorectal cancer are diagnosed annually, and approximately 52,980 Americans die from CRC, making it the third leading cause of cancer deaths in both men and women

[33] . Approximately 50%-60% of patients with colorectal cancer have metastatic disease at the time of diagnosis, with common sites of involvement including the liver, lymph nodes, lungs, peritoneum and soft tissues [34, 35, 36], and approximately half of early stage patients eventually develop metastatic or progressive disease

[37] . Survival outcomes for patients with metastatic CRC are dismal, with a 5-year survival rate of approximately 14%

[24] .

[0012] Compared with HNSCC, patients with CRC have only seen modest improvements in survival with new therapies, mainly due to the lack of efficacy of anti-PD-(L)1 therapy, except for a small group of patients with microsatellite instability-high (MSI-H) CRC. The exact way in which microsatellite stable (MSS) CRC evades immune detection and destruction is still not fully understood. MSS CRC can be classified into two patterns representative of non-inflammatory tumors. The first is through immunological neglect, where tumors are poorly infiltrated by lymphocytes; the second is through an immunosuppressive microenvironment created to evade immune surveillance by recruiting myeloid-derived suppressor cells or by secreting factors, including TGFβ, to suppress chemokines and cytokines required to facilitate T cell infiltration into tumors

[38] .

[0013] Improvements in traditional therapies, such as chemotherapy, radiation therapy, surgery, and targeted therapy, as well as recent advances in immunotherapy, have improved outcomes for patients with advanced solid tumors. In the last few years, the FDA and the European Medicines Agency have approved one monoclonal antibody targeting six CPI:CTLA-4 pathways, ipilimumab, and six antibodies targeting PD-L / PD-L1, including atezolizumab, avelumab, durvalumab, nivolumab, cemiplimab, and pembrolizumab, for the treatment of patients with multiple cancer types, mainly solid tumors [39, 40]. These approvals have dramatically changed the face of cancer treatment. However, the majority of cancer patients, including the majority with tumors considered to be "sensitive" to these agents, either do not respond or become resistant to these agents

[41] . Moreover, some of the most prevalent tumors, colorectal, breast and prostate cancer, have proven largely refractory to checkpoint inhibition

[42] .

[0014] Many immunotherapy treatments are only effective in selected subgroups of cancers, such as those with microsatellite instability (MSI) / mismatch repair (MMR) deficiency or high tumor mutation burden

[43] , those expressing PD-L1 [Tecentriq® 2019], and even novel targeted agents address only very small subpopulations across tumor types [Vitrakvi® 2019]. Indeed, despite considerable early success and relatively few side effects compared to all other therapies, the majority of cancer patients do not respond to CPIs or novel targeted therapies

[41] . [Prior art documents] [Patent documents]

[0015] [Patent Document 1] US9987500 [Patent Document 2] US8008449 [Patent Document 3] US8168757 [Patent Document 4] US20110008369 [Patent Document 5] US20130017199 [Patent Document 6] US20130022595 [Patent Document 7] WO2006121168 [Patent Document 8] WO20091154335 [Patent Document 9] WO2012145493 [Patent Document 10] WO2013014668 [Patent Document 11] WO2009101611 [Patent Document 12] EP2262837 [Patent Document 13] EP2504028 [Non-patent literature]

[0016] [Non-licensed document 1] Naik 2012 [Non-licensed document 2] Barber 2004 [Non-licensed document 3] Lichty 2004) [Non-licensed document 4] Obuchi 2003

Non-licensed Document 5

Non-licensed Document 6

Non-licensed Document 7

Non-licensed literature 9

Non-licensed literature 10

Non-licensed Document 11

Non-licensed Document 12

Non-licensed Document 13

Non-licensed Document 14

Non-licensed Document 15

Non-licensed Document 16

Non-licensed Document 17

Non-licensed Document 18

Non-licensed Document 19

Non-licensed Document 20

Non-licensed Document 21

Non-licensed Document 22

Non-licensed Document 23

Non-licensed Document 24

Non-licensed Document 25

Non-licensed Document 26

Non-licensed Document 27

Non-licensed Document 28

Non-licensed Document 29

Non-licensed Document 30

Non-licensed Document 31

Non-licensed Document 32

Non-licensed Document 33

Non-licensed Document 34

Non-licensed Document 35

Non-licensed Document 36

[0017] As treatments for patients with advanced solid tumors remain scarce, there is an unmet medical need for more effective and less toxic therapies, such as combination therapies with synergistic mechanisms of action with immune CPIs as disclosed herein. [Means for solving the problem]

[0018] In one aspect, the disclosed technology relates to a method of treating or inhibiting tumor growth comprising: (a) selecting a cancer patient; and (b) administering to a patient in need thereof a combination therapy comprising: (i) a therapeutically effective amount of an oncolytic virus; and (ii) a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds to Programmed Death 1 (PD-1), wherein the anti-PD-1 antibody or antigen-binding fragment thereof comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) of the heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:1, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) of the light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:2.

[0019] In some embodiments, the oncolytic vesiculovirus comprises an oncolytic vesicular stomatitis virus (VSV). In some embodiments, the VSV comprises a recombinant VSV. In some embodiments, the recombinant VSV expresses a cytokine. In some embodiments, the cytokine comprises interferon beta (IFNb). In some embodiments, a nucleic acid sequence encoding IFNb is located between the M viral gene and the G viral gene. In some embodiments, the recombinant VSV further expresses a sodium / iodine symporter (NIS). In some embodiments, a nucleic acid sequence encoding NIS is located between the G viral gene and the L viral gene. In some embodiments, the oncolytic virus is Voyager V1. In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof comprises an HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 3, 4, 5, 6, 7, and 8, respectively. In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 1; and an LCVR comprising the amino acid sequence of SEQ ID NO: 2. In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 9 and a light chain comprising the amino acid sequence of SEQ ID NO: 10. In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof comprises cemiplimab or a biological equivalent thereof.

[0020] In some embodiments, the oncolytic virus is 10 to 10 14 TCID 50 In some embodiments, the oncolytic virus is administered to the patient in one or more doses of 10 9 TCID 50 or 10 11 TCID 50 In some embodiments, the patient is administered one or more doses of 10 9 TCID 50 A single dose of the oncolytic virus is administered to the patient, followed by 10 11 TCID 50Another dose of the oncolytic virus is administered to the patient. In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is administered to the patient in one or more doses of about 0.1 mg / kg to about 20 mg / kg of patient body weight. In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is administered to the patient in one or more doses of about 1 mg to about 1000 mg. In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is administered to the patient in one or more doses of 350 mg.

[0021] In some embodiments, the oncolytic virus is administered to the patient intratumorally or intravenously. In some embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is administered to the patient intravenously, subcutaneously, or intraperitoneally.

[0022] In some embodiments, the method further comprises administering a therapeutically effective amount of a CTLA-4 inhibitor, including ipilimumab or a biological equivalent thereof. In some embodiments, the CTLA-4 inhibitor is ipilimumab.

[0023] In some embodiments, the method comprises administering one or more doses of an oncolytic virus and one or more doses of a CTLA-4 inhibitor simultaneously. In some embodiments, the method comprises administering one or more doses of an oncolytic virus and one or more doses of a CTLA-4 inhibitor prior to administering one or more doses of an anti-PD-1 antibody or antigen-binding fragment thereof. In some embodiments, the method comprises administering one or more doses of an oncolytic virus and one or more doses of a CTLA-4 inhibitor 7 days prior to administering one or more doses of an anti-PD-1 antibody or antigen-binding fragment thereof.

[0024] In some embodiments, the CTLA-4 inhibitor is administered to the patient in one or more doses of about 0.1 mg / kg to about 15 mg / kg of patient body weight. In some embodiments, the CTLA-4 inhibitor is administered to the patient in one or more doses of about 1 mg to about 600 mg. In some embodiments, the CTLA-4 inhibitor is administered to the patient in one or more doses of 50 mg. In some embodiments, the CTLA-4 inhibitor is administered to the patient intravenously, subcutaneously, or intraperitoneally.

[0025] In some embodiments, the cancer is selected from adrenal gland tumors, biliary tract cancer, bladder cancer, brain cancer, breast cancer, carcinoma, central or peripheral nervous system tissue cancer, cervical cancer, colon cancer, endocrine or neuroendocrine or hematopoietic cancer, esophageal cancer, fibroma, gastrointestinal cancer, glioma, head and neck cancer, Li-Fraumeni tumor, liver cancer, lung cancer, lymphoma, melanoma, meningioma, multiple neuroendocrine type I and type II tumors, nasopharyngeal cancer, oral cavity cancer, oropharyngeal cancer, osteogenic sarcoma tumor, ovarian cancer, pancreatic cancer, islet cell cancer, parathyroid cancer, pheochromocytoma, pituitary tumor, prostate cancer, rectal cancer, kidney cancer, respiratory cancer, sarcoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer, tracheal cancer, genitourinary cancer, and uterine cancer.

[0026] In some embodiments, administration of the combination therapy leads to at least one therapeutic effect selected from the following: a reduction in tumor cell count, tumor regression, partial response, and complete response. In some embodiments, administration of the combination therapy leads to at least one improvement selected from the following: long-distance effect, tumor growth delay, increased overall survival, increased progression-free survival, increased overall response rate, increased complete response, increased partial response, and increased stable disease compared to patients treated with monotherapy selected from the following: an oncolytic virus and an anti-PD-1 antibody or antigen-binding fragment thereof. In some embodiments, administration of the combination therapy leads to at least one improvement selected from the following: long-distance effect, tumor growth delay, increased overall survival, increased progression-free survival, increased overall response rate, increased complete response, increased partial response, and increased stable disease compared to patients treated with oncolytic virus monotherapy. In some embodiments, tumor growth is inhibited by at least 50% compared to untreated patients.

[0027] In some embodiments, administration of the combination therapy leads to at least one improvement selected from the following: long-distance effect, delayed tumor growth, increased overall survival, increased progression-free survival, increased overall response rate, increased complete response, increased partial response, and increased stable disease, compared to patients treated with a monotherapy selected from the following: oncolytic virus, anti-PD-1 antibody or antigen-binding fragment thereof, and CTLA-4 inhibitor. In some embodiments, tumor growth is inhibited by at least 50% compared to patients administered an oncolytic virus, anti-PD-1 antibody or antigen-binding fragment thereof, or CTLA-4 inhibitor as a monotherapy. In some embodiments, tumor growth is inhibited by at least 50% compared to patients administered any two of an oncolytic virus, anti-PD-1 antibody or antigen-binding fragment thereof, and CTLA-4 inhibitor. In some embodiments, the method further comprises administering to the patient an additional therapeutic agent or therapy.In some embodiments, the additional therapeutic agent or therapy is selected from the following: radiation, surgery, chemotherapy, a cancer vaccine, a B7-H3 inhibitor, a B7-H4 inhibitor, a lymphocyte activation gene 3 (LAG3) inhibitor, a T-cell immunoglobulin and mucin domain-containing 3 (TIM3) inhibitor, a galectin 9 (GAL9) inhibitor, a V-domain immunoglobulin (Ig)-containing suppressor of T-cell activation (VISTA) inhibitor, a killer cell immunoglobulin-like receptor (KIR) inhibitor, a B- and T-lymphocyte attenuator (BTLA) inhibitor, an inhibitor of T-cell immunoreceptor with Ig and ITIM domains (TIGIT), a CD47 inhibitor, an inhibitor of IFN-γ ... doleamine-2,3-dioxygenase (IDO) inhibitors, vascular endothelial growth factor (VEGF) antagonists, angiopoietin-2 (Ang2) inhibitors, transforming growth factor beta (TGFβ) inhibitors, epidermal growth factor receptor (EGFR) inhibitors, antibodies against tumor-specific antigens, Bacillus Calmette-Guerin vaccines, granulocyte-macrophage colony-stimulating factor (GM-CSF), cytotoxins, interleukin-6 receptor (IL-6R) inhibitors, interleukin-4 receptor (IL-4R) inhibitors, IL-10 inhibitors, IL-2, IL-7, IL-12, IL-21, IL-15, antibody-drug conjugates, anti-inflammatory drugs and combinations thereof.

[0028] In another aspect, the disclosed technology provides a combination of an oncolytic virus and an antibody or antigen-binding fragment thereof that specifically binds to programmed death 1 (PD-1) and a CTLA-4 inhibitor, including ipilimumab or a biological equivalent thereof, for use in a method of treating or inhibiting tumor growth, the method comprising: (a) selecting a cancer patient; and (b) administering to a patient a therapeutically effective amount of: (i) an oncolytic virus; and (ii) an anti-PD-1 antibody or antigen-binding fragment thereof, the anti-PD-1 antibody or antigen-binding fragment thereof comprising the three heavy chain complementarity determining regions (HCRs) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:1. The present invention relates to a combination therapy comprising a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds to programmed death 1 (PD-1), and a CTLA-4 inhibitor, comprising ipilimumab or a biological equivalent thereof, the combination therapy comprising: administering to a patient in need thereof a combination therapy comprising a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds to programmed death 1 (PD-1), the antibody or antigen-binding fragment thereof comprising three light chain complementarity determining regions (LCDR1, LCDR2 and LCDR3) and a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:2.

[0029] In another aspect, the disclosed technology provides a combination of an oncolytic virus and an antibody or antigen-binding fragment thereof that specifically binds to programmed death 1 (PD-1) and a CTLA-4 inhibitor, including ipilimumab or a biological equivalent thereof, for use in a method of treating or inhibiting tumor growth, the method comprising: (a) selecting a cancer patient; and (b) administering to a patient a therapeutically effective amount of an oncolytic virus; (ii) an anti-PD-1 antibody or antigen-binding fragment thereof, the anti-PD-1 antibody or antigen-binding fragment thereof comprising three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:1, and a CTLA-4 inhibitor comprising ipilimumab or a biological equivalent thereof, the combination comprising: (a) selecting a cancer patient; and (b) administering to a patient a therapeutically effective amount of an oncolytic virus; The present invention relates to a combination therapy comprising: (i) a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds to programmed death 1 (PD-1), and a CTLA-4 inhibitor, including ipilimumab or a biological equivalent thereof, the combination comprising administering to a patient in need thereof a combination therapy comprising: (iii) a therapeutically effective amount of a CTLA-4 inhibitor, including ipilimumab or a biological equivalent thereof; and (iv) an antibody or antigen-binding fragment thereof that specifically binds to programmed death 1 (PD-1), and a CTLA-4 inhibitor, including ipilimumab or a biological equivalent thereof.

[0030] In another aspect, the disclosed technology relates to a kit comprising an oncolytic virus and an antibody or antigen-binding fragment thereof that specifically binds to programmed death 1 (PD-1), together with instructions describing the use of the combination of the oncolytic virus and a therapeutically effective amount of an anti-PD-1 antibody or antigen-binding fragment thereof to treat or inhibit the growth of a tumor in a patient.

[0031] In another aspect, the disclosed technology relates to a kit comprising an oncolytic virus, an antibody or antigen-binding fragment thereof that specifically binds to programmed death 1 (PD-1), and a CTLA-4 inhibitor, including ipilimumab or a biological equivalent thereof, together with instructions describing the use of the combination of the oncolytic virus, an anti-PD-1 antibody or antigen-binding fragment thereof, and a therapeutically effective amount of a CTLA-4 inhibitor to treat or inhibit the growth of a tumor in a patient.

[0032] The foregoing summary is not intended to define all aspects of the disclosure, and further aspects are described in other sections, for example, in the detailed description below. It should be understood that the entire document is intended to be linked as an integrated disclosure, and that all combinations of features described herein are contemplated, even if the combinations of features are not found together in the same sentence or paragraph or section of this document. Other features and advantages of the present invention will become apparent from the detailed description that follows. However, it should be understood that the detailed description and specific examples, while showing specific embodiments of the disclosure, are given by way of example only, since various changes and modifications within the spirit and scope of the disclosure will become apparent to those skilled in the art from this detailed description. [Brief description of the drawings]

[0033] [Figure 1] FIG. 1 is a schematic diagram of the study design described herein in Example 1. Abbreviations: 1L=first line; 2L-second line; CPI=checkpoint inhibitor; DRC=data review committee; IV=intravenous; NSCLC=non-small cell lung cancer; ORR=overall response rate; PD-(L)1=programmed death-(ligand)1; TCID50=50% tissue culture infectious dose; VV1=Voyager V1. [Diagram 2] FIG. 1 is a schematic diagram of the study design described in Example 2 herein. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0034] It should be understood that the present disclosure is not limited to the specific methods and experimental conditions described, because such methods and conditions may vary. It should also be understood that the terms used herein are intended to describe only specific embodiments and are not intended to be limiting, and that the scope of the present disclosure is limited only by the appended claims. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which the present disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, the preferred methods and materials are now described. All publications mentioned herein are incorporated herein by reference in their entirety unless otherwise specified.

[0035] The present disclosure generally relates to novel dual or triple combination therapies of an oncolytic virus, a PD-1 pathway inhibitor and a CTLA-4 inhibitor that exhibit superior activity of tumor growth inhibition compared to either monotherapy or, for triple combinations, compared to either dual combination therapies of an oncolytic virus, a PD-1 pathway inhibitor and a CTLA-4 inhibitor.

[0036] In one aspect, the present disclosure relates to the use of a dual or triple combination therapy in a method of treating or inhibiting tumor growth, the method comprising; selecting a cancer patient; and administering to the patient in need thereof: (i) a therapeutically effective amount of an oncolytic virus in combination with (ii) a therapeutically effective amount of a PD-1 pathway inhibitor (e.g., cemiplimab or a biological equivalent thereof) and / or (iii) a therapeutically effective amount of a CTLA-4 inhibitor (e.g., ipilimumab or a biological equivalent thereof).

[0037] As used herein, the term "patient" can be used interchangeably with the term "subject." The phrase "subject in need thereof" refers to a human or non-human mammal that exhibits one or more symptoms or signs of cancer and / or has been diagnosed with cancer. In some embodiments, the human subject is diagnosed with a primary or metastatic tumor and / or one or more symptoms or signs, such as, but not limited to, enlarged lymph nodes, abdominal swelling, chest pain / pressure, unexplained weight loss, fever, night sweats, persistent fatigue, loss of appetite, enlarged spleen, itching. The phrase includes patients who have undergone one or more cycles of chemotherapy with toxic side effects. In some embodiments, the phrase "subject in need thereof" includes cancer patients who have undergone treatment but have subsequently relapsed or metastasized. For example, patients who have been treated with one or more anti-cancer agents that led to tumor regression, but who subsequently relapse with cancer that is resistant to the one or more anti-cancer agents (e.g., chemotherapy-resistant cancer) are treated with the methods of the present disclosure.

[0038] As used herein, the terms "treating," "treat," and the like mean alleviating or reducing the severity of at least one symptom or sign, eliminating the cause of a symptom either temporarily or permanently, slowing or inhibiting tumor growth, reducing tumor cell burden or tumor mass, promoting tumor regression, causing tumor shrinkage, necrosis, and / or disappearance, preventing tumor recurrence, preventing or inhibiting metastasis, inhibiting metastatic tumor growth, eliminating the need for radiation or surgery, and / or prolonging patient survival.

[0039] In many embodiments, the terms "tumor", "lesion", "tumor lesion", "cancer" and "malignant tumor" are used interchangeably to refer to one or more cancerous growths. In some embodiments, the cancer is selected from adrenal tumor, biliary tract cancer, bladder cancer, brain cancer, breast cancer, carcinoma, central or peripheral nervous system tissue cancer, cervical cancer, colon cancer, endocrine or neuroendocrine or hematopoietic cancer, esophageal cancer, fibroma, gastrointestinal cancer, glioma, head and neck cancer, Li-Fraumeni tumor, liver cancer, lung cancer, lymphoma, melanoma, meningioma, multiple neuroendocrine type I and type II tumors, nasopharyngeal cancer, oral cancer, oropharyngeal cancer, osteogenic sarcoma tumor, ovarian cancer, pancreatic cancer, pancreatic islet cell cancer, parathyroid cancer, pheochromocytoma, pituitary tumor, prostate cancer, rectal cancer, kidney cancer, respiratory cancer, sarcoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer, tracheal cancer, genitourinary cancer and uterine cancer.

[0040] According to some embodiments, the disclosure includes methods for treating, slowing or inhibiting tumor growth. In some embodiments, the disclosure includes methods for promoting tumor regression. In some embodiments, the disclosure includes methods for reducing tumor cell burden or reducing tumor mass. In some embodiments, the disclosure includes methods for preventing tumor recurrence.

[0041] In some embodiments, the methods include administering to the patient one or more doses of an oncolytic virus before, after, or concomitantly with administration to the patient of one or more doses of a PD-1 pathway inhibitor and / or one or more doses of a CTLA-4 inhibitor. In some embodiments, the one or more doses of a PD-1 pathway inhibitor can be administered in combination with one or more doses of a CTLA-4 inhibitor.

[0042] As used herein, the term "in combination with" includes sequential or simultaneous administration of two or more of an oncolytic virus, a PD-1 pathway inhibitor (e.g., an anti-PD-1 antibody or antigen-binding fragment thereof), and a CTLA-4 inhibitor (e.g., an anti-CTLA-4 antibody or antigen-binding fragment thereof). In some embodiments, one therapy disclosed herein can be administered to a patient before or after the simultaneous administration of the other two therapies disclosed herein.

[0043] For example, when a first therapy is administered "before" a second and / or third therapy, one or more doses of the first therapy can be administered about 12 weeks, about 11 weeks, about 10 weeks, about 9 weeks, about 8 weeks, about 7 weeks, about 6 weeks, about 5 weeks, about 4 weeks, about 3 weeks, about 2 weeks, about 7 days, about 150 hours, about 100 hours, about 72 hours, about 60 hours, about 48 hours, about 36 hours, about 24 hours, about 12 hours, about 10 hours, about 8 hours, about 6 hours, about 4 hours, about 2 hours, about 1 hour, about 30 minutes, about 15 minutes, or more than about 10 minutes before administration of one or more doses of the second and / or third therapy.

[0044] For example, when a first therapy is administered "after" a second and / or third therapy, one or more doses of the first therapy can be administered about 12 weeks, about 11 weeks, about 10 weeks, about 9 weeks, about 8 weeks, about 7 weeks, about 6 weeks, about 5 weeks, about 4 weeks, about 3 weeks, about 2 weeks, about 7 days, about 150 hours, about 150 hours, about 100 hours, about 72 hours, about 60 hours, about 48 hours, about 36 hours, about 24 hours, about 12 hours, about 10 hours, about 8 hours, about 6 hours, about 4 hours, about 2 hours, about 1 hour, about 30 minutes, about 15 minutes, or about 10 minutes after administration of one or more doses of the second and / or third therapy.

[0045] As used herein, "concurrent" administration means that two or more of an oncolytic virus, a PD-1 pathway inhibitor (e.g., an anti-PD-1 antibody or antigen-binding fragment thereof), and a CTLA-4 inhibitor (e.g., an anti-CTLA-4 antibody or antigen-binding fragment thereof) are administered to a patient in separate dosage forms on the same day (e.g., within a 10 minute period) or are administered to a patient in a single combined dosage formulation including two or more of an oncolytic virus, a PD-1 pathway inhibitor, and a CTLA-4 inhibitor.

[0046] In some embodiments, the disclosed methods can further include administering an anti-tumor therapy, including but not limited to conventional anti-tumor therapies such as chemotherapy, radiation, surgery, or as described elsewhere herein.

[0047] In some embodiments, the treatment results in a therapeutic effect selected from one or more of the following: delayed tumor growth, reduction in tumor cell count, tumor regression, extended survival, partial response, and complete response. In some embodiments, the patient's tumor growth is delayed by at least 10 days compared to tumor growth in untreated patients. In some embodiments, tumor growth is inhibited by at least 20% (e.g., at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 200%, at least 300%) compared to untreated patients. In some embodiments, tumor growth is inhibited by at least 20% (e.g., at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 200%, at least 300%) compared to patients to whom an oncolytic virus, a PD-1 pathway inhibitor, or a CTLA-4 inhibitor is administered as monotherapy. In some embodiments, tumor growth is inhibited by at least 20% (e.g., at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 200%, at least 300%) compared to a patient administered the oncolytic virus alone, or only two of the oncolytic virus, the PD-1 pathway inhibitor, and the CTLA-4 inhibitor.

[0048] In some embodiments, administration of the combination therapy (oncolytic virus and PD-1 pathway inhibitor and / or CTLA-4 inhibitor) leads to at least one therapeutic effect selected from the following: reduction in tumor cell count, tumor regression, partial response, and complete response. In some embodiments, administration of the combination therapy (oncolytic virus and PD-1 pathway inhibitor and / or CTLA-4 inhibitor) leads to at least one improvement selected from the following: long-distance effect, delayed tumor growth, increased overall survival, increased progression-free survival, increased overall response rate, increased complete response, increased partial response, and increased stable disease compared to patients treated with an oncolytic virus as a monotherapy, or compared to patients treated with a PD-1 pathway inhibitor as a monotherapy, and / or compared to patients treated with a CTLA-4 inhibitor as a monotherapy.

[0049] In some embodiments, administration of the combination therapy (oncolytic virus in combination with a PD-1 pathway inhibitor and / or a CTLA-4 inhibitor) involves selecting a subject having a first tumor lesion and a second tumor lesion, and administering one or more doses of the combination therapy to the first tumor lesion but not to the second tumor lesion, whereby the administration leads to tumor regression of both the first and second (i.e., treated and untreated) tumor lesions, thereby achieving a long-range effect.

[0050] Oncolytic viruses Oncolytic viruses are cancer therapies that use engineered or naturally evolved cancer-tropic viruses to induce tumor cell death in treated patients.Generally, when replicative oncolytic viruses are administered, the infected tumor cells can produce progeny viruses, which allows destructive infection to spread to adjacent tumor cells.The possibility of viral replication is determined by the ability of cells to sense and respond to viral infection.

[0051] In some embodiments, the oncolytic virus is a recombinant VSV (e.g., VV1). In some embodiments, the oncolytic virus is a replication-competent oncolytic rhabdovirus. Such oncolytic rhabdoviruses include, but are not limited to, wild-type or genetically modified Arajas virus, Chandipura virus, Kokar virus, Isfahan virus, Maraba virus, Pirie virus, Vesicular stomatitis Alagoas virus, Vesicular stomatitis virus (VSV), BeAn157575 virus, Boteke virus, Calchaqui virus, American eel virus, Gray Lodge virus, Jurona virus, Klamath virus, Kwatta virus, La Joya virus, Malpais Spring virus, Mt. Elgon bat virus, Perinet virus, Tupaia virus, Farmington, Bahia Grande virus, Muir Springs virus, Reed Ranch virus, and the like. Ranch virus, Hart Park virus, Flanders virus, Kamese virus, Mosqueiro virus, Mossuril virus, Barur virus, Fukuoka virus, Cairn Canyon virus, Nkolbisson virus, Le Dantec virus, Keuraliba virus, Connecticut virus, New Minto virus, Sawgrass virus, Chaco virus, Sena Madureira virus, Timbo virus, Almpiwar virus, Aruac virus, Bangoran virus, Bimbo virus, Bivens Arm virus, Blue crab virus, Charleville virus, Coastal Plains virus,Plains virus, DakArK7292 virus, Entamoeba virus, Garba virus, Gossas virus, Humpty Doo virus, Joinjakaka virus, Kannamangalam virus, Kolongo virus, Koolpinyah virus, Kotonkan virus, Landjia virus, Manitoba virus, Marco virus, Nasoule virus, Navarro virus, Ngaingan virus, Oak-Vale virus, Obodihan virus, Oita virus, Ouango virus, Parry Creek virus, Rio Grande cichlid virus, chlid virus, Sandjimba virus, Sigma virus, Sripur virus, Sweetwater Branch virus, Tibrogargan virus, Xibrema virus, Yata virus, Rhode Island virus, Adelaide River virus, Berrimah virus, Kimberley virus or Bovine ephemeral fever virus.

[0052] Vesicular stomatitis virus (VSV), as previously mentioned, is a member of the Rhabdoviridae family. The VSV genome is a single molecule of negative-stranded RNA that encodes five major polypeptides: a nucleocapsid (N) polypeptide, a phosphoprotein (P) polypeptide, a matrix (M) polypeptide, a glycoprotein (G) polypeptide, and a viral polymerase (L) polypeptide.

[0053] In some embodiments, the oncolytic virus is a wild-type or recombinant VSV. In some embodiments, the recombinant VSV contains one or more mutations, such as an M51R substitution (also referred to herein as VSV-M51R).

[0054] In some embodiments, the oncolytic virus can be engineered to express one or more cytokines, such as interferon beta (IFNb). In some embodiments, the IFNb (e.g., interferon beta-1a) can be human or murine IFNb or a variant thereof. In some embodiments, the IFNb comprises an amino acid sequence having at least 90% (e.g., 90%, 95%, 96%, 97%, 98%, 99%) sequence identity to the amino acid sequence of SEQ ID NO: 23 or 24, or comprises the amino acid sequence of SEQ ID NO: 23 or 24. In some embodiments, the nucleic acid sequence encoding the IFNb is located between the M and G viral genes. Such a location allows the virus to express an amount of IFNb polypeptide effective to activate an antiviral immune response in non-cancerous tissues, thereby reducing potential viral toxicity, without preventing efficient viral replication in cancer cells.

[0055] In some embodiments, the recombinant VSV further expresses a sodium / iodine symporter (NIS) or a variant thereof. In some embodiments, the NIS comprises an amino acid sequence having at least 90% (e.g., 90%, 95%, 96%, 97%, 98%, 99%) sequence identity to the amino acid sequence of SEQ ID NO: 25, or comprises the amino acid sequence of SEQ ID NO: 25. In some embodiments, the nucleic acid sequence encoding NIS is located between the G and L viral genes, which allows for appropriate expression levels of the NIS polypeptide.

[0056] In certain embodiments, the oncolytic virus is a recombinant VSV known in the art as Voyager V1, for example, as described in US9428736, the contents of which are incorporated herein by reference in their entirety.

[0057] PD-1 pathway inhibitors In some embodiments, the methods disclosed herein include administering a therapeutically effective amount of a PD-1 pathway inhibitor. As used herein, a "PD-1 pathway inhibitor" refers to any molecule that can inhibit, block, abrogate or interfere with the activity or expression of PD-1. In some embodiments, the PD-1 pathway inhibitor may be an antibody, a small molecule compound, a nucleic acid, a polypeptide, or a functional fragment or variant thereof. Non-limiting examples of suitable PD-1 pathway inhibitors include anti-PD-1 antibodies and antigen-binding fragments thereof, anti-PD-L1 antibodies and antigen-binding fragments thereof, and anti-PD-L2 antibodies and antigen-binding fragments thereof.

[0058] Other non-limiting examples of suitable PD-1 pathway inhibitors include RNAi molecules, such as anti-PD-1 RNAi molecules, anti-PD-L1 RNAi and anti-PD-L2 RNAi, antisense molecules, such as anti-PD-1 antisense RNA, anti-PD-L1 antisense RNA and anti-PD-L2 antisense RNA, and dominant negative proteins, such as dominant negative PD-1 protein, dominant negative PD-L1 protein and dominant negative PD-L2 protein. Some examples of the aforementioned PD-1 pathway inhibitors are described, for example, in US9308236, US10011656 and US20170290808, the portions identifying the PD-1 pathway inhibitors are incorporated herein by reference.

[0059] As used herein, the term "antibody" is intended to refer to an immunoglobulin molecule (i.e., "intact antibody molecule") composed of four polypeptide chains, two heavy (H) chains and two light (L) chains interconnected by disulfide bonds, as well as multimers thereof (e.g., IgM) or antigen-binding fragments thereof. Each heavy chain comprises a heavy chain variable region ("HCVR" or "VH") and a heavy chain constant region (composed of domains CH1, CH2 and CH3). Each light chain comprises a light chain variable region ("LCVR" or "VL") and a light chain constant region (CL). The VH and VL regions can be further subdivided into regions of hypervariability called complementarity determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In some embodiments, the FRs of an antibody (or an antigen-binding fragment thereof) may be identical to human germline sequences or may be naturally or artificially modified. An amino acid consensus sequence may be defined based on side-by-side analysis of two or more CDRs. As used herein, the term "antibody" also includes antigen-binding fragments of complete antibody molecules.

[0060] As used herein, the terms "antigen-binding fragment" of an antibody, "antigen-binding portion" of an antibody, etc., include any naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds to an antigen to form a complex. Antigen-binding fragments of antibodies can be generated from intact antibody molecules, for example, using any suitable standard technique, such as proteolytic digestion or recombinant genetic engineering techniques involving the manipulation and expression of DNA encoding the variable and optionally constant domains of the antibody. Such DNA is known and / or readily available, for example, from commercial sources, DNA libraries (including, for example, phage antibody libraries), or can be synthesized. The DNA can be sequenced and engineered, for example, by using chemical or molecular biology techniques to place one or more variable and / or constant domains in the appropriate configuration, or to introduce codons, create cysteine ​​residues, modify, add, or delete amino acids, etc.

[0061] Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single chain Fv (scFv) molecules; (vi) dAb fragments; and (vii) minimal recognition units consisting of amino acid residues that mimic the hypervariable regions of an antibody (e.g., isolated complementarity determining regions (CDRs), such as CDR3 peptides), or constrained FR3-CDR3-FR4 peptides. Other engineered molecules, such as domain-specific antibodies, single domain antibodies, domain deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains, are also encompassed within the expression "antigen-binding fragments" as used herein.

[0062] Antigen-binding fragments of antibodies typically contain at least one variable domain. The variable domain may be of any size or amino acid composition and generally contains at least one CDR adjacent to or in frame with one or more framework sequences. L V bound to the domain H For antigen-binding fragments containing domains, V H and V L The domains may be positioned relative to each other in any suitable configuration. For example, the variable region may be a dimer, with the V H -V H , V H -V L or V L -V L Alternatively, the antigen-binding fragment of the antibody may comprise a dimer of monomeric V H or V L It may include a domain.

[0063] In some embodiments, an antigen-binding fragment of an antibody can comprise at least one variable domain covalently linked to at least one constant domain. Non-limiting exemplary configurations of variable and constant domains that can be found within an antigen-binding fragment of an antibody of the present disclosure include: (i) a V H -C H 1;(ii)V H -C H 2;(iii)V H -C H 3;(iv)V H -C H 1-C H 2;(v)V H -C H 1-C H 2-C H 3;(vi)V H -C H 2-C H 3;(vii)V H -C L ;(viii)V L -C H 1;(ix)V L -CH 2;(x)V L -C H 3;(xi)V L -C H 1-C H 2;(xii)V L -C H 1-C H 2-C H 3;(xiii)V L -C H 2-C H 3; and (xiv) V L -C L In any configuration of variable and constant domains, including any of the exemplary configurations listed above, the variable and constant domains are directly linked to each other or are linked by a complete or partial hinge or linker region. The hinge region can consist of at least two (e.g., 5, 10, 15, 20, 40, 60 or more) amino acids that provide a flexible or semi-flexible linkage between adjacent variable and / or constant domains in a single polypeptide molecule. Furthermore, antigen-binding fragments of antibodies of the present disclosure can be linked to each other and / or to one or more monomeric V H Domain or V L The variable and constant domains may comprise homodimers or heterodimers (or other multimers) of any of the configurations listed above, in which the variable and constant domains are non-covalently associated (e.g., by disulfide bonds).

[0064] The antibody used in the methods disclosed herein may be a human antibody. As used herein, the term "human antibody" refers to an antibody having variable and constant regions derived from human germline immunoglobulin sequences. The human antibody of the present disclosure may nevertheless contain, for example, in the CDRs, particularly CDR3, amino acid residues that are not encoded in human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, as used herein, the term "human antibody" is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, are grafted onto human framework sequences.

[0065] The antibody used in the methods disclosed herein may be a recombinant human antibody. As used herein, the term "recombinant human antibody" includes all human antibodies produced, expressed, created or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell (described further below), antibodies isolated from a recombinant combinatorial human antibody library (described further below), antibodies isolated from an animal (e.g., a mouse) that is transgenic for human immunoglobulin genes (see, e.g., Taylor et al. (1992) Nucl. Acids Res. 20:6287-6295), or antibodies produced, expressed, created or isolated by any other means involving splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from germline immunoglobulin sequences. However, in some embodiments, such recombinant human antibodies are subjected to in vitro mutagenesis (or, when animals transgenic for human Ig sequences are used, in vivo somatic mutagenesis), thereby modifying the V H and V L The amino acid sequence of the region is H and V LA sequence that is derived from and related to a sequence, but which may not naturally occur within the human antibody germline repertoire in vivo.

[0066] Anti-PD-1 antibodies and antigen-binding fragments thereof In some embodiments, the PD-1 pathway inhibitor used in the methods disclosed herein is an antibody or antigen-binding fragment thereof that specifically binds to PD-1 (e.g., an anti-PD-1 antibody). The term "specifically binds" and the like means that the antibody or antigen-binding fragment thereof forms a complex with the antigen that is relatively stable under physiological conditions. Methods for determining whether an antibody specifically binds to an antigen are well known in the art, and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. For example, an antibody that "specifically binds" to PD-1, as used in this disclosure, has a K of less than about 500 nM, less than about 300 nM, less than about 200 nM, less than about 100 nM, less than about 90 nM, less than about 80 nM, less than about 70 nM, less than about 60 nM, less than about 50 nM, less than about 40 nM, less than about 30 nM, less than about 20 nM, less than about 10 nM, less than about 5 nM, less than about 4 nM, less than about 3 nM, less than about 2 nM, less than about 1 nM, or less than about 0.5 nM, as measured by a surface plasmon resonance assay. D and binds to PD-1 or a portion thereof. However, an isolated antibody that specifically binds human PD-1 may have cross-reactivity to other antigens, for example, PD-1 molecules from other (non-human) species.

[0067] According to certain exemplary embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (HCVR), a light chain variable region (LCVR) and / or a complementarity determining region (CDR) comprising the amino acid sequence of any of the anti-PD-1 antibodies described in US9987500, the contents of which are incorporated herein by reference in their entirety.

[0068] In certain exemplary embodiments, an anti-PD-1 antibody or antigen-binding fragment thereof that can be used in the context of the present disclosure comprises a heavy chain complementarity determining region (HCDR) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:1 and a light chain complementarity determining region (LCDR) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:2.

[0069] According to some embodiments, the anti-PD-1 antibody, or antigen-binding fragment thereof, comprises three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDRs (LCDR1, LCDR2, and LCDR3), where HCDR1 comprises the amino acid sequence of SEQ ID NO:3; HCDR2 comprises the amino acid sequence of SEQ ID NO:4; HCDR3 comprises the amino acid sequence of SEQ ID NO:5; LCDR1 comprises the amino acid sequence of SEQ ID NO:6; LCDR2 comprises the amino acid sequence of SEQ ID NO:7; and LCDR3 comprises the amino acid sequence of SEQ ID NO:8.

[0070] In yet other embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof comprises a HCVR comprising SEQ ID NO: 1 and a LCVR comprising SEQ ID NO: 2. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 9. In some embodiments, the anti-PD-1 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 10.

[0071] An exemplary antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:1 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:2 is the fully human anti-PD-1 antibody known as cemiplimab (REGN2810; also known as LIBTAYO®).

[0072] According to certain exemplary embodiments, the method of the present disclosure includes the use of cemiplimab or a bioequivalent thereof. As used herein, the term "bioequivalent" in reference to a PD-1 pathway inhibitor refers to an anti-PD-1 antibody or a PD-1 binding protein or fragment thereof that is a pharmaceutical equivalent or pharmaceutical substitute that exhibits no significant difference in rate and / or extent of absorption from a reference antibody (e.g., cemiplimab) when administered in either a single dose or multiple doses at the same molar dose under similar experimental conditions. In the context of the present disclosure, the term "bioequivalent" includes an antigen binding protein that binds to PD-1 and has no clinically meaningful difference from cemiplimab in terms of safety, purity and / or potency.

[0073] According to some embodiments of the present disclosure, the anti-human PD-1 or antigen-binding fragment thereof comprises a HCVR having at least 90% (e.g., 90%, 95%, 96%, 97%, 98%, 99%) sequence identity to SEQ ID NO:1.

[0074] According to some embodiments of the present disclosure, the anti-human PD-1 or antigen-binding fragment thereof comprises an LCVR having sequence identity (e.g., 90%, 95%, 96%, 97%, 98%, 99%) to SEQ ID NO: 2. Sequence identity can be measured by methods known in the art (e.g., GAP, BESTFIT and BLAST).

[0075] According to some embodiments of the present disclosure, the anti-human PD-1 or antigen-binding fragment thereof comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 1 with no more than 10 amino acid substitutions. According to some embodiments of the present disclosure, the anti-human PD-1 or antigen-binding fragment thereof comprises a LCVR comprising the amino acid sequence of SEQ ID NO: 2 with no more than 10 amino acid substitutions.

[0076] Variants of any of the HCVR, LCVR and / or CDR amino acid sequences disclosed herein that have one or more conservative amino acid substitutions are also within the scope of the disclosure. For example, the disclosure includes the use of anti-PD-L1 antibodies having HCVR, LCVR and / or CDR amino acid sequences that have, for example, 10 or less, 8 or less, 6 or less, 4 or less, etc., conservative amino acid substitutions relative to any of the HCVR, LCVR and / or CDR amino acid sequences disclosed herein.

[0077] Other anti-PD-1 antibodies or antigen-binding fragments thereof that can be used in the context of the methods of the present disclosure include those referred to in the art as, for example, nivolumab, pembrolizumab, MEDI0608, pidilizumab, BI754091, spartalizumab (also known as PDR001), camrelizumab (also known as SHR-1210), JNJ-63723283, and MCLA-134. Anti-PD-1 antibodies include any of the known antibodies or those described in U.S. Patent Nos. 6,808,710, 7,488,802, 8,008,449, 8,168,757, 8,354,509, 8,609,089, 8,686,119, 8,779,105, 8,900,587, and 9,987,500, and patent publications WO2006 / 121168, WO2009 / 114335. All portions of the foregoing publications identifying anti-PD-1 antibodies are incorporated herein by reference.

[0078] Anti-PD-1 antibodies used in the context of the methods of the present disclosure may have pH-dependent binding properties. For example, anti-PD-1 antibodies for use in the methods of the present disclosure may exhibit reduced binding to PD-1 at acidic pH compared to neutral pH. Alternatively, anti-PD-1 antibodies of the present disclosure may exhibit enhanced binding to their antigen at acidic pH compared to neutral pH. The phrase "acidic pH" includes pH values ​​less than about 6.2, e.g., about 6.0, 5.95, 5.9, 5.85, 5.8, 5.75, 5.7, 5.65, 5.6, 5.55, 5.5, 5.45, 5.4, 5.35, 5.3, 5.25, 5.2, 5.15, 5.1, 5.05, 5.0 or less. As used herein, the phrase "neutral pH" refers to a pH of about 7.0 to about 7.4. The expression "neutral pH" includes pH values ​​of about 7.0, 7.05, 7.1, 7.15, 7.2, 7.25, 7.3, 7.35 and 7.4.

[0079] In certain instances, "reduced binding to PD-1 at acidic pH compared to neutral pH" refers to the K D K value vs. K for antibody binding to PD-1 at neutral pH D For example, an antibody or antigen-binding fragment thereof may have an acidic / neutral K of about 3.0 or greater. D When a ratio is presented, it can be considered for purposes of this disclosure to indicate "reduced binding to PD-1 at acidic pH compared to neutral pH." In certain exemplary embodiments, the acidic / neutral K D The ratio can be about 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, 15.0, 20.0, 25.0, 30.0, 40.0, 50.0, 60.0, 70.0, 100.0 or more.

[0080] Antibodies with pH-dependent binding properties can be obtained, for example, by screening a population of antibodies for reduced (or enhanced) binding to a particular antigen at acidic pH compared to neutral pH. Furthermore, modification of the antigen-binding domain at the amino acid level can result in antibodies with pH-dependent properties. For example, by replacing one or more amino acids in the antigen-binding domain (e.g., in the CDR) with histidine residues, an antibody can be obtained that has reduced antigen binding at acidic pH compared to neutral pH. As used herein, the expression "acidic pH" refers to a pH of 6.0 or less.

[0081] Anti-PD-L1 antibodies and antigen-binding fragments thereof In some embodiments, the PD-1 pathway inhibitor used in the methods disclosed herein is an antibody or antigen-binding fragment thereof that specifically binds to PD-L1 (e.g., an anti-PD-L1 antibody). For example, as used in the context of this disclosure, an antibody that "specifically binds to PD-L1" includes an antibody that specifically binds to PD-L1, such as ... -8 K below M D (For example, K D A "high affinity" anti-PD-L1 antibody includes an antibody that binds to PD-L1 or a portion thereof with an affinity of at least 10 as measured by surface plasmon resonance, e.g., BIACORE™, or solution affinity ELISA. -8 M, for example, 10 -9 M, 10 -10 M, 10 -11 M or 10 -12 K of M D " refers to a mAb that has binding affinity for PD-L1 expressed as ". However, an isolated antibody that specifically binds human PD-L1 may have cross-reactivity to other antigens, such as PD-L1 molecules from other (non-human) species.

[0082] According to certain exemplary embodiments, the anti-PD-L1 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (HCVR), a light chain variable region (LCVR) and / or a complementarity determining region (CDR) comprising the amino acid sequence of any of the anti-PD-L1 antibodies described in US9938345, the contents of which are incorporated herein by reference in their entirety.

[0083] In certain exemplary embodiments, an anti-PD-L1 antibody or antigen-binding fragment thereof that can be used in the context of the present disclosure comprises a heavy chain complementarity determining region (HCDR) of a heavy chain variable region (HCVR) comprising SEQ ID NO: 11 and a light chain complementarity determining region (LCDR) of a light chain variable region (LCVR) comprising SEQ ID NO: 12. An exemplary anti-PD-L1 antibody comprising the HCVR of SEQ ID NO: 11 and the LCVR of SEQ ID NO: 12 is REGN3504.

[0084] According to some embodiments of the present disclosure, the anti-human PD-L1 antibody or antigen-binding fragment thereof comprises a HCVR that has at least 90% (e.g., 90%, 95%, 96%, 97%, 98%, 99%) sequence identity to SEQ ID NO: 11. According to some embodiments of the present disclosure, the anti-human PD-L1 antibody or antigen-binding fragment thereof comprises a LCVR that has at least 90% (e.g., 90%, 95%, 96%, 97%, 98%, 99%) sequence identity to SEQ ID NO: 12.

[0085] According to some embodiments of the present disclosure, the anti-human PD-L1 antibody or antigen-binding fragment thereof comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 11 with no more than 10 amino acid substitutions. According to some embodiments of the present disclosure, the anti-human PD-L1 antibody or antigen-binding fragment thereof comprises a LCVR comprising the amino acid sequence of SEQ ID NO: 12 with no more than 10 amino acid substitutions.

[0086] Variants of any of the HCVR, LCVR and / or CDR amino acid sequences disclosed herein that have one or more conservative amino acid substitutions are also within the scope of the disclosure. For example, the disclosure includes the use of anti-PD-L1 antibodies having HCVR, LCVR and / or CDR amino acid sequences that have, for example, 10 or less, 8 or less, 6 or less, 4 or less, etc., conservative amino acid substitutions relative to any of the HCVR, LCVR and / or CDR amino acid sequences disclosed herein.

[0087] Other anti-PD-L1 antibodies that can be used in the context of the methods of the disclosure include, for example, MDX-1105, atezolizumab (TECENTRIQ™), durvalumab (IMFINZI™), avelumab (BAVENCIO™), LY3300054, FAZ053, STI-1014, CX-072, KN035 (Zhang et al., Cell Discovery, 3, 170004 (March 2017)), CK-301 (Gorelik et al., American Association for Cancer Research Annual Meeting, 2014). Anti-PD-L1 antibodies include those known in the art as PD-L1 antibodies (Patent Document 1, Abstract 4606 at the American Association of Clinical Chemists Conference (AACR) 2016-04-04), or any of the other anti-PD-L1 antibodies described in U.S. Patent Nos. 7,943,743, 8,217,149, 9,402,899, 9,624,298, and 9,938,345, and patent publications WO2007 / 005874, WO2010 / 077634, WO2013 / 181452, WO2013 / 181634, WO2016 / 149201, WO2017 / 034916, or EP3177649. All portions of the foregoing publications identifying anti-PD-L1 antibodies are incorporated herein by reference.

[0088] Anti-PD-L2 antibodies and antigen-binding fragments thereof In some embodiments, the PD-1 pathway inhibitor used in the methods disclosed herein is an antibody or antigen-binding fragment thereof that specifically binds to PD-L2 (e.g., an anti-PD-L2 antibody). For example, an antibody that "specifically binds" to PD-L2, as used in the context of this disclosure, is an antibody that specifically binds to PD-L2, such as an antibody having a binding activity of about 1×10 -8 K below M D (For example, K D A "high affinity" anti-PD-L2 antibody is one that binds to PD-L2 or a portion thereof with an affinity of at least 10 as measured by surface plasmon resonance, e.g., BIACORE™, or solution affinity ELISA. -8 M, for example, 10 -9 M, 10 -10 M, 10 -11 M or 10 -12 K of M D " refers to a mAb that has binding affinity for PD-L2 expressed as ". However, an isolated antibody that specifically binds human PD-L2 may have cross-reactivity to other antigens, such as PD-L2 molecules from other (non-human) species.

[0089] Anti-PD-L2 antibodies that may be used in the context of the methods of the present disclosure include, for example, the anti-PD-L2 antibodies described in U.S. Patent Nos. 8,552,154 and 10,647,771. All portions of the aforementioned publications that identify anti-PD-L2 antibodies are incorporated herein by reference.

[0090] CTLA-4 inhibitors In some embodiments, the methods disclosed herein include administering a therapeutically effective amount of a CTLA-4 inhibitor. As used herein, "CTLA-4 inhibitor" refers to any molecule that can inhibit, block, abrogate or interfere with the activity or expression of CTLA-4. In some embodiments, the CTLA-4 inhibitor may be an antibody, a small molecule compound, a nucleic acid, a polypeptide, or a functional fragment or variant thereof. Non-limiting examples of suitable CTLA-4 inhibitors include anti-CTLA-4 antibodies and antigen-binding fragments thereof. Other non-limiting examples of suitable CTLA-4 inhibitors include RNAi molecules, e.g., anti-CTLA-4 RNAi molecules, and dominant negative proteins, e.g., dominant negative CTLA-4 proteins.

[0091] Anti-CTLA-4 antibodies and antigen-binding fragments thereof In some embodiments, the CTLA-4 inhibitor used in the methods disclosed herein is an antibody or antigen-binding fragment thereof that specifically binds to CTLA-4 (e.g., anti-CTLA-4 antibody). The term "specifically bind" and the like means that the antibody or antigen-binding fragment thereof forms a complex with the antigen that is relatively stable under physiological conditions. Methods for determining whether an antibody specifically binds to an antigen are well known in the art, and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. For example, an antibody that "specifically binds" to CTLA-4, as used in the context of this disclosure, includes an antibody that has a K of less than about 500 nM, less than about 300 nM, less than about 200 nM, less than about 100 nM, less than about 90 nM, less than about 80 nM, less than about 70 nM, less than about 60 nM, less than about 50 nM, less than about 40 nM, less than about 30 nM, less than about 20 nM, less than about 10 nM, less than about 5 nM, less than about 4 nM, less than about 3 nM, less than about 2 nM, less than about 1 nM, or less than about 0.5 nM as measured by a surface plasmon resonance assay. D and binds to a CTLA-4 antibody or portion thereof. However, an isolated antibody that specifically binds human CTLA-4 may have cross-reactivity to other antigens, such as CTLA-4 molecules from other (non-human) species.

[0092] In certain exemplary embodiments, an anti-CTLA-4 antibody or antigen-binding fragment thereof that can be used in the context of the present disclosure comprises a heavy chain complementarity determining region (HCDR) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 13 and a light chain complementarity determining region (LCDR) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 14.

[0093] According to some embodiments, the anti-CTLA-4 antibody or antigen-binding fragment thereof comprises three HCDRs (HCDR1, HCDR2 and HCDR3) and three LCDRs (LCDR1, LCDR2 and LCDR3), where HCDR1 comprises the amino acid sequence of SEQ ID NO: 15; HCDR2 comprises the amino acid sequence of SEQ ID NO: 16; HCDR3 comprises the amino acid sequence of SEQ ID NO: 17; LCDR1 comprises the amino acid sequence of SEQ ID NO: 18; LCDR2 comprises the amino acid sequence of SEQ ID NO: 19; and LCDR3 comprises the amino acid sequence of SEQ ID NO: 20.

[0094] In still other embodiments, the anti-CTLA-4 antibody or antigen-binding fragment thereof comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 13 and a LCVR comprising the amino acid sequence of SEQ ID NO: 14. In some embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 21. In some embodiments, the anti-CTLA-4 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 22.

[0095] An exemplary antibody comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:13 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:14 is the fully human anti-CTLA-4 antibody known as REGN4659.

[0096] According to certain exemplary embodiments, the method of the present disclosure includes the use of REGN4659 or its bioequivalent.As used herein, the term "bioequivalent" in relation to CTLA-4 inhibitor refers to an anti-CTLA-4 antibody or CTLA-4 binding protein or fragment thereof that is a pharmaceutical equivalent or pharmaceutical substitute that shows no significant difference in the rate and / or extent of absorption with reference antibody (e.g., REGN4659) when administered in single or multiple doses at the same molar dose under similar experimental conditions.In the context of the present disclosure, the term "bioequivalent" includes antigen binding protein that binds to CTLA-4 and has no clinically meaningful difference from REGN4659 in terms of safety, purity and / or potency.

[0097] According to some embodiments of the present disclosure, the anti-human CTLA-4 or antigen-binding fragment thereof comprises a HCVR having at least 90% (e.g., 90%, 95%, 96%, 97%, 98%, 99%) sequence identity to the amino acid sequence of SEQ ID NO:13.

[0098] According to some embodiments of the present disclosure, the anti-human CTLA-4 or antigen-binding fragment thereof comprises an LCVR having sequence identity (e.g., 90%, 95%, 96%, 97%, 98%, 99%) to the amino acid sequence of SEQ ID NO:14.

[0099] According to some embodiments of the present disclosure, the anti-human CTLA-4 or antigen-binding fragment thereof comprises an HCVR comprising the amino acid sequence of SEQ ID NO: 13 with no more than 10 amino acid substitutions. According to some embodiments of the present disclosure, the anti-human CTLA-4 or antigen-binding fragment thereof comprises an LCVR comprising the amino acid sequence of SEQ ID NO: 14 with no more than 10 amino acid substitutions.

[0100] Variants of any of the HCVR, LCVR and / or CDR amino acid sequences disclosed herein that have one or more conservative amino acid substitutions are also within the scope of the disclosure. For example, the disclosure includes the use of anti-PD-L1 antibodies having HCVR, LCVR and / or CDR amino acid sequences that have, for example, 10 or less, 8 or less, 6 or less, 4 or less, etc., conservative amino acid substitutions relative to any of the HCVR, LCVR and / or CDR amino acid sequences disclosed herein.

[0101] Other anti-CTLA-4 antibodies or antigen-binding fragments thereof that can be used in the context of the methods of the present disclosure can include, for example, the known antibodies referred to in the art as ipilimumab, tremelimumab, or any of the anti-CTLA-4 antibodies described in U.S. Patent Nos. 7527969, 8779098, 7666424, 7737258, 7740845, 8148154, 8414892, 8501471 and 9062110; and patent publications US2013 / 0078234, US2010 / 0143245, WO2017062615A2, WO2004 / 001381 and WO2012 / 147713. All portions of the aforementioned publications that identify anti-CTLA-4 antibodies are incorporated herein by reference.

[0102] Pharmaceutical Compositions and Administration The present disclosure includes methods comprising administering an oncolytic virus, a PD-1 pathway inhibitor, and / or a CTLA-4 inhibitor to a subject, where one or more antibodies are included in a separate or combined (single) pharmaceutical composition. The pharmaceutical compositions of the present disclosure can be formulated with suitable carriers, excipients and other agents that provide suitable entry, delivery, tolerability, etc. Numerous suitable formulations can be found in the formulary known to every pharmacist: Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa. These formulations include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, lipid (cationic or anionic)-containing vesicles (such as lipofectin), DNA conjugates, anhydrous absorbent pastes, oil-in-water and water-in-oil emulsions, emulsions carbowax (polyethylene glycol of various molecular weights), semi-solid gels, and semi-solid mixtures containing carbowax. See also Powell et al., PDA (1998) J Pharm Sci Technol 52:238-311.

[0103] A variety of delivery systems are known and can be used to administer the pharmaceutical compositions of the present disclosure, such as encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing mutant viruses, receptor-mediated endocytosis (see, e.g., Wu et al., 1987, J.Biol.Chem.262:4429-4432). Methods of administration include, but are not limited to, intradermal, intramuscular, intratumoral, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compositions can be administered by any convenient route, such as by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.), and can be administered together with other bioactive substances.

[0104] The pharmaceutical composition comprising the oncolytic virus, the PD-1 pathway inhibitor or the CTLA-4 inhibitor can be delivered intratumorally, subcutaneously or intravenously using a standard needle and syringe. In some embodiments, one or more of the oncolytic virus, the PD-1 pathway inhibitor and / or the CTLA-4 inhibitor can be delivered by more than one route of administration - for example, one or more doses are administered intratumorally and one or more doses are administered intravenously or by some other route of administration. Furthermore, with respect to subcutaneous delivery, a pen delivery device is easily applied in the delivery of the pharmaceutical composition of the present disclosure. Such a pen delivery device can be reusable or disposable. A reusable pen delivery device generally utilizes a replaceable cartridge containing the pharmaceutical composition. Once all of the pharmaceutical composition in the cartridge has been administered and the cartridge is empty, the empty cartridge can be easily discarded and replaced with a new cartridge containing the pharmaceutical composition. The pen delivery device can then be reused. In a disposable pen delivery device, there is no replaceable cartridge. Rather, the disposable pen delivery device is pre-filled with the pharmaceutical composition held in a reservoir within the device. Once the reservoir is emptied of the pharmaceutical composition, the entire device is discarded.

[0105] In certain circumstances, pharmaceutical compositions can be delivered in controlled release systems. In one embodiment, pumps can be used. In another embodiment, polymeric materials can be used; see, for example, Medical Applications of Controlled Release, Langer and Wise (eds.), 1974, CRC Pres., Boca Raton, Fla. In yet another embodiment, controlled release systems can be placed close to the target of the composition, so that only a fraction of the systemic dose is required (see Goodson, 1984, Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138). Other controlled release systems are discussed in the review by Langer, 1990, Science 249:1527-1533.

[0106] The injectable preparations may include dosage forms for intravenous, subcutaneous, intradermal and intramuscular injections, drip infusions, and the like. These injectable preparations can be manufactured by known methods. For example, the injectable preparations can be manufactured, for example, by dissolving, suspending or emulsifying the above-mentioned antibody or its salt in a sterile aqueous or oily medium that is conventionally used for injections. The aqueous medium for injection includes, for example, physiological saline, isotonic solutions containing glucose and other auxiliary agents, and these can be used in combination with appropriate solubilizers, for example, alcohols (e.g., ethanol), polyalcohols (e.g., propylene glycol, polyethylene glycol), nonionic surfactants [e.g., polysorbate 80, HCO-50 (polyoxyethylene (50 moles) adduct of hydrogenated castor oil)], and the like. As the oily medium, for example, sesame oil, soybean oil, and the like can be used in combination with solubilizers, for example, benzyl benzoate, benzyl alcohol, and the like. The injectable preparations thus manufactured are preferably filled into suitable ampoules.

[0107] Conveniently, the pharmaceutical composition for oral or parenteral use is prepared in a dosage form of a unit dose adapted to the dosage of the active ingredient.Such dosage forms of unit dose include, for example, tablets, pills, capsules, injections (ampoules), suppositories, etc.

[0108] The disclosure also provides kits comprising an oncolytic virus and a PD-1 pathway inhibitor, along with instructions describing the use of a combination of a therapeutically effective amount of the oncolytic virus and a PD-1 pathway inhibitor to treat or inhibit the growth of a tumor in a patient.

[0109] The present disclosure also provides kits comprising an oncolytic virus and a CTLA-4 inhibitor, along with instructions describing the use of a combination of a therapeutically effective amount of the oncolytic virus and a CTLA-4 inhibitor to treat or inhibit the growth of a tumor in a patient.

[0110] The disclosure also provides kits including an oncolytic virus, a PD-1 pathway inhibitor, and a CTLA-4 inhibitor, along with instructions describing the use of a therapeutically effective amount of the combination of an oncolytic virus, a PD-1 pathway inhibitor, and a CTLA-4 inhibitor to treat or inhibit the growth of a tumor in a patient.

[0111] Dosing regimen The disclosed method can include administering to a subject an oncolytic virus, a PD-1 pathway inhibitor (e.g., an anti-PD-1, anti-PD-L1 or anti-PD-L2 antibody or antigen-binding fragment thereof) or a CTLA-4 inhibitor (e.g., an anti-CTLA-4 antibody or antigen-binding fragment thereof) at a dosing frequency of about 4 times per week, twice per week, once per week, once per 2 weeks, once per 3 weeks, once per 4 weeks, once per 5 weeks, once per 6 weeks, once per 8 weeks, once per 12 weeks, or less frequently as long as a therapeutic response is achieved. The disclosed method can also include administering a single dose of each of the oncolytic virus, the PD-1 pathway inhibitor or the CTLA-4 inhibitor.

[0112] In some embodiments, at least one of the oncolytic virus, the PD-1 pathway inhibitor, or the CTLA-4 inhibitor is administered to the patient once a day, once every two days, once every three days, once every four days, once every five days, once a week, once every two weeks, or once every three weeks.

[0113] In some embodiments, the oncolytic virus, the PD-1 pathway inhibitor and the CTLA-4 inhibitor are administered to the patient simultaneously.

[0114] In some embodiments, the method can include sequentially administering to a subject two or more of an oncolytic virus, a PD-1 pathway inhibitor, and a CTLA-4 inhibitor. In some embodiments, the oncolytic virus is administered to a patient before or after the PD-1 pathway inhibitor and the CTLA-4 inhibitor. In some embodiments, the PD-1 pathway inhibitor is administered to a patient before or after the oncolytic virus and the CTLA-4 inhibitor. In some embodiments, the CTLA-4 inhibitor is administered to a patient before or after the oncolytic virus and the PD-1 pathway inhibitor.

[0115] In some embodiments, the method can include sequentially administering an oncolytic virus and a PD-1 pathway inhibitor (or a CTLA-4 inhibitor) to a subject. In some embodiments, the oncolytic virus is administered to the patient before or after the PD-1 pathway inhibitor. In some embodiments, the PD-1 pathway inhibitor is administered to the patient before or after the oncolytic virus.

[0116] As used herein, "sequentially administering" means that each dose of an oncolytic virus, a PD-1 pathway inhibitor, or a CTLA-4 inhibitor is administered to a subject at different times, e.g., on different days separated by a predetermined interval (e.g., hours, days, weeks, or months). The present disclosure includes methods that include sequentially administering to a patient a single initial dose of an oncolytic virus, a PD-1 pathway inhibitor, or a CTLA-4 inhibitor, followed by one or more secondary doses of an oncolytic virus, a PD-1 pathway inhibitor, or a CTLA-4 inhibitor, optionally followed by one or more tertiary doses of an oncolytic virus, a PD-1 pathway inhibitor, or a CTLA-4 inhibitor. In some embodiments, the method further comprises sequentially administering to the patient a single initial dose of the oncolytic virus, PD-1 pathway inhibitor or CTLA-4 inhibitor, followed by one or more secondary doses of the oncolytic virus, PD-1 pathway inhibitor or CTLA-4 inhibitor, optionally followed by one or more tertiary doses of the oncolytic virus, PD-1 pathway inhibitor or CTLA-4 inhibitor.

[0117] The terms "initial dose", "secondary dose" and "tertiary dose" refer to the temporal order of administration. Thus, an "initial dose" is a dose administered at the beginning of a treatment regimen (also referred to as a "baseline dose"); a "secondary dose" is a dose administered after the initial dose; and a "tertiary dose" is a dose administered after the secondary dose. The initial, secondary and tertiary doses may all contain the same amount of oncolytic virus, PD-1 pathway inhibitor or CTLA-4 inhibitor. However, in some embodiments, the amounts contained in the initial, secondary and / or tertiary doses differ from each other (e.g., adjusted up or down as appropriate) during the course of treatment. In some embodiments, one or more doses (e.g., 1, 2, 3, 4 or 5) are administered at the beginning of a treatment regimen as a "loading dose", followed by subsequent doses (e.g., "maintenance doses") that are administered less frequently.

[0118] In an exemplary embodiment of the present disclosure, each secondary and / or tertiary dose is administered ½ to 14 (e.g., ½, 1, 1½, 2, 2½, 3, 3½, 4, 4½, 5, 5½, 6, 6½, 7, 7½, 8, 8½, 9, 9½, 10, 10½, 11, 11½, 12, 12½, 13, 13½, 14, 14½ or more) weeks after the immediately preceding dose. As used herein, the phrase “immediately preceding dose” refers to a dose of oncolytic virus, PD-1 pathway inhibitor or CTLA-4 inhibitor that is administered to a patient in a multiple administration sequence prior to administration of the immediately following dose in a sequence with no intervening dose.

[0119] In some embodiments, the method can include administering any number of secondary and / or tertiary doses of an oncolytic virus, a PD-1 pathway inhibitor (e.g., an anti-PD-1 antibody or antigen-binding fragment thereof), and / or a CTLA-4 inhibitor (e.g., an anti-CTLA-4 antibody or antigen-binding fragment thereof) to the patient. For example, in some embodiments, only a single secondary dose is administered to the patient. In other embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8 or more) secondary doses are administered to the patient. Similarly, in some embodiments, only a single tertiary dose is administered to the patient. In other embodiments, two or more (e.g., 2, 3, 4, 5, 6, 7, 8 or more) tertiary doses are administered to the patient.

[0120] In embodiments with multiple secondary doses, each secondary dose can be administered at the same frequency as the other secondary doses. For example, each secondary dose can be administered to the patient 1-2 weeks after the immediately preceding dose. Similarly, in embodiments with multiple tertiary doses, each tertiary dose can be administered at the same frequency as the other tertiary doses. For example, each tertiary dose can be administered to the patient 2-4 weeks after the immediately preceding dose. Alternatively, the frequency with which the secondary and / or tertiary doses are administered to the patient can vary over the course of the treatment regimen. The frequency of administration can also be adjusted by the physician over the course of treatment depending on the needs of the individual patient after clinical testing.

[0121] In some embodiments, one or more doses of the oncolytic virus, the PD-1 pathway inhibitor and / or the CTLA-4 inhibitor are administered in one or more cycles. The "first treatment cycle" is the treatment cycle administered at the beginning of the treatment regimen; the "second treatment cycle" is the treatment cycle administered after the first treatment cycle; and the "third treatment cycle" is the treatment cycle administered after the second treatment. In the context of the present disclosure, the treatment cycles may be the same or different from each other. In some embodiments, any of the aforementioned treatment cycles may have a duration of 1 week (7 days), 2 weeks (14 days), 3 weeks (21 days), 4 weeks (28 days), 5 weeks (35 days), 6 weeks (42 days), 7 weeks (49 days), 8 weeks (56 days) or more.

[0122] In some embodiments, one or more doses of the oncolytic virus, PD-1 pathway inhibitor and / or CTLA-4 inhibitor are administered more frequently (e.g., twice a week, once a week, or once every two weeks) at the beginning of a treatment regimen as an "induction dose", followed by subsequent doses ("consolidation doses" or "maintenance doses") administered less frequently (e.g., once every 4-12 weeks).

[0123] In some embodiments, the dosing regimen for the combination includes administering an oncolytic virus (e.g., a recombinant VSV, such as VV1) and optionally a CTLA-4 inhibitor (e.g., an anti-CTLA-4 antibody or antigen-binding fragment thereof, such as ipilimumab or a biological equivalent thereof) simultaneously, followed by administration of a PD-1 pathway inhibitor (e.g., an anti-PD-1 antibody or antigen-binding fragment thereof, such as cemiplimab or a biological equivalent thereof). For example, the dosing regimen for the combination can include administering an oncolytic virus and optionally a CTLA-4 inhibitor on day 1, followed by administration of a PD-1 pathway inhibitor 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days later.

[0124] In some embodiments, the dosing regimen for the combination includes one or more of the following: intratumor administration of an oncolytic virus (e.g., a recombinant VSV, such as VV1), intravenous administration of an oncolytic virus (e.g., a recombinant VSV, such as VV1), intravenous administration of a CTLA-4 inhibitor (e.g., an anti-CTLA-4 antibody or antigen-binding fragment thereof, such as ipilimumab or a biological equivalent thereof), and intravenous administration of a PD-1 pathway inhibitor (e.g., an anti-PD-1 antibody or antigen-binding fragment thereof, such as cemiplimab or a biological equivalent thereof).

[0125] For example, the dosing regimen can include administering a therapeutically effective amount of an oncolytic virus intratumorally on the first day of a first 28-day treatment cycle, administering a therapeutically effective amount of an oncolytic virus intravenously on the first day of the first treatment cycle, administering a therapeutically effective amount of a CTLA-4 inhibitor intravenously on the first day of the first treatment cycle, administering a therapeutically effective amount of a PD-1 pathway inhibitor intravenously on day 8 of the first treatment cycle, and administering a therapeutically effective amount of a PD-1 pathway inhibitor intravenously on the first day of a second treatment cycle and each treatment cycle thereafter.

[0126] In another example, the dosing regimen can include administering a therapeutically effective amount of a PD-1 pathway inhibitor (e.g., an anti-PD-1 antibody or antigen-binding fragment thereof, e.g., cemiplimab or a biological equivalent thereof) intravenously on the first day of a first treatment cycle during a 21-day period, and administering a therapeutically effective amount of an oncolytic virus intratumorally on the first day of the first treatment cycle.

[0127] Dosage The amount of oncolytic virus, PD-1 pathway inhibitor (e.g., an anti-PD-1 antibody or antigen-binding fragment thereof), or CTLA-4 inhibitor (e.g., an anti-CTLA-4 antibody or antigen-binding fragment thereof) administered to a subject in accordance with the methods disclosed herein is generally a therapeutically effective amount. As used herein, the term "therapeutically effective amount" means an amount of an oncolytic virus, a PD-1 pathway inhibitor and / or a CTLA-4 inhibitor that results in one or more of the following, as compared to an untreated subject, a subject treated with a monotherapy, or a subject treated with any two of the three therapeutic agents disclosed herein (a PD-1 pathway inhibitor, a CTLA-4 inhibitor and an oncolytic virus): (a) a reduction in the severity or duration of a symptom or sign of cancer, e.g., a tumor lesion; (b) an inhibition of tumor growth or an increase in tumor necrosis, tumor shrinkage and / or tumor disappearance; (c) a delay in tumor growth and development; (d) an inhibition of tumor metastasis; (e) a prevention of recurrence of tumor growth; (f) an extension of survival of a subject with cancer; and / or (g) a reduction in the use or need for conventional anti-cancer therapy (e.g., elimination of the need for surgery or a reduction or elimination of the use of chemotherapeutic or cytotoxic agents).

[0128] In some embodiments, the combination of oncolytic viruses is 10, 100, 10 3 , 10 4 , 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 , 10 11 , 10 12 , 10 13 , 10 14 In some embodiments, the oncolytic virus is an oncolytic rhabdovirus (e.g., wild-type or genetically modified VSV) and can be administered in doses of 10 or more viral particles (vp) or plaque forming units (pfu). 4 ~10 14 p.f.u., 10 6 ~10 14 p.f.u., 108 ~10 14 p.f.u., 10 4 ~10 12 p.f.u., 10 6 ~10 12 p.f.u., 10 8 ~10 12 p.f.u., 10 9 ~10 11 pfu or 10 10 ~10 12 One or more dosages of pfu or any range therebetween are administered to a human with cancer.

[0129] In some embodiments, the combination of oncolytic viruses is 10, 100, 10 3 , 10 4 , 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 , 10 11 , 10 12 , 10 13 , 10 14 or more than 50% tissue culture infectious dose (TCID 50 In some embodiments, the oncolytic virus is an oncolytic rhabdovirus (e.g., wild-type or genetically modified VSV) and can be administered in one or more unit doses of 10 4 ~10 14 TCID 50 , 10 6 ~10 14 TCID 50 , 10 8 ~10 14 TCID 50 , 10 4 ~10 12 TCID 50 , 10 6 ~10 12 TCID 50 , 10 8 ~10 12 TCID 50 , 10 9 ~10 11 TCID 50 Or 1010 ~10 12 TCID 50 or any range therebetween. In some embodiments, the oncolytic virus is a recombinant VSV, e.g., VV1, and is administered to a human having cancer in one or more doses. 4 ~10 14 TCID 50 , 10 6 ~10 14 TCID 50 , 10 8 ~10 14 TCID 50 , 10 4 ~10 12 TCID 50 , 10 6 ~10 12 TCID 50 , 10 8 ~10 12 TCID 50 , 10 9 ~10 11 TCID 50 , 10 10 ~10 12 TCID 50 or any range therebetween. In some embodiments, a therapeutically effective amount of an oncolytic virus (e.g., a recombinant VSV, e.g., VV1) is administered to a human having cancer in one or more doses of 10, 100, 10 3 , 10 4 , 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 , 10 11 , 10 12 , 10 13 , 10 14 or higher TCID 50 It could be.

[0130] In some embodiments, a therapeutically effective amount of a PD-1 pathway inhibitor (e.g., an anti-PD-1 antibody or antigen-binding fragment thereof, e.g., cemiplimab, or a biological equivalent thereof) can be about 0.05 mg to about 1500 mg, about 1 mg to about 800 mg, about 5 mg to about 600 mg, about 10 mg to about 550 mg, about 50 mg to about 400 mg, about 75 mg to about 350 mg, or about 100 mg to about 300 mg of antibody. For example, in various embodiments, the amount of PD-1 pathway inhibitor is about 0.05 mg, about 0.1 mg, about 1.0 mg, about 1.5 mg, about 2.0 mg, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 400 mg, about 500 mg, about 600 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 35 ...500 mg, about 600 mg, about 70 mg, about 80 mg, about g, about 210mg, about 220mg, about 230mg, about 240mg, about 250mg, about 260mg, about 270mg, about 280mg, about 290mg, about 300mg, about 310mg, about 320mg, about 330mg, about 340mg, about 3 50mg, about 360mg, about 370mg, about 380mg, about 390mg, about 400mg, about 410mg, about 420mg, about 430mg, about 440mg, about 450mg, about 460mg, about 470mg, about 480mg, about 490mg , about 500mg, about 510mg, about 520mg, about 530mg, about 540mg, about 550mg, about 560mg, about 570mg, about 580mg, about 590mg, about 600mg, about 610mg, about 620mg, about 630mg, about 6 40mg, about 650mg, about 660mg, about 670mg, about 680mg, about 690mg, about 700mg, about 710mg, about 720mg, about 730mg, about 740mg, about 750mg, about 760mg, about 770mg, about 780mg , about 790mg, about 800mg, about 810mg, about 820mg, about 830mg, about 840mg, about 850mg, about 860mg, about 870mg, about 880mg, about 890mg, about 900mg, about 910mg, about 920mg, about 930mg, about 940mg, about 950mg, about 960mg, about 970mg, about 980mg, about 990mg, about 1000mg, about 1010mg, about 1020mg, about 1030mg, about 1040mg, about 1050mg, about 1060mg,About 1070 mg, about 1080 mg, about 1090 mg, about 1200 mg, about 1210 mg, about 1220 mg, about 1230 mg, about 1240 mg, about 1250 mg, about 1260 mg, about 1270 mg, about 1280 mg, about 1290 mg, about 1300 mg, about 1310 mg, about 1320 mg, about 1330 mg, about 1340 mg, about 1350 mg, about 1360 mg, about 1370 mg, about 1380 mg, about 1390 mg, about 1400 mg, about 1410 mg, about 1420 mg, about 1430 mg, about 1440 mg, about 1450 mg, about 1460 mg, about 1470 mg, about 1480 mg, about 1490 mg, or about 1500 mg.

[0131] The amount of PD-1 pathway inhibitor (e.g., anti-PD-1 antibody or antigen-binding fragment thereof) included in each dose is expressed in terms of milligrams antibody per kilogram of subject body weight (i.e., mg / kg). In some embodiments, the PD-1 pathway inhibitor used in the methods disclosed herein can be administered to the subject at a dose of about 0.0001 to about 100 mg / kg of subject body weight. In some embodiments, the anti-PD-1 antibody can be administered at a dose of about 0.1 mg / kg to about 20 mg / kg of patient body weight. In some embodiments, the methods of the disclosure include administration of a PD-1 pathway inhibitor (e.g., anti-PD-1 antibody or antigen-binding fragment thereof) at a dose of about 1 mg / kg to 3 mg / kg, 1 mg / kg to 5 mg / kg, 1 mg / kg to 10 mg / kg, 3 mg / kg to 10 mg / kg, 5 mg / kg to 10 mg / kg, 1 mg / kg, 3 mg / kg, 5 mg / kg, or 10 mg / kg of patient body weight.

[0132] In some embodiments, each dose of the PD-1 pathway inhibitor comprises 0.1-10 mg / kg of the subject's body weight (e.g., 0.3 mg / kg, 1 mg / kg, 3 mg / kg, or 10 mg / kg). In certain other embodiments, each dose contains between 5 and 1500 mg of a PD-1 pathway inhibitor (such as an anti-PD-1 antibody or antigen-binding fragment thereof), e.g., 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 40 mg, 45 mg, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, 1000 mg, 1050 mg, 1100 mg, 1150 mg, 1200 mg, 1550 mg, 1300 mg, 1350 mg, 1400 mg, 1450 mg, or 1500 mg of a PD-1 pathway inhibitor.

[0133] In some embodiments, a therapeutically effective amount of a CTLA-4 inhibitor (e.g., an anti-CTLA-4 antibody or antigen-binding fragment thereof, e.g., ipilimumab or a biological equivalent thereof) can be about 0.05 mg to about 1000 mg, about 1 mg to about 800 mg, about 5 mg to about 600 mg, about 10 mg to about 550 mg, about 50 mg to about 400 mg, about 75 mg to about 350 mg, or about 100 mg to about 300 mg of antibody. For example, in various embodiments, the amount of CTLA-4 inhibitor is about 0.05 mg, about 0.1 mg, about 1.0 mg, about 1.5 mg, about 2.0 mg, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190mg, about 200mg, about 210mg, about 220mg, about 230mg, about 240mg, about 250mg, about 260mg, about 270mg, about 280mg, about 290mg, about 300mg, about 310mg, about 320mg , about 330mg, about 340mg, about 350mg, about 360mg, about 370mg, about 380mg, about 390mg, about 400mg, about 410mg, about 420mg, about 430mg, about 440mg, about 450mg, about 460m g, about 470mg, about 480mg, about 490mg, about 500mg, about 510mg, about 520mg, about 530mg, about 540mg, about 550mg, about 560mg, about 570mg, about 580mg, about 590mg, about 60 0mg, about 610mg, about 620mg, about 630mg, about 640mg, about 650mg, about 660mg, about 670mg, about 680mg, about 690mg, about 700mg, about 710mg, about 720mg, about 730mg, about 7 40 mg, about 750 mg, about 760 mg, about 770 mg, about 780 mg, about 790 mg, about 800 mg, about 810 mg, about 820 mg, about 830 mg, about 840 mg, about 850 mg, about 860 mg, about 870 mg, about 880 mg, about 890 mg, about 900 mg, about 910 mg, about 920 mg, about 930 mg, about 940 mg, about 950 mg, about 960 mg, about 970 mg, about 980 mg, about 990 mg or about 1000 mg.

[0134] The amount of CTLA-4 inhibitor (e.g., anti-CTLA-4 antibody or antigen-binding fragment thereof) included in each dose can be expressed in terms of milligrams antibody per kilogram of subject body weight (i.e., mg / kg). In some embodiments, the anti-CTLA-4 antibody can be administered at a dose of about 0.1 mg / kg to about 20 mg / kg patient body weight. In some embodiments, the methods of the disclosure include administration of a CTLA-4 inhibitor (e.g., anti-CTLA-4 antibody or antigen-binding fragment thereof) at a dose of about 1 mg / kg to 3 mg / kg, 1 mg / kg to 5 mg / kg, 1 mg / kg to 10 mg / kg, 1 mg / kg, 3 mg / kg, 5 mg / kg, 10 mg / kg, or 15 mg / kg patient body weight.

[0135] In some embodiments, each dose of the CTLA-4 inhibitor comprises 0.1-10 mg / kg of subject body weight (e.g., 0.3 mg / kg, 1 mg / kg, 3 mg / kg, or 10 mg / kg). In certain other embodiments, each dose comprises 5-1000 mg of the CTLA-4 inhibitor (such as an anti-CTLA-4 antibody or antigen-binding fragment thereof), e.g., 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 40 mg, 45 mg, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, or 1000 mg of the CTLA-4 inhibitor.

[0136] In some embodiments, the methods of the disclosure further include administering to the subject an additional therapeutic agent or therapy. The additional therapeutic agent or therapy can be administered to enhance antitumor efficacy, to reduce toxic effects of one or more therapies, and / or to reduce the dosage of one or more therapies. In various embodiments, the additional therapeutic agent or therapy can include one or more of the following: radiation, surgery, cancer vaccines, imiquimod, antiviral agents (e.g., cidofovir), photodynamic therapy, lymphocyte activation gene 3 (LAG3) inhibitors (e.g., anti-LAG3 antibodies, glucocorticoid-induced tumor necrosis factor receptor (GITR) agonists (e.g., anti-GITR antibodies), T cell immunoglobulin and mucin-containing 3 (TIM3) inhibitors, B and T lymphocyte attenuator (BTLA) inhibitors, T cell immunoreceptor with Ig and ITIM domains (TIGIT) inhibitors, CD38 inhibitors, CD47 inhibitors, indoleamine-2,3-dioxygenase (IDO) inhibitors, CD28 activators, vascular endothelial growth factor (VEGF) antagonists (e.g., "VEGF-Trap" such as aflibercept, or anti-VEGF antibodies or antigen-binding fragments thereof (e.g., bevacizumab or ranibizumab) or small molecule kinase inhibitors of the VEGF receptor (e.g., sunitinib, sorafenib or pazopanib)), angiopoietin-2 (Ang2) inhibitors, transforming growth factor beta (TGFβ) inhibitors, epidermal growth factor receptor (EGFR) inhibitors, antibodies against tumor-specific antigens (e.g., CA9, CA125, melanoma-associated antigen 3 (MAGE3), carcinoembryonic antigen (CEA), vimentin, tumor-M2-PK, prostate-specific antigen (PSA), mucin-1, MART-1 and CA19-9), vaccines (e.g., Calmette-Guerin bacillus), granulocyte-macrophage colony stimulating factor (GM-CSF), a second oncolytic virus, a cytotoxin, a chemotherapeutic agent (e.g., pemetrexed, dacarbazine, temozolomide, cyclophosphamide, docetaxel, doxorubicin, daunorubicin, cisplatin, carboplatin, gemcitabine, methotrexate, mitoxantrone, oxaliplatin, paclitaxel, topotecan, irinotecan, vinorelbine and vincristine), IL-6R inhibitors, IL-4R inhibitors, IL-10 inhibitors, cytokines such as IL-2, IL-7, IL-12, IL-21 and IL-15, antibody drug conjugates, anti-inflammatory drugs such as corticosteroids, nonsteroidal anti-inflammatory drugs (NSAIDs), cryotherapy, anti-HPV therapy, laser therapy, electrosurgical removal of HPV-bearing cells and combinations thereof.

[0137] In some embodiments, the method further comprises administering an additional therapeutic agent, for example, an anti-cancer agent. As used herein, "anti-cancer agent" refers to any agent useful for treating cancer, including, but not limited to, cytotoxins, and agents such as antimetabolites, alkylating agents, anthracyclines, antibiotics, antimitotic agents, procarbazine, hydroxyurea, asparaginase, corticosteroids, mitotane (O,P'-(DDD)), biopharmaceuticals (e.g., antibodies and interferons), and radioactive agents. As used herein, "cytotoxins or cytotoxic agents" also refers to chemotherapeutic agents, and refers to any agent that is harmful to cells. Examples include taxol (paclitaxel), temozolomide, cytochalasin B, gramicidin D, ethidium bromide, emetine, cisplatin, mitomycin, etoposide, teniposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxyanthracenedione, mitoxantrone, mithramycin, actinomycin D, 1-dihydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin, as well as analogs or homologs thereof.

[0138] Further definitions To aid in the understanding of the detailed description of the compositions and methods according to the present disclosure, some clear definitions are provided to facilitate a clear disclosure of the various aspects of the present disclosure. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0139] As used herein, the term "agent" refers to a chemical compound, a mixture of chemical compounds, a biological macromolecule (such as a nucleic acid, an antibody, a protein or a portion thereof, e.g., a peptide, etc.), or an extract made from biological material, e.g., a bacterial, plant, fungus, or animal (especially mammalian) cell or tissue. The activity of such an agent may qualify it as a "therapeutic agent", which is a biologically, physiologically, or pharmacologically active substance (or substances) that acts locally or systemically in a subject. In the context of this disclosure, the term "therapeutic agent" refers to any of the PD-1 pathway inhibitors, CTLA-4 inhibitors, or oncolytic viruses disclosed herein.

[0140] As used herein, the terms "therapeutic agent", "therapeutic agent" or "treatment agent" are used interchangeably and refer to a molecule or compound that provides some beneficial effect upon administration to a subject. Beneficial effects include enablement of a diagnostic determination; amelioration of a disease, symptom, disorder or condition; reduction or prevention of the onset of a disease, symptom, disorder or condition; and generally attenuating a disease, symptom, disorder or condition.

[0141] As used herein, the term "pharmaceutical acceptable" refers to a material, such as a carrier or diluent, that does not abrogate the biological activity or properties of the composition and is relatively non-toxic, i.e., the material can be administered to an individual without causing undesired biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.

[0142] As used herein, the term "pharmaceutical acceptable carrier" includes pharmaceutical acceptable salts, pharmaceutical acceptable materials, compositions or carriers, such as liquid or solid fillers, diluents, excipients, solvents or encapsulating materials, that are involved in carrying or transporting a compound of the present disclosure in or to a subject so that it can perform its intended function. Typically, such compounds are carried or transported from one organ or part of the body to another organ or part of the body. Each salt or carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the subject. Some examples of materials which can serve as pharma- ceutically acceptable carriers include: sugars, such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository wax; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols, such as , propylene glycol; polyols such as glycerin, sorbitol, mannitol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer solution; diluents; granulating agents; lubricants; binders; disintegrating agents; wetting agents; emulsifying agents; coloring agents; release agents; coating agents; sweeteners; flavoring agents; fragrances; preservatives; antioxidants; plasticizers; gelling agents; thickening agents; hardening agents; setting agents; suspending agents; surfactants; wetting agents; carriers; stabilizers; and other non-toxic compatible substances used in pharmaceutical formulations, or any combination thereof. As used herein, "pharmaceutical acceptable carriers" also include any and all coatings, antibacterial and antifungal agents, and absorption delaying agents, etc., that are compatible with the activity of one or more components of the present disclosure and are physiologically acceptable to the subject. Supplementary active compounds can also be incorporated into the compositions.

[0143] Doses are often expressed in relation to body weight. Thus, a dose expressed as [g, mg or other unit] / kg (or g, mg, etc.) usually refers to [g, mg or other unit] "per kg (or g, mg, etc.) body weight" even if the term "body weight" is not explicitly mentioned. Treatments may include various "unit doses". A unit dose is defined as containing a predetermined amount of a therapeutic composition. A unit dose need not be administered as a single injection, but may include continuous infusion over a set period of time. For oncolytic viruses, the unit dose is described in terms of plaque forming units (pfu) or viral particles for the viral construct. A unit dose is about 10 3 , 10 4 , 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 , 10 11 , 10 12 , 10 13 pfu or vp and up. Alternatively, depending on the virus type and achievable titer, 1-100, 10-50, 100-1000, or about 1 x 10 4 , 1×10 5 , 1×10 6 , 1×10 7 , 1×10 8 , 1×10 9 , 1×10 10 , 1×10 11 , 1×10 12 , 1×10 13 , 1×10 14 Or 1×10 15 or more infectious viral particles (vp) to a patient or to a patient's cells. Alternatively, a unit dose for an oncolytic virus is 50 It is represented by "TCID 50" refers to "tissue culture infectious dose" and is defined as the dilution of virus required to infect 50% of a given batch of seeded cell cultures. TCID 50 For a description of the Spearman-Carver method, see BW Mahy & H. O. Kangro, Virology Methods Manual 25-46 (1996). In some embodiments, the unit dose is 10 3 , 10 4 , 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , 10 10 , 10 11 , 10 12 , 10 13 TCID 50 and above or any range therebetween.

[0144] As used herein, the term "disease" is intended to be synonymous with, and used interchangeably with, the terms "disorder" and "condition" (as well as medical condition), in that all generally reflect an abnormal condition of the human or animal body or one of its parts that impairs normal functioning (e.g., cancer), is typically manifested by prominent signs and symptoms, and causes a reduction in lifespan or quality of life in humans or animals.

[0145] As used herein, the term "in vitro" refers to events that take place not within a multicellular organism, but in an artificial environment, e.g., a test tube or reactor, cell culture, etc.

[0146] As used herein, the term "in vivo" refers to events that take place within a multicellular organism, e.g., a non-human animal.

[0147] As used herein, the singular forms "a," "an," and "the" include the plural references unless the context clearly indicates otherwise.

[0148] As used herein, the terms "including," "comprising," "containing," or "having" and variations thereof are intended to encompass the items listed thereafter and equivalents thereof, as well as additional subject matter, unless otherwise specified.

[0149] As used herein, the phrases "in one embodiment," "in various embodiments," "in some embodiments," etc. are used repeatedly. Such phrases do not necessarily refer to the same embodiment, but may refer to the same embodiment unless the context indicates otherwise. There is.

[0150] As used herein, the term "and / or" or " / " means any one of the items, any combination of the items, or all of the items with which this term is associated.

[0151] As used herein, the word "substantially" does not exclude "completely", e.g., a composition that is "substantially free" of Y may be completely free of Y. Where necessary, the word "substantially" may be omitted from the definitions of the present disclosure.

[0152] As used herein, the term "each," when used in reference to a collection of items, is intended to identify each individual item in the collection, but does not necessarily refer to every item in the collection. Exceptions may occur where express disclosure or context indicates otherwise.

[0153] As used herein, the term "approximately" or "about" refers to a value that is similar to the stated reference value when applied to one or more values ​​of interest. In some embodiments, the term "approximately" or "about" refers to a range of values ​​that falls within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less in either direction (greater or smaller) of the stated reference value, unless otherwise stated or clear from the context (except when such number exceeds 100% of possible values). Unless otherwise indicated herein, the term "about" is intended to include the closest value, e.g., weight percent, to the recited range that is equivalent in terms of the functionality of the individual component, composition or embodiment.

[0154] As disclosed herein, several ranges of values ​​are provided. It is understood that each intervening value between the upper and lower limits of the range, to the tenth of the unit of the lower limit, unless otherwise specified in the context, is also specifically disclosed. Each smaller range between any stated or intervening value in a stated range and any other stated or intervening value in that stated range is encompassed within the disclosure. The upper and lower limits of these smaller ranges can be independently included or excluded, and each range in which either limit is included, neither limit is included, or both limits are included in the smaller range is also encompassed within the disclosure, subject to any specifically excluded limit in the stated range. When a stated range includes one or both limits, ranges excluding either or both of those included limits are also included in the disclosure.

[0155] The use of any and all examples or exemplary language (e.g., "such as") provided herein is intended merely to better illustrate the disclosure and does not pose a limitation on the scope of the disclosure unless specifically claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the disclosure.

[0156] All methods described herein may be performed in any suitable order unless otherwise indicated herein or clearly contradicted by context. For any of the methods provided, the steps of the method may be performed simultaneously or sequentially. When the steps of the method are performed sequentially, the steps may be performed in any order unless otherwise specified. When the method includes a combination of steps, any and all combinations or subcombinations of the steps are included within the scope of the present disclosure, unless otherwise specified herein.

[0157] Each publication, patent application, patent and other reference cited herein is incorporated by reference in its entirety to the extent that it does not contradict this disclosure. Publications disclosed herein are provided solely for their disclosure prior to the filing date of this disclosure. Nothing herein should be construed as an admission that the present disclosure is not entitled to antedate such publication by virtue of prior disclosure. Moreover, the publication dates provided may differ from the actual publication dates, which may need to be confirmed independently.

[0158] It will be understood that the examples and embodiments described herein are for illustrative purposes only, and that various modifications or alterations will be suggested to those skilled in the art in light thereof, and that such various modifications or alterations are within the spirit and scope of this application and the scope of the appended claims. EXAMPLES

[0159] The following examples are set forth to provide those skilled in the art with a complete disclosure and description of how to make and use the disclosed methods and compositions, and are not intended to limit the scope of what the inventors regard as their invention. Likewise, the disclosure is not limited to any particular preferred embodiment described herein. Indeed, modifications and variations of the embodiments may be apparent to those skilled in the art upon reading this specification, and may be made without departing from the spirit and scope thereof. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperatures, etc.), but some experimental error and deviation should be accounted for. Unless otherwise indicated, parts are parts by weight, molecular weight is average molecular weight, temperature is degrees Celsius, and room temperature is about 25°C. o C and the pressure is at or near atmospheric pressure. EXAMPLES

[0160] A phase 2 trial of Voyager V1 (vesicular stomatitis virus expressing NIS and human interferon beta, VV1) in combination with cemiplimab in patients with hepatocellular carcinoma, non-small cell lung cancer, melanoma, or endometrial cancer. This example relates to an open-label, Simon2, Phase 2 clinical trial to determine the antitumor activity of Voyager V1 (VV1) in combination with cemiplimab alone and with ipilimumab in patients with hepatocellular carcinoma, non-small cell lung cancer (NSCLC), melanoma or endometrial cancer.

[0161] Background: VV1 is an oncolytic vesicular stomatitis virus engineered to express human interferon beta (IFNβ) to enhance cellular antitumor immune responses and tumor selectivity. Novel immunotherapies are needed to address patient populations who have never responded or no longer respond to immune checkpoint inhibitors (CPIs). Phase 1 studies have demonstrated antitumor activity of VV1 in several malignancies with or without CPIs. This example is concerned with optimizing the efficacy of VV1 in combination with cemiplimab, an anti-PD1 antibody approved for lung, basal and squamous cell skin cancer, in two ways: at higher doses and using triple combinations in a representative patient population.

[0162] Patients with advanced melanoma (after progression on anti-PD1) are being enrolled in this study, which will eventually include first-line NSCLC patients whose disease is amenable to immunotherapy. The study consists of a single 1.0 × 10 11 Intravenous (IV) VV1 at a dose of 1.0 × 10 followed by IV cemiplimab Q3W starting on D8, or VV1 once on D1 at a dose of 1.0 × 10 9 TCID 50The preliminary antitumor activity, safety and immunogenicity activity of the same regimen with additional intratumoral injections of ipilimumab will be investigated first. Patients will receive IV cemiplimab Q3W until disease progression or intolerable toxicity is confirmed. Once at least six patients are safely treated across the two melanoma doublet cohorts, a third (triplet) cohort will be initiated in the same patient population to add a single dose of ipilimumab 50 mg on D1. Once at least six melanoma patients have safely received the triplet, the first-line NSCLC cohort will begin triplet therapy. The cohorts will be expanded based on a Simon2 staged design using a type I error rate of 0.05 and a power of 85%. The null overall response rate (ORR) is 10% for second-line melanoma with a target of 35% and a null ORR of 40% for first-line NSCLC with a true target of 70%. The melanoma cohort requires a response in ≥2 of 10 patients in phase 1 to add another 11 in phase 2, while in NSCLC, 5 / 9 of evaluable patients must respond initially, followed by 13 more to complete the design. Responses will be assessed at week 7 and then Q9W according to RECIST v1.1. The study includes serial biopsies in ≥3 / 10 patients in stage 1 of each of the IV melanoma cohorts (doublet and triplet therapy), all patients in stage 2 of these IV melanoma cohorts, and all patients in both stage 1 and stage 2 of the intravenous / intratumoral (IV / IT) melanoma cohort to allow for a thorough investigation of the impact of the three immunotherapies under investigation.

[0163] Rationale for Study: Cancer immunotherapy has shown significant clinical success, resulting in durable responses in patients with advanced, previously refractory cancers. In particular, immuno-CPI agents, which block signals that tumors exploit to dampen immune responses that allow activation of T cell-mediated antitumor immunity, have shown clinical efficacy and have been approved for the treatment of several indications, including melanoma, NSCLC, urothelial, head and neck, gastric, hepatocellular carcinoma (HCC), and other tumors. The majority of patients remain resistant to CPI therapy, with response rates ranging from 10-40% depending on the indication. The population of patients whose disease is resistant or refractory to CPIs is currently emerging as a significant unmet medical need. Not all NSCLC or melanoma patients for whom anti-PD-(L)1 therapy has become standard of care derive long-term sustained benefit from such treatment. Once resistant, these patients have limited treatment options. Additionally, patients who are unresponsive and refractory to CPI therapy within these indications continue to represent an unmet medical need.

[0164] It is predicted that combination therapy with oncolytic VSV-IFNβ-NIS and CPIs will enhance the infiltration and activation of intratumoral CD8+ T cells, enhancing therapeutic responses in cancer patients previously resistant to CPI monotherapy. This study, conducted in a population of melanoma patients who have failed single-agent CPIs, is designed to provide insight into the putative mechanism of resensitization to CPIs via injection of a single dose of oncolytic virus, with the hypothesis that cell infection expands the available endogenous antitumor T cell repertoire, enhances tumor cell antigenicity for T cell recognition, enhances antigen presentation, and ultimately converts a noninflammatory or immune-exclusionary phenotype into a highly inflammatory phenotype.

[0165] Study Objectives: The primary objective of the study is to determine the preliminary antitumor activity in each study cohort, measured by overall response rate (ORR) according to standard criteria (RECIST 1.1) [Eisenhauer 2009]. Secondary objectives of the study include: to determine the safety and tolerability of VV1 in combination with cemiplimab; to evaluate the systemic pharmacokinetics (PK) of VV1 and cemiplimab; to investigate the pharmacodynamics of VV1 in blood by measuring serum IFNβ expression; to determine the preliminary clinical activity of each cohort [e.g., progression-free survival (PFS), duration of response (DOR), disease control rate (DCR) and overall survival (OS)]; and to evaluate the safety and efficacy of administration of VV1, cemiplimab and ipilimumab. Other objectives include evaluation of: immunocompetent cell infiltration in selected tumor specimens pre- and post-treatment; time course of viremia, viral shedding and viral persistence; efficacy of expansion / suppression of host peripheral blood immunocompetent cells; immunogenicity as measured by anti-drug antibodies against cemiplimab+VV1; and the ability of ruxolitinib to reduce the risk of IFNβ toxicity.

[0166] Overall Study Design: This is a Phase 2 study designed to determine preliminary antitumor activity and confirm the safety of VV1 in combination with cemiplimab alone and then with ipilimumab. The hepatocellular carcinoma and endometrial cancer cohorts have been completed, and the melanoma and NSCLC cohorts will be modified as follows: the dose of VV1 will be increased to 1.0 x 10 in combination with cemiplimab (doublet). 11 TCID 50to IV VV1, followed by the addition of ipilimumab to the combination (triplet). The NSCLC cohort will enroll CPI treatment-naïve patients. The study design is illustrated in Figure 1. Patients with advanced melanoma will be enrolled in two parallel doublet cohorts; in one cohort (IV melanoma cohort), patients will receive IV VV1 and in the other cohort (IV / IT melanoma cohort), patients will receive both IV VV1 and IT VV1; both cohorts will receive IV cemiplimab on day 8 after receiving VV1 treatment. After completion of the high-dose doublet melanoma cohort, the subsequent triplet melanoma cohort will receive combination therapy of IV VV1, IV cemiplimab, and ipilimumab (one dose). Finally, a cohort of NSCLC patients will receive the triplet combination.

[0167] 1.7×10 11 TCID 50 Doses up to VSV-IFNβ-NIS (IV monotherapy) were found to be safe and potentially more effective than lower doses.

[0168] All patients enrolled in this study will be required to have an overnight hospital stay on Day 1 and, if clinically stable, may be discharged for outpatient follow-up on Day 2. Patients will then be required to undergo outpatient safety evaluations on Days 3, 4, and 5. Laboratory evaluations performed on Days 2, 3, 4, and 5 must be evaluated and assessed for potential IFNβ toxicity on these days before patients are discharged home.

[0169] Patients will be enrolled simultaneously into each melanoma doublet cohort. A safety lead-in of three patients in the IV cohort will be dosed and observed for 28 days, and their safety data will be reviewed before the IT / IV melanoma cohort will be enrolled. A total of six patients across both cohorts will be dosed and observed for one cycle. Once the combination is deemed safe, enrollment of the IV melanoma triplet cohort will follow.

[0170] For all cohorts in the study, a two-stage Simon design [Simon 1989] will be used. In the first stage, 10 evaluable patients will be recruited in each melanoma tumor cohort; this cohort will progress to the second stage based on a combination of safety and efficacy. If there are fewer than two objective responses in each 10-patient melanoma cohort, the cohort will be discontinued; otherwise, 11 additional patients will be recruited, for a total of 21 patients in each melanoma cohort. A similar design will be used for NSCLC, with the first stage enrolling 9 evaluable patients. In this NSCLC cohort, a higher efficacy bar of at least 5 of the 9 patients having an objective response is required to initiate stage 2 for a total of 22 evaluable patients.

[0171] Patients in the IV melanoma cohort will receive IV VV1, and patients in the IV / IT melanoma cohort will receive both IV VV1 and IT VV1 on day 1 of cycle 1; both cohorts will receive IV cemiplimab on day 8 after receiving VV1 treatment. After cycle 1 (a 28-day cycle), cemiplimab treatment will continue Q3W according to the Libtayo® US package insert, and after cycle 2, there will then be efficacy evaluations every 9 weeks until progressive disease (PD). After completion of the high-dose melanoma cohort, the subsequent triplet melanoma cohort will receive a combination therapy of IV VV1, ipilimumab (one dose) and IV cemiplimab. Finally, a cohort of NSCLC patients will receive the triplet combination.

[0172] Overall toxicity will be graded using NCI CTCAE, v5.0, and CRS and neurological toxicity will be graded using ASTCT consensus grading [Lee 2019]. Safety assessments will include AEs, SAEs, PEs, vital sign measurements, ECOG status, clinical safety laboratory assessments (hematology, serum chemistry and liver panel, coagulation and urinalysis), and ECGs.

[0173] Study Population: Only patients who meet all of the inclusion criteria and none of the exclusion criteria will be eligible to receive the drug.

[0174] Inclusion Criteria: Patients who meet all of the following inclusion criteria at screening / day 1 of first dose will be eligible to participate in the study: (1) Age ≥ 18 years on the date of signing the informed consent. (2) Specifics by tumor cohort: (a) For the NSCLC cohort, histologically confirmed diagnosis of advanced and / or metastatic NSCLC suitable for first-line immunotherapy. NSCLC harboring activating EGFR mutations or anaplastic lymphoma kinase (ALK) rearrangements must have progressed after available EGFR or ALK targeted therapy in addition to treatment with platinum-based chemotherapy (unless ineligible for platinum therapy). (i) Ability to provide archival (or fresh) formalin-fixed, paraffin-embedded tumor tissue collected within 6 months prior to enrollment for determination of programmed death-ligand 1 (PD-L1) status. (ii) PD-L1 status ≥ 50% according to local standardized testing. Samples should be provided to a central laboratory for post-hoc centralized testing. (b) For the melanoma cohort, histologically confirmed diagnosis of progressive and / or cutaneous metastatic melanoma with radiological progression during therapy with a PD-(L)1 immune checkpoint inhibitor and where existing options are not considered to provide clinical benefit (only one line of PD-(L)1 therapy is permitted). Progression on ipilimumab is not required. Patients with BRAF V600 mutations must have progressed during or be intolerant to BRAF+ / -MEK inhibitor therapy. i. Patients with BRAF V600 positive tumors should have been pretreated with a BRAF inhibitor (alone or in combination with a MEK inhibitor) in addition to treatment with pembrolizumab or nivolumab with or without ipilimumab, or should have declined targeted therapy. Note: Patients with BRAF V600 positive tumors without clinically significant tumor-related symptoms or evidence of rapidly progressive disease do not need to be treated with a BRAF inhibitor (alone or in combination with a MEK inhibitor) based on the investigator's discretion. For the IV / IT melanoma cohort: (i) at least one tumor lesion amenable to IT injection by palpation or ultrasound;Injection of deep visceral lesions is not permitted; (ii) consent to provide a freshly obtained biopsy of the injected lesion prior to the start of study treatment, to repeat the biopsy twice during study treatment, and to provide the obtained tissue for biomarker analysis. Tissue obtained from biopsy must not have been previously irradiated, although new or progressing lesions within the radiation field are permitted. (3) For patients previously treated with anti-PD-(L)1 therapy: (a) the last dose of anti-PD-(L)1 must have been within 16 weeks of the start of study treatment; (b) patients must have received at least 4 doses on a q2w, 3 doses on a q3w, or 2 doses on a q4w schedule on their previous anti-PD-(L)1 therapy. (c) Progression on prior anti-PD-(L)1 therapy must be defined by: (i) radiographic progression documented on a single radiographic scan if treatment with anti-PD-(L)1 was ≥ 16 weeks; (ii) radiographic progression documented on two consecutive radiographic scans at least 4 weeks apart if treatment with anti-PD-(L)1 therapy was between 8 and 16 weeks; if radiographic progression is accompanied by clinical progression, a single scan assessment may be used; (iii) if progression is exclusively within lymph nodes, a biopsy is required to histologically confirm progression in lymph nodes; (4) measurable disease per RECIST 1.1; (5) performance status of 0 or 1 on the ECOG performance scale; (6) life expectancy > 3 months; (7) willingness to provide biopsy samples as required for the duration of the study, including fresh tumor biopsy samples. At least 3 of 10 patients in stage 1 of each of the IV melanoma cohorts (doublet and triplet therapy) must provide fresh tumor biopsy samples before and during treatment. In stage 2 of these cohorts, all patients must provide fresh tumor biopsy samples before and during treatment. All patients in both stage 1 and stage 2 of the IT / IV melanoma cohorts must provide fresh tumor biopsy samples before and during treatment.(8) adequate organ function, defined as laboratory values ​​obtained ≤14 days prior to enrollment as shown in Table 1; (9) negative pregnancy test for female patients of childbearing potential; (10) absence of active CNS complications. Note: pre-enrollment imaging is not required for asymptomatic patients; (11) ability to provide written informed consent.

[0175] [Table 1]

[0176] Exclusion Criteria: Patients who meet any of the following exclusion criteria at screening / day 1 of first dose will not be enrolled in the study: (1) availability and patient acceptance of alternative curative treatment options; (2) recent or ongoing serious infection, including any active viral, bacterial or fungal infection of grade 3 or higher per NCI CTCAE, v5.0, within 2 weeks of enrollment; (3) patients diagnosed with ocular, mucosal or acral melanoma; (4) known seropositivity for HIV and active HIV infection. a. Patients who are seropositive for HIV but receiving antiviral therapy and have undetectable viral loads and normal CD4 T-cell counts for at least 6 months are eligible; (5) seropositive for HBV and have evidence of active viral infection with HBV. Patients who are HBsAg negative and HBV viral DNA negative are eligible. (a) Patients who have had HBV but have received antiviral treatment and have demonstrated undetectable viral DNA for 6 months are eligible; (b) patients who are seropositive for HBV vaccine are eligible; (6) patients who are seropositive for HCV and have active HCV viral infection.a. Patients who have had HCV but have received antiviral treatment and have demonstrated undetectable HCV viral RNA for 6 months are eligible;(7) known history of active or latent TB;(8) any concomitant serious health condition that, in the opinion of the investigator, would place the patient at undue risk from the study, e.g., uncontrolled hypertension and / or diabetes, clinically significant pulmonary disease (e.g., chronic obstructive pulmonary disease requiring hospitalization within 3 months) or neurological disorder (e.g., active seizure disorder within 3 months);(9) prior therapy within the following timeframes prior to the planned initiation of study treatment:(a) small molecule inhibitors and / or other investigational drugs: ≦2 weeks or 5 half-lives, whichever is shorter;(b) chemotherapy, other monoclonal antibodies, antibody-drug conjugates or other similar experimental therapies: ≦3 weeks or 5 half-lives, whichever is shorter;(c) radioimmunoconjugates or other similar experimental therapies, ≦6 weeks or 5 half-lives, whichever is shorter;(10 ) NYHA Class III or IV, known symptomatic coronary artery disease, or symptoms of coronary artery disease at the time of systems review, or known arrhythmias (atrial fibrillation or SVT);(11) Any known or suspected active organ-threatening autoimmune disease, except treatment-controlled hypothyroidism and type 1 diabetes, e.g., inflammatory bowel disease, autoimmune hepatitis, lupus, or interstitial pneumonia;(12) Immunodeficiency or immunosuppression, including systemic corticosteroids at >10 mg / day prednisone or equivalent within 1 week prior to the planned start of study treatment;(13) History of Grade 3 or 4 immune-mediated adverse reaction to an immune CPI;(14) Toxicity from prior therapy that has not resolved to Grade 1 or less;(15) Current or past noninfectious interstitial pneumonia requiring steroids, or current interstitial pneumonia;(16) High-volume disease as assessed clinically by medical monitors via radiological findings and parameters such as tumor markers or LDH. volume disease);(17) portal vein thrombosis involving more than the intrahepatic portal vein branches: thrombosis of the right or left portal vein branch or bifurcation, partial or complete occlusion of the portal vein trunk;(18) known concurrent malignancy.Exclusions: Basal cell carcinoma of the skin, squamous cell carcinoma of the skin, or cervical carcinoma in situ treated with curative intent, prostate cancer confined to the prostate with a Gleason score <6 or PSA <1, and any stage I cancer treated with curative intent or any previous cancer with a disease-free interval of ≥3 years;(19) Any other concomitant anticancer therapy (chemotherapy, immunotherapy, radiation therapy, or any adjuvant therapy considered investigational [used for non-FDA approved indications and used in the context of a research study]);(20) Receipt of a live vaccination within 30 days of the planned start of study treatment. Injectable seasonal influenza vaccines are generally inactivated influenza vaccines and are permitted. COVID-19 mRNA vaccines are also allowed; intranasal influenza vaccines (e.g., Flu-Mist®) are live attenuated vaccines and are not allowed 4 weeks prior to the first dose and 12 weeks after the last dose of VV1 administration; (21) because this study involves an investigational drug with unknown genotoxic, mutagenic, and teratogenic effects on the developing fetus and newborn, any of the following: (a) WOCBP* or sexually active males who are unwilling to practice highly effective contraception prior to the start of the first dose / treatment, during the study, and for at least 4 weeks after the last dose. Highly effective contraceptive measures included: stable use of combined (estrogen- and progestogen-containing) hormonal contraception (oral, intravaginal, transdermal) or progestogen-only hormonal contraception (oral, injectable, implantable) associated with ovulation inhibition that had begun at least two menstrual cycles prior to screening; intrauterine device (IUD); intrauterine hormone-releasing system (IUS); bilateral tubal ligation; vasectomized partner; and / or sexual abstinence†‡.(i) stable use of combined (estrogen and progestogen-containing) hormonal contraception (oral, vaginal, transdermal) or progestogen-only hormonal contraception (oral, injectable, implantable) associated with inhibition of ovulation that has initiated at least two menstrual cycles prior to screening; (ii) IUD; IUS; (iii) bilateral tubal ligation; (iv) vasectomized partner (provided that the vasectomized male partner is the WOCBP study participant's only sexual partner and the vasectomized partner has undergone a medical evaluation for surgical success for the procedure); and / or sexual abstinence†,‡. *WOCBP are defined as women who are capable of pregnancy from menarche until postmenopause, unless permanently infertile. Permanent sterilization methods include hysterectomy, bilateral salpingectomy, and bilateral oophorectomy. Postmenopausal women are defined as having no menses for 12 months without another medical cause. High follicle-stimulating hormone (FSH) levels in the late menopausal period can be used to confirm the postmenopausal state in women not using hormonal contraception or hormone replacement therapy. However, in the absence of amenorrhea for 12 months, a single FSH measurement is insufficient to determine the emergence of the postmenopausal state. Pregnancy testing and contraception are not required for women with documented hysterectomy or tubal ligation. †Sexual abstinence is considered a highly effective method only if it is defined as abstinence from heterosexual intercourse during the entire period of risk associated with the study drug. The reliability of sexual abstinence needs to be evaluated in the context of the duration of the clinical trial and the patient's preferred and usual lifestyle. ‡Periodic abstinence (calendar method, symptomatic temperature method, postovulatory method), withdrawal (abortion intercourse), spermicide alone, and lactational amenorrhea method (LAM) are not acceptable methods of contraception.Female condoms and male condoms should not be used together;(22) Pregnant or breastfeeding women;(23) Men who prefer not to use condoms (even if they have had a previous vasectomy) during vaginal intercourse, while taking the drug, and for 4 weeks after stopping treatment;(24) Patients planning to receive ipilimumab: a. Hypersensitivity to ipilimumab excipients: If hypersensitivity is present to the product excipients (Tris hydrochloride (2-amino-2-hydroxymethyl-1,3-propanediol hydrochloride), sodium chloride, mannitol (E421), pentetic acid (diethylenetriaminepentaacetic acid), polysorbate 80, sodium hydroxide (to adjust pH), hydrochloric acid (to adjust pH).

[0177] Study treatment: As-prepared (i.e., undiluted) VSV-IFNβ-NIS was diluted in 5% sucrose, 50 mM Tris (pH 7.4), 2 mM MgCl 2 After manufacture, the virus is stored frozen at ≦−65° C. until it is prepared for use immediately prior to administration. Cemiplimab (Libtayo®) is prepared, stored, and handled according to its package insert. Ipilimumab (Yervoy®) is prepared, stored, and handled according to its package insert.

[0178] Dosage Schedule and Administration: The first cycle will be 28 days. Subsequent cycles will be 21 days. Both IV and IT VV1 will be given on day 1. In the triplet cohort, ipilimumab must also be given on day 1. Cemiplimab will be given on day 8 of cycle 1 and on day 1 of cycle 2 (21 day cycle). The treatment schedule for the combination therapy is shown in Table 2. Cemiplimab should be given on day 8 of cycle 1 (28 day cycle), then day 1 of each subsequent 21 day cycle for up to 2 years. Ipilimumab should be given on day 1 for the triplet cohort.

[0179] [Table 2]

[0180] Cemiplimab Administration: Cemiplimab will be administered as an IV infusion over 30 minutes under close observation according to the institution's standard of care. Cemiplimab should be given on day 8 of cycle 1 (28-day cycle) and on day 1 of each 21-day cycle thereafter for up to 2 years.

[0181] Intravenous VV1 administration: The dose of VV1 is 1.0 × 10 administered IV on day 1. 11 TCID 50 Virus will be diluted in 100 mL of normal saline containing 1% HSA and administered by slow IV infusion (30 min) under close observation.

[0182] Dose Administration for IV / IT Melanoma Cohort: In this cohort, all patients will receive both IV VV1 and IT VV1. The order of administration should be arranged for convenience. VV1 will be administered at 1 x 10 doses into one or more tumor lesions on day 1 using a TB syringe (or equivalent) fitted with a 20-23 gauge needle. 9 TCID 50 A single dose of VV1 is administered intratumorally (IT) by injection at a total dose of 100 mg / kg. Needle type can also be at the investigator's discretion to best distribute virus within the lesion. VV1 can be administered IT to multiple accessible lesions using a syringe and needle type appropriate for the location of the lesion. The rationale for dosing multiple lesions is that it would result in an immune response to a broader range of genetic alterations and antigenic diversity than can occur within a single lesion, which in turn would be expected to reduce the chance of immune escape by the tumor.

[0183] Administration will only be permitted directly into the tumor or accessible lymph nodes. Injections of skin, soft tissue or deep visceral lesions (e.g. liver or lung metastases) that require radiological control via CT or MRI or referral to an interventional radiologist will not be permitted. If multiple lesions are designated for injection, the lesions should be in as essentially different sites as possible within the body. Once lesions are identified, the maximum volume to be injected is 4 mL distributed across the different injectable tumors. Lesions must be larger than 0.5 cm in size to qualify for IT VV1 administration for a maximum of 8 lesions in total. Distribution of the injection volume will be determined based on the size of the lesions to be injected (Table 3):

[0184] [Table 3]

[0185] IV VV1 administration will be in the same manner as all other cohorts. Patients will receive 1.0 x 10 11 TCID 50 Patients will receive a single IV dose of VSV. All concomitant measures described above for mitigation of infusion-related reactions (IRR) should be initiated prior to IT administration for patients in this cohort. If a patient experiences an IRR with the first dose of VV1, the second modality of VV1 can be delayed until days 2-4.

[0186] Ipilimumab Dosing for the Triplet Melanoma and NSCLC Cohort: All patients in the triplet cohort will receive both IV VV1 and ipilimumab on day 1 and cemiplimab on day 8 of cycle 1. Ipilimumab will be administered as a single 50mg IV infusion over 90 minutes. Ipilimumab should be given only on day 1 of cycle 1. The order of administration on day 1 should be ipilimumab followed by VV1.

[0187] Duration of Treatment: In the absence of treatment delays due to AEs, treatment may be continued for up to 2 years or until one of the following criteria applies: (i) disease progression; (ii) intercurrent illness preventing further administration of treatment; (iii) AEs requiring treatment discontinuation: Any dose interruption lasting >12 weeks, except for the following: (a) dose interruptions of >12 weeks occurring for non-drug related reasons may be allowed if approved by the PI. The PI must be consulted prior to reinitiating treatment in patients with a dose interruption lasting >12 weeks; (b) tumor evaluation should continue according to the protocol even when dosing is interrupted; (iv) patient decision to withdraw from the study; (v) general or specific changes in the patient's condition that, in the opinion of the Investigator, make the patient unacceptable for further treatment. Clinical progression will be defined as occurring when the patient's condition deteriorates and the patient is unable to continue study treatment according to the investigator's judgment, regardless of imaging findings; (vi) patient noncompliance; (vii) pregnancy; (viii) termination of the study by the sponsor; (ix) the drug manufacturer is no longer able to provide the study drug; (x) if a patient has been treated with cemiplimab for at least 24 weeks and has been confirmed to have a complete CR lasting at least two cycles beyond CR, discussions may be considered between the treating physician and the investigator regarding discontinuation of study treatment.

[0188] Prior and Concomitant Illnesses and Medications: Patients should receive full supportive care during the study, including blood products, antibiotics, and treatment of concurrent medical conditions and other newly diagnosed illnesses.

[0189] All prior significant illnesses related to patient safety or experienced by the patient prior to screening should be recorded. Any additional illness present at the time of informed consent will be considered a concomitant illness. Illnesses that first arise or are detected during the study and / or worsening of a concomitant illness during the study will be recorded as an AE. If a patient enrolled in this study becomes infected with COVID-19, record any difficulties in administering or using the investigational product or performing the procedures specified in the protocol, including adherence to protocol-specified clinic visits and laboratory / diagnostic tests. Reasons for failure to obtain or delaying an assessment must be recorded (e.g., identify specific limitations imposed by COVID-19 that lead to the inability to perform protocol-specified assessments). COVID-19 positive test results must be collected as an AE. Imaging of patients with early COVID-19 pneumonia may be confused with interstitial pneumonia, which has been described with immune checkpoint inhibitors. If an interstitial pneumonia AE is diagnosed, COVID-19 should be placed in the differential. Once the patient has clinically recovered from actual COVID-19 infection or sufficient time has passed since exposure to high-risk situations (i.e., deemed low risk) and the patient is clinically stable, protocol-specific procedures and planned treatments may be continued.

[0190] Prohibited Procedures: Patients should not receive any other investigational drugs within 3 weeks prior to study entry, should not participate in device studies, should not receive systemic anti-cancer therapy, and will make every effort not to start any other investigational product or device studies within 28 days after the last drug administration, according to the eligibility criteria and during the study. Enrollment in non-therapeutic studies is permitted. Within 1 week prior to and during study drug administration, the following are not permitted: systemic steroids >10 mg / day prednisone or equivalent unless required for treatment of an AE; systemic immunosuppressants; other systemic anti-cancer therapy; live vaccines (which should also be avoided for 3 months after the last dose of VV1 administration unless the risks outweigh the benefits).

[0191] Description of Evaluation: Medical history will be recorded during screening to ensure patient eligibility and will focus on current or past relevant abnormalities or diseases of the following systems: gynecological, cardiovascular, respiratory, gastrointestinal, hepatic, biliary, renal, endocrine / metabolic, musculoskeletal, hematological / lymphatic, neurological / psychiatric, dermatological, immunological, infectious diseases, bleeding tendency, and allergy / drug sensitivity.

[0192] Demographics: Information regarding nativity, sex, race / ethnicity, detailed smoking and alcohol history data will be recorded during screening.

[0193] Physical Examination: A complete physical examination will include a general appearance, examination of the skin, head, ears, eyes, nose, pharynx, heart, lungs, abdomen, lymph nodes, extremities, genitals (if indicated based on symptomatology and medical history), and nervous system, as well as measurement of weight. An abbreviated (focused) physical examination should include a cardiopulmonary evaluation and abdominal examination, but will focus on any new symptoms and will include examination of relevant systems identified by the investigator. Height and weight will be assessed at screening, and then weight only.

[0194] ECOG performance status and vital signs: ECOG performance status will be recorded according to the investigator's assessment of the patient's performance status. ECOG PS: 0 = fully active and able to continue all pre-disease performance without limitation; 1 = physically strenuous activity is limited, but ambulatory and can perform tasks of a light or sedentary nature, e.g., light housework, clerical work; 2 = ambulatory and all self-care, but unable to perform any work activities; out of bed more than 50% of waking hours; 3 = limited self-care; in bed or chair more than 50% of waking hours; 4 = completely disabled; unable to continue any self-care; completely in bed or chair; 5 = death. Vital signs will include temperature, respiratory rate, heart rate, and systolic and diastolic blood pressure. Blood pressure and heart rate will be measured after 5 minutes in a sitting or semi-recumbent position by oscillometry using a blood pressure measuring device. Measurements will be repeated if systolic blood pressure is below 100 mmHg or above 150 mmHg and / or diastolic blood pressure is below 50 mmHg or above 90 mmHg. Heart rate measurements will be repeated if below 50 beats per minute (bpm) or above 100 bpm. If measurements are still outside the normal range, it is up to the investigator to decide whether measurements should be repeated. Vital sign measurements outside the normal range will be assessed by the investigator as "abnormal but not clinically significant" or "abnormal and clinically significant." In the latter case, abnormal vital sign measurements will be reported as an AE and, if clinically indicated, will be investigated further.

[0195] Electrocardiogram: A single standard 12-lead ECG will be performed at screening and one time point after VV1 administration. If the ECG is abnormal, it should be repeated. Thereafter, an ECG should be performed if clinically indicated and according to standard technique. ECG parameters recorded are: rhythm, PR interval, heart rate, QRS interval, QT interval, QTcF interval, and QRS axis. The ECG will be recorded while the patient is resting in the supine or semi-recumbent position. The ECG will be read extemporaneously and categorized as "normal", "abnormal but not clinically significant" or "abnormal and clinically significant".

[0196] Pharmacokinetic evaluation: Viral PK will be assessed by RT-PCR for VSV-IFNβ-NIS. Collection of the first whole blood sample (baseline) will occur any time within the first 24 hours prior to VV1 dosing. PK of cemiplimab in serum will be assessed by ELISA. Blood samples will be drawn pre-dose (generally within 2 hours prior to study treatment) and at the end of infusion for the first cycle and less frequently thereafter.

[0197] Tumor Biopsy: All patients will have baseline FFPE tissue identified for correlative studies at screening. In addition, mandatory fresh pre-treatment core biopsies (fine needle aspirates are insufficient) are required for at least 3 of 10 patients in stage 1 of each of the IV melanoma cohorts (doublet and triplet therapy), all stage 2 patients in these IV melanoma cohorts, and all patients in both stage 1 and stage 2 in the IV / IT melanoma cohorts. Pre-treatment biopsies must be obtained within 28 days prior to enrollment and will be sent to a central laboratory for correlative studies, which will be performed retrospectively. All patients required to provide fresh pre-treatment biopsy samples will also be required to undergo in-treatment biopsies of the same lesions on days 3-8 of cycle 1 and day 1 of cycle 2. Further optional biopsies of any other lesions may be performed if the patient consents. If the initially biopsied lesion becomes too small to allow for a repeat biopsy, another lesion may be biopsied instead. For all cohorts, both stage 1 and stage 2, upon disease progression at EOT, further optional biopsies of progressing lesions can be performed if patients consent. IHC of current FFPE biopsies will be used to evaluate markers including PD-L1, CD3+ T cells and CD8+ T cells. Other markers can be evaluated including expression of TIM3, LAG3 and IDO1 in the injected tumor microenvironment, as well as cytotoxic cells, NK cells, Th-1 cells or other immune cell markers. IHC results will be compared to baseline.

[0198] Efficacy Assessment: RECIST 1.1 will be used throughout the study for evaluation of tumor response. Either CT or MRI may be utilized per RECIST 1.1, with CT being the preferred imaging technique in this study. The same imaging modality will have to be used for each patient throughout the duration of the study. For cutaneous melanoma lesions in the melanoma cohort, digital photography will be used for evaluation per RECIST 1.1. Imaging assessment will be performed until PD. Radiologic assessment of CR or PR will require confirmatory imaging at least 4 weeks after the first assessment of response is observed. Assessment of DCR, DOR, and PFS will be based on RECIST measurements. PFS will be determined by the time from first study treatment administration to first documented disease progression or death. DOR will be determined by the time from first observed response to first documented disease progression or death. OS will be extrapolated from the time of first study treatment administration. Patients will be followed every 3 months after the end of study treatment.

[0199] Table 4 outlines baseline tumor assessment requirements.

[0200] [Table 4]

[0201] Tumor assessments and target lesion assessments during the study will be as follows:

[0202] Complete Response (CR): Disappearance of all target lesions. Any pathological lymph nodes (target or non-target) must have a short axis reduction to <10mm. Tumor marker results must normalize.

[0203] Partial response (PR): At least a 30% reduction in the sum of the diameters of the target lesions, taking the baseline sum diameter as reference.

[0204] Stable Disease (SD): Neither sufficient shrinkage to qualify for PR nor sufficient increase to qualify for PD, as referenced by the smallest (nadir) sum of diameters since the start of treatment.

[0205] Progressive Disease (PD): At least a 20% increase in the sum of the diameters of the target lesions or the appearance of one or more new lesions, taken from the smallest (nadir) sum since the start of treatment. Not only a 20% increase is required, but also an absolute increase in the sum of at least 5 mm.

[0206] On-study tumor evaluation of non-target lesions, evaluation will be as follows:

[0207] Complete Response (CR): Disappearance of all non-target lesions and normalization of tumor markers. All lymph nodes must be of non-pathological size (<10 mm short axis).

[0208] Stable Disease (SD): Persistence of one or more non-target lesions and / or persistence of tumor marker levels above the normal range.

[0209] Progressive Disease (PD): Appearance of one or more new lesions and / or unsuspected progression of existing non-target lesions. If the patient also has measurable disease, there must be a substantial worsening of the overall level of non-target disease such that the overall tumor burden has increased sufficiently to merit cessation of therapy, even if there is SD or PR in target disease.

[0210] Table 5 outlines the method for assessing best overall response. If nodal disease is included in the target lesion totals and the lymph nodes are reduced to a "normal" size (<10 mm), the lymph nodes may still have a measurement reported on the scan. This measurement should be recorded to avoid exaggerating progression, even if the lymph nodes are normal, if based on an increase in lymph node size. As mentioned above, this means that patients with a CR may not have a "zero" total. Unplanned evaluations should be performed if there is suspicion of disease progression based on clinical or laboratory findings prior to the planned evaluation. Patients with a general deterioration in health status requiring discontinuation of treatment without objective evidence of disease progression at that time should be reported as "symptomatic deterioration." Every effort should be made to document objective progression, even after cessation of treatment.

[0211] [Table 5]

[0212] Adverse Event: An AE is any symptom, physical sign, syndrome or disease that either appears during the study or, if present at screening, worsens during the study, regardless of the suspected cause of the event. All medical and psychiatric conditions present at screening (except those associated with the indication under study) will be recorded. Changes in these conditions and any new symptoms, physical signs, syndromes or diseases should be noted for the remainder of the study. Abnormalities in laboratory tests, vital signs and ECGs should also be recorded as AEs if considered clinically significant. AEs may be stated spontaneously by the patient, discovered as a result of general questioning by study staff, or determined by physical examination. During each visit to the study clinic, patients will be asked "Have you experienced any problems since your last visit?" All AEs will be recorded.

[0213] Primary endpoint / analysis, tumor response rate: The primary efficacy endpoint is the tumor response rate (ORR) by RECIST 1.1 (investigator assessed). Unless otherwise stated, tumor response is considered synonymous with success. A confirmed tumor response is defined as an objective CR or PR on two consecutive assessments at least 4 weeks apart. Tumor response will be evaluated using all cycles of treatment. ORR will be calculated overall and by cohort / subgroup using RECIST 1.1. Success rates will be estimated by dividing the number of successes by the total number of evaluable patients in each cohort. Confidence intervals for true success rates will be calculated in each cohort according to the approach of Duffy and Santner (Duffy, 1987). If more than the target number of patients are recruited in an expansion cohort, the additional patients from that cohort will not be used in any decision making; however, the additional patients will be included in the final endpoint estimate and confidence interval.

[0214] Rationale: In this study, combination therapy with oncolytic VV1, anti-PD-1 cemiplimab and anti-CTLA4 monoclonal antibody ipilimumab is predicted to enhance IT CD8+ T cell infiltration and activation to improve treatment responses in patients whose cancers are resistant to anti-PD-1 / PD-L1 alone. In this study, conducted in a melanoma patient population who failed single-agent CPIs, it is predicted that cellular infection will expand the available endogenous antitumor T cell repertoire, increase tumor cell antigenicity for T cell recognition, enhance antigen presentation, and ultimately convert a non-inflammatory or immune-exclusionary phenotype into a highly inflammatory phenotype. EXAMPLES

[0215] A Phase 2 Study of Voyager V1 (Vesicular Stomatitis Virus Expressing NIS and Human Interferon Beta, VV1) in Combination with Cemiplimab in Patients with Selected Solid Tumors This example relates to an open-label, non-randomized Phase 2 study designed to determine the antitumor activity and confirm safety of VV1 in combination with cemiplimab (350 mg IV). This study is a modification of the study described in Example 1, in which the hepatocellular, endometrial and NSCLC cohorts have been completed, the melanoma cohort has been modified, and head and neck squamous cell carcinoma (HNSCC) and colorectal cancer (CRC) cohorts have been added. Thus, the study described in Example 2 includes melanoma, HNSCC and CRC cohorts; the study design is illustrated in Figure 2.

[0216] As-prepared (i.e., undiluted) VSV-IFNβ-NIS was diluted in 5% sucrose, 50 mM Tris (pH 7.4), 2 mM MgCl 2 After manufacture, the virus will be stored frozen at ≦-65°C until it is prepared for use immediately prior to administration. The virus will be thawed and diluted in 100 mL of normal saline containing 1% HSA and administered under close observation.

[0217] The dose of VV1 was 1.0 × 10 on day 1 (D1) in combination with IV cemiplimab (also given on D1). 9 TCID 50 / m (up to 5 mL total volume; up to 5 lesions) will be administered as an intratumoral (IT) injection. Data from an ongoing clinical trial (VYR-VSV2-101) and investigator-initiated studies suggest that 1.7 × 10 11 TCID 50 (IV monotherapy and combination) and 3.0×10 9 TCID 50 It has been shown that doses up to (IT of the combination) are safe and may be more effective than lower doses of VV1. Based on previous experience with VV1 and other oncolytic viruses in combination with CPIs, no amplification of toxicity of cemiplimab or VV1 is expected.

[0218] The IT route of VV1 administration offers the advantage of significantly reducing systemic toxicity compared to IV VV1 and having a more favorable safety profile without compromising antitumor activity. In the clinical development program, there have been no observations of sustained high circulating concentrations of virally encoded IFNβ following IT VV1 administration. IT administration of VV1 is better tolerated than IV administration, with fewer systemic symptoms and fewer pre- and post-pharmacotherapy requirements.

[0219] It is expected that combination therapy with oncolytic VSV-IFNβ-NIS and cemiplimab will enhance the infiltration and activation of intratumoral CD8+ T cells, enhancing therapeutic responses in cancer patients previously resistant to CPI monotherapy. Clinical trials will include correlative studies to monitor the PK and pharmacodynamics (PD) of the virus, including monitoring local and systemic immune responses, particularly CD8+ T cell responses. These studies will help to establish the safety of combination therapy with VSV and checkpoint blockade; provide preliminary evidence of clinical efficacy in patients with advanced cancer; and elucidate the clinical mechanism of action and biomarkers of response to combination therapy.

[0220] This study, conducted in a population of melanoma patients who have failed single-agent CPIs, is designed to provide insight into the putative mechanism of resensitization to CPIs via injection of a single dose of oncolytic virus, with the hypothesis that cell infection expands the available endogenous antitumor T cell repertoire, increases the antigenicity of tumor cells for T cell recognition, enhances antigen presentation, and ultimately converts a noninflammatory or immune-exclusionary phenotype into a highly inflammatory one. Preliminary studies of immune responses in patients undergoing VSV-IFNβ-NIS therapy suggest that some patients develop T cell responses against viral and known tumor antigens.

[0221] Study Objectives: The primary objective of the study is to determine the preliminary antitumor activity in each study cohort, as measured by ORR, both in the injected lesion and according to standard criteria (RECIST 1.1) [Eisenhauer 2009]. Secondary objectives of the study include: to determine the safety and tolerability of IT VV1 in combination with cemiplimab; to evaluate the systemic PK of IT VV1 and cemiplimab; to investigate the PD of IT VV1 in blood by measuring serum IFNβ expression; and to determine the preliminary clinical activity [e.g., PFS, DOR, DCR, and OS] of each cohort.

[0222] Overall Study Design: Approximately 30-87 patients will be enrolled in the study. For the purposes of cohort expansion, response will be considered as 30% shrinkage of STL of injected lesions in the absence of RECIST 1.1-defined disease progression in uninjected lesions. Patients will be enrolled in three parallel doublet cohorts, each with an optimal Simon two-stage design; in all cohorts (melanoma, HNSCC, and CRC), patients will receive the same IT VV1 / IV cemiplimab doublet. After cycle 1, cemiplimab will be administered on D1 of each 3-week cycle. IT VV1 will be injected twice on D1 of cycle 2, and if possible, IT VV1 may be given repeatedly every 3 weeks (Q3W) until clinical benefit or limiting toxicity. Efficacy assessments will be performed every 6 weeks (Q6W) (at the end of every other cycle).

[0223] Data from an ongoing clinical trial (VYR-VSV2-101) and investigator-initiated studies indicate that 1.7 × 10 11 TCID 50 (IV monotherapy and combination) and 3.0×10 9 TCID 50It has been shown that doses up to 1000 mg / kg / day (IT of the combination) are safe and potentially more effective than lower doses of VV1 (see section 1.1.4). The IT route of VV1 administration offers the advantage of significantly reducing systemic toxicity compared to IV VV1 and having a more favorable safety profile without compromising antitumor activity. In the clinical development program, there have been no observations of sustained high circulating concentrations of virally encoded IFNβ following IT VV1 administration.

[0224] All patients enrolled in this study will receive IT VV1 only, which will no longer require an overnight hospital stay on day 1 and, if clinically stable, can be discharged for outpatient follow-up after 6 hours of observation. Patients will then need to undergo outpatient safety evaluations on days 2, 3, 4, and 5. Laboratory evaluations performed on days 2, 3, 4, and 5 must be evaluated and assessed for possible IFNβ toxicity on these days.

[0225] An optimal Simon two-stage design [Simon 1989] will be used for all cohorts in the study. In the first stage, 10 evaluable patients will be recruited in each tumor cohort; this cohort will be advanced to the second stage based on a combination of safety and efficacy. If there are fewer than three objective responses in the HNSCC cohort, fewer than two responses in the melanoma cohort, or fewer than one response in the CRC cohort among the first 10 patients, the cohort will be discontinued; otherwise, in the second stage, 11–19 additional patients will be recruited, for a total of 21–29 evaluable patients in each cohort.

[0226] Patients must have evaluable disease per RECIST 1.1. Patients must agree to provide freshly obtained biopsies prior to the start of study treatment and to repeat biopsies during study treatment, as well as to provide tissue obtained and other blood samples and buccal swabs for biomarker analysis.

[0227] Overall toxicity will be graded using NCI CTCAE, v5.0, and CRS and neurological toxicity will be graded using ASTCT consensus grading [Lee 2019]. Safety assessments will include AEs, SAEs, PEs, vital sign measurements, ECOG status, clinical safety laboratory assessments (hematology, serum chemistry and liver panel, coagulation and urinalysis), and ECGs.

[0228] For a given patient, the end of participation usually corresponds to the 90-day safety visit after the last dose of cemiplimab. However, if a patient discontinues treatment for reasons other than PD and efficacy evaluations continue after the 90-day safety visit, the end of participation is the date of the last study evaluation.

[0229] Inclusion Criteria: Patients who meet all of the following inclusion criteria at screening / day 1 of first dose will be eligible to participate in the study: (1) Age ≥ 18 years on the date of signing informed consent. (2) Specifics by tumor cohort: (a) For the HNSCC cohort, histologically confirmed diagnosis of advanced and / or metastatic HNSCC suitable for first-line immunotherapy: (i) HPV+ and HPV+ patients are allowed; (ii) Primary tumor location in the oropharynx, oral cavity, hypopharynx or larynx. Participants must not have a primary tumor site (any histology) in the nasopharynx; (iii) PD-L1 status ≥ 10% according to the local CPS score. Samples should be provided to a central laboratory for post-hoc centralized testing; (iv) at least 12 months between the last dose of prior adjuvant therapy and the date of recurrence diagnosis (if given); (v) no prior anti-PD-(L)1 treatment for HNSCC; (b) for the melanoma cohort, histologically confirmed diagnosis of progressive and / or metastatic cutaneous melanoma where existing options are not considered to provide clinical benefit: (i) best response of uPR, SD or PD to an anti-PD-(L)1-containing regimen; (ii) prior anti-PD-(L)1 therapy must have lasted ≥ 12 weeks; (iii) radiological progression during or after therapy with a PD-(L)1 immune CPI (only one prior line of PD-(L)1 therapy is permitted); (iv) if the patient received prior anti-PD-1 as adjuvant therapy, the patient should have relapsed during therapy or within the subsequent 6 months after the last dose. NOTE: Progression while receiving ipilimumab is not required; (v) Patients with BRAF V600-positive tumors should have been pretreated with a BRAF inhibitor (alone or in combination with a MEK inhibitor) in addition to treatment with anti-PD-1 or have declined targeted therapy.NOTE: Patients with BRAF V600-positive tumors without clinically significant tumor-related symptoms or evidence of rapidly progressive disease should not be treated with a BRAF inhibitor (alone or in combination with a MEK inhibitor) based on the investigator's judgment; (c) for the CRC cohort, histologically confirmed diagnosis of progressive and / or metastatic CRC: (i) received or not suitable for standard of care fluoropyrimidines, oxaliplatin, irinotecan, anti-VEGF and EGFR targeted therapy; (ii) non-microsatellite instability-high (non-MSI-high); (iii) progression on prior systemic therapy.

[0230] (3) At least one tumor lesion and biopsy suitable for IT injection that has not been previously irradiated. (a) If the lesion to be injected has not been previously biopsied and confirmed as malignant, the clinical and / or radiological configuration of the lesion must be consistent with a malignant lesion. (4) Measurable disease per RECIST 1.1, including ≥1 measurable lesion to be injected. (5) Performance status of 0 or 1 on the ECOG performance scale. (6) Life expectancy >3 months. (7) Willingness to provide biosamples as required for the duration of the study, including fresh tumor biopsy samples during the study. Note: At baseline, samples may be fresh with no intervening systemic therapy prior to enrollment, or archival. Tissue obtained from biopsy must not have been previously irradiated, although new or ongoing lesions in the radiation field are acceptable. (8) Adequate organ function, defined as laboratory values ​​obtained ≤14 days prior to enrollment, as shown in Table 6.

[0231] [Table 6]

[0232] (9) Negative pregnancy test in female patients of childbearing potential. (10) Absence of active CNS complications. Note: Pre-enrollment imaging is not required for asymptomatic patients. (11) Ability to provide written informed consent.

[0233] Exclusion Criteria: Patients who meet any of the following exclusion criteria at screening / day 1 of first dose will not be enrolled in the study: (1) Availability and patient acceptance of alternative curative treatment options. (2) Recent or ongoing serious infection, including any active viral, bacterial or fungal infection of grade 3 or higher per NCI CTCAE, v5.0, within 2 weeks of enrollment. (3) Patients diagnosed with ocular, mucosal or acral melanoma. (4) Known seropositivity for HIV and active HIV infection; (a) Patients who are seropositive for HIV but receiving antiviral therapy and who demonstrate undetectable viral load and normal CD4 T-cell counts for at least 6 months are eligible.

[0234] (5) Seropositive for HBV and evidence of active viral infection with HBV. Patients who are HBsAg negative and HBV viral DNA negative are eligible; (a) Patients who had HBV but received antiviral treatment and demonstrate undetectable viral DNA for 6 months are eligible. (b) Patients who are seropositive with HBV vaccine are eligible. (6) Seropositive for HCV and have active viral infection with HCV; (a) Patients who had HCV but received antiviral treatment and demonstrate undetectable HCV viral RNA for 6 months are eligible. (7) Known history of active or latent TB. (8) Any concomitant serious health condition that, in the opinion of the investigator, would place the patient at undue risk from the study, e.g., uncontrolled hypertension and / or diabetes, clinically significant pulmonary disease (e.g., chronic obstructive pulmonary disease requiring hospitalization within 3 months) or neurological disorder (e.g., active seizure disorder within 3 months).

[0235] (9) Previous therapy within the following timeframes prior to the planned initiation of study treatment: (a) small molecule inhibitors and / or other investigational drugs: ≤2 weeks or 5 half-lives, whichever is shorter; (b) chemotherapy, other monoclonal antibodies, antibody-drug conjugates or other similar experimental therapies: ≤3 weeks or 5 half-lives, whichever is shorter; (c) radioimmunoconjugates or other similar experimental therapies, ≤6 weeks or 5 half-lives, whichever is shorter. (10) NYHA class III or IV, known symptomatic coronary artery disease, or symptoms of coronary artery disease at the time of systems review, or known arrhythmias (atrial fibrillation or SVT). (11) Any known or suspected active organ-threatening autoimmune disease, e.g., inflammatory bowel disease, autoimmune hepatitis, lupus, or interstitial pneumonia, except for treatment-controlled hypothyroidism and type 1 diabetes. (12) Immunodeficiency or immunosuppression, including systemic corticosteroids at >10 mg / day prednisone or equivalent, within 1 week prior to the planned initiation of study treatment.

[0236] (13) History of grade 3 or 4 immune-mediated adverse reaction to an immune CPI. (14) Toxicity from previous therapy that has not resolved to grade 1 or less. (15) History of non-infectious interstitial pneumonia or current interstitial pneumonia requiring steroids. (16) High volume disease assessed clinically by medical monitor via radiological findings and parameters such as tumor markers or LDH. (17) Portal vein thrombosis involving more than the intrahepatic portal vein branch: thrombosis of right or left portal vein branch or bifurcation, partial or complete obstruction of portal vein trunk. (18) Known concurrent malignancy. Exclusions: Basal cell carcinoma of the skin, squamous cell carcinoma of the skin, or cervical carcinoma in situ treated with curative intent, prostate cancer confined to the prostate with Gleason score <6 or PSA <1, and any stage I cancer treated with curative intent or any previous cancer with a disease-free interval of ≥3 years. (19) Any other concomitant anticancer therapy (chemotherapy, immunotherapy, radiation therapy, or any adjuvant therapy considered investigational [used for non-FDA approved indications and used in the context of a research study]).

[0237] (20) Have received a live vaccination within 30 days of the planned start of study treatment. Injectable seasonal influenza vaccines are generally inactivated influenza vaccines and are permitted. COVID-19 mRNA vaccines are also permitted, following the following guidance: Patients undergoing screening who have not started study treatment should plan to receive the second dose at least 2 weeks prior to day 1 (assuming the vaccine is a two-dose vaccine). If a single-dose vaccine is approved, the single dose should be at least 2 weeks prior to day 1. Vaccine administration during cycle 1 between the first and second doses of cemiplimab is not recommended. From cycle 2 onwards, a minimum of 7 days should elapse between the last treatment and the CoVID-19 vaccine administration, and patients should have 7 days elapse between the vaccine administration and the next study treatment. Efforts should be made to avoid treatment delays, but if necessary, a delay of up to 7 days for the next treatment with the study drug will be permitted. Intranasal influenza vaccines (e.g., FLU-MIST) are live attenuated vaccines and should not be administered for 4 weeks prior to the first dose of VV1 or for 12 weeks after the last dose.

[0238] (21) Because this study involves an investigational drug with unknown genotoxic, mutagenic, and teratogenic effects on the developing fetus and newborn, any of the following: (a) WOCBP* or sexually active men unwilling to practice highly effective contraception prior to the start of the first dose / treatment, during the study, and for at least 4 weeks after the last dose. Highly effective contraceptive measures include: stable use of combined (estrogen- and progestogen-containing) hormonal contraception (oral, intravaginal, transdermal) or progestogen-only hormonal contraception (oral, injectable, implantable) associated with ovulation inhibition that has begun 2 or more menstrual cycles prior to screening; intrauterine device (IUD); intrauterine hormone-releasing system (IUS); bilateral tubal ligation; vasectomized partner; and / or sexual abstinence†‡. (i) stable use of combined (estrogen- and progestogen-containing) hormonal contraception (oral, vaginal, transdermal) or progestogen-only hormonal contraception (oral, injectable, implant) associated with ovulation inhibition that has initiated two or more menstrual cycles prior to screening; (ii) IUD; IUS; (iii) bilateral tubal ligation; (iv) vasectomized partner (provided that the vasectomized male partner was the WOCBP study participant's only sexual partner and the vasectomized partner had undergone a medical evaluation for surgical success of the procedure); and / or sexual abstinence†,‡.

[0239] *WOCBP are defined as women who are fertile from menarche until postmenopause, unless permanently infertile. Permanent sterilization methods include hysterectomy, bilateral salpingectomy, and bilateral oophorectomy. Postmenopausal women are defined as the absence of menses for 12 months without another medical cause. Elevated follicle-stimulating hormone (FSH) levels in the late menopausal period can be used to confirm the postmenopausal state in women not using hormonal contraception or hormone replacement therapy. However, in the absence of 12 months of amenorrhea, a single FSH measurement is insufficient to determine the emergence of the postmenopausal state. Pregnancy testing and contraception are not required for women with documented hysterectomy or tubal ligation.

[0240] †Sexual abstinence is considered to be highly effective only if it is defined as abstinence from heterosexual intercourse during the entire period of risk associated with the study drug. The reliability of sexual abstinence needs to be evaluated in the context of the duration of the clinical trial and the patient's preferred and usual lifestyle.

[0241] ‡Periodic abstinence (calendar, symptomatic-thermal, or postovulatory methods), withdrawal (abortive intercourse), spermicide alone, and lactational amenorrhea method (LAM) are not acceptable methods of contraception. Female condoms and male condoms should not be used together.

[0242] (22) Pregnant or lactating women. (23) Men who prefer not to use condoms (even if they have had a previous vasectomy) during vaginal intercourse, while taking the drug, and for 4 weeks after stopping treatment. (24) Previous toxicity of grade ≥ 3 on anti-PD-1 therapy. (25) Intent to inject into lesions with major vascular involvement (e.g., carotid arteries, large hepatic vessels). (26) Progressive disease within 3 months of completion of curative-intent systemic treatment for locoregionally advanced head and neck cancer.

[0243] Dosage schedule and administration: A cycle is defined as 3 weeks (21 days). Both IT VV1 and cemiplimab will be administered on day 1 of cycle 1 and after day 1 of cycle 2. VV1 IT injections should continue Q3W if possible, and cemiplimab infusions will continue Q3W until limiting toxicity or clinical benefit. End of treatment is defined as the last administration of either study drug.

[0244] The treatment schedule for the combination therapy is shown in Table 7. The dose of VV1 is based on previous clinical experience and data (see Investigator Brochure). The dose of cemiplimab is as recommended in the package insert [Libtayo® 2020]. Cemiplimab should be given on day 1 of each 21-day cycle for up to 2 years.

[0245] [Table 7]

[0246] VV1 IT injections should be continued Q3W if possible, and cemiplimab infusions will continue Q3W until limiting toxicity or clinical benefit. End of treatment will be defined as the last administration of either study drug.

[0247] Cemiplimab Administration: Cemiplimab should be prepared according to the package insert [Libtayo® 2020]. Cemiplimab is administered as an IV infusion over 30 minutes prior to IT VV1 with close observation according to the institution's standard of care. Patients should then be observed for IRR for 60 minutes before receiving VV1. Cemiplimab should be given on day 1 of each subsequent 21-day cycle for up to 2 years.

[0248] IT VV1 Dose Administration: VV1 will be administered as above into one or more tumor lesions on day 1 using a TB syringe (or equivalent) fitted with a 20-23 gauge needle. Needle type can also be at the investigator's discretion to best distribute virus within the lesion.

[0249] VV1 can be administered IT into multiple reachable lesions using a syringe and needle type appropriate for the location of the lesion. For VV1 treatment, 1 to 5 lesions can be injected; starting with the largest and progressing to the smallest, a maximum volume of 5 mL (5.0 x 10 total). 9 TCID 50) Each tumor was treated with at least 1.0 × 10 9 TCID 50 1 mL / tumor will be injected IT. Newly formed lesions should be treated first in the next cycle if the patient meets retreatment criteria. If there are only 1 or 2 large lesions, more than 1 mL per tumor can be given per physician's discretion and with the consent of the medical monitor.

[0250] Administration is permitted only directly into the tumor or into accessible lymph nodes. If multiple lesions are designated for injection, the lesions should be in as disparate sites in the body as possible.

[0251] Once a lesion is identified, the maximum volume injected is 5 mL distributed across the different tumors that can be injected. Lesions must be larger than 1.0 cm in size at baseline to be eligible for IT VV1 dosing. Distribution of injection volume will be determined based on the size of the lesion to be injected, as shown in Table 8.

[0252] [Table 8]

[0253] *Lesions ≤1.0 cm may be injected only if clinically responsive to a previous VV1 injection. The treating physician has the discretion to identify and treat lesions, but these must be recorded on the injection worksheet. VV1 is administered in doses of 1.0 x 10 9 TCID 50 / mL. The total dose is up to 5.0 × 10 9 TCID 50 Each tumor can receive at least 1.0 × 10 9 TCID 50 / mL were injected IT starting from the largest to the smallest, with a maximum volume of 5 mL (5.0 × 10 9 TCID 50 Inject between 3-5 or multiple lesions as needed until a total volume of 1000 mg / kg is reached or until all injectable lesions have been treated. Newly formed lesions should be treated first in the next cycle. If there are only one or two large lesions, more than 1 mL per tumor may be given, according to the physician's discretion. All concomitant measures for relief of virus-associated systemic symptoms should be initiated prior to IT VV1 administration.

[0254] Patients may continue to receive IT VV1 treatment if the requirements set out in Table 9 are met.

[0255] [Table 9]

[0256] Description of Evaluation: Medical history will be recorded during screening to ensure patient eligibility and will focus on current or past relevant abnormalities or diseases of the following systems: gynecological, cardiovascular, respiratory, gastrointestinal, hepatic, biliary, renal, endocrine / metabolic, musculoskeletal, hematological / lymphatic, neurological / psychiatric, dermatological, immunological, infectious diseases, bleeding tendency, and allergy / drug sensitivity.

[0257] Demographics: Information regarding nativity, sex, race / ethnicity, detailed smoking and alcohol history data will be recorded during screening.

[0258] Physical Examination: A complete physical examination will include a general appearance, examination of the skin, head, ears, eyes, nose, pharynx, heart, lungs, abdomen, lymph nodes, extremities, genitals (if indicated based on symptomatology and medical history), and nervous system, as well as a measurement of weight. An abbreviated (focused) physical examination should include a cardiopulmonary evaluation and abdominal examination, but will focus on new symptoms and will include examination of relevant systems identified by the investigator. AE reporting should be completed for all changes identified as clinically important. Height and weight will be assessed at screening, and then weight only.

[0259] ECOG performance status and vital signs: ECOG performance status will be recorded according to the investigator's assessment of the patient's performance status. ECOG PS: 0 = fully active and able to continue all pre-disease performance without limitation; 1 = physically strenuous activity is limited, but ambulatory and can perform tasks of a light or sedentary nature, e.g., light housework, clerical work; 2 = ambulatory and all self-care, but unable to perform any work activities; out of bed more than 50% of waking hours; 3 = limited self-care; in bed or chair more than 50% of waking hours; 4 = completely disabled; unable to continue any self-care; completely in bed or chair; 5 = death. Vital signs will include temperature, respiratory rate, heart rate, and systolic and diastolic blood pressure. Blood pressure and heart rate will be measured after 5 minutes in a sitting or semi-recumbent position by oscillometry using a blood pressure measuring device. Measurements will be repeated if systolic blood pressure is below 100 mmHg or above 150 mmHg and / or diastolic blood pressure is below 50 mmHg or above 90 mmHg. Heart rate measurements will be repeated if below 50 beats per minute (bpm) or above 100 bpm. If measurements are still outside the normal range, it is up to the investigator to decide whether measurements should be repeated. Vital sign measurements outside the normal range will be assessed by the investigator as "abnormal but not clinically significant" or "abnormal and clinically significant." In the latter case, abnormal vital sign measurements will be reported as an AE and, if clinically indicated, will be investigated further.

[0260] Laboratory Parameters: The following laboratory tests will be performed: Serology / Virology, Urinalysis, Electrocardiogram (ECG), Drug Dosing, Pharmacokinetic Assessment, Viral Pharmacokinetics, Cemiplimab Pharmacokinetics, PD Assessment, Anti-VSV IgG Assay, Cemiplimab Immunogenicity, Serum IFN-beta and Cytokine Profile, Peripheral Blood Mononuclear Cells, Tumor Biopsy, Efficacy Assessment and Baseline Assessment.

[0261] Serology and virology measurements will be performed at screening, although recent previously reported results may be used for eligibility purposes at the discretion of the medical monitor. HIV(1 / 2) testing is required if the patient has never been tested for HIV, or if testing predates the primary cancer diagnosis, or testing was performed ≥12 months prior to the study. Virology testing at screening will be as follows: HbsAg; HCV RNA; HIV-1 / HIV-2 antibodies. Patients with HIV, HBV or HCV infection and who have received antiviral treatment should demonstrate undetectable viral RNA / load for 6 months prior to enrollment in this study.

[0262] Standard dipstick urine testing is sufficient for study purposes; however, investigational sites may perform urinalysis according to the investigational site's typical procedure. Any clinically significant abnormalities should be investigated according to standard medical practice, and any AEs that are diagnosed should be recorded.

[0263] A single standard 12-lead ECG will be performed at screening and one time point after VV1 administration. If the ECG is abnormal, it should be repeated. Thereafter, if clinically indicated, an ECG should be performed according to standard technique. ECG parameters recorded are: rhythm, PR interval, heart rate, QRS interval, QT interval, QTcF interval and QRS axis. The ECG will be recorded while the patient is resting in the supine or semi-recumbent position. The ECG will be read ex situ and categorized as "normal", "abnormal but not clinically significant" or "abnormal and clinically significant". If the post-study drug ECG is determined to be abnormal and clinically significant, a corresponding AE report should be completed for the clinically significant finding.

[0264] Viral PK will be assessed by RT-PCR for VSV-IFNβ-NIS. Collection of the first whole blood sample (baseline) will occur any time within the first 24 hours prior to VV1 dosing. PK of cemiplimab in serum will be assessed by ELISA. Blood samples will be drawn pre-dose (generally within 2 hours prior to study treatment) and at the end of infusion for the first cycle and less frequently thereafter.

[0265] Anti-VSV IgG assays are required to determine the patient's immune response to the oncolytic virus. IFNβ will be evaluated in conjunction with PK data. Other proinflammatory cytokines and safety related ones can be assayed in the same samples if dictated by the safety profile and ongoing findings in the study. Peripheral blood mononuclear cells (PBMCs) will be collected at baseline and at designated time points post-treatment for T cell assays, including sequencing.

[0266] Tumor Biopsy: All patients will have baseline formalin-fixed paraffin-embedded (FFPE) tissue identified for correlative studies at screening. In addition, a fresh pre-treatment core biopsy (fine-needle aspirate is not sufficient) will be requested if the patient has received systemic therapy since the previous biopsy. Pre-treatment biopsy tissue must not have been previously irradiated, although new or progressing lesions in the radiation field are acceptable. This pre-treatment biopsy is optional in phase 1 of the study, but mandatory in phase 2. All patients will also be required to undergo an in-treatment biopsy of the same lesion as per the plan of evaluation. If the patient consents, a further optional biopsy of any other lesion may be performed. If the initially biopsied lesion becomes too small to allow for a repeat biopsy, another lesion may be biopsied instead. If a previously injected lesion is not suitable for biopsy, this information will be recorded. For all cohorts, both phase 1 and phase 2, upon disease progression at EOT, a further optional biopsy of a progressing lesion may be performed if the patient consents. IHC of current FFPE biopsies will be used to assess markers including PD-L1, CD3+ T cells and CD8+ T cells. Other markers can be assessed including expression of TIM3, LAG3 and IDO1 in the injected tumor microenvironment, as well as cytotoxic cells, NK cells, Th-1 cells or other immune cell markers. IHC results will be compared to baseline.

[0267] Efficacy Assessment: For evaluation of tumor response, both the sole measurement of shrinkage of injected lesions and RECIST 1.1 will be used throughout the study. Either CT or MRI may be utilized per RECIST 1.1, but CT is the preferred imaging technique in this study. The same imaging modality will have to be used for each patient throughout the duration of the study. For cutaneous melanoma lesions, digital photography will be used for evaluation per RECIST 1.1. Imaging assessment will be performed until PD. Radiologic assessment of CR or PR will require confirmatory imaging at least 4 weeks after the first assessment of response is observed. Measurement of injected lesions will follow RECIST 1.1, but total target lesions will include only injected lesions. Assessment of DCR, DOR, and PFS will be based on RECIST measurements. PFS will be determined by the time from first study treatment administration to first documented disease progression or death. DOR will be determined by the time from first observed response to first documented disease progression or death. OS will be estimated from the time of first study treatment administration. Patients will be followed up every 3 months after the end of study treatment.

[0268] Baseline eligibility tumor assessments will be performed according to Table 10.

[0269] [Table 10]

[0270] Tumor evaluations during the study, including evaluation of target and non-target lesions, will be the same as those described in Example 1 and Table 5.

[0271] Assessment of Concomitant Medications: Any medications or blood products (including non-prescription medications and nutritional supplements) used within 4 weeks prior to the first dose of investigational drug through 28 days (for all concomitant medications) or 90 days (for concomitant medications used to treat an investigational drug-related AE) after the last dose will be recorded along with the primary reason for its prescription. Any new anti-cancer treatments started within 90 days after the last dose of study drug should be recorded.

[0272] Primary endpoint / analysis, tumor response rate: The primary efficacy endpoint is the rate of tumor response (ORR) of injected lesions and overall by RECIST 1.1 (investigator assessed). Unless otherwise stated, tumor response is considered synonymous with success. A confirmed tumor response is defined as an objective status of CR or PR on two consecutive assessments at least 4 weeks apart. Tumor response will be assessed using all cycles of treatment. ORR will be calculated overall and by cohort / subgroup using RECIST 1.1. Success rates will be estimated by dividing the number of successes by the total number of evaluable patients in each cohort.

[0273] Efficacy parameters will include the following: DCR will be defined as the percentage of patients with CR, PR, or SD (i.e., confirmed ≥12-week CR / PR or SD) on at least two consecutive tumor assessments. DOR will be defined as the date the patient's earliest best objective status is first documented as either CR or PR, for all evaluable patients who achieved an objective response, to the earliest date of documented progression. DOR will be estimated using the Kaplan-Meier method. PFS will be defined as the time from first administration of study drug to the earliest date of documented disease progression or death from any cause. Distribution of PFS will be estimated using the Kaplan-Meier [Kaplan and Meier 1958] method overall and, for appropriate subgroups and cohorts, using RECIST 1.1 as above. Progression-free percentages for time points such as 6, 12, 18, and 24 months may also be reported. Survival will be defined as the time from first administration of study drug to death from any cause. Survival distributions will be estimated using the Kaplan-Meier method overall [Kaplan and Meier 1958] and, for appropriate subgroups and cohorts, using RECIST 1.1 as above. Percent survival for time points such as 6, 12, 18, and 24 months can also be reported.

[0274] SEQUENCE LISTING: The sequence listing of the present application is submitted electronically as an xml file in ST.26 format with the filename "11163_seqlist_ST26", creation date of November 23, 2022, and size of 30.8. This submitted sequence listing is part of the specification and is incorporated herein by reference in its entirety. The following sequences have lengths less than the minimum length permitted under the ST.26 format: SEQ ID NO:7 (Ala Ala Ser) and SEQ ID NO:19 (Ala Ala Ser).

[0275] The present disclosure is not limited in scope by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications are intended to fall within the scope of the appended claims.

[0276] References ACS: American Cancer Society. Cancer Facts and Figures 2018 and 2019. Antonia et al., N Engl J Med 2017;377(20):1919-1929. Arora et al., Adv Ther 2019;36(10):2638-2678. Barber, Viral Immunol 2004;17(4):516-527. Borcherding et al., J Mol Biol 2018;430(14):2014-2029. Bray et al., CA Cancer J Clin 2018;68:394-424. Buchbinder et al., Am J Clin Oncol. 201;39(1):98-106. Burtness et al., Ann Oncol 2018;29:viii729. Chaturvedi et al., J Clin Oncol 2011;29(32):4294-301. Chow et al, J Clin Oncol 2016;34(32):3838-45. Clinical Guidelines Network. Role of biological agents in first-line treatment of metastatic colorectal cancer, available at: https: / / wiki.cancer.org.au / australia / Guidelines:Colorectal_cancer / Biological_agents_first-line_treatment_of_metastatic_CRC Dranoff, Immunol Rev 2002;188:147-54. Duffy et al., Biometrics 1987;43(1):81-93. Durham et al., Mol Ther 2017;25(8):1917-1932. Eisenhauer et al., Eur J Cancer 2009;45(2):228-47. Garassino et al., Ann Oncol 2017;28(suppl_2):ii28-ii51. Garon et al., Lancet 2014;384(9944):665-673. Gentzler et al., Immunother 2016;8(5):583-600. doi: 10.2217 / imt-2015-0029. Govindan et al., J Clin Oncol. 2017;35(30):3449-3457. doi: 10.1200 / JCO.2016.71.7629. Hegde et al., Clin Cancer Res. 2016;22(8):1865-1874. Jakafi(R) US prescribing information available at https: / / www.jakafi.com / pdf / prescribing-information.pdf (1 Jul 2021). Jiang et al., EJNMMI Res 2017;7(1):90. Kaplan et al., J Am Statistical Assoc 1958;53:457-481. Kulangara et al., Arch Pathol Lab Med 2019;143(3):330-7. Kurisetty et al., Head Neck 2014;36(11):1619-1627. Lee et al., Int J Colorectal Dis. 2007;22(6):699-704. Lee et al., Biol Blood Marrow Transplant 2019;25:625-638. Levey et al., Ann Intern Med 2009;150(9):604-612. Libtayo(R) US prescribing information available at https: / / packageinserts.bms.com / pi / pi_yervoy.pdf (19 Oct 2021). Lichty et al.,Trends Mol Med 2004;10(5):210-216. Long et al., J Clin Oncol 2016;34(15):9568-9568. Luchini et al.,Ann Oncol 2019;30(8):1232-1243 Luke et al., Nat Rev Clin Oncol 2017;14(8):463-482. Martini et al., World J Gastroenterol. 2017;23(26):4675-4688. Miller et al., Mol Ther Oncolytics 2014;1:14005. Miller et al., Expert Opin Biol Ther 2016;16(1):15-32. Miura et al., J Thoracic Oncol 2018;13:S692 (P2.01-71). Naik et al., Leukemia 2012;26(8):1870-1878. Obuchi et al., J Virol 2003;77(16):8843-56. O'Day et al., Ann Oncol. 2010;21(8):1712-1717. Patel et al., Oncotarget 2015;6(32):33165-33177. Puzanov et al., J Clin Oncol 2016;34(22):2619-26. Ramalingam et al., J Clin Oncol 2020;38(15_suppl):9500-9500. Reck et al., ESMO Open 2021;6(5):100273. Reck et al., N Engl J Med 2016;375(19):1823-1833. Ribas, Cancer Discov 2015;5:915-919. Ribas et al., Cell 2017;170(6):1109-1119. Rischin et al., J Clin Oncol (ASCO Meeting Abstracts) 2019;37(15_suppl). Abstract 6000. Rojas et al., Clin Cancer Res. 2015;21(24):5543-51 Saloura et al., Hum Gene Ther 2010;21(1):51-64. Sampson et al., J Allergy Clin Immunol 2006;117(2):391-397. Sezer et al., Lancet 2021;397(10274):592-604. Shen W et al., Blood 2016;127(11):1449-58. Siegel et al., CA Cancer J Clin. 2019;69(1):7-34. Siegel et al., CA Cancer J Clin 2021; 71:7-33. Simon, Control Clin Trials. 1989;10(1):1-10. Spranger et al., Sci Transl Med 2013;5(200):200ra116. Spigel et al. Impower110: Interim overall survival (OS) analysis of a Phase III study of atezolizumab (atezo) vs platinum-based chemotherapy (chemo) as first-line (1L) treatment (tx) in PD-L1-selected NSCLC. Presented at ESMO 2019; September 27-October 1, 2019; Barcelona, Spain. Abstract LBA78. Sung et al., CA Cancer J Clin. 2021 May;71(3):209-249. Van, Oncologist. 2006;11(9):1010-1017. Willmon et al., Cancer Res 2009; 69(19):7713-7720. Wolchok et al., J Clin Oncol 2021;39(15_suppl):9506-9506. Yervoy(R) US prescribing information available at https: / / packageinserts.bms.com / pi / pi_yervoy.pdf (19 Oct 2021). Yoo et al., Clin Colorectal Cancer. 2006;6(3):202-207. Zhang et al., Mol Ther 2016;24(12):2109-17.

Claims

1. 1. A method of treating or inhibiting tumor growth comprising: (a) selecting cancer patients; and (b) the following: (i) a therapeutically effective amount of an oncolytic virus; and (ii) a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds to programmed death 1 (PD-1), wherein the anti-PD-1 antibody or antigen-binding fragment thereof comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:1, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:2; to a patient in need thereof.

2. The method of claim 1 , wherein the oncolytic vesiculovirus comprises an oncolytic vesicular stomatitis virus (VSV).

3. The method of claim 2 , wherein the VSV comprises a recombinant VSV.

4. The method of claim 3 , wherein the recombinant VSV expresses a cytokine.

5. 5. The method of claim 4, wherein the cytokine comprises interferon beta (IFNb).

6. The method of claim 5, wherein the nucleic acid sequence encoding IFNb is located between the M viral gene and the G viral gene.

7. The method of any one of claims 3 to 6, wherein the recombinant VSV further expresses a sodium / iodide symporter (NIS).

8. The method of claim 7, wherein the nucleic acid sequence encoding the NIS is located between the G and L viral genes.

9. The method according to any one of claims 1 to 8, wherein the oncolytic virus is Voyager V1.

10. The method of any one of claims 1 to 9, wherein the anti-PD-1 antibody or antigen-binding fragment thereof comprises HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 3, 4, 5, 6, 7 and 8, respectively.

11. The method of any one of claims 1 to 11, wherein the anti-PD-1 antibody or antigen-binding fragment thereof comprises an HCVR comprising the amino acid sequence of SEQ ID NO:1; and an LCVR comprising the amino acid sequence of SEQ ID NO:

2.

12. The method of any one of claims 1 to 11, wherein the anti-PD-1 antibody or antigen-binding fragment thereof comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain comprising the amino acid sequence of SEQ ID NO:

10.

13. The method of any one of claims 1 to 12, wherein the anti-PD-1 antibody or antigen-binding fragment thereof comprises cemiplimab or a biological equivalent thereof.

14. Oncolytic viruses are 10-10 １４ T.C.I.D. ５０ The method of any one of claims 1 to 13, wherein the compound is administered to the patient in one or more doses of

15. Oncolytic viruses are ９ T.C.I.D. ５０ or 10 １１ T.C.I.D. ５０ 15. The method of claim 14, wherein the compound is administered to the patient in one or more doses of

16. 10 ９ T.C.I.D. ５０ A single dose of the oncolytic virus is administered to the patient, and １１ T.C.I.D. ５０ 15. The method of claim 13 or 14, wherein another dose of the oncolytic virus is administered to the patient.

17. 17. The method of any one of claims 1-16, wherein the anti-PD-1 antibody or antigen-binding fragment thereof is administered to the patient in one or more doses of about 0.1 mg / kg to about 20 mg / kg of the patient's body weight.

18. The method of any one of claims 1 to 17, wherein the anti-PD-1 antibody or antigen-binding fragment thereof is administered to the patient in one or more doses of from about 1 mg to about 1000 mg.

19. 20. The method of claim 18, wherein the anti-PD-1 antibody or antigen-binding fragment thereof is administered to the patient in one or more doses of 350 mg.

20. The method of any one of claims 1 to 19, wherein the oncolytic virus is administered intratumorally or intravenously to the patient.

21. The method of any one of claims 1 to 20, wherein the anti-PD-1 antibody or antigen-binding fragment thereof is administered to the patient intravenously, subcutaneously, or intraperitoneally.

22. 22. The method of any one of claims 1 to 21, further comprising administering a therapeutically effective amount of a CTLA-4 inhibitor comprising ipilimumab or a biological equivalent thereof.

23. 23. The method of claim 22, wherein the CTLA-4 inhibitor is ipilimumab.

24. 24. The method of claim 22 or 23, comprising administering one or more doses of an oncolytic virus and one or more doses of a CTLA-4 inhibitor simultaneously.

25. 25. The method of any one of claims 22-24, comprising administering one or more doses of an oncolytic virus and one or more doses of a CTLA-4 inhibitor prior to administering the one or more doses of an anti-PD-1 antibody or antigen-binding fragment thereof.

26. 26. The method of any one of claims 22-25, comprising administering one or more doses of an oncolytic virus and one or more doses of a CTLA-4 inhibitor 7 days prior to administering the one or more doses of an anti-PD-1 antibody or antigen-binding fragment thereof.

27. (blank)

28. 28. The method of any one of claims 22-27, wherein the CTLA-4 inhibitor is administered to the patient in one or more doses of from about 0.1 mg / kg to about 15 mg / kg of the patient's body weight.

29. The method of any one of claims 22 to 28, wherein the CTLA-4 inhibitor is administered to the patient in one or more doses of from about 1 mg to about 600 mg.

30. 30. The method of claim 29, wherein the CTLA-4 inhibitor is administered to the patient in one or more doses of 50 mg.

31. The method of any one of claims 22 to 30, wherein the CTLA-4 inhibitor is administered to the patient intravenously, subcutaneously, or intraperitoneally.

32. The method according to any one of claims 1 to 29, wherein the cancer is selected from adrenal tumor, biliary tract cancer, bladder cancer, brain cancer, breast cancer, carcinoma, central or peripheral nervous system tissue cancer, cervical cancer, colon cancer, endocrine or neuroendocrine or hematopoietic cancer, esophageal cancer, fibroma, gastrointestinal cancer, glioma, head and neck cancer, Li-Fraumeni tumor, liver cancer, lung cancer, lymphoma, melanoma, meningioma, multiple neuroendocrine type I and type II tumors, nasopharyngeal cancer, oral cancer, oropharyngeal cancer, osteogenic sarcoma tumor, ovarian cancer, pancreatic cancer, pancreatic islet cell cancer, parathyroid cancer, pheochromocytoma, pituitary tumor, prostate cancer, rectal cancer, kidney cancer, respiratory cancer, sarcoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer, tracheal cancer, genitourinary cancer and uterine cancer.

33. The method of any one of claims 1 to 30, wherein administration of the combination therapy leads to at least one therapeutic effect selected from the following: reduction in tumor cell count, tumor regression, partial response, and complete response.

34. The method of any one of claims 1 to 33, wherein administration of the combination therapy leads to at least one improvement selected from the following: long-range effect, delayed tumor growth, increased overall survival, increased progression-free survival, increased overall response rate, increased complete response, increased partial response, and increased stable disease compared to patients treated with monotherapy selected from the following: an oncolytic virus and an anti-PD-1 antibody or antigen-binding fragment thereof.

35. The method of claim 34, wherein administration of the combination therapy leads to at least one improvement selected from the following: long-distance response, delayed tumor growth, increased overall survival, increased progression-free survival, increased overall response rate, increased complete response, increased partial response, and increased stable disease compared to patients treated with oncolytic virus monotherapy.

36. The method of any one of claims 1 to 35, wherein tumor growth is inhibited by at least 50% compared to untreated patients.

37. The method of any one of claims 22-33, wherein administration of the combination therapy leads to at least one improvement selected from the following: long-distance effect, delayed tumor growth, increased overall survival, increased progression-free survival, increased overall response rate, increased complete response, increased partial response, and increased stable disease compared to patients treated with monotherapy selected from the following: an oncolytic virus, an anti-PD-1 antibody or antigen-binding fragment thereof, and a CTLA-4 inhibitor.

38. 38. The method of claim 37, wherein tumor growth is inhibited by at least 50% compared to a patient administered an oncolytic virus, an anti-PD-1 antibody or antigen-binding fragment thereof, or a CTLA-4 inhibitor as monotherapy.

39. The method of claim 37 or 38, wherein tumor growth is inhibited by at least 50% compared to a patient administered any two of an oncolytic virus, an anti-PD-1 antibody or antigen-binding fragment thereof, and a CTLA-4 inhibitor.

40. 40. The method of any one of claims 1 to 39, further comprising administering to the patient an additional therapeutic agent or therapy.

41. Additional therapeutic agents or therapies include radiation, surgery, chemotherapy agents, cancer vaccines, B7-H3 inhibitors, B7-H4 inhibitors, lymphocyte activation gene 3 (LAG3) inhibitors, T-cell immunoglobulin and mucin domain-containing 3 (TIM3) inhibitors, galectin 9 (GAL9) inhibitors, V-domain immunoglobulin (Ig)-containing suppressor of T-cell activation (VISTA) inhibitors, killer cell immunoglobulin-like receptor (KIR) inhibitors, B- and T-lymphocyte attenuator (BTLA) inhibitors, inhibitors of Ig and T-cell immunoreceptor with ITIM domains (TIGIT), CD47 inhibitors, indoleamine-2,3-dioxygenase (I 41. The method of claim 40, wherein the therapeutic agent is selected from an anti-inflammatory drug (IDD) inhibitor, a vascular endothelial growth factor (VEGF) antagonist, angiopoietin-2 (Ang2) inhibitor, a transforming growth factor beta (TGFβ) inhibitor, an epidermal growth factor receptor (EGFR) inhibitor, an antibody against a tumor specific antigen, a Bacillus Calmette-Guerin vaccine, a granulocyte-macrophage colony stimulating factor (GM-CSF), a cytotoxin, an interleukin 6 receptor (IL-6R) inhibitor, an interleukin 4 receptor (IL-4R) inhibitor, an IL-10 inhibitor, IL-2, IL-7, IL-12, IL-21, IL-15, an antibody-drug conjugate, an anti-inflammatory drug, and combinations thereof.

42. 1. A combination of an oncolytic virus and an antibody or antigen-binding fragment thereof that specifically binds to Programmed Death 1 (PD-1) for use in a method of treating or inhibiting tumor growth, the method comprising: (a) selecting cancer patients; and (b) the following: (i) a therapeutically effective amount of an oncolytic virus; and (ii) a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds to programmed death 1 (PD-1), wherein the anti-PD-1 antibody or antigen-binding fragment thereof comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:1, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:2; administering to a patient in need thereof a combination therapy comprising:

43. 1. A combination of an oncolytic virus, an antibody or antigen-binding fragment thereof that specifically binds to programmed death 1 (PD-1), and a CTLA-4 inhibitor, including ipilimumab or a biological equivalent thereof, for use in a method of treating or inhibiting tumor growth, the method comprising: (a) selecting cancer patients; and (c) the following: (i) a therapeutically effective amount of an oncolytic virus; (ii) a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds to programmed death 1 (PD-1), wherein the anti-PD-1 antibody or antigen-binding fragment thereof comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:1, and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:2; and (iii) a therapeutically effective amount of a CTLA-4 inhibitor, including ipilimumab or a biological equivalent thereof. administering to a patient in need thereof a combination therapy comprising: an oncolytic virus; an antibody or antigen-binding fragment thereof that specifically binds to programmed death 1 (PD-1); and a CTLA-4 inhibitor, comprising ipilimumab or a biological equivalent thereof.

44. A kit comprising an oncolytic virus and an antibody or antigen-binding fragment thereof that specifically binds to programmed death 1 (PD-1), together with instructions describing the use of the combination of the oncolytic virus and a therapeutically effective amount of an anti-PD-1 antibody or antigen-binding fragment thereof to treat or inhibit the growth of a tumor in a patient.

45. A kit comprising an oncolytic virus, an antibody or antigen-binding fragment thereof that specifically binds to programmed death 1 (PD-1), and a CTLA-4 inhibitor, including ipilimumab or a biological equivalent thereof, together with instructions describing the use of the combination of the oncolytic virus, an anti-PD-1 antibody or antigen-binding fragment thereof, and a therapeutically effective amount of a CTLA-4 inhibitor to treat or inhibit the growth of a tumor in a patient.

Images