Methods of Treating Cancer and Other Conditions with Mu Opioid Receptor Antagonists

Abstract

Compositions and methods are provided that use a μ-opioid receptor (MOR) antagonist (e.g., axeroplan) alone or in combination with a checkpoint inhibitor (e.g., a PD-1 / PD-L1 pathway inhibitor (e.g., an antibody, e.g., pembrolizumab)) and / or a VEGF inhibitor (e.g., bevacizumab) to reduce angiogenesis or treat cancer. Also provided are kits that include a MOR antagonist (e.g., axeroplan) alone or in combination with a checkpoint inhibitor and / or a VEGF inhibitor to reduce angiogenesis or treat cancer.

Description

[Technical field]

[0001] (CROSS REFERENCE TO RELATED APPLICATIONS) This application claims the benefit of U.S. Provisional Application No. 63 / 318,465, filed March 10, 2022, the contents of which are incorporated herein by reference in their entirety.

[0002] FIELD OF THEINVENTION The present disclosure relates to the use of mu opioid receptor (MOR) antagonists, particularly axelopran, alone or in combination with PD-1 / PD-L1 pathway inhibitors and / or VEGF inhibitors, to treat angiogenesis and cancer. [Background technology]

[0003] Mu opioid receptor (MOR) antagonists are commonly used to treat opioid and alcohol dependence. They are also used to treat side effects associated with the treatment of mu opioid agonists, such as morphine. Some of these side effects include intestinal dysfunction, for example in cancer patients and postoperative ileus.

[0004] MOR antagonists also show other promising uses.For example, WO 2016 / 061531 discloses, in its abstract, "a method for preventing or treating tumor growth, tumor metastasis and / or abnormal proliferation of tumor cells in a subject, comprising administering a pharmaceutical composition comprising methylnaltrexone."MOR antagonists are widely used and, in light of WO 2016 / 061531, show relatively high resistance in patients, especially when compared with conventional chemotherapy, making MOR antagonists an attractive target for treating cancer.

[0005] There is a need for additional treatments against angiogenesis and cancer. Summary of the Invention

[0006] The present disclosure relates to the use of mu opioid receptor (MOR) antagonists, particularly axelopran, alone or in combination with PD-1 / PD-L1 pathway inhibitors and / or VEGF inhibitors, to treat angiogenesis and cancer.

[0007] One aspect of the disclosure relates to reducing and inhibiting angiogenesis through the use of MOR antagonists (e.g., Axeroplan). One embodiment relates to a method of reducing or inhibiting endothelial cell proliferation, comprising contacting an endothelial cell with a MOR antagonist. Another embodiment relates to a method of reducing angiogenesis in a patient, comprising administering a MOR antagonist to a patient in need of reduced angiogenesis. Yet another embodiment relates to a method of reducing angiogenesis in a cancer patient, comprising administering a MOR antagonist to the cancer patient.

[0008] Another aspect of the present disclosure relates to the use of MOR antagonists to treat cancer or to treat certain aspects of cancer.Accordingly, one embodiment relates to a method of reducing tumor growth in a cancer patient, comprising administering a MOR antagonist to the cancer patient.Another embodiment relates to a method of reducing metastasis in a cancer patient by administering a MOR antagonist to the cancer patient.

[0009] Yet another embodiment relates to a method of improving an immune response to a tumor comprising administering to a cancer patient a MOR antagonist. In certain embodiments, the method comprises infiltration of immune cells, such as NK cells, lymphocytes and / or monocytes / macrophages, into the tumor. For example, the method comprises administering to a patient a MOR antagonist, such as CD3 + , CD244 + and MMD + Including immune cell infiltration.

[0010] In certain embodiments, the present disclosure relates to a method for treating cancer by administering axeroplan, naloxegol, or a combination thereof to a cancer patient.In one embodiment, the method reduces angiogenesis, tumor growth and / or metastasis.Alternatively, the method inhibits angiogenesis, tumor growth and / or metastasis.In some embodiments, the cancer is melanoma, colon cancer, pancreatic cancer, or breast cancer.

[0011] The present disclosure also provides combination therapies using MOR antagonists in combination with checkpoint inhibitors and / or VEGF inhibitors.

[0012] Yet another embodiment of the present disclosure relates to a method of treating cancer, comprising administering to a cancer patient a MOR antagonist and a checkpoint inhibitor and / or a VEGF inhibitor, in which the MOR antagonist and the checkpoint inhibitor and / or the VEGF inhibitor synergistically treat the cancer.

[0013] Yet another embodiment of the present disclosure relates to a method for improving an immune response to a tumor, comprising administering to a cancer patient a MOR antagonist and a checkpoint inhibitor and / or a VEGF inhibitor. In certain embodiments, the method comprises infiltration of immune cells (e.g., NK cells, lymphocytes, and / or monocytes / macrophages) into the tumor. In certain embodiments, the method comprises infiltration of CD3 + , CD244 + and MMD + In some embodiments of the method, the MOR antagonist and the checkpoint inhibitor synergistically improve the immune response to the tumor.

[0014] In some embodiments, the methods and combination therapies disclosed herein are particularly useful for treating patients who are not receiving opioid therapy. In other embodiments, the methods and combination therapies disclosed herein are particularly useful for treating patients who are receiving opioid therapy.

[0015] In certain embodiments of the methods and combination treatments, the MOR antagonist comprises axeroplan, naloxegol, methylnaltrexone, or a combination thereof. In certain preferred embodiments, the MOR antagonist is axeroplan. In some embodiments, the MOR antagonist does not include (i.e., excludes) methylnaltrexone, naltrexone, and / or naloxone.

[0016] In some embodiments of the combination therapy, the checkpoint inhibitor is a PD-1 / PD-L1 pathway inhibitor. For example, the PD-1 / PD-L1 pathway inhibitor is an antibody, such as an antibody that binds to PD-1. In some embodiments, the antibody is a humanized antibody, such as pembrolizumab.

[0017] In another embodiment of the combination therapy, the VEGF inhibitor is an anti-VEGF antibody, for example, an anti-VEGF-A antibody.In one embodiment, the anti-VEGF antibody is a humanized antibody.In a particular embodiment, the VEGF inhibitor is bevacizumab.In a particular embodiment of the combination therapy, the VEGF inhibitor is injected intravenously.

[0018] Various routes of administration are used in the methods or combination treatments. In one embodiment, the methods or combination treatments include administering the MOR antagonist orally or subcutaneously.

[0019] The methods and combination treatments of the present disclosure include administering a pharmaceutical composition comprising a MOR antagonist and a pharma- ceutical acceptable carrier. In one embodiment, such a composition is formulated for modified release.

[0020] When used in combination therapy, the checkpoint inhibitor is administered simultaneously with, before, or after the MOR antagonist. Similarly, the VEGF antagonist is administered simultaneously with, before, or after the MOR antagonist. Additionally, the VEGF inhibitor is administered simultaneously with, before, or after the PD-1 / PD-L1 pathway inhibitor.

[0021] Yet another embodiment of the present disclosure relates to a method of reducing tumor size comprising contacting the tumor with axeroplan, naloxegol, or a combination thereof. The present disclosure also relates to kits comprising a MOR antagonist (e.g., axeroplan), a checkpoint inhibitor, and / or a VEGF inhibitor.

[0022] Other features and advantages of the present invention will be apparent from the following detailed description and examples. [Brief description of the drawings]

[0023] For the purpose of illustrating the invention, there is shown in the drawings certain embodiments of the invention. However, the invention is not limited to the precise arrangements and instrumentalities of the embodiments shown in the drawings.

[0024] [Figure 1] Figure 1A and Figure 1B show tube formation of HUVEC cells under increasing concentrations of MOR antagonists (0-1000 nM). Comparing axelopran to naloxegol and methylnaltrexone (MNTX).

[0025] [Diagram 2] Figures 2A and 2B show angiogenesis under increasing concentrations of the MOR antagonist (acceleroplan) (0, 0.1, 0.5, 1.0 mg / kg) in the chick CAM assay compared to docetaxel. Figure 2A shows images of CAM angiogenesis. Panels A)-D) of Figure 2B show graphs testing the effects of accceleroplan, bevacizumab and accceleroplan + bevacizumab on CAM angiogenesis.

[0026] [Diagram 3] Figures 3A-3C show schematic diagrams of three model systems used to evaluate the contribution of endogenous MOR activity to tumor progression: Figure 3A shows a schematic diagram of the melanoma model system; Figure 3B shows a schematic diagram of the breast cancer model system; and Figure 3C shows a schematic diagram of the colon cancer model system.

[0027] [Figure 4] Figures 4A-C show that zebrafish larvae survive up to 72 hours under increasing concentrations of acceleroplan in the incubation water. Figures 4A and 4B show graphs measuring survival as a function of time. Figure 4C shows photographs of zebrafish larvae exposed to vehicle, 100 μM acceleroplan, and 500 μM acceleroplan at 24, 48, and 72 hours. Doses up to 100 μM showed survival and pathological lesion development at levels comparable to vehicle controls.

[0028] [Diagram 5] FIG. 5 shows a schematic of the time course of zebrafish larval development.

[0029] [Figure 6] Figures 6A and 6B show representative plots of melanoma tumor growth size (y-axis) (normalized as % of vehicle control) versus number of metastases (x-axis) in zebrafish treated with 10 μM Acceleroplan (n=18) (Figure 6A) or vehicle control (n=32) (Figure 6B).

[0030] [Figure 7] Figure 7 shows an overview of melanoma tumor progression (tumor size x number of metastases) in the presence of increasing concentrations of Acceroplan or the checkpoint inhibitor pembrolizumab. In experiments with pembrolizumab, exogenous tumor infiltrating lymphocytes (TILs) were injected. Further controls included the effect of TILs alone and in combination with Acceroplan (500 nM) and pembrolizumab, and TILs alone and in combination with Acceroplan (500 nM). *=<0.05, **=<0.005, ***=<0.0005.

[0031] [Figure 8] FIG. 8 shows mortality of zebrafish larvae over a 72 hour period following exposure to test substances at 48 hours post-fertilization.

[0032] [Figure 9] FIG. 9 shows an overview of tumor progression in the CAM assay.

[0033] [Figure 10A] Figure 10A shows a representative two-dimensional plot of breast cancer tumor growth size (normalized as % of vehicle control) versus number of metastases (normalized as % of vehicle control) in the CAM assay with the following treatments: negative control = vehicle (n = 15), A = 1.0 mg / kg axelopran (n = 13), P = pembrolizumab (2.5 mg / kg), B = bevacizumab (4 mg / kg) or relevant combination. Circles indicate the mean ± standard deviation for both dimensions.

[0034] [Figure 10B] FIG. 10B shows tumor weight x metastases for pembrolizumab, axeroplan, bevacizumab, axeroplan + bevacizumab, pembrolizumab + bevacizumab, and axeroplan + pembrolizumab + bevacizumab.

[0035] [Figure 11] FIG. 11 shows infiltration of chicken CD3+, CD244+ and MMD+ immune cells into breast cancer tumors grown on the upper CAM in the presence of axeroplan, pembrolizumab, bevacizumab and their combinations.

[0036] [Figure 12] Figures 12A and 12B show tumor growth of MC38 colon carcinoma in syngenic mice treated with vehicle (control), Acceleropran (1 mg / kg, orally, daily), mouse anti-PD-1 (12.5 mg / kg, intraperitoneally, three times a week), or Acceleropran + anti-PD-1. Figure 12A shows tumor size (mm3) as a function of days of treatment. Figure 12B shows individual tumor size distribution on day 13.

[0037] [Figure 13]FIG. 13 shows Kaplan-Meier survival curves of mice injected with human colon cancer MC-38 cells and treated with Accelerome (1 mg / kg), anti-PD-1 (2.5 mg / kg) or both drugs.

[0038] [Figure 14] Figures 14A and 14B show inhibition of tumor progression.

[0039] FIG. 14C shows the distribution of tumor volumes. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0040] The general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, which is defined in the appended claims. Other aspects of the invention will be apparent to those skilled in the art from a consideration of the detailed description of the invention provided herein.

[0041] The present disclosure is based on the discovery that MOR antagonists, particularly axeroplan, can treat angiogenesis and cancer. The present disclosure is also based on the surprising discovery that certain MOR antagonists (e.g., axeroplan) act synergistically when used in combination with PD-1 / PD-L1 pathway inhibitors and / or VEGF inhibitors to treat angiogenesis or cancer.

[0042] (definition) Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention pertains. Although methods and materials similar or equivalent to those described herein can be used to carry out the testing of the present invention, the preferred materials and methods are described herein. In describing and claiming the present invention, the following terms are used:

[0043] It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0044] As used herein, the term "MOR antagonist" refers to a μ opioid receptor antagonist.

[0045] As used herein, the term "acceloplan" refers to the compound having the structure shown below: [ka] and also known as 3-[(1R,3r,5S)-8-(2-{(cyclohexylmethyl)[(2S)-2,3-dihydroxypropanoyl]amino}ethyl)-8-azabicyclo[3.2.1]octan-3-yl]benzamide or 3-[(3-endo)-8-(2-{(cyclohexylmethyl)[(2S)-2,3-dihydroxypropanoyl]amino}ethyl)-8-azabicyclo[3.2.1]oct-3-yl]benzamide. The term axeropran also includes pharma- ceutically acceptable salts of axeropran and stereoisomers of axeropran.

[0046] Furthermore, the term "acceloplan" also includes derivatives of axeloplan. Examples of suitable derivatives of axeloplan are disclosed in U.S. Pat. No. 7,622,508, the disclosure of which is incorporated herein as it relates to axeloplan derivatives. U.S. Pat. No. 7,622,508 discloses a compound of formula (I): [ka] Compounds having a core structure shown in

[0047] The term "salt thereof" refers to a compound formed by replacing the proton of an acid with a cation, such as a metal cation or an organic cation. Preferably, the salt is a pharma- ceutically acceptable salt. By way of example, salts of the compounds of the present invention include those that are protonated with an inorganic or organic acid to form a cation, with the conjugate base of the inorganic or organic acid being the anion component of the salt. Salts of interest include, but are not limited to, aluminum, ammonium, arginine, barium, benzathine, calcium, cesium, cholinate, ethylenediamine, lithium, magnesium, meglumine, procaine, N-methylglucamine, piperazine, potassium, sodium, tromethamine, zinc, N,N'-dibenzylethylene-diamine, chloroprocaine, diethanolamine, ethanolamine, piperazine, diisopropylamine, diisopropylethylamine, triethylamine, and triethanolamine salts.

[0048] "Stereoisomer" refers to a compound in which the atomic bonds are the same but the atomic arrangement in space is different. Stereoisomers include, for example, cis-trans isomers, E and Z isomers, enantiomers and diastereomers. For any group that contains one or more substituents disclosed herein, it is of course understood that such groups do not include sterically impractical and / or synthetically unfeasible substitutions or substitution patterns. All stereoisomers are intended to be included within the scope of this disclosure.

[0049] As used herein, the articles "a" and "an" are used to refer to one or to more than one (at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

[0050] As used herein, when referring to a measurable value, such as an amount, time period, etc., the term "about" is meant to encompass variations of ±20% or ±10%, more preferably ±5%, even more preferably ±1%, and even more preferably ±0.1% from the given value, as such variations are appropriate for carrying out the disclosed methods. One of skill in the art can select an appropriate variation based on the context of the value.

[0051] The term "biological" or "biological sample" refers to a sample obtained from an organism, or from components (e.g., cells) of an organism. A biological sample may be obtained from tumor cells or tumor tissue. A sample may be any biological tissue or body fluid. Often, a sample is a "clinical sample" that is a sample derived from a patient. Such samples include, but are not limited to, bone marrow, cardiac tissue, sputum, blood, lymph, blood cells (e.g., white blood cells), tissue or fine needle biopsy samples, urine, peritoneal and pleural fluids, or cells therefrom. A biological sample may also include tissue sections, such as frozen sections taken for histological purposes.

[0052] As used herein, the terms "comprising," "including," "containing," "having," and "characterized by" are interchangeable, inclusive, and open-ended and do not exclude additional, unrecited elements or method steps. When the term "comprising" appears herein, particularly in describing a component of a composition or in describing an element of a device, it is understood to encompass compositions and methods that consist essentially of and consist of the recited components or elements.

[0053] As used herein, the term "consisting of" excludes any element, step, or ingredient not specifically recited in the claim element.

[0054] As used herein, "combination therapy" means that a first drug is administered in combination with another drug. "In combination with" or "In conjunction with" refers to administering one treatment method in addition to another treatment method. Thus, "combination" refers to administering one treatment method to an individual before, during or after the administration of the other treatment method. Such a combination is considered to be a part of one treatment regimen or regimen. For the purposes of this specification, combination therapy can include treatment regimens that include administering a MOR antagonist and a checkpoint inhibitor, respectively, to treat the same disease or condition, such as the same tumor or cancer. Combination therapy can include treatment regimens that include administering a MOR antagonist and a checkpoint inhibitor and / or a VEGF antagonist, respectively, to treat the same disease or condition, such as the same tumor or cancer.

[0055] The term "treatment" as used in the context of the present invention is meant to include therapeutic treatment as well as preventive or suppressive measures of a disease or disorder. As used herein, the term "treatment" and related terms, such as "treat" and "treating", refer to the alleviation of the progression, severity and / or duration of a disease state or at least one of its symptoms. Thus, the term "treatment" refers to any regimen that can benefit a subject. Treatment may be for an existing condition or may be preventive (prophylactic treatment). Treatment may include a curative effect, a palliative effect or a preventive effect. References herein to "therapeutic" and "preventive" treatment are considered in the broadest context. The term "therapeutic" does not necessarily mean that a subject is treated until it is completely cured. Similarly, "prophylactic" does not necessarily mean that a subject will not eventually suffer from a disease state. Thus, for example, the term treatment includes administering a drug before or after the onset of a disease or disorder, thereby preventing or eliminating all signs of a disease or disorder. As another example, administration of a drug after clinical manifestation of a disease to combat the symptoms of the disease constitutes "treatment" of the disease.

[0056] The term "pharmaceutical composition" as used herein includes a mixture of at least one compound useful in the present invention with other chemical components, such as carriers, stabilizers, diluents, adjuvants, dispersants, suspending agents, thickeners and / or additives. Pharmaceutical compositions facilitate the administration of a compound to an organism. There are multiple techniques of administering a compound in the art, including but not limited to intratumoral, intravenous, intrapleural, oral, aerosol, parenteral, intraocular, intrapulmonary and topical administration.

[0057] The term "pharmaceutical acceptable carrier" includes pharmaceutical acceptable salts, pharmaceutical acceptable materials, compositions or carriers, such as liquid or solid excipients, diluents, additives, solvents or encapsulating materials, that are involved in carrying or transporting the compounds of the present invention (e.g., MOR antagonists and / or checkpoint inhibitors and / or VEGF antagonists) in or to a subject so that they can perform their 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 components of the formulation and not harmful to the subject. Some examples of substances which may function 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; additives, 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, sorb ... Examples of suitable carriers include ethanol 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; diluents; granulating agents; lubricants; binders; disintegrating agents; wetting agents; emulsifying agents; coloring agents; release agents; coating agents; sweeteners; flavoring agents; flavorings; preservatives; antioxidants; plasticizers; gelling agents; thickening agents; hardening agents; solidifying agents; suspending agents; surfactants; wetting agents; carriers; stabilizers; and other non-toxic compatible substances used in pharmaceutical preparations, or any combination thereof. As used herein, "pharmaceutical acceptable carriers" also include any coatings, antibacterial agents, antifungal agents, and absorption retardants that are compatible with the activity of the compound and physiologically acceptable to the subject. Supplementary active substances can also be incorporated into the composition.

[0058] As used herein, the term "effective amount" or "therapeutically effective amount" refers to the amount of a MOR antagonist required to prevent a particular disease state or to reduce and / or ameliorate the severity of a disease state or at least one symptom thereof or a condition associated therewith.

[0059] As used herein, a "subject" or "patient" can be a human or non-human mammal. Non-human mammals include, for example, farm animals and pets, such as sheep, cows, pigs, dogs, cats and marine mammals. Preferably, the subject is a human.

[0060] Ranges: Throughout this disclosure, various aspects of the invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as a burdensome limitation on the scope of the invention. Thus, the description of a range should be considered to have specifically disclosed all possible subranges as well as each individual numerical value within that range. For example, the description of a range such as 1-6 should be considered to have specifically disclosed subranges such as 1-3, 1-4, 1-5, 2-4, 2-6, 3-6, etc., and each individual numerical value within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.

[0061] Therapeutic Uses of MOR Antagonists The present disclosure provides new therapeutic uses for μ-opioid receptor (MOR) antagonists. As the following examples show, MOR antagonists can inhibit angiogenesis in models of angiogenesis that are used as models for a variety of conditions, such as: In addition, based on the studies, MOR antagonists alone or in combination with other therapeutic agents can treat cancer or related conditions.

[0062] Thus, one embodiment of the present disclosure relates to a method of reducing or inhibiting endothelial cell proliferation, comprising contacting an endothelial cell with a MOR antagonist. In one embodiment, the method inhibits endothelial cell proliferation. In another embodiment, the method reduces endothelial cell proliferation.

[0063] Another embodiment of the present disclosure relates to a method for reducing angiogenesis in a patient, comprising administering a MOR antagonist to a patient in need of reducing angiogenesis.In one embodiment, the method reduces angiogenesis.In another embodiment, the method inhibits angiogenesis.In one embodiment, the method reduces or inhibits ocular angiogenesis.In another embodiment, the patient has psoriasis.

[0064] Another embodiment of the present disclosure relates to a method of reducing angiogenesis in a patient suffering from an angiogenesis-associated disease, comprising administering a MOR antagonist to a patient in need of reduced angiogenesis. As used herein, an "angiogenesis-associated disease" is a disease associated with abnormal angiogenesis. Examples of suitable angiogenesis-associated diseases include, but are not limited to, skin psoriasis, rheumatoid arthritis, neurodegenerative diseases, neuropathic pain, hemangiomas, collagen synthesis diseases such as Ehlers-Danlos syndrome and portal hypertension. Further examples of angiogenesis-associated diseases include retinopathies (multiple conditions including diabetic, wet AMD, sickle cell retinopathy), psoriasis, rheumatoid arthritis, hemangiomas, hereditary hemorrhagic telangiectasia, von Hippel-Lindau disease, uncontrolled vascular remodeling in ischemic diseases (including myocardial infarction, etc.), chronic kidney disease and arteriovenous fistulas.

[0065] Further embodiments of the present disclosure relate to methods of treating angiogenesis-associated diseases, comprising administering a MOR antagonist to a patient suffering from an angiogenesis-associated disease. Examples of such diseases are the angiogenesis-associated diseases mentioned above.

[0066] Yet another embodiment of the present disclosure relates to a method of reducing angiogenesis in a cancer patient, comprising administering to the cancer patient a MOR antagonist. In one embodiment, the method reduces angiogenesis. In another embodiment, the method inhibits angiogenesis.

[0067] Another embodiment of the present disclosure relates to a method of reducing tumor growth in a cancer patient comprising administering to the cancer patient a MOR antagonist, the method reducing or inhibiting tumor growth.

[0068] Yet another embodiment of the present disclosure relates to a method of reducing metastasis in a cancer patient, comprising administering to the cancer patient a MOR antagonist. In one embodiment, the method inhibits metastasis. In another embodiment, the method reduces metastasis.

[0069] Another embodiment of the present disclosure relates to a method for improving an immune response to a tumor, comprising administering a MOR antagonist to a cancer patient. In one embodiment, the method increases the infiltration of immune cells into the tumor. In another embodiment, the method increases the infiltration of NK cells, lymphocytes and / or monocytes / macrophages. For example, the method increases the infiltration of CD3 + , CD244 + and MMD + Immune cells, e.g. CD3 + Lymphocytes, CD244 + Natural Killer Cells and MMD + Increases macrophage infiltration.

[0070] The method of the present disclosure can be used in various cancers and tumors. In certain embodiments, the cancer is melanoma, colon cancer or breast cancer. In other embodiments, the tumor is a tumor in a patient who also suffers from melanoma, pancreatic cancer, colon cancer or breast cancer.

[0071] In a preferred embodiment of the method, the patient is not receiving opioid treatment. Thus, in a preferred embodiment, the method does not include administration of an opioid. In another embodiment, the patient is receiving opioid treatment.

[0072] A variety of MOR antagonists can be used in the methods of the present disclosure. For example, in one embodiment, the μ receptor opioid (MOR) antagonist is axelopran, naloxegol, naldemedine, alvimopan, or a combination thereof. [ka] In another embodiment, the MOR antagonist is axelopran, naloxegol, or a combination thereof. In a preferred embodiment, the MOR antagonist is axelopran. In a particular embodiment, the MOR antagonist is a peripheral MOR antagonist. In one embodiment, the MOR antagonist does not include methylnaltrexone. In another embodiment, the MOR antagonist is a peripheral MOR antagonist other than methylnaltrexone. In another embodiment, the MOR antagonist does not include methylnaltrexone, naltrexone and / or naloxone.

[0073] Yet another aspect of the present disclosure relates to a method for treating cancer, comprising administering axeroplan, naloxegol, or a combination thereof to a cancer patient. In one embodiment, the method comprises administering axeroplan. In certain embodiments, the method reduces angiogenesis, tumor growth, and / or metastasis. In other embodiments, the method inhibits angiogenesis, tumor growth, and / or metastasis. In other embodiments, the method improves the immune response to cancer. In another embodiment, the method excludes or does not include administration of exogenous opioids. Thus, in some embodiments, the patient is not receiving opioid treatment. These methods can be used to treat various cancers. In some embodiments, the cancer is melanoma, colon cancer, pancreatic cancer, or breast cancer.

[0074] In certain embodiments, the method requires administering a pharmaceutical composition comprising a MOR antagonist (e.g., axelopran). Examples of suitable pharmaceutical compositions are described below. In one embodiment, the pharmaceutical composition is formulated for controlled release. Alternatively, the pharmaceutical composition is formulated for controlled release, local administration or intravitreal injection. For example, the pharmaceutical composition is formulated for "controlled release" when it is formulated for rapid / accelerated release, site-specific release, controlled / pulsatile release, delayed release into the lower gastrointestinal tract or sustained release.

[0075] The method includes administration of the MOR antagonist (e.g., axeroplan) by various routes, including any of the administration routes described below. In one embodiment, the method includes orally administering the MOR antagonist. In another embodiment, the method includes subcutaneously administering the MOR antagonist. In yet another embodiment, the method includes intravitreally administering the MOR antagonist. In another embodiment, the method includes topically administering the MOR antagonist.

[0076] In further embodiments, the methods include screening a patient for the presence of cancer. These embodiments include obtaining a biological sample from the patient and testing the sample for the presence of cancer cells.

[0077] Yet another embodiment of the present disclosure relates to a method of reducing tumor size comprising contacting the tumor with axeroplan, naloxegol, or a combination thereof, hi one embodiment, the method comprises contacting the tumor with axeroplan.

[0078] Other embodiments of the present disclosure relate to the use of MOR antagonists (e.g., axeroplan, naloxegol, or combinations thereof) in the manufacture of a medicament for treating cancer. Further embodiments relate to the use of MOR antagonists (e.g., axeroplan, naloxegol, or combinations thereof) for treating cancer.

[0079] (Combination treatment) The present disclosure also provides combination therapy, including using MOR antagonists in combination with checkpoint inhibitors and / or VEGF antagonists.In some embodiments, the combination therapy is used to treat angiogenesis or cancer.

[0080] When two or three drugs are administered as part of the same therapeutic course, these drugs are administered to a subject in "combination." A therapeutic course refers to the administration of a combination of drugs that a medical professional believes act additively, complementary, synergistically or in other ways to produce a more favorable result than would be expected from administration of a single drug. A therapeutic course can last one or several days, but often lasts for several weeks.

[0081] When two drugs are administered in combination, various schedules can be used.In one example, for example, but not limited to, drug 1 (e.g., MOR antagonist) is first administered before drug 2 (e.g., checkpoint inhibitor and / or VEGF antagonist) is administered, and drug 1 treatment is continued throughout the course of drug 2 administration; alternatively, drug 1 is administered after drug 2 therapy is started or completed; alternatively, drug 1 is first administered simultaneously with the start of other cancer therapy.In this context, "concurrently" means that two drugs are administered on the same day or on consecutive days.

[0082] In principle, certain drugs may be formulated together, but are generally administered in separate compositions. Similarly, certain drugs may be administered simultaneously, but in many cases (particularly in the case of drugs administered by infusion), the drugs will be administered at different times on the same day, on consecutive days, or according to another schedule.

[0083] Thus, one embodiment of the present disclosure relates to a method of treating cancer, comprising administering to a cancer patient a MOR antagonist and a checkpoint inhibitor and / or a VEGF inhibitor, such as a combination of a MOR antagonist and a checkpoint inhibitor and / or a VEGF inhibitor, such as bevacizumab. In a preferred embodiment of these methods, the MOR antagonist and the checkpoint inhibitor and / or the VEGF inhibitor synergistically treat the cancer. In one embodiment, the MOR antagonist and the checkpoint inhibitor (anti-PD1 antibody) synergistically treat the cancer. One particular embodiment is a method of treating cancer with a synergistic combination of axeroplan and pembrolizumab (anti-PD1 antibody). Another particular embodiment is a method of treating cancer with a synergistic combination of axeroplan and bevacizumab (anti-VEGF antibody) and pembrolizumab (anti-PD1 antibody). In other embodiments, the method comprises administering a MOR antagonist in combination with a checkpoint inhibitor. In another embodiment, the method comprises administering a MOR antagonist in combination with a VEGF antagonist.

[0084] Another embodiment of the present disclosure relates to a method for improving immune response to a tumor, comprising administering a MOR antagonist and a checkpoint inhibitor and / or a VEGF inhibitor to a cancer patient. In some embodiments, the method comprises administering a MOR antagonist in combination with a checkpoint inhibitor. In other embodiments, the method comprises administering a MOR antagonist in combination with a VEGF antagonist. In certain embodiments, the method increases the infiltration of immune cells into the tumor. In certain embodiments, the method increases the infiltration of NK cells, lymphocytes and / or monocytes / macrophages. Alternatively, the method increases the infiltration of CD3 + , CD244 + and MMD + Increases immune cell infiltration. In certain embodiments, the MOR antagonist and the checkpoint inhibitor synergistically improve the immune response to tumors.

[0085] A variety of MOR antagonists can be used. In some embodiments, the MOR antagonist comprises axeroprand, naloxegol, methylnaltrexone, or a combination thereof. In other embodiments, the MOR antagonist is axeroprand.

[0086] In certain embodiments of the combination treatment, the MOR antagonist does not include methylnaltrexone and / or naloxone.

[0087] Various checkpoint inhibitors may be used in combination with MOR antagonists, such as axeroplan. In certain embodiments, the checkpoint inhibitor is a PD-1 / PD-L1 pathway inhibitor, such as an antibody. In other embodiments, the checkpoint inhibitor is an inhibitor of CTLA-4; LAG-3; TIM-3; B7-H3; B7-H4; A2aR; NKG21; PVRIG; PVRL2; CEACAM 1; CEACAM 5; CEACAM 6; FAK; CCL2 / CCR2; LIF; CD45; CSF-1; SEMA4D; CLEVER-1; OX-40; IL-1; IL-6; or IL-8. In certain embodiments, the checkpoint inhibitor is an antibody. Other examples of suitable checkpoint inhibitors are disclosed in Marin-Acevedo et al. J Hematol Oncol (2021) 14:45 (incorporated herein as relating to checkpoint inhibitors). In some embodiments, the checkpoint inhibitor is an antibody, e.g., a humanized antibody, that binds to PD-1. In one embodiment, the checkpoint inhibitor is pembrolizumab.

[0088] In some embodiments of the combination therapy, the method comprises administering the MOR antagonist orally or subcutaneously. In other embodiments, the method comprises administering a pharmaceutical composition comprising the MOR antagonist described herein and a pharma- ceutical acceptable carrier. For example, the method comprises administering a pharmaceutical composition formulated for modulated release. For example, a pharmaceutical composition is formulated for "modulated release" if it is formulated for rapid / accelerated release, site-specific release, controlled / pulsatile release, delayed release into the lower gastrointestinal tract or sustained release. When used, the checkpoint inhibitor is administered simultaneously with, before or after the MOR antagonist (e.g., axeroplan).

[0089] In certain embodiments, the method comprises administering an angiogenesis inhibitor. The angiogenesis inhibitor can be used alone or in combination with a checkpoint inhibitor. In some embodiments, the angiogenesis inhibitor is a VEGF inhibitor. The VEGF inhibitor can be used alone or in combination with a checkpoint inhibitor (e.g., an antibody that binds to PD-1, e.g., pembrolizumab). In some embodiments, the VEGF inhibitor is an anti-VEGF antibody, e.g., an anti-VEGF-A antibody. In some embodiments, the VEGF inhibitor is a humanized antibody. The antibody is an anti-VEGF-A antibody. In other embodiments, the VEGF inhibitor is bevacizumab. Other suitable VEGF inhibitors include, but are not limited to, Ziv-aflibercept, aflibercept, ranibizumab, pegaptanib or faricimab. In some embodiments of the method (combination therapy), the VEGF inhibitor is intravenously injected. When used, the VEGF inhibitor is administered simultaneously with, before or after the MOR antagonist. In other embodiments, when used in combination with a checkpoint inhibitor, the VEGF inhibitor is administered simultaneously with, before or after the checkpoint inhibitor, e.g., a PD-1 / PD-L1 pathway inhibitor.

[0090] In some embodiments, the present disclosure relates to the use of the above MOR antagonist and the above PD-1 / PD-L1 pathway inhibitor in the manufacture of a medicament for treating cancer.The present disclosure also relates to the use of the above MOR antagonist and the above PD-1 / PD-L1 pathway inhibitor in the manufacture of a kit for treating cancer.Furthermore, the present disclosure relates to the use of the above MOR antagonist and the above PD-1 / PD-L1 pathway inhibitor for treating cancer.

[0091] In further embodiments, the present disclosure relates to the use of the above MOR antagonist and the above VEGF inhibitor in the manufacture of a medicament for treating cancer. Alternatively, the present disclosure relates to the use of the above MOR antagonist and the above VEGF inhibitor in the manufacture of a medicament for treating cancer. Also, the present disclosure relates to the use of the above MOR antagonist and the above VEGF inhibitor in the manufacture of a medicament for treating cancer. Furthermore, the present disclosure relates to the use of the above MOR antagonist and the above VEGF inhibitor in the manufacture of a kit for treating cancer.

[0092] The present disclosure also relates to the use of the above MOR antagonist, the above PD-1 / PD-L1 pathway inhibitor, and the above VEGF inhibitor in the manufacture of a medicament for treating cancer.The present disclosure further relates to the use of the above MOR antagonist, the above PD-1 / PD-L1 pathway inhibitor, and the above VEGF inhibitor for treating cancer.

[0093] Also, in one embodiment, the present disclosure relates to a combination comprising the above-mentioned MOR antagonist (e.g., axeroplan) and the above-mentioned PD-1 / PD-L1 pathway inhibitor (e.g., pembrolizumab) for use in a method for treating cancer, the method comprising administering the combination to a subject in need of treatment. In certain embodiments, the combination also comprises the above-mentioned VEGF inhibitor (e.g., bevacizumab).

[0094] In certain embodiments of the above methods and uses, the MOR antagonist is axeroplan, and the checkpoint inhibitor is an anti-PD1 antibody, such as pembrolizumab.In another embodiment of the above methods and uses, axeroplan and pembrolizumab act synergistically.In another embodiment of the above methods and uses, the MOR antagonist is axeroplan, the checkpoint inhibitor is an anti-PD1 antibody, such as pembrolizumab, and the VEGF inhibitor is bevacizumab.In a further embodiment of the above methods and uses, axeroplan, pembrolizumab and bevacizumab act synergistically.

[0095] (Compositions and pharmaceutical compositions) One embodiment is a composition comprising a synergistic amount of a MOR antagonist and a PD-1 / PD-L1 pathway inhibitor. In certain embodiments of the composition, the MOR antagonist comprises axeroplan, naloxegol, methylnaltrexone, or a combination thereof. In one embodiment, the MOR antagonist is axeroplan. The PD-1 / PD-L1 pathway inhibitor is an antibody, such as an antibody that binds to PD-1 (e.g., pembrolizumab). The antibody may be humanized.

[0096] In some embodiments, the present invention relates to pharmaceutical compositions comprising a MOR antagonist configured for use in the methods described herein. In one preferred embodiment, the MOR antagonist is axelopran.

[0097] In a particular embodiment, the invention relates to a pharmaceutical composition comprising a MOR antagonist and a checkpoint inhibitor and / or a VEGF antagonist. In another embodiment, the invention relates to a pharmaceutical composition comprising a MOR antagonist and another pharmaceutical composition comprising a checkpoint inhibitor and / or a VEGF antagonist.

[0098] Provided is a pharmaceutical composition for treating cancer in a subject in need of treatment.The pharmaceutical composition comprises MOR antagonist and pharma- ceutical acceptable carrier.In certain embodiments, the pharmaceutical composition comprises checkpoint inhibitor and / or VEGF antagonist.

[0099] In one embodiment, the pharmaceutical composition comprises axeroplan, pembrolizumab, and / or bevacizumab. In another embodiment, the pharmaceutical composition comprises axeroplan and pembrolizumab.

[0100] Such pharmaceutical compositions are in a form suitable for administration to a subject, or the pharmaceutical compositions may further comprise one or more pharma- ceutically acceptable carriers, one or more additional components, or some combination thereof.Various components of the pharmaceutical compositions may be present in a physiologically acceptable salt form, e.g., in combination with a biologically acceptable cation or anion, as is well known in the art.

[0101] Pharmaceutical compositions useful in the methods of the present invention may be suitably developed for inhalation, oral, rectal, vaginal, parenteral, topical, transdermal, pulmonary, intranasal, buccal, ocular, intrathecal, intravenous, or other routes of administration. Other contemplated formulations include projected nanoparticles, liposomal formulations, resealed erythrocytes containing the active ingredient, immunologically-based formulations. The route of administration is readily apparent to those skilled in the art and depends on a variety of factors, including the type and severity of the disease being treated, the type and age of the veterinary or human patient being treated, and the like.

[0102] The formulations of the pharmaceutical compositions described herein can be prepared by any method known or hereafter developed in the field of pharmacology. In general, such preparation methods include the step of combining the active ingredient with the carrier or one or more other accessory ingredients, and then, if necessary or desired, shaping or packaging the product into the desired single or multiple dosage unit.

[0103] The amount of active ingredient is generally equal to the dosage of active ingredient administered to a subject, or a convenient fraction of such a dosage, such as one-half or one-third of such a dosage. The unit dosage form can be for a single daily administration, or for multiple daily administrations (e.g., about 1 to 4 or more times per day). When multiple daily administrations are used, the unit dosage form can be the same or different for each administration.

[0104] The description of pharmaceutical compositions provided herein is primarily directed to pharmaceutical compositions suitable for ethical administration to humans, but those skilled in the art will understand that such compositions are generally suitable for administration to any type of animal. Modifications of pharmaceutical compositions suitable for administration to humans to make them suitable for administration to various animals are well understood, and an ordinary skilled veterinary pharmacologist can design and implement such modifications with no more than routine experimentation, if any. Subjects to which the pharmaceutical compositions of the present invention are intended to be administered include, but are not limited to, mammals, including commercially relevant mammals, including humans and other primates, cows, pigs, horses, sheep, cats, and dogs. In one embodiment, the subject is a human or non-human mammal, such as, but not limited to, horses, sheep, cows, pigs, dogs, cats, and mice. In one embodiment, the subject is a human.

[0105] In one embodiment, the composition is formulated with one or more pharma- ceutical acceptable additives or carriers. In one embodiment, the pharmaceutical composition is disclosed for treating cancer in a subject. The pharmaceutical composition comprises a MOR antagonist. In another embodiment, the pharmaceutical composition also comprises a checkpoint inhibitor, a VEGF antagonist, or a combination thereof. Useful pharma-ceutical acceptable carriers include, but are not limited to, glycerol, water, saline, ethanol, and other pharma-ceutical acceptable salt solutions, such as phosphates and other organic acid salts. The carrier can be, for example, a solvent or dispersion medium, including water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, etc.), suitable mixtures thereof, and vegetable oils. The proper fluidity can be maintained, for example, by using a coating such as lecithin, maintaining the required particle size in the case of dispersion, and using surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it is preferable to include isotonic agents in the composition, for example, sugars, sodium chloride, or polyalcohols, such as mannitol and sorbitol. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate or gelatin.

[0106] The preparations may be used in admixture with conventional additives, i.e., pharma- ceutically acceptable organic or inorganic carrier substances suitable for oral, parenteral, nasal, intravenous, subcutaneous, enteral, or other modes of administration well known in the art. The pharmaceutical preparations may be sterilized and, if desired, adjuvants, such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic buffers, colorants, flavors, and / or aromatic substances, etc. Also, if desired, they may be combined with other active substances, such as other analgesics.

[0107] The composition may contain about 0.005%-2.0% preservative by total weight of the composition. Preservatives are used to prevent spoilage when exposed to contaminating bacteria in the environment. Examples of preservatives useful according to the present invention include, but are not limited to, those selected from the group consisting of benzyl alcohol, sorbic acid, parabens, imidurea, and combinations thereof. A particularly preferred preservative is a combination of about 0.5%-2.0% benzyl alcohol and 0.05%-0.5% sorbic acid.

[0108] The composition may include antioxidants and chelating agents that prevent degradation of the compounds. Preferred antioxidants for some compounds are BHT, BHA, α-tocopherol and ascorbic acid in the preferred range of about 0.01%-0.3%, more preferably BHT in the range of 0.03%-0.1% by weight based on the total weight of the composition. Preferably, the chelating agent is present in an amount of 0.01%-0.5% by weight based on the total weight of the composition. Particularly preferred chelating agents include edetate (e.g., disodium edetate) and citric acid in the range of about 0.01%-0.20% by weight based on the total weight of the composition, more preferably in the range of 0.02%-0.10% by weight based on the total weight of the composition. Chelating agents are useful for chelating metal ions in the composition that may adversely affect the shelf life of the formulation. BHT and disodium edetate are particularly preferred antioxidants and chelating agents, respectively, for some compounds, but may be substituted with other suitable and equivalent antioxidants and chelating agents as known to those skilled in the art.

[0109] The pharmaceutical compositions described herein may be used in combination with additional therapeutic agents, such as antitumor agents, including, but not limited to, chemotherapeutic agents, anti-cell proliferation agents, or any combination thereof. For example, any conventional chemotherapeutic agent from the non-limiting exemplary classes of alkylating agents, nitrosoureas, antimetabolites, antitumor antibiotics, plant alkyloids, taxanes, hormonal agents, and other drugs are included in the present invention. In another aspect, the pharmaceutical compositions described herein may be used in combination with radiation therapy.

[0110] (Administration) In certain embodiments of the present invention, MOR antagonist and checkpoint inhibitor and / or VEGF antagonist are administered simultaneously.In other embodiments, checkpoint inhibitor and / or VEGF antagonist are administered before MOR antagonist.In another embodiment, checkpoint inhibitor and / or VEGF antagonist are administered after MOR antagonist.

[0111] Dosage regimen can affect what constitutes an effective amount.For example, the therapeutic formulation can be administered to the patient subject before or after a surgical intervention related to cancer, or immediately after the patient is diagnosed with cancer.Also, several divided doses and staggered doses can be administered daily or continuously, or the dose can be continuously infused or bolus injected.Furthermore, the dosage of the therapeutic formulation can be proportionally increased or decreased according to the exigencies of the therapeutic or prophylactic situation.

[0112] The volume of the composition can be any volume and can be for single or multiple doses, including, but not limited to, the ranges of from or about 0.01 mL to 100 mL, 0.1 mL to 100 mL, 1 mL to 100 mL, 10 mL to 100 mL, 0.01 mL to 10 mL, 0.1 mL to 10 mL, 1 mL to 10 mL, 0.02 mL to 20 mL, 0.05 mL to 5 mL, 0.5 mL to 50 mL, or 0.5 mL to 5 mL (each inclusive).

[0113] Administration of the compositions of the present invention to a patient subject, preferably a mammal, more preferably a human, may be carried out using known procedures at dosages and for periods of time effective to treat cancer in the subject. The effective amount of the therapeutic compound required to achieve a therapeutic effect may vary depending on factors such as: the activity of the particular compound used; the time of administration; the rate of excretion of the compound; the duration of treatment; other drugs, compounds or materials used in combination with the compound; the state of the disease or disorder, age, sex, weight, condition, general health and previous medical history of the patient being treated, and similar factors well known in the medical field. The administration regimen may be adjusted to obtain the optimal therapeutic effect. For example, the composition may be administered in several divided doses daily, or the dosage may be proportionally reduced depending on the exigencies of the therapeutic situation. A non-limiting example of an effective dose range of the therapeutic compound is about 0.01 to about 50 mg / kg body weight / day.

[0114] MOR antagonist and checkpoint inhibitor and / or VEGF receptor antagonist may be administered to a subject several times a day, or less frequently, for example, once a day, once a week, once every two weeks, once a month, or even less frequently, for example, once every few months, or once a year or less. It is understood that the amount of compound administered daily can be administered, in non-limiting examples, every day, every other day, every third day, every fourth day, or every fifth day. For example, every other day administration can start with a 5 mg / day dose on Monday, followed by a first 5 mg / day dose on Wednesday, a second 5 mg / day dose on Friday, and so on. The frequency of administration is readily apparent to those skilled in the art and depends on a number of factors, including, but not limited to, the type and severity of the disease being treated, and the type and age of the animal. The actual dosage level of the active ingredient in the pharmaceutical composition of the present invention can be changed so as to obtain the amount of active ingredient effective for achieving the desired therapeutic response for a specific patient, composition and administration mode without causing toxicity to the patient.A doctor, such as a physician or veterinarian, who has ordinary skill in the art, can easily determine and prescribe the effective amount of pharmaceutical composition required.For example, a doctor or veterinarian can start the dosage of the compound of the present invention used in the pharmaceutical composition at a level lower than that required to achieve the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved.

[0115] In certain embodiments, it is particularly advantageous to formulate compound in dosage unit form for ease of administration and uniformity of dosage.Dosage unit form as used herein refers to a physically separate unit suitable as a unitary dosage for the patient to be treated, each unit containing a predetermined amount of therapeutic compound calculated to produce desired therapeutic effect in combination with required pharmaceutical vehicle.The dosage unit form of the present invention is determined and directly depends on (a) the inherent characteristics of therapeutic compound and the specific therapeutic effect to be achieved, and (b) the inherent limitations of the technology for compounding / formulating such therapeutic compound to treat cancer or other conditions in patients.

[0116] (Route of Administration) Those skilled in the art will recognize that multiple routes of administration can be used, but certain routes can provide more immediate and effective responses than other routes.In certain embodiments of the present invention, MOR antagonist and checkpoint inhibitor and / or VEGF antagonist are administered by the same administration route.In other embodiments, MOR antagonist and checkpoint inhibitor and / or VEGF antagonist are administered by different administration routes.

[0117] Routes of administration of the disclosed compositions (containing a MOR antagonist or a MOR antagonist, and a checkpoint inhibitor and / or a VEGF antagonist) include inhalation, oral, nasal, rectal, parenteral, sublingual, transdermal, transmucosal (e.g., sublingual, lingual, (trans)buccal, (trans)urethral, ​​vaginal (e.g., vaginal and perivaginal), (intramyosal), and (trans)rectal), intravesical, intrapulmonary, intraduodenal, intragastric, intrathecal, subcutaneous, intramuscular, intradermal, intraarterial, intravenous, intrabronchial, inhalation, and topical administration. Suitable compositions and dosage forms include, for example, tablets, capsules, caplets, pills, gel capsules, troches, dispersions, suspensions, liquids, syrups, granules, beads, transdermal patches, gels, powders, pellets, magmas, lozenges, creams, pastes, plasters, lotions, disks, suppositories, liquid sprays for nasal or oral administration, dry powders or aerosolized formulations for inhalation, compositions and formulations for intravesical administration, etc. It should be understood that the formulations and compositions useful in the present invention are not limited to the specific formulations and compositions described herein. In one embodiment, the MOR antagonist treatment and / or checkpoint inhibitor and / or VEGF antagonist treatment comprises a route of administration selected from the group consisting of inhalation, oral, rectal, vaginal, parenteral, topical, transdermal, pulmonary, intranasal, buccal, ocular, intrahepatic artery, intrapleural, intrathecal, intratumoral, intravenous, and any combination thereof.

[0118] (kit) The present invention also includes a kit comprising an MOR antagonist, a checkpoint inhibitor and / or a VEGF antagonist, the kit being used to treat cancer. In one embodiment, the kit comprises a pharmaceutical composition comprising an MOR antagonist and a pharma- ceutical acceptable carrier. In one embodiment, the kit comprises a pharmaceutical composition comprising a μ receptor (e.g., axelopran) antagonist and a checkpoint inhibitor (e.g., pembrolizumab). In certain embodiments, the pharmaceutical composition also comprises a VEGF inhibitor. The pharmaceutical composition may be formulated for controlled release, local administration, or intravitreal injection.

[0119] In one embodiment, the kit includes a pharmaceutical composition comprising a μ receptor (e.g., axelopran) antagonist and a checkpoint inhibitor (e.g., pembrolizumab). The kit also includes another pharmaceutical composition comprising a VEGF inhibitor (e.g., pembrolizumab). EXAMPLES

[0120] The present invention will now be described with reference to the following examples, which are provided for illustrative purposes only and should not be construed as limiting the invention to these examples, but rather as embracing any variations that become evident as a result of the teachings provided herein.

[0121] Without further description, it is believed that one of ordinary skill in the art can, using the preceding description and the following illustrative examples, make and utilize the compounds of the present invention and practice the claimed methods. Accordingly, the following examples specifically point out preferred embodiments of the present invention, and are not to be construed as limiting in any way the remainder of the disclosure.

[0122] Example 1 Effect of MOR antagonists on angiogenesis We investigated the role of MOR and endogenous opioids in angiogenesis using two independent model systems. Human umbilical vein endothelial cells (HUVECs) in culture initiate angiogenesis by tube formation when cultured on an appropriate extracellular matrix. In a living system, new blood vessel formation can be directly visualized by the chicken egg chorioallantoic membrane assay (CAM).

[0123] Specifically, tube formation of HUVEC cells was evaluated in the presence of various doses (0–1000 nM) of three different μ-opioid receptor antagonists (axeroprane, naloxegol and methylnaltrexone (MNTX)). In this study, HUVEC cells were exposed to various MOR antagonist concentrations and incubated under the following conditions; briefly, 0.01 µL of Matrigel was loaded onto a 96-well plate to create a monolayer of extracellular matrix. The plate was kept at 37 °C for 1 h. HUVEC cells were seeded (0.02 × 10 6 Cells were plated at 1000 x g / well (100 x 100 cells / well) and waited 4 hours to allow tubes to form. After 4 hours, the plates were observed for tube formation. Once tubes had formed, the cells were treated with increasing amounts of test compounds (accelerogram, naloxegol and MNTX) and incubated for 18 hours. Cells were stained with calcein AM and images were taken. Data analysis was performed using NIH imageJ software. Tube lengths were measured and data were plotted using Prism software. The results are shown in Figure 1A and Figure 1B.

[0124] Additionally, a chicken egg chorioallantoic membrane assay was performed to determine angiogenesis under increasing concentrations (0, 0.1, 0.5, 1.0 mg / kg) of the MOR antagonist (Acceleroplan). Docetaxel was used as a target. In this study, CAM membranes were incubated in the presence of MOR antagonist or docetaxel under the following conditions: Fertilized White Leghorn eggs were incubated at 37.5°C and 50% relative humidity for 9 days. At that time point (E9), the CAM was dropped from the eggshell through a small hole in the air sac, and a 1 cm window was cut in the eggshell above the CAM. At least 20 eggs (which may exceed 20 per group depending on the survival rate of the embryos after 9 days of development) were implanted in each group. MDA-MB-231 tumor cell line was cultured in D-MEM medium supplemented with 10% FBS and 1% penicillin / streptomycin. On day E9, cells were detached with trypsin, washed with complete medium, and suspended in transplantation medium. 6 A cell was inoculated into the CAM of each egg (E9) and the eggs were then randomized. On day E16, the upper CAM (with tumor) was photographed (n=8 per group). Tumor angiogenesis was assessed by counting the number of blood vessels reaching the tumor and feeding the tumor growth in triplicate. Tumor blood vessel counts were performed on 8 eggs per group. The results of this study are shown in Figure 2A and Figure 2B.

[0125] Using both of these assays, the present study shows that axelopran, naloxegol, and methylnaltrexone inhibit angiogenesis in the absence of exogenous opioids. In contrast, Feng et al. state that "our findings that (the MOR antagonist) naloxone does not antagonize opioid-induced angiogenesis and does not induce angiogenesis by itself were also observed in human microvascular endothelial cells exposed to morphine and naloxone" (Feng et al. BMC Anesthesiol (2021) 21:257, 9-10).

[0126] According to Goel et al., "Given that hypoxia, inflammation and dysregulation of the extracellular matrix can all lead to abnormalities in vascular morphology and function, it is not surprising that vascular tortuosity and 'dysbiosis' have also been reported in many other non-malignant diseases. These include skin psoriasis, rheumatoid arthritis, neurodegenerative diseases and neuropathic pain models, hemangiomas, collagen synthesis disorders such as Ehlers-Danlos syndrome and portal hypertension." (Goel et al., Physiol Rev. 2011 July; 91(3): 1071-1121) Thus, the present study establishes that MOR antagonists can treat a variety of diseases involving angiogenesis.

[0127] Example 2 Effect of MOR antagonists on tumor progression Three unique model systems were selected to test the relevance of endogenous MOR activity to tumor progression, each allowing the examination of different aspects of the host-tumor interaction (Figures 3A-3C).

[0128] Zebrafish larvae (Figure 3A) allow direct visualization of injected tumor cell growth and metastasis (collectively tumor progression) using fluorescently labeled tumor cells. At this developmental stage, both the zebrafish immune system and gut microbiota are immature or absent. By co-injecting exogenous tumor-infiltrating lymphocytes (ex-TILs), immune surveillance in the absence of microbiota can be separately tested.

[0129] The chicken egg chorioallantoic membrane assay (Figure 3B) allows the examination of tumor progression of injected tumor cells in the presence of active endogenous immune surveillance but in the absence of active gut flora.

[0130] The syngenic mouse model assay (Figure 3B) allows for a complete system to quantify tumor progression in the presence of endogenous immune surveillance and an active gut microbiota.

[0131] (zebrafish larvae) The MOR antagonist, Acceleroplan, was first tested for toxicity on zebrafish development to ensure that doses tested were within the range that would not disrupt normal fish ontogeny. Transgenic Tg(fli1:EGFP)y1 zebrafish embryos were used for these studies. The results of this study are shown in Figure 4A and Figure 4B, which show that zebrafish larvae survive up to 72 hours under increasing concentrations of Acceleroplan in the incubation water. Figure 4A and Figure 4B show graphs measuring survival as a function of time. Figure 4C shows photographs of zebrafish larvae exposed to vehicle, 100 μM Acceleroplan, and 500 μM Acceleroplan at 24, 48, and 72 hours. Doses up to 100 μM resulted in survival and pathological lesion development at levels comparable to vehicle controls.

[0132] Thus, toxicity screening studies indicate that increasing doses of Acceleroplan in the incubation water for larvae up to 100 μM did not result in larval death and only limited pathological abnormalities. Based on the toxicity screening, doses up to 10 μM were used in the tumor progression assay.

[0133] After establishing the appropriate dose for testing of Axelroplan, a tumor progression assay was performed. The general procedure for the tumor progression assay is shown in Figure 5. Specifically, Figure 5 shows a schematic diagram of the time course of zebrafish larval development.

[0134] The toxicity screening period was the first 72 h of development. Efficacy evaluation of Acceloplan was performed with M-001 melanoma cells and matched TILs. Approximately 700 M-001 cells labeled with Dil red fluorescent dye were injected subcutaneously into the perivitelline space of 2-day-old zebrafish embryos, and TILs were injected intravenously with or without pembrolizumab antibody. Simultaneously, a test dose of MOR antagonist was added to the water. After an additional 72 h of growth, fish were analyzed for tumor growth (size of original tumor implant) and metastatic spread to the distal caudal vein plexus (CVP) 3 days after implantation.

[0135] The results of this study are shown in Figures 6A, 6B, 7 and 8 and Table 2-1. Figures 6A and 6B show representative plots of melanoma tumor growth size (y-axis) (normalized as % of vehicle control) versus number of metastases (x-axis) in zebrafish treated with 10 μM Acceleropran (n=18) (Figure 6A) or vehicle control (n=32) (Figure 6B). As can be seen in Figures 6A and 6B, exposure to 10 μM Acceleropran significantly reduced metastases, indicating that Acceleropran is effective in treating cancer in this model.

[0136] The above study was repeated to test the effect of various doses of axeroplan (0, 50nM, 100nM, 500nM and 1000nM) alone, pembrolizumab alone, or the combination of pembrolizumab and axeroplan. The results of this study are shown in Figure 7, which summarizes the progression of melanoma tumors (tumor size x number of metastases) in the presence of increasing concentrations of axeroplan or the checkpoint inhibitor pembrolizumab. In experiments with pembrolizumab, exogenous tumor infiltrating lymphocytes (TILs) were also injected. Further controls included the effect of TILs alone and in combination with axeroplan (500nM) and pembrolizumab, and TILs alone and in combination with axeroplan (500nM). *=<0.05, **=<0.005, ***=<0.0005. The results are also summarized in Table 2-1 below. [Table 1]

[0137] In studies examining combination treatments, zebrafish larval mortality was also examined. While no significant mortality was observed with Acceleroplan alone, mortality was significantly increased (up to 63%) in the presence of exogenous TILs and the combination of TILs with pembrolizumab, whereas Acceleroplan significantly reduced mortality when added with TILS and TILS with pembrolizumab (Figure 8).

[0138] (Chicken Egg Chorioallantoic Membrane (CAM) Assay) Tumor progression of breast cancer cells was assessed in chicken egg chorioallantoic membrane assay. The timing of the experiment is shown in Figure 9 below. Fertilized chicken eggs were incubated for 9 days. MDA-MB-231 tumor cell line was cultured in D-MEM medium supplemented with 10% FBS and 1% penicillin / streptomycin. On day E9, cells were detached with trypsin, washed with complete medium, and suspended in transplantation medium. 1×10 6 10 cells were inoculated onto the CAM of each egg (E9), and then the eggs were randomly separated. Then, tumor cells (breast cancer MDA-MB231 cells (10 6 )) were seeded onto the top of the CAM and incubated for an additional 3 days. Test drugs were then added at selected concentrations to the same sites. After an additional 5 days of incubation, grown tumors were excised, and the CAM tissue was removed and weighed. An equivalent area of ​​the bottom CAM was harvested and screened for human Alu sequences by quantitative PCR to determine the relative metastatic value of human cells migrating from the top to the bottom CAM.

[0139] In this study, chicken eggs were treated with Axeroplan; Pembrolizumab (Pembro); Axeroplan + Pembrolizumab; Bevacizumab (Beva); Axeroplan + Bevacizumab; Pembrolizumab + Bevacizumab; and Axeroplan + Pembrolizumab + Bevacizumab. Axeroplan was used at a dose of 1.0 mg / kg; Pembrolizumab was used at a dose of 2.5 mg / kg; and Bevacizumab was used at a dose of 4 mg / kg. The results of this study are shown in Figure 10A, Figure 10B, Figure 11, and Table 2-2.

[0140] Figure 10A shows a representative two-dimensional plot of breast cancer tumor growth size (normalized as % of vehicle control) versus number of metastases (normalized as % of vehicle control) in the CAM assay with the following treatments: negative control = vehicle (n = 15), A = 1.0 mg / kg axelopran (n = 13), P = pembrolizumab (2.5 mg / kg), B = bevacizumab (4 mg / kg) or relevant combination. Circles indicate the mean ± standard deviation for both dimensions.

[0141] FIG. 10B shows tumor weight x metastases for pembrolizumab, axeroplan, bevacizumab, axeroplan + bevacizumab, pembrolizumab + bevacizumab, and axeroplan + pembrolizumab + bevacizumab. [Table 2]

[0142] Infiltration of host immune cells into the tumor environment is necessary for the host to attenuate tumor progression. Of the tumors harvested at E18, 10 tumors per group (n=10) were used for RNA extraction in six groups (groups 1, 2, 3, 4, 7 and 8) after tumor weight measurement. The extracted RNA was then analyzed by RT-qPCR using primers specific for chicken CD3, MMD and CD244 sequences. For all points performed by qPCR, the expression of human GAPDH was also analyzed as a reference gene expression and used to normalize immune biomarker expression between samples. Calculation of Cq for each sample, mean Cq and relative amount of immune cells in each group were handled directly by Bio-Rad CFXMaestro software (see Figure 11).

[0143] FIG. 11 shows the effect of chicken CD3 on breast cancer tumors growing on the upper CAM in the presence of axeroplan, pembrolizumab, bevacizumab, and their combinations. + , CD244 + and MMD + In all cases, Axeroplan, when used in the presence of bevacizumab and pembrolizumab, promoted the uptake of CD3+ cells into the tumor tissue. + Lymphocytes, CD244 + Natural Killer Cells and MMD + It synergistically increased macrophage infiltration.

[0144] (syngenic mice + colon cancer) The effect of axelopran alone or in combination with an anti-PD1 antagonist (pembro) was tested in a model without gut flora, and the effect of axelopran alone or in combination with an anti-PD1 antagonist was further tested in a model with gut flora.

[0145] Murine colon carcinoma (MC38) cells were expanded in vitro prior to implantation. On the day of injection, cells were harvested, counted by adding trypan blue viability dye (cutoff 80%), and resuspended in PBS at the appropriate concentration. Cells were injected subcutaneously into the right flank of 5-week-old (body weight 18-22 g) female mice (C57BL6) at 5.106 cells / mouse within 30 min after harvest. After 15-18 days of tumor growth, mice were randomized into treatment groups, 10 mice / group. Animals were weighed three times a week and tumor growth was measured using calipers. Tumor volume (TV) was calculated by calculating the mean radius from the two measurements, using the following formula:

number

[0146] The results of these studies are shown in Figure 12A, Figure 12B, Figure 13 and Tables 2-3. Tumor growth curves for animals treated with MOR antagonist, Acceleropran, mouse anti-PD-1 or a combination of both treatments are shown in Figure 12A and Figure 12B. Figures 12A and 12B show tumor growth of MC38 colon carcinoma in syngenic mice treated with vehicle (control), Acceleropran (1 mg / kg, orally, daily), mouse anti-PD-1 (12.5 mg / kg, intraperitoneally, three times a week) or Acceleropran + anti-PD-1. Figure 12A shows tumor size (mm 3 ) is shown. Figure 12B shows the individual tumor size distribution on day 13.

[0147] FIG. 13 shows Kaplan-Meier survival curves of mice injected with human colon cancer MC-38 cells and treated with Accelerome (1 mg / kg), anti-PD-1 (2.5 mg / kg) or both drugs.

[0148] Although both Accelerogramin and anti-PD-1 each slowed tumor progression numerically (by 10-16%) (not statistically significant), the combination of both drugs significantly reduced tumor size by 53% on day 13 of treatment. These effects are reflected in the average growth rate during the first 8 days of treatment (Tables 2-3). Associated with the reduction in tumor progression was a 64% increase in survival time from 12.5 days in vehicle-treated control animals to 20.5 days in animals treated with Accelerogramin + anti-PD-1. [Table 3]

[0149] The study in this example shows that the MOR antagonist Axeroplan inhibits tumor growth in the absence of exogenous opioids, at least in melanoma or breast cancer. The study further supports that Axeroplan inhibits metastasis in the absence of exogenous opioids, at least in melanoma or breast cancer. The study also supports that the MOR antagonist activity of Axeroplan is additive with anti-PD-1 treatment in the absence of bacterial flora. The study further supports that the MOR antagonist activity of Axeroplan is synergistic with anti-PD-1 treatment in the presence of bacterial flora, at least in colon cancer.

[0150] (syngenic mouse + pancreatic cancer) Mouse pancreatic tumor cells (PAN02 cells) were maintained in RPMI 1640 medium supplemented with 10% FBS and 2 mmol / L L-glutamine and expanded in vitro before implantation. On the day of injection, cells were harvested, counted with trypan blue viability dye (cutoff 80%), and resuspended in PBS at the appropriate concentration. Cells (5 × 10 in 200 μL PBS) were cultured in 100 mL of PBS for 1 h. 61000 mAbs) were injected subcutaneously into the right flank of 5-week-old (body weight 18-22 g) female mice (C57BL6) within 30 minutes of harvest. After 9 days of tumor growth, mice were randomized into treatment groups, 10 mice / group. Treatments included vehicle (PBS), Acceroplan (1 mg / kg, given orally daily), mouse anti-PD-1 (12.5 mg / kg, given intraperitoneally three times per week), and Acceroplan + mouse anti-PD-1. Animals were weighed three times per week, and tumor growth was measured using calipers as described above in the colon cancer study. Results are shown in Figure 14A-C. At 23 days of measurement, inhibition of tumor progression was most pronounced in mice treated with Acceroplan (Figure 14A,B) and when Acceroplan was used in the presence of anti-PD-1. As shown in FIG. 14C, the distribution of tumor volumes was lower and more uniform in animals treated with Acceleropran compared to animals not treated with Acceleropran.

[0151] ILLUSTRATIVE EMBODIMENTS Exemplary embodiments of the disclosed technology are provided, which are merely illustrative and not limiting to the scope of the disclosure and the claims.

[0152] Embodiment 1. A method of reducing or inhibiting endothelial cell proliferation comprising contacting an endothelial cell with a μ-opioid receptor (MOR) antagonist.

[0153] Embodiment 2. The method of embodiment 1, wherein the method inhibits endothelial cell proliferation.

[0154] Embodiment 3. A method of reducing angiogenesis in a patient comprising administering to a patient in need thereof a MOR antagonist.

[0155] Embodiment 4. The method of embodiment 3, wherein the method inhibits angiogenesis.

[0156] Embodiment 5. The method of embodiment 3 or 4, wherein the method reduces or inhibits ocular neovascularization.

[0157] Embodiment 6. The method of embodiment 3 or 4, wherein the patient has psoriasis.

[0158] Embodiment 7. A method of reducing angiogenesis in a cancer patient comprising administering to the cancer patient a MOR antagonist.

[0159] Embodiment 8. The method of embodiment 7, wherein the method inhibits angiogenesis.

[0160] Embodiment 9. A method of reducing tumor growth in a cancer patient comprising administering to the cancer patient a MOR antagonist.

[0161] Embodiment 10. The method of embodiment 9, wherein the method inhibits tumor growth.

[0162] Embodiment 11. A method of reducing metastasis in a cancer patient comprising administering to the cancer patient a MOR antagonist.

[0163] Embodiment 12. The method of embodiment 11, wherein the method inhibits metastasis.

[0164] Embodiment 13. A method of improving the immune response to a tumor, comprising administering to a cancer patient a MOR antagonist.

[0165] Embodiment 14. The method of embodiment 13, wherein the method increases infiltration of immune cells into the tumor.

[0166] Embodiment 15. The method of embodiment 13, wherein the method increases the infiltration of NK cells, lymphocytes and / or monocytes / macrophages.

[0167] 16. The method comprises: + , CD244 + and MMD + 14. The method of embodiment 13, which increases immune cell infiltration.

[0168] Embodiment 17. The method of any one of embodiments 1 to 16, wherein the cancer is melanoma, colon cancer or breast cancer.

[0169] Embodiment 18. The method of any one of embodiments 1 to 17, wherein the patient is opioid naive.

[0170] Embodiment 19. The method of any one of embodiments 1 to 18, wherein the method does not include administering an opioid.

[0171] Embodiment 20. The method of any one of embodiments 1 to 19, wherein the MOR antagonist does not include methylnaltrexone.

[0172] Embodiment 21. The method of any one of embodiments 1 to 20, wherein the MOR antagonist is axelopran, naloxegol, naldemedine, alvimopan, or a combination thereof.

[0173] Embodiment 22. The method of embodiment 21, wherein the MOR antagonist is axelopran.

[0174] Embodiment 23. The method of any one of embodiments 1 to 23, wherein the method comprises orally administering the MOR antagonist.

[0175] Embodiment 24. The method of any one of embodiments 1 to 23, wherein the method comprises administering the MOR antagonist subcutaneously.

[0176] Embodiment 25. The method of any one of embodiments 1 to 23, wherein the method comprises intravitreally administering the MOR antagonist.

[0177] Embodiment 26. The method of any one of embodiments 1 to 23, wherein the method comprises locally administering a MOR antagonist.

[0178] Embodiment 27. The method of any of embodiments 1-23, wherein the method comprises administering a pharmaceutical composition comprising a MOR antagonist and a pharma- ceutically acceptable carrier.

[0179] Embodiment 28. The method of embodiment 19, wherein the pharmaceutical composition is formulated for modified release.

[0180] Embodiment 29. A method of treating cancer comprising administering axeroplan, naloxegol, or a combination thereof to a cancer patient.

[0181] Embodiment 30. The method of embodiment 29, wherein the method comprises administering axeroplan.

[0182] Embodiment 31. The method of embodiment 29 or 30, wherein the method reduces angiogenesis, tumor growth and / or metastasis.

[0183] Embodiment 32. The method of embodiment 32, wherein the method inhibits angiogenesis, tumor growth and / or metastasis.

[0184] Embodiment 33. The method of any one of embodiments 29 to 32, wherein the method does not include administering an exogenous opioid.

[0185] Embodiment 34. The method of any one of embodiments 29 to 32, wherein the patient is not receiving opioid treatment.

[0186] Embodiment 35. The method of any one of embodiments 29 to 32, wherein the cancer is melanoma, colon cancer, pancreatic cancer or breast cancer.

[0187] Embodiment 36. The method of any one of embodiments 29 to 35, wherein the method comprises orally administering the MOR antagonist.

[0188] Embodiment 37. The method of any one of embodiments 29 to 35, wherein the method comprises administering the MOR antagonist subcutaneously.

[0189] Embodiment 38. The method of any of embodiments 29-35, wherein the method comprises administering a pharmaceutical composition comprising a MOR antagonist and a pharma- ceutically acceptable carrier.

[0190] Embodiment 39. The method of embodiment 38, wherein the pharmaceutical composition is formulated for modified release, topical administration, or intravitreal injection.

[0191] Embodiment 40. The method of any one of embodiments 7 to 39, wherein the cancer is resistant to treatment with a checkpoint inhibitor.

[0192] The method according to any of the preceding embodiments, which may comprise administering a checkpoint inhibitor and / or an angiogenesis inhibitor, such as a VEGF inhibitor.

[0193] Embodiment 41. A method of reducing tumor size comprising contacting the tumor with axeroplan, naloxegol, or a combination thereof.

[0194] Embodiment 42. A method of treating cancer comprising administering to a cancer patient a MOR antagonist and a checkpoint inhibitor and / or a VEGF inhibitor.

[0195] Embodiment 43. The method of embodiment 42, wherein the MOR antagonist and the checkpoint inhibitor and / or VEGF inhibitor synergistically treat the cancer.

[0196] Embodiment 44. A method of improving the immune response to a tumor, comprising administering to a cancer patient a MOR antagonist and a checkpoint inhibitor and / or a VEGF inhibitor.

[0197] Embodiment 45. The method of embodiment 44, wherein the method increases infiltration of immune cells into the tumor.

[0198] Embodiment 46. The method of embodiment 45, wherein the method increases the infiltration of NK cells, lymphocytes and / or monocytes / macrophages.

[0199] 47. The method comprises: + , CD244 + and MMD + The method of embodiment 45, which increases immune cell infiltration.

[0200] Embodiment 48. The method of embodiment 45, wherein the MOR antagonist and the checkpoint inhibitor synergistically improve the immune response to the tumor.

[0201] Embodiment 49. The method of any one of embodiments 42 to 49, wherein the MOR antagonist comprises axelopran, naloxegol, methylnaltrexone, or a combination thereof.

[0202] Embodiment 50. The method of embodiment 49, wherein the MOR antagonist is axelopran.

[0203] Embodiment 51. The method of any one of embodiments 42 to 50, wherein the checkpoint inhibitor is a PD-1 / PD-L1 pathway inhibitor.

[0204] Embodiment 52. The method of embodiment 51, wherein the PD-1 / PD-L1 pathway inhibitor is an antibody.

[0205] Embodiment 53. The method of embodiment 52, wherein the antibody is an antibody that binds to PD-1.

[0206] Embodiment 54. The method of embodiment 53, wherein the antibody is a humanized antibody.

[0207] Embodiment 55. The method of embodiment 53, wherein the human antibody is pembrolizumab.

[0208] Embodiment 56. The method of any one of embodiments 42 to 55, wherein the method comprises administering the MOR antagonist orally or subcutaneously.

[0209] Embodiment 57. The method of any of embodiments 42-56, wherein the method comprises administering a pharmaceutical composition comprising a MOR antagonist and a pharma- ceutically acceptable carrier.

[0210] Embodiment 58. The method of embodiment 57, wherein the pharmaceutical composition is formulated for modified release.

[0211] Embodiment 59. The method of any of embodiments 42 to 58, wherein the checkpoint inhibitor is administered simultaneously with, before or after the MOR antagonist.

[0212] Embodiment 60. The method of any one of embodiments 42 to 59, wherein the method comprises administering a checkpoint inhibitor.

[0213] Embodiment 61. The method of any one of embodiments 42 to 60, wherein the method comprises administering a VEGF inhibitor.

[0214] Embodiment 62. The method of any one of embodiments 42 to 60, wherein the VEGF inhibitor is an anti-VEGF antibody.

[0215] Embodiment 63. The method of embodiment 62, wherein the antibody is a humanized antibody.

[0216] Embodiment 64. The method of embodiment 62 or 63, wherein the antibody is an anti-VEGF-A antibody.

[0217] Embodiment 65. The method of any one of embodiments 61 to 64, wherein the VEGF inhibitor is bevacizumab.

[0218] Embodiment 66. The method of any one of embodiments 61 to 65, comprising intravenously injecting a VEGF inhibitor.

[0219] Embodiment 67. The method of any of embodiments 61-66, wherein the VEGF inhibitor is administered simultaneously with, before or after the MOR antagonist.

[0220] Embodiment 68. The method of any one of embodiments 61 to 67, wherein the VEGF inhibitor is administered simultaneously with, before or after the checkpoint inhibitor.

[0221] Embodiment 69. A kit comprising a MOR antagonist and a checkpoint inhibitor and / or a VEGF inhibitor.

[0222] Embodiment 70. The kit of embodiment 69, wherein the checkpoint inhibitor is a PD-1 / PD-L1 pathway inhibitor.

[0223] Embodiment 71. The kit of embodiment 69, wherein the kit comprises a pharmaceutical composition comprising a MOR antagonist and a pharma- ceutically acceptable carrier.

[0224] Embodiment 72. The kit of embodiment 71, wherein the pharmaceutical composition further comprises a VEGF inhibitor.

[0225] Embodiment 73. The kit of embodiment 71 or 72, wherein the pharmaceutical composition is formulated for controlled release, topical administration, or intravitreal injection.

[0226] Embodiment 74. A composition comprising synergistic amounts of a MOR antagonist and a PD-1 / PD-L1 pathway inhibitor.

[0227] Embodiment 75. The composition of embodiment 74, wherein the MOR antagonist comprises axelopran, naloxegol, methylnaltrexone, or a combination thereof.

[0228] Embodiment 76. The composition of embodiment 74, wherein the MOR antagonist is axelopran.

[0229] Embodiment 77. The composition of any one of embodiments 74 to 76, wherein the PD-1 / PD-L1 pathway inhibitor is an antibody.

[0230] Embodiment 78. The composition described in any one of embodiments 74 to 77, wherein the antibody is an antibody that binds to PD-1.

[0231] Embodiment 79. The composition described in embodiment 78, wherein the antibody is a humanized antibody.

[0232] Embodiment 80. The method of embodiment 79, wherein the human antibody is pembrolizumab.

[0233] Embodiment 81. Use of axelopran, naloxegol, or a combination thereof in the manufacture of a medicament for treating cancer.

[0234] Embodiment 82. Use of axeroplan, naloxegol, or a combination thereof to treat cancer.

[0235] Embodiment 83. Use of a MOR antagonist and a PD-1 / PD-L1 pathway inhibitor in the manufacture of a medicament for treating cancer.

[0236] Embodiment 84. Use of a MOR antagonist and a PD-1 / PD-L1 pathway inhibitor to treat cancer.

[0237] Embodiment 85. Use of a MOR antagonist and a VEGF inhibitor in the manufacture of a medicament for treating cancer.

[0238] Embodiment 86. Use of a MOR antagonist and a VEGF inhibitor to treat cancer.

[0239] Embodiment 87. Use of a MOR antagonist, a PD-1 / PD-L1 pathway inhibitor, and a VEGF inhibitor in the manufacture of a medicament for treating cancer.

[0240] Embodiment 88. Use of a MOR antagonist, a PD-1 / PD-L1 pathway inhibitor and a VEGF inhibitor to treat cancer.

[0241] Embodiment 89. The use of any one of embodiments 85 to 88, wherein the VEGF inhibitor is an anti-VEGF-A antibody.

[0242] Embodiment 90. The use according to embodiment 89, wherein the VEGF inhibitor is bevacizumab.

[0243] Embodiment 91. The use of any one of embodiments 83 to 90, wherein the PD-1 / PD-L1 pathway inhibitor is an antibody.

[0244] Embodiment 92. The use of any one of embodiments 83 to 91, wherein the antibody is an antibody that binds to PD-1.

[0245] Embodiment 93. The use described in embodiment 92, wherein the antibody is a humanized antibody.

[0246] Embodiment 94. The use described in embodiment 93, wherein the human antibody is pembrolizumab.

[0247] Embodiment 95. Use of a MOR antagonist to treat cancer in a patient undergoing therapy that inhibits the PD-1 / PD-L1 pathway.

[0248] Embodiment 96. The use of any of embodiments 83 to 95, wherein the MOR antagonist comprises axelopran, naloxegol, methylnaltrexone, or a combination thereof.

[0249] Embodiment 97. The use according to embodiment 96, wherein the MOR antagonist is axelopran.

[0250] Embodiment 98. A combination comprising a MOR antagonist and a PD-1 / PD-L1 pathway inhibitor for use in a method of treating cancer, the method comprising administering the combination to a subject in need of treatment.

[0251] Embodiment 99. The combination described in embodiment 98, wherein the MOR antagonist is axelopran.

[0252] Embodiment 100. A combination according to embodiment 98 or 99, wherein the PD-1 / PD-L1 pathway inhibitor is pembrolizumab.

[0253] Embodiment 101. The combination described in embodiment 98 or 99, wherein the combination further comprises a VEGF inhibitor.

[0254] A combination according to any of embodiments 98 to 101, wherein the VEGF inhibitor is bevacizumab.

[0255] The disclosures of each patent, patent application, and publication cited or described in this specification are hereby incorporated by reference in their entirety for all purposes.

Claims

1. A method of reducing or inhibiting endothelial cell proliferation comprising contacting an endothelial cell with a μ opioid receptor (MOR) antagonist.

2. The method of claim 1, wherein the method inhibits endothelial cell proliferation.

3. A method of reducing angiogenesis in a patient in need thereof comprising administering to the patient a MOR antagonist.

4. The method of claim 3, wherein the method inhibits angiogenesis.

5. 5. The method of claim 3 or 4, wherein the method reduces or inhibits ocular neovascularization.

6. 5. The method of claim 3 or 4, wherein the patient has psoriasis.

7. A method of reducing angiogenesis in a cancer patient comprising administering to the cancer patient a MOR antagonist.

8. The method of claim 7, wherein the method inhibits angiogenesis.

9. A method of reducing tumor growth in a cancer patient comprising administering to the cancer patient a MOR antagonist.

10. The method of claim 9, wherein the method inhibits tumor growth.

11. A method of reducing metastasis in a cancer patient comprising administering to the cancer patient a MOR antagonist.

12. The method of claim 11 , wherein the method inhibits metastasis.

13. A method of improving the immune response to a tumor comprising administering to a cancer patient a MOR antagonist.

14. The method of claim 13, wherein the method increases infiltration of immune cells into the tumor.

15. The method of claim 13, wherein the method increases the infiltration of NK cells, lymphocytes and / or monocytes / macrophages.

16. The method is CD3 + , CD244 + and MMD + The method of claim 13, which increases immune cell infiltration.

17. The method of any one of claims 1 to 16, wherein the cancer is melanoma, colon cancer, pancreatic cancer or breast cancer.

18. The method of any one of claims 1 to 17, wherein the patient is opioid naive.

19. 19. The method of any one of claims 1 to 18, wherein the method does not include administering an opioid.

20. The method of any one of claims 1 to 19, wherein the MOR antagonist does not include methylnaltrexone.

21. The method of any one of claims 1 to 20, wherein the MOR antagonist is axelopran, naloxegol, naldemedine, alvimopan, or a combination thereof.

22. 22. The method of claim 21, wherein the MOR antagonist is axelopran.

23. The method of any one of claims 1 to 23, wherein the method comprises orally administering the MOR antagonist.

24. 24. The method of any one of claims 1 to 23, wherein the method comprises administering the MOR antagonist subcutaneously.

25. The method of any one of claims 1 to 23, wherein the method comprises intravitreally administering the MOR antagonist.

26. The method of any one of claims 1 to 23, wherein the method comprises locally administering the MOR antagonist.

27. The method of any one of claims 1 to 23, wherein the method comprises administering a pharmaceutical composition comprising a MOR antagonist and a pharma- ceutically acceptable carrier.

28. 20. The method of claim 19, wherein the pharmaceutical composition is formulated for modified release.

29. 23. A method of treating cancer comprising administering axeroplan, naloxegol, or a combination thereof to a cancer patient.

30. 30. The method of claim 29, wherein the method comprises administering axeroplan.

31. 31. The method of claim 29 or 30, wherein the method reduces angiogenesis, tumor growth and / or metastasis.

32. 32. The method of claim 31 , wherein the method inhibits angiogenesis, tumor growth and / or metastasis.

33. 33. The method of any one of claims 29 to 32, wherein the method does not include administering an exogenous opioid.

34. The method of any one of claims 29 to 32, wherein the patient is opioid naive.

35. The method of any one of claims 29 to 32, wherein the cancer is melanoma, colon cancer, pancreatic cancer or breast cancer.

36. The method of any one of claims 29 to 35, wherein the method comprises orally administering the MOR antagonist.

37. The method of any one of claims 29 to 35, wherein the method comprises administering the MOR antagonist subcutaneously.

38. The method of any one of claims 29 to 35, wherein the method comprises administering a pharmaceutical composition comprising a MOR antagonist and a pharma- ceutically acceptable carrier.

39. 39. The method of claim 38, wherein the pharmaceutical composition is formulated for controlled release, topical administration, or intravitreal injection.

40. The method of any one of claims 7 to 39, wherein the cancer is resistant to treatment with a checkpoint inhibitor.

41. A method of reducing tumor size comprising contacting the tumor with axeroplan, naloxegol, or a combination thereof.

42. A method of treating cancer comprising administering to a cancer patient a MOR antagonist and a checkpoint inhibitor and / or a VEGF inhibitor.

43. 43. The method of claim 42, wherein the MOR antagonist and the checkpoint inhibitor and / or VEGF inhibitor synergistically treat the cancer.

44. A method of improving the immune response to a tumor comprising administering to a cancer patient a MOR antagonist and a checkpoint inhibitor and / or a VEGF inhibitor.

45. 45. The method of claim 44, wherein the method increases immune cell infiltration into the tumor.

46. 46. ​​The method of claim 45, wherein the method increases the infiltration of NK cells, lymphocytes and / or monocytes / macrophages.

47. The method is CD3 + , CD244 + and MMD + 46. ​​The method of claim 45, which increases immune cell infiltration.

48. 46. ​​The method of claim 45, wherein the MOR antagonist and the checkpoint inhibitor synergistically improve the immune response to the tumor.

49. The method of any one of claims 42-48, wherein the MOR antagonist comprises axelopran, naloxegol, methylnaltrexone, or a combination thereof.

50. 50. The method of claim 49, wherein the MOR antagonist is axelopran.

51. The method of any one of claims 42 to 50, wherein the checkpoint inhibitor is a PD-1 / PD-L1 pathway inhibitor.

52. 52. The method of claim 51, wherein the PD-1 / PD-L1 pathway inhibitor is an antibody.

53. 53. The method of claim 52, wherein the antibody is an antibody that binds to PD-1.

54. The method of claim 53, wherein the antibody is a humanized antibody.

55. 54. The method of claim 53, wherein the human antibody is pembrolizumab.

56. 56. The method of any one of claims 42-55, wherein the method comprises administering the MOR antagonist orally or subcutaneously.

57. The method of any one of claims 42 to 56, wherein the method comprises administering a pharmaceutical composition comprising a MOR antagonist and a pharma- ceutically acceptable carrier.

58. 58. The method of claim 57, wherein the pharmaceutical composition is formulated for modified release.

59. The method of any one of claims 42-58, wherein the checkpoint inhibitor is administered simultaneously with, prior to, or after the MOR antagonist.

60. 60. The method of any one of claims 42-59, wherein the method comprises administering a checkpoint inhibitor.

61. 61. The method of any one of claims 42 to 60, wherein the method comprises administering a VEGF inhibitor.

62. The method of any one of claims 42 to 60, wherein the VEGF inhibitor is an anti-VEGF antibody.

63. The method of claim 62, wherein the antibody is a humanized antibody.

64. The method of claim 62 or 63, wherein the antibody is an anti-VEGF-A antibody.

65. The method of any one of claims 61 to 64, wherein the VEGF inhibitor is bevacizumab.

66. 66. The method of any one of claims 61 to 65, comprising intravenously injecting the VEGF inhibitor.

67. The method of any one of claims 61 to 66, wherein the VEGF inhibitor is administered simultaneously with, prior to or after the MOR antagonist.

68. 68. The method of any one of claims 61 to 67, wherein the VEGF inhibitor is administered simultaneously with, prior to or after the checkpoint inhibitor.

69. A kit comprising a MOR antagonist and a checkpoint inhibitor and / or a VEGF inhibitor.

70. The kit of claim 69, wherein the checkpoint inhibitor is a PD-1 / PD-L1 pathway inhibitor.

71. 70. The kit of claim 69, wherein the kit comprises a pharmaceutical composition comprising the MOR antagonist and a pharma- ceutically acceptable carrier.

72. 72. The kit of claim 71, wherein the pharmaceutical composition further comprises a VEGF inhibitor.

73. 73. The kit of claim 71 or 72, wherein the pharmaceutical composition is formulated for controlled release, topical administration, or intravitreal injection.

74. A composition comprising a synergistic amount of a MOR antagonist and a PD-1 / PD-L1 pathway inhibitor.

75. 75. The composition of claim 74, wherein the MOR antagonist comprises axelopran, naloxegol, methylnaltrexone, or a combination thereof.

76. The composition of claim 74, wherein the MOR antagonist is axelopran.

77. The composition of any one of claims 74 to 76, wherein the PD-1 / PD-L1 pathway inhibitor is an antibody.

78. The composition of any one of claims 74 to 77, wherein the antibody is an antibody that binds to PD-1.

79. The composition of claim 78, wherein the antibody is a humanized antibody.

80. 80. The method of claim 79, wherein the human antibody is pembrolizumab.

81. 23. The use of axelopran, naloxegol, or a combination thereof in the manufacture of a medicament for treating cancer.

82. Use of axeroplan, naloxegol, or a combination thereof to treat cancer.

83. Use of a MOR antagonist and a PD-1 / PD-L1 pathway inhibitor in the manufacture of a medicament for treating cancer.

84. Use of MOR antagonists and PD-1 / PD-L1 pathway inhibitors to treat cancer.

85. Use of a MOR antagonist and a VEGF inhibitor in the manufacture of a medicament for treating cancer.

86. Use of a MOR antagonist and a VEGF inhibitor to treat cancer.

87. Use of a MOR antagonist, a PD-1 / PD-L1 pathway inhibitor and a VEGF inhibitor in the manufacture of a medicament for treating cancer.

88. Use of MOR antagonists, PD-1 / PD-L1 pathway inhibitors and VEGF inhibitors to treat cancer.

89. 89. The use according to any one of claims 85 to 88, wherein the VEGF inhibitor is an anti-VEGF-A antibody.

90. 90. The use of claim 89, wherein the VEGF inhibitor is bevacizumab.

91. The use of any one of claims 83 to 90, wherein the PD-1 / PD-L1 pathway inhibitor is an antibody.

92. 92. The use according to any one of claims 83 to 91, wherein the antibody is an antibody that binds to PD-1.

93. 93. The use according to claim 92, wherein the antibody is a humanized antibody.

94. 94. The use of claim 93, wherein the human antibody is pembrolizumab.

95. Use of MOR antagonists to treat cancer in patients undergoing therapy that inhibits the PD-1 / PD-L1 pathway.

96. The use of any one of claims 83 to 95, wherein the MOR antagonist comprises axelopran, naloxegol, methylnaltrexone, or a combination thereof.

97. 97. The use of claim 96, wherein the MOR antagonist is axelopran.

98. 1. A combination comprising a MOR antagonist and a PD-1 / PD-L1 pathway inhibitor for use in a method of treating cancer, the method comprising administering the combination to a subject in need of treatment.

99. 99. The combination of claim 98, wherein the MOR antagonist is axelopran.

100. 100. The combination of claim 98 or 99, wherein the PD-1 / PD-L1 pathway inhibitor is pembrolizumab.

101. 100. The combination of claim 98 or 99, wherein the combination further comprises a VEGF inhibitor.

102. 102. The combination according to any one of claims 98 to 101, wherein the VEGF inhibitor is bevacizumab.

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