Methods and compositions for the treatment of cancer

Abstract

To provide a novel effective, uniform and successful treatment strategy for cancer.SOLUTION: Based on the surprising discovery that naloxone selectively induces cytotoxicity in tumor cells compared to non-tumor cells, methods and compositions are provided for administering OGFR locally adjacent to tumor or cancer cells in a patient. These methods and compositions can inhibit the proliferation of tumor cells while sparing surrounding normal cells.SELECTED DRAWING: Figure 4

Description

Technical Field

[0001] Cross-Reference to Related Applications This application claims the benefit of priority to U.S. Provisional Application No. 62 / 874,037, filed Jul. 15, 2019, the disclosure of which is hereby incorporated by reference in its entirety.

[0002] Technical Field The present invention relates to methods and compositions for treating cancer.

Background Art

[0003] Cancer is a group of diseases involving abnormal cell growth that can invade or spread to other parts of the body. In 2015, approximately 90.5 million people had cancer. Approximately 14.1 million new cases occur each year (excluding skin cancers other than melanoma), and cancer causes approximately 8.8 million deaths per year, or about 15.7% of all deaths. Overall, cancer is the second leading cause of death in the United States today. The risk of cancer increases significantly with age, and many cancers are more common in developed countries.

[0004] Cancer is thought to exhibit six hallmarks that underlie the ability of cancer cells to form tumors and progress to a malignant state that allows cells to proliferate and invade tissues other than the original tissue. These six hallmarks are cell growth and division in the absence of appropriate signals, continued growth and division even when opposing signals are given, avoidance of programmed cell death, unlimited numbers of cell divisions, promotion of angiogenesis, invasion of tissues, and formation of metastases.

[0005] While our understanding of cancer characteristics has improved significantly, there is no single, universally recognized trait or characteristic that has enabled effective, uniform, and successful cancer treatment strategies. Cancer can be treated with surgery, chemotherapy, radiation therapy, hormone therapy, and targeted therapies (including immunotherapy such as monoclonal antibody therapy). Treatment options vary depending on the location and grade of the tumor, the stage of the disease, and the patient's overall health.

[0006] Despite a rapidly advancing understanding of the underlying mechanisms of cancer development, many, if not most, cancers remain incurable, and there is still a great medical need for new cancer treatment strategies. [Overview of the project]

[0007] One embodiment is a method for treating a target cancer in need of such treatment, comprising topically administering an opioid growth factor receptor (OGFR) antagonist to a target site containing cancer cells. In some embodiments, the OGFR antagonist includes naloxone, naltrexone, functional derivatives or analogs thereof, or combinations thereof. Another embodiment is a composition for use in a method for treating a target cancer in need of such treatment, comprising topically administering an opioid growth factor receptor (OGFR) antagonist to a target site containing cancer cells.

[0008] In some embodiments, cancers include soft tissue non-osteogenic sarcoma, bone sarcoma, osteosarcoma, Ewing's sarcoma, chondrosarcoma, benign bone tumors, basal cell carcinoma, melanoma, thyroid cancer, glial blastoma, pituitary tumors, oligodendrocytoma, bladder cancer, triple-negative breast cancer, breast cancer, neuroblastoma, or astrocytoma. In some embodiments, the OGFR antagonist is administered in a local dosage of approximately 1 mM to approximately 10 mM.

[0009] In one embodiment, the OGFR antagonist is administered locally by intratumor injection.

[0010] In another embodiment, the OGFR antagonist is administered with a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutically acceptable carrier includes a collagen sponge, powdered collagen, or a collagen-based gelatin hydrogel. In some embodiments, the pharmaceutically acceptable carrier includes a hydrogel-based carrier. In one particular embodiment, the pharmaceutically acceptable carrier is a collagen sponge. In some embodiments, the hydrogel-based carrier includes a poly-(butyric acid) / poly-(ethylene glycol) / poly-(butyric acid) hydrogel, a poly-(ethylene glycol) / poly-(butyric acid) / poly-(ethylene glycol) hydrogel, or a combination thereof. In one particular embodiment, the hydrogel-based carrier is poly-(ethylene glycol) / poly-(butyric acid) / poly-(ethylene glycol).

[0011] In some embodiments, the OGFR antagonist is administered with a diluent, which enhances the solubility of the OGFR antagonist. In some embodiments, the diluent comprises a polar organic solvent. Preferably, the polar organic solvent is DMSO, including dimethyl sulfoxide (DMSO) based saline solution. In other embodiments, the solubility-enhancing diluent is an acidified saline solution, an alcohol, a polyol, an alkane, a fatty acid, an ester, an amine, an amide, a terpene, a cyclodextrin, or a surfactant. In one embodiment, the diluent comprises a dimethyl sulfoxide (DMSO) based saline solution. In some embodiments, the DMSO-based saline solution contains about 5% v / v DMSO to about 80% v / v DMSO. In some embodiments, the DMSO-based saline solution contains about 50% v / v DMSO. In some embodiments, the acidified saline solution has a pH of about 4.5 to about 7.4. [Brief explanation of the drawing]

[0012] [Figure 1] This graph shows the expression of lactate dehydrogenase (LDH) in triple-negative breast cancer cells (BT474), osteosarcoma cells (SaOS2), and breast cancer cells (BT20) in the treatment of these cells compared to non-tumor cells. LDH expression is shown in comparison to LDH release in non-tumor cells upon naloxone treatment. [Figure 2] The graph shows the change in the number of triple-negative breast cancer cells (BT474) compared to normal non-tumor cells (HUMECs) and normal vascular endothelial cells (HUVECs) in response to an increase in naloxone dosage. [Figure 3]Figure 3A shows a spheroid assay demonstrating that naloxone (an OGFR antagonist) treatment reduces tumor size in vitro. Figure 3A shows tumor spheroids formed in SaOS2 (osteosarcoma) and BT474 (triple-negative breast cancer) cells in the presence of naloxone treatment at doses of 1 mM, 1.5 mM, 2 mM, 2.5 mM, and 5 mM, and in an untreated control. Figure 3B is a graph quantifying the size of spheroids formed in the spheroid assay described in Figure 3A as the percentage change in the percentage of spheroid area from 0 to 120 hours after treatment. Figure 3C is a graph quantifying the size of spheroids formed in the spheroid assay described in Figure 3A as the percentage change in the percentage of cell area of ​​each spheroid at 120 hours after treatment. [Figure 4] This study demonstrates the effect of a naloxone preparation on tumor growth in mice transplanted with MDA-MB-231 triple-negative breast cancer cells. The preparation was implanted in the tibia of 8-week-old female Cr nude mice. Figure 4A shows tumor volume measured with calipers over 60 days. Tumor volume was half the size in the preparation-implanted group. Regression analysis revealed significantly different slopes for the two treatment groups (p<0.001). Figure 4B shows the survival rate assessment using Kaplan-Meier statistics. Survival was twice as high in the preparation-treated group (p<0.0328). [Modes for carrying out the invention]

[0013] This disclosure is based on the remarkable finding that naloxone selectively induces cytotoxicity in tumor cells compared to non-tumor cells. Therefore, this disclosure provides methods and compositions useful for inhibiting tumor cell proliferation while preserving surrounding normal cells. In some embodiments, this disclosure provides methods and compositions for administering OGFR locally adjacent to a patient's tumor or cancer cells.

[0014] The exemplary examples herein demonstrate that naloxone has a selective inhibitory effect on the proliferation of breast cancer cells, including osteosarcoma and triple-negative breast cancer cells. In some embodiments, naloxone treatment selectively induced cytotoxicity in tumor cells compared to corresponding non-tumor cells, as identified by increased lactate dehydrogenase (LDH) expression. In some embodiments, naloxone induced a 6-fold increase in LDH expression in osteosarcoma and breast cancer cells compared to non-tumor cells. In some embodiments, naloxone induced a 3-fold increase in LDH expression in triple-negative breast cancer cells. In some embodiments, naloxone treatment selectively inhibited cancer cell proliferation compared to non-cancer cells, as identified by counting cell numbers 72 hours after naloxone treatment. See Figures 1 and 2. In some embodiments, naloxone treatment reduced tumor growth in triple-negative breast cancer cells, as identified by a spheroid assay. See Figure 3. In some embodiments, the naloxone formulation Nalovent® inhibited tumor growth in vivo when transplanted adjacent to tumors formed by mouse triple-negative breast cancer cells. See Figure 4.

[0015] While exemplary examples illustrate the use of the methods and compositions disclosed herein for the treatment of osteosarcoma and breast cancer, this disclosure is considered useful for treating all types of cancer. Generally speaking, cancer refers to a condition in which abnormal cells divide uncontrollably and can invade nearby tissues. There are several main types of cancer. Carcinomas are cancers that occur in the skin or tissues that line or cover internal organs. Sarcomas are cancers that occur in the bone, cartilage, fat, muscle, blood vessels, or other connective or supporting tissues. Leukemia is a cancer that originates in hematopoietic tissues such as bone marrow and invades the bloodstream by producing large numbers of abnormal blood cells. Lymphoma and multiple myeloma are cancers that originate in the cells of the immune system. Central nervous system cancers are cancers that originate in the tissues of the brain and spinal cord. In some embodiments, cancer is one or more of the following: pancreatic cancer, kidney cancer, small cell lung cancer, brain cancer, nerve cancer, bone cancer, lymphoma, myeloma, gastrointestinal cancer, uterine cancer, breast cancer, leukemia, liver cancer, prostate cancer, skin cancer, and melanoma. In some embodiments, cancer is soft tissue non-osteogenic sarcoma, bone sarcoma, osteosarcoma, Ewing's sarcoma, chondrosarcoma, benign bone tumor, basal cell carcinoma, melanoma, thyroid cancer, glial blastoma, pituitary tumor, oligodendrocytoma, bladder cancer, triple-negative breast cancer, breast cancer, neuroblastoma, or astrocytoma.

[0016] definition The following definitions are provided to facilitate understanding of the specific terms used throughout this specification.

[0017] The technical and scientific terms used herein have the meanings generally understood by those skilled in the art unless otherwise defined. Any suitable materials and / or methodologies known to those skilled in the art may be used in carrying out the methods described herein.

[0018] As used in the specification and appended claims of this invention, the singular forms "a," "an," and "the" are used interchangeably and are intended to include the plural forms, each encompassing their respective meanings, unless the context explicitly indicates otherwise. Furthermore, as used herein, "and / or" refers to and encompasses one or any possible combination of the enumerated items, as well as the absence of any combination when interpreted as an alternative ("or").

[0019] All numerical specifications, including ranges, such as pH, temperature, time, concentration, volume, and molecular weight, are approximations that may vary by 10%, 1%, or 0.1% (+) or (-) as needed. It should be understood that, although not always explicitly stated, the term "approximately" may be used before any numerical specification. It should also be understood that, although not always explicitly stated, the reagents described herein are merely illustrative, and such equivalents may be known in the art.

[0020] The terms “comprising” or “comprises” are intended to mean that the compositions and methods described include the elements but do not exclude others. “Consisting essentially of,” when used to define compositions and methods, means excluding other elements that are essentially important to the combination. For example, a composition essentially consisting of the elements defined herein would not exclude other elements that do not substantially affect the basic and novel features of the claimed invention. “Consisting of” means excluding trace amounts of other components and the substantial steps of the methods described in the claims. Embodiments defined by each of these transition terms are within the scope of the present invention.

[0021] As used herein, the term "antagonist" is used interchangeably with "inhibitor" and refers to a substrate that blocks or suppresses the activity, function, effect, or expression of a target. In some embodiments, the target is a compound, protein, gene, cell, or agent. As used herein, the term "expression" refers to the amount produced by a living cell from a target. In some embodiments, the inhibitor suppresses the expression of a target gene or protein. In some embodiments, the inhibitor comprises a compound that prevents the binding of another molecule to an enzyme or molecular pump. In some embodiments, the inhibitor is a compound that causes downregulation of an enzyme. In some embodiments, the inhibitor may be a competitive or non-competitive inhibitor.

[0022] As used herein, the term "administer" includes a prescription for administration as well as the actual administration, and includes physical administration by the subject being treated or another.

[0023] As used herein, the terms "subject", "patient", or "individual" refer to any subject, patient, or individual, and these terms are used interchangeably herein. In this regard, the terms "subject", "patient", and "individual" include mammals, particularly humans. When used in combination with "needing it", the terms "subject", "patient", or "individual" mean any subject, patient, or individual having or at risk of having a particular symptom or disorder.

[0024] As used herein, the terms "therapeutically effective" or "effective" in the context of "dosage" or "amount" mean the dosage or amount that provides a particular pharmacological effect for which the compound is administered. Even if such a dosage is considered by one of ordinary skill in the art to be a therapeutically effective amount, it is emphasized that a therapeutically effective amount is not always effective to achieve the intended effect in a given subject. By way of convenience, exemplary dosages are provided herein. One of ordinary skill in the art can adjust such amounts according to the methods disclosed herein for treating a particular subject suffering from a particular condition or disorder. A therapeutically effective amount can vary based on the route of administration and the dosage form.

[0025] The terms "treat" or "treatment" cover the treatment of cancer as described herein in subjects such as humans and include the following: (i) suppressing cancer, i.e., slowing the progression of cancer; (ii) reducing cancer or a disorder, i.e., causing regression of cancer; (iii) slowing the progression of cancer; and / or (iv) inhibiting, alleviating, or slowing the progression of one or more symptoms of cancer. For example, treatment of cancer includes, but is not limited to, elimination of cancer or a condition caused by cancer, remission of a tumor, suppression of cancer, or reduction or elimination of at least one symptom of a tumor.

[0026] The term "analog" refers to a compound in which one or more individual atoms or functional groups are replaced by either a different atom or a different functional group, generally resulting in a compound having similar properties. In some embodiments, an analog refers to a structure that is similar to another but has one or two different components.

[0027] The term "derivative" refers to a compound formed from a similar starting compound by bonding another molecule or atom to the starting compound. Further, derivatives according to the present invention include one or more compounds formed from a precursor compound by addition of one or more atoms or molecules or by combining two or more precursor compounds.

[0028] Opioid growth factor receptor (OGFR) antagonist An "opioid growth factor receptor (OGFR) antagonist" refers to a molecule that inhibits, suppresses, or causes the termination of biological activity mediated by at least one OGFR, such as naloxone or its functional derivatives.

[0029] In some embodiments, OGFR antagonists are molecules that interfere with, block, or otherwise prevent the interaction or binding of met5-ligand (OGF) to OGFR. Met-5 is derived from the prohormone proenkephalin (PENK).

[0030] OGFR-binding antagonists may compete with met5-ligands for binding to OGFRs on the surface of the nuclear membrane, thereby interfering, blocking, or otherwise preventing the binding of met5-ligands to OGFRs without inducing downstream signaling otherwise induced by the binding of met5-ligands to OGFRs. Alternatively, OGFR-binding antagonists may bind to or sequester proenkephalin (PENK) or met5-ligands, substantially interfering, blocking, or otherwise preventing the binding of met5-ligands to OGFRs, thereby causing inhibition, repression, or termination of biological activity mediated by at least one OGFR. Generally speaking, OGFR-binding antagonists may be large molecules (e.g., antibodies) or small molecules (e.g., compounds with molecular weights of 15-kD, 12-kD, 10-kD, or even less than 8-kD), and may be polypeptides, nucleic acids, or synthetic small molecule compounds. OGFR-binding antagonists can be identified in any in vitro assay readily selected by those skilled in the art. For example, OGFR antagonists can be identified using the methods described in U.S. Patent No. 5,882,944, U.S. Patent No. 6,007,986, or U.S. Patent No. 6,270,979.

[0031] In one embodiment, the OGFR-binding antagonist is naloxone or a functional derivative thereof, naltrexone or a functional derivative thereof, or a combination thereof.

[0032] As used herein, “functional derivative” refers to a derivative or analog that is structurally and functionally similar to the original molecule (for example, maintaining the function of naltrexone or naloxone as an OGFR antagonist). Naloxone and naltrexone analogs can be synthesized using standard synthetic procedures, such as those described in March J., Advanced Organic Chemistry, 3rd Ed. (1985). Examples of functional derivatives of naltrexone and naloxone include salt forms, e.g., naloxone hydrochloride dihydrate or naltrexone hydrochloride. Additional examples of functional derivatives of naltrexone and naloxone suitable for use in the methods of the present invention include, for example, the naltrexone and naloxone analogs disclosed in U.S. Patent Application Publication No. 2007 / 0197573 A1, U.S. Patent No. 6,713,488.

[0033] In another embodiment, the OGFR-binding antagonist is derived from oxymorphone, binds to OGFR, and includes naloxone, naltrexone, nalorphine, naloxonazine, levallorphan, nalmefene, cyprodime, cyclorphan, cyclazosine, oxilorphan, LY113878, MR2266, diprenorphine, WIN 44,441-3, naltindole, or norbinaltolfimine.

[0034] In yet another embodiment, the OGFR-binding antagonist is derived from trans-3,4-dimethyl-4-phenylpiperidine and binds to OGFRs including LY99335, LY25506, LY117413, or LY255582.

[0035] In another embodiment, the OGFR-binding antagonist is derived from a met5-enkephalin or leu-enkephalin peptide and contains at least the following amino acid sequence as a means of binding to and targeting OGFR: Tyr-Gly-Gly-Phe-Met (SEQ ID NO: 1) if derived from met5-enkephalin, or Tyr-Gly-Gly-Phe-Leu (SEQ ID NO: 2) if derived from leu-enkephalin.

[0036] In yet another embodiment, the OGFR-binding antagonist is derived from the peptide antagonist 101174864 (N,N-diallyl-Tyr-Aib-Aib-Phe-Leu-OH, SEQ ID NO: 3; Aib = aminoisobutyric acid) or the somatostatin analog CTP (D-Phe-Cys-Tyr-D-Trp-Lys-Thr-Pen-Thr-NH.sub.2, SEQ ID NO: 4).

[0037] In other embodiments, the OGFR antagonist is a molecule that disrupts the nuclear localization sequence found within OGFR:251 QSALDYFMFAVRCRHQRRQLVHFAWEHFRPRCKFVWGPQDKLRRFKPSSL (SEQ ID NO: 5), rather than being an OGFR-binding antagonist.

[0038] In yet another embodiment, the OGFR antagonist used in the method of the present invention is a small hairpin RNA (shRNA) or small interfering RNA (siRNA) that is produced against the OGFR gene and is effective in disrupting OGFR gene expression.

[0039] The OGFR antagonists described herein may be administered individually or in combination. Suitable combinations include, for example, naloxone and naltrexone; naloxone and / or naltrexone in combination with another OGFR-binding antagonist or another OGFR antagonist.

[0040] Delivery systems and carriers for local administration In one embodiment of this disclosure, the OGFR antagonist is administered locally adjacent to the tumor site. In some embodiments, the OGFR antagonist is administered in a local dose of about 1 μM to about 1 mM. In some embodiments, the OGFR antagonist is administered in a local dose of about 1 mM to about 10 mM. In some embodiments, the local doses are 1 mM, 1.5 mM, 2.0 mM, 3.0 mM, 4.0 mM, 5.0 mM, 6.0 mM, 7.0 mM, 8.0 mM, 9.0 mM, and / or 10.0 mM.

[0041] In some embodiments, the OGFR antagonist is administered together with a carrier, the volume of which may be about 0.1 cubic centimeters (cc) to about 20 cc, about 0.25 cc to about 15 cc, about 0.5 cc to about 10 cc, about 1 cc to about 10 cc, or about 2 cc to about 10 cc. The corresponding amount of OGFR antagonist in the carrier may be about 0.2 mg / cc, 0.5 mg / cc, 1.0 mg / cc, 2.0 mg / cc, 3.0 mg / cc, 4.0 mg / cc, 5.0 mg / cc, 6.0 mg / cc, 7.0 mg / cc, 8.0 mg / cc, 9.0 mg / cc, and / or 10.0 mg / cc.

[0042] In some embodiments, OGFR antagonists are administered locally by intratumor injection.

[0043] In some embodiments, the OGFR antagonist is combined with a carrier for administration or encapsulated within the carrier.

[0044] Suitable carriers may be in the form of beads, microspheres, or nanoparticles and can be made from natural and / or synthetic biocompatible polymers. Examples of suitable biocompatible polymers include hyaluronic acid, collagen, tricalcium phosphate, chondroitin sulfate, polybutyrate, polylactide, polyglycolide, and lactide / glycolide copolymers, and mixtures or copolymers thereof. Suitable carriers also include on-polymer systems such as carboxylic acids, fatty acids, phospholipids, amino acids, lipids such as sterols, hydrogel-releasing systems; silastic systems; peptide-based systems; implants, etc.

[0045] In one embodiment, the carrier is a hygroscopic collagen-based carrier such as a collagen sponge, collagen scaffold, collagen powder, or collagen-based gelatin hydrogel.

[0046] In another embodiment, the carrier is a hydrophilic hydrogel-based carrier (e.g., polylactic acid, polyglycolic acid) which can release an OGFR antagonist (e.g., naloxone or naltrexone or a functional derivative thereof) injected therein for a certain period of time.

[0047] In another embodiment, the carrier is a triblock copolymer comprising a central block of PLA (two blocks of PEG-(poly-(ethylene glycol) adjacent to poly-(lactic acid)).

[0048] In yet another embodiment, the carrier is albumin, a derivative of albumin, or a fragment that maintains a naloxone / morphine binding site located at the interface between the IA domain and the IIA domain, and / or maintains a naloxone binding site around tryptophan (Trp)-214 that binds to an OGFR antagonist such as naloxone or naltrexone or a functional derivative thereof, enabling the sustained release of the OGFR antagonist.

[0049] In yet another embodiment, the gel is inert, which binds methylcellulose to an OGFR antagonist such as naloxone or naltrexone or a functional derivative thereof, thereby enabling the sustained release of the antagonist.

[0050] In further embodiments, the carrier is a bovine collagen implant. OGFR antagonists, such as naloxone or naltrexone or their functional derivatives, can be combined with a bovine collagen implant in a manner similar to that of INFUSE (BMP2) or OP1 (bone morphogenetic protein 1 or BMP7)-putty or OP1-implant, which are supplemented with bovine collagen sponge, powdered bovine collagen, or collagen-based gelatin constructs. Administration of naloxone, naltrexone or their functional derivatives can be achieved, for example, by reconstituting powdered naloxone or powdered naltrexone or their functional derivatives with sterile saline, and then adding the OGFR antagonist-saline mixture to the collagen implant; the implant can then be delivered topically to the surgical intervention site.

[0051] In further embodiments, the carrier comprises cement (polymethyl methacrylate or "PMMA") or an injectable formulation.

[0052] In another embodiment, the carrier is composed of PGA(poly-(glycolic acid)-PLGA(poly-(glycolic acid lactate)) spheres and can encapsulate an OGFR antagonist to provide immediate release, delayed release, or sustained release.

[0053] Pharmaceutical composition and administration In some embodiments, the pharmaceutical compositions disclosed herein include one or more "pharmaceutically acceptable carriers," such as aqueous carriers, buffers, antioxidants, and / or diluents. In some embodiments, the pharmaceutical composition includes an OGFR antagonist in a dimethyl sulfoxide (DMSO)-based saline. In some embodiments, the pharmaceutical composition includes an OGFR antagonist in a dimethyl sulfoxide (DMSO)-based saline at a concentration greater than approximately 100 mM. In some embodiments, the pharmaceutical composition includes an OGFR antagonist in a dimethyl sulfoxide (DMSO)-based saline at a concentration of approximately 10 mM to approximately 150 mM. In some embodiments, the pharmaceutical composition includes an OGFR antagonist in a dimethyl sulfoxide (DMSO)-based saline at a concentration of approximately 20 mM to approximately 140 mM. In some embodiments, the pharmaceutical composition includes an OGFR antagonist in a dimethyl sulfoxide (DMSO)-based saline at a concentration of approximately 30 mM to approximately 130 mM. In some embodiments, the pharmaceutical composition contains about 40 mM to about 120 mM of an OGFR antagonist in a dimethyl sulfoxide (DMSO)-based saline. In some embodiments, the pharmaceutical composition contains about 50 mM to about 110 mM of an OGFR antagonist in a dimethyl sulfoxide (DMSO)-based saline. In some embodiments, the pharmaceutical composition contains about 50 mM to about 110 mM of an OGFR antagonist in a dimethyl sulfoxide (DMSO)-based saline. In some embodiments, the pharmaceutical composition contains more than 100 mM of an OGFR antagonist in a dimethyl sulfoxide (DMSO)-based saline. In some embodiments, the OGFR antagonist contains about 100 mM of naloxone, naltrexone, or their functional derivatives or analogs in a dimethyl sulfoxide (DMSO)-based saline.

[0054] In some embodiments, the acidified saline solution-based solution has a pH of approximately 4.5 to approximately 7.4. In some embodiments, the acidified saline solution-based solution has a pH of approximately 5.5 to approximately 7.4. In some embodiments, the acidified saline solution-based solution has a pH of approximately 6.5 to approximately 7.4.

[0055] In some embodiments, the diluent is a dimethyl sulfoxide (DMSO)-based saline. In some embodiments, the DMSO-based saline contains about 1% volume / volume (v / v) DMSO to about 80% v / v DMSO. In some embodiments, the DMSO-based saline contains about 5% volume / volume (v / v) DMSO to about 80% v / v DMSO. In some embodiments, the DMSO-based saline contains about 10% volume / volume (v / v) DMSO to about 80% v / v DMSO. In some embodiments, the DMSO-based saline contains about 15% volume / volume (v / v) DMSO to about 80% v / v DMSO. In some embodiments, the DMSO-based saline contains about 25% volume / volume (v / v) DMSO to about 80% v / v DMSO. In some embodiments, the DMSO-based saline contains about 30% volume / volume (v / v) DMSO to about 80% v / v DMSO. In some embodiments, the DMSO-based saline contains approximately 35% volume / volume (v / v) DMSO to approximately 80% v / v DMSO. In some embodiments, the DMSO-based saline contains approximately 40% volume / volume (v / v) DMSO to approximately 80% v / v DMSO. In some embodiments, the DMSO-based saline contains approximately 40% volume / volume (v / v) DMSO to approximately 70% v / v DMSO.

[0056] In some embodiments, the DMSO-based saline contains about 1% volume / volume (v / v) of DMSO. In some embodiments, the DMSO-based saline contains about 2% volume / volume (v / v) of DMSO. In some embodiments, the DMSO-based saline contains about 3% volume / volume (v / v) of DMSO. In some embodiments, the DMSO-based saline contains about 4% volume / volume (v / v) of DMSO. In some embodiments, the DMSO-based saline contains about 5% volume / volume (v / v) of DMSO. In some embodiments, the DMSO-based saline contains about 6% volume / volume (v / v) of DMSO. In some embodiments, the DMSO-based saline contains about 7% volume / volume (v / v) of DMSO. In some embodiments, the DMSO-based saline contains about 8% volume / volume (v / v) of DMSO. In some embodiments, the DMSO-based saline contains about 9% volume / volume (v / v) of DMSO. In some embodiments, the DMSO-based saline contains about 10% volume / volume (v / v) of DMSO. In some embodiments, the DMSO-based saline contains about 20% volume / volume (v / v) of DMSO. In some embodiments, the DMSO-based saline contains about 30% volume / volume (v / v) of DMSO. In some embodiments, the DMSO-based saline contains about 40% volume / volume (v / v) of DMSO. In some embodiments, the DMSO-based saline contains about 50% volume / volume (v / v) of DMSO. In some embodiments, the DMSO-based saline contains about 60% volume / volume (v / v) of DMSO. In some embodiments, the DMSO-based saline contains about 70% volume / volume (v / v) of DMSO. In some embodiments, the DMSO-based saline contains about 80% volume / volume (v / v) of DMSO.

[0057] In some embodiments, the dimethyl sulfoxide (DMSO)-based saline solution includes phosphate-buffered saline, borate-buffered saline, Tris-buffered saline, or carbonate-buffered saline.

[0058] In some embodiments, physiological saline consists of salt and water. In some embodiments, the salt in physiological saline contains sodium chloride or potassium chloride. In some embodiments, physiological saline contains about 0.7% w / w salt to about 1.5% w / w salt. In some embodiments, physiological saline contains about 0.7% salt. In some embodiments, physiological saline contains about 0.8% salt. In some embodiments, physiological saline contains about 0.9% salt. In some embodiments, physiological saline contains 1.0% salt. In some embodiments, physiological saline contains about 1.1% salt. In some embodiments, physiological saline contains about 1.2% salt. In some embodiments, physiological saline contains about 1.3% salt. In some embodiments, physiological saline contains about 1.4% salt. In some embodiments, physiological saline contains about 1.5% salt.

[0059] OGFR antagonists may be administered in combination with or in conjunction with an appropriate carrier or medium, depending on the route of administration. The term “pharmaceutically acceptable carrier” refers to a carrier that has been conventionally used in the art to store, administer, and / or enhance the therapeutic effects of the active ingredient of a pharmaceutical composition.

[0060] The aqueous liquid carrier may contain pharmaceutically acceptable additives such as acidifying agents, alkalizing agents, antimicrobial and antiseptic agents, antioxidants, buffering agents, chelating agents, complexing agents, solubilizing agents, humectants, solvents, suspending agents and / or thickeners, isotonic agents, wetting agents, or other biocompatible materials. A table of the components listed in the above categories is found in the United States Pharmacopeia, National Collection of Drugs, 1857–1859 and (1990). Some examples of materials that function as pharmaceutically acceptable carriers include sugars (lactose, glucose, and sucrose, etc.); cyclodextrins, including alpha-cyclodextrin, beta-cyclodextrin, and gamma-cyclodextrin; starches (corn starch and potato starch, etc.); cellulose and its derivatives (sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate, etc.); powdered tragacanth; malt; gelatin; talc; excipients (cocoa butter and suppository wax, etc.); oils (peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil). Oils, etc.; glycols such as propylene glycol; polyols (glycerin, sorbitol, mannitol, and polyethylene glycol, etc.); esters (ethyl oleate and ethyl laurate, etc.); agar; buffers (magnesium hydroxide and aluminum hydroxide, etc.); alginic acid; pyrogen-free water; isotonic saline; Ringer's solution, ethyl alcohol and phosphate buffer, and other non-toxic, suitable substances used in pharmaceutical formulations. Wetting agents, emulsifiers, lubricants, and colorants, release agents, coating agents, sweeteners, flavoring agents, and fragrances such as sodium lauryl sulfate and magnesium stearate.

[0061] In some embodiments, the pharmaceutical composition includes preservatives and antioxidants. Examples of pharmaceutically acceptable antioxidants include water-soluble antioxidants (such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfite, sodium metabisulfite, and sodium sulfite); oil-soluble antioxidants (such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, and α-tocopherol); and metal chelating agents (such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, and phosphoric acid).

[0062] The pharmaceutical compositions according to the present invention may also include one or more binders, fillers, lubricants, suspending agents, sweeteners, flavoring agents, preservatives, buffers, wetting agents, disintegrants, foaming agents, and other excipients. Such excipients are known in the art. Examples of fillers include lactose monohydrate, anhydrous lactose, and various starches; examples of binders include various celluloses and crosslinked polyvinylpyrrolidone, microcrystalline cellulose (Avicel® PH101 microcrystalline cellulose and / or Avicel® PH102 microcrystalline cellulose, and silicified microcrystalline cellulose such as ProSolv SMCC®). Suitable lubricants, including agents that act on the flowability of the powder to be tableted, may include colloidal silicon dioxide such as Aerosil® 200 (colloidal silicon dioxide), talc, stearic acid, magnesium stearate, calcium stearate, and silica gel. Examples of sweeteners may include any natural or artificial sweetener such as sucrose, xylitol, sodium saccharin, cyclamate, aspartame, and acesulfame. Examples of flavorings include monoammonium glycylliginates such as Magnasweet® (a flavor composition containing monoammonium glycylliginate and the MAFCO trademark), bubblegum flavors, and fruit flavors. Examples of preservatives include potassium sorbate, methylparaben, propylparaben, benzoic acid and its salts, other esters of parahydroxybenzoic acid such as butylparaben, alcohols such as ethyl or benzyl alcohol, phenolic compounds such as phenol, or quaternary compounds such as benzalkonium chloride.

[0063] Any pharmaceutical product used for therapeutic administration may be sterile. Sterilization can be easily achieved, for example, by filtration using a sterile filtration membrane (e.g., a 0.2 micron membrane). Any pharmaceutically acceptable sterilization method can be used in the compositions of the present invention.

[0064] Pharmaceutical compositions containing OGFR antagonist derivatives or salts thereof are formulated and administered in a manner consistent with good medical practice, taking into account the individual patient's clinical condition, method of administration, schedule of administration, and other factors known to the practicing physician.

[0065] Various routes of administration can be utilized. The pharmaceutical compositions of the present invention can be administered using any medically acceptable mode of administration, i.e., any mode of administration that produces an effective level of the active ingredient without causing clinically unacceptable side effects. Accordingly, the pharmaceutical compositions can be administered to a subject parenterally, orally, intraperitoneally, intravenously, intra-arterially, percutaneously, sublingually, intramuscularly, rectally, transbuccally, intranasally, intraliposomally, via minicells, via antibody binding, via cell-targeted peptides, via inhalation, via vaginal, intraocular, via local delivery by catheter or stent, subcutaneously, intrafatally, intra-articularly, or intrathecally.

[0066] The mode of administration includes oral, rectal, topical, nasal, intradermal, or parenteral routes. The term "parenteral" includes subcutaneous, intravenous, intramuscular, or infusion routes. Intravenous or intramuscular routes are not particularly suitable for long-term treatment and prophylaxis. Oral administration is used for prophylactic treatment due to patient convenience and medication scheduling.

[0067] Parenteral formulations include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcohol / aqueous solutions, emulsions, or suspensions containing physiological saline and buffer medium. Parenteral media include sodium chloride solution, Ringer's glucose, glucose and sodium chloride, Ringer's lactate solution, or fixed oil. Intravenous administration media include liquids and nutrient replacement solutions, electrolyte replacement solutions (such as those based on Ringer's glucose), etc. Preservatives and other additives, such as antibacterial agents, antioxidants, chelating agents, and inert gases, may also be present. Lower doses will result from other forms of administration, such as intravenous administration. If the subject's response is insufficient with the initial dose applied, higher doses (or effectively higher doses via different, more localized delivery routes) may be used, to the extent that the patient's tolerance allows. Multiple daily doses are intended to achieve appropriate systemic levels of the compound.

[0068] Compositions suitable for oral administration may be presented as individual units such as capsules, tablets, or lozenges, each containing a predetermined amount of the active ingredient. Other compositions may include suspensions in aqueous liquids or non-aqueous liquids such as syrups, elixirs, or emulsions.

[0069] Other delivery systems may include time-release, delayed-release, or sustained-release delivery systems. Such systems can avoid repeated administration of the pharmaceutical composition of the present invention, improving convenience for both the subject and the physician. Many types of release delivery systems are available and known to those skilled in the art. They include polymer-based systems such as poly(lactide-glycolides), copolyoxalates, polycaprolactones, polyesteramides, polyorthoesters, polyhydroxybutyric acid, and polyanhydrides. Microcapsules of the aforementioned polymers containing drugs are described, for example, in U.S. Patent No. 5,075,109. Delivery systems also include non-polymer systems such as: lipids containing sterols such as cholesterol, cholesterol esters, and fatty acids, or neutral fats such as monoglycerides, diglycerides, and triglycerides; hydrogel-release systems; silastic systems; peptide-based systems; wax coatings; compressed tablets using conventional binders and excipients; partially fused implants; etc.

[0070] The methods and compositions described herein may be provided in the form of kits. “Kit” is defined herein as a package and includes several separate parts that exhibit complementary effects when applied together. In this embodiment, the effects achieved by the kit and the pharmaceutical composition are similar. The kit may optionally include instructions for using the pharmaceutical composition.

[0071] The present invention will be further illustrated by the following embodiments, but will not be limited thereto. [Examples]

[0072] Example 1 - Naloxone exhibits a selective cytotoxic effect against tumor cells compared to non-tumor cells. The objective of this experiment was to measure the antitumor growth activity of naloxone solubilized in acidified saline containing 50% (v / v) DMSO compared to other non-tumor cell phenotypes likely to be located adjacent to tumors. The naloxone preparation used in this example is referred to as "naloxone" in this specification.

[0073] The effects of naloxone treatment at doses of 1 mM, 1.5 mM, 2 mM, 2.5 mM, and 5 mM on lactate dehydrogenase (LDH) in triple-negative breast cancer cells (BT474), osteosarcoma cells (SaOS2), and breast cancer cells (BT20) were compared with those in non-tumor cells (HUMEC) and normal vascular endothelial cells (HUVEC), as shown in Figure 1. Naloxone was added for 3 hours, and LDH release was assessed at 24 hours post-treatment. At 24 hours, a significant increase in LDH, adjusted for changes in cell number caused by treatment, was observed in triple-negative breast cancer cells (BT474), osteosarcoma cells (SaOS2), and breast cancer cells (BT20) compared to normal cells. As shown in Figure 1, a 6-fold increase in LDH levels was observed in SaOS2 and BT20 cells, and a 3-fold increase in cellular LDH levels was observed in BT474 cells (*p<0.001). At the highest dose, the number of SaOS2 cells decreased by 60%, while the number of BT20 and BT90 cells decreased by 90% after 72 hours. See Figure 2.

[0074] As shown in Figure 1, 72-hour exposure of triple-negative breast cancer (TNBC) cells BT474 resulted in a dose-dependent, significantly reduced number of metabolically active cells compared to control non-tumor cells, namely breast epithelial (HUMEC) cells or vascular endothelial (HUVEC) cells. These data suggest that naloxone exhibits significant antitumor activity and may provide a non-tumor cell-preserving effect up to a dose of 2.5 mM.

[0075] Naloxone treatment was also demonstrated to reduce tumor size in vitro. Malignant tumors are characterized by invasion into surrounding normal tissue, and the spheroid assay is a simple microplate method (based on homogeneous self-organizing 3D tumor spheroids) routinely used to assess tumor growth. The antitumor effect of naloxone was evaluated using the spheroid assay, as shown in Figure 3. The antitumor effect was determined by quantifying the size of the spheroids formed and calculating the percentage change in spheroid area from 0 to 120 hours after treatment, as shown in Figure 3B. The antitumor effect was also determined by quantifying the percentage change in the cell area fraction of each spheroid at 120 hours after treatment, as determined by staining the cells with crystal violet, as shown in Figure 3C. The results shown in Figure 3 indicate that naloxone dose-dependently reduced the size and cell solidity of tumor masses in SaOS2 osteosarcoma cells and BT474 TNBC cells.

[0076] Therefore, the results herein demonstrate that naloxone has a selective cytotoxic effect on tumor cells while preserving non-tumor cells. Furthermore, the results show that naloxone can inhibit tumor cell growth in vitro.

[0077] Example 2 - This example demonstrates that topical administration of a 50% DMSO (50%) formulation in approximately 0.9% saline solution in a collagen sponge adjacent to a growing tumor effectively inhibits in vivo tumor growth and increases the survival rate of mice inoculated with mammary cancer.

[0078] To study the effect of this formulation on in vivo tumor growth, MDA-MB-231 triple-negative breast cancer cells were transplanted into female NCr nude mice. Three weeks after inoculation with MDA-MB-231 triple-negative breast cancer cells in mice in which tumor growth was confirmed to be active, the formulation was transplanted into the mice. The formulation was prepared by dissolving naloxone in acidified saline containing 50% (v / v) DMSO in a collagen gel-based carrier. The formulation was implanted into the tibia of 8-week-old female NCr nude mice with actively growing tumors formed by the transplanted MDA-MB-231 triple-negative breast cancer cells. Figure 4 shows the effect of the formulation on tumor growth in mice transplanted with MDA-MB-231 triple-negative breast cancer cells. Figure 4A shows that tumor volume was measured with calipers over 60 days. Tumor volume was half the size in the formulation implant group. Regression analysis found that the slopes of the two treatment groups were significantly different (p<0.001). Figure 4B shows the assessment of survival rates using Kaplan-Meier statistics. Survival rates were twice as high in the drug treatment group (p<0.0328).

[0079] All references cited herein are incorporated by reference to the same extent as they are incorporated individually. While preferred embodiments have been illustrated and described herein, it should be understood that modifications and alterations can be made by those skilled in the art without departing from the invention in broader embodiments as defined herein.

[0080] Example 3 - Local injection of an OGFR antagonist directly into or adjacent to a tumor inhibits tumor growth in human patients. This embodiment demonstrates that delivering OGFR antagonists directly to or adjacent to tumors may suppress tumor growth in human patients with cancer.

[0081] OGFR antagonists are administered to human patients with cancers such as breast cancer, liver cancer, lung cancer, skin cancer, brain cancer, or prostate cancer. OGFR antagonists are administered as pharmaceutical formulations of naloxone, dimethyl sulfide, saline, pharmaceutically acceptable excipients, and a hydrogel-based carrier, as described herein. The volume of the pharmaceutical formulation is between 0.25 cubic centimeters (cc) and 10 cc; or between 10% and 150% of the tumor volume. The dose of naloxone as an OGFR antagonist ranges from 2 milligrams (mg) per cc of carrier to approximately 8 mg per cc of carrier. Specifically, the doses of naloxone as an OGFR antagonist are 2 mg per cc of carrier, 4 mg per cc of carrier, or 8 mg per cc of carrier. The amount of carrier and the amount of OGFR antagonist vary depending on the type of cancer and the method of delivery, as shown below.

[0082] For some patients, pharmaceutical formulations containing OGFR antagonists are suitable for injection. Injectable OGFR antagonist compositions are injected directly into solid primary or metastatic tumors. For some patients with lung cancer, formulations containing OGFR antagonists are suitable for inhalation, and these formulations are delivered via inhalation.

[0083] In liver cancer patients, pharmaceutical formulations containing OGFR antagonists are injected into the viscera and deep tissue of the liver of patients with liver cancer using fluorescence fluoroscopy-guided injection. Alternatively, to treat liver cancer patients, pharmaceutical formulations containing OGFR antagonists are delivered to the tumor via local perfusion through a catheter. Furthermore, once the liver tumor is removed, pharmaceutical formulations containing OGFR antagonists are delivered to the tissue layers adjacent to the tumor site to control tumor recurrence.

[0084] Pharmaceutical formulations containing OGFR antagonists are also used in the treatment of lung cancer. These formulations are injected into the visceral and deep tissues of the lungs of patients with lung cancer. Therefore, in patients with lung cancer, OGFR antagonist formulations are injected directly into or adjacent to the tumor during tumor resection or lobectomy. In some cases, OGFR antagonist formulations are adapted for inhalation and are delivered to patients with lung cancer via inhalation.

[0085] In patients with brain tumors, pharmaceutical formulations containing OGFR antagonists are administered by injection during tumor resection or via a port or catheter before or after tumor resection.

[0086] Pharmaceutical formulations containing OGFR antagonists are also used in the treatment of skin cancer. These formulations are injected into or adjacent to skin tumors to treat patients with skin cancer. In patents where the tumor is a skin tumor located in the skin, the skin tumor is treated by scraping or excision of the tumor, after which the pharmaceutical formulation containing the OGFR antagonist is delivered via conventional means that do not involve injection (i.e., placement of the hand to the excision and / or packing by hand). During incisional biopsy or partial excision of skin tumors such as melanoma, squamous cell carcinoma, or basal cell carcinoma, the pharmaceutical formulation containing the OGFR antagonist is injected into the excision space and / or adjacent to the tumor. For the treatment of skin cancer, the volume of the pharmaceutical composition containing the OGFR antagonist is between 0.25cc and 2.5cc, or between 10% and 150% of the tumor volume. The dosage of naloxone, an OGFR antagonist, ranges from 2 milligrams (mg) per cc of carrier to approximately 8 mg per cc of carrier. In one patient with skin cancer, the corresponding dose of naloxone as an OGFR antagonist is 2 mg per cc of carrier. In another skin cancer patient, the corresponding dose of naloxone as an OGFR antagonist is 4 mg per cc of carrier. In yet another skin cancer patient, the corresponding dose of naloxone as an OGFR antagonist is 8 mg per cc of carrier.

[0087] To treat breast tumors, pharmaceutical formulations containing OGFR antagonists are delivered directly to the breast tumor during a wire localization or needle localization procedure (i.e., stereotactic wire localization) in which a thin, hollow needle is guided to the tumor using mammography or ultrasound. In this procedure, a portion of breast tissue is removed for biopsy, and then a second needle containing the preferred formulation is injected into and adjacent to the tumor mass. To treat breast cancer, the volume of the pharmaceutical composition containing the OGFR antagonist is between 1 cc and 10 cc, or between 10% and 150% of the tumor volume. The dose of naloxone, which is an OGFR antagonist, ranges from 2 milligrams (mg) per cc of carrier to approximately 8 mg per cc of carrier. For a single patient with breast cancer, the corresponding dose of naloxone, which is an OGFR antagonist, is 2 mg per cc of carrier. In another breast cancer patient, the corresponding dose of naloxone, an OGFR antagonist, was 4 mg per cc of carrier. In yet another breast cancer patient, the corresponding dose of naloxone, an OGFR antagonist, was 8 mg per cc of carrier.

Claims

1. A pharmaceutical composition for treating target cancers requiring the same, comprising (a) at least 1 mM opioid growth factor receptor (OGFR) antagonist selected from the group consisting of naloxone, naltrexone, and combinations thereof, and (b) a diluent comprising acidified DMSO-based saline, which is administered topically to a target site including a cancerous tumor.

2. The pharmaceutical composition according to claim 1, wherein the cancer is soft tissue non-osteogenic sarcoma, bone sarcoma, osteosarcoma, Ewing's sarcoma, chondrosarcoma, benign bone tumor, basal cell carcinoma, melanoma, thyroid cancer, gliablastoma, pituitary tumor, oligodendroglioma, bladder cancer, triple-negative breast cancer, breast cancer, neuroblastoma, or astrocytoma.

3. The pharmaceutical composition according to claim 1, wherein the cancer includes triple-negative breast cancer.

4. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition is administered locally by intratumor injection.

5. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition comprises a pharmaceutically acceptable carrier.

6. The pharmaceutical composition according to claim 5, wherein the pharmaceutically acceptable carrier comprises a collagen-based carrier in the form of a collagen sponge, powdered collagen, or a collagen-based gelatin hydrogel.

7. The pharmaceutical composition according to claim 6, wherein the pharmaceutically acceptable carrier is the collagen sponge.

8. The pharmaceutical composition according to claim 5, wherein the pharmaceutically acceptable carrier comprises a hydrogel-based carrier.

9. The pharmaceutical composition according to claim 8, wherein the hydrogel-based carrier comprises a poly-(butyric acid) / poly-(ethylene glycol) / poly-(butyric acid) hydrogel, a poly-(ethylene glycol) / poly-(butyric acid) / poly-(ethylene glycol) hydrogel, or a combination thereof.

10. The pharmaceutical composition according to claim 9, wherein the hydrogel-based carrier is poly(ethylene glycol) / poly(butyric acid) / poly(ethylene glycol).

11. The pharmaceutical composition according to claim 1, wherein the acidified DMSO-based physiological saline comprises 1% volume / volume (v / v) DMSO to 80% v / v DMSO.

12. The pharmaceutical composition according to claim 1, wherein the acidified DMSO-based physiological saline contains 50% v / v DMSO.

13. The pharmaceutical composition according to claim 1, wherein the OGFR antagonist is naloxone, and the diluent enhances the solubility of the naloxone to 150 mM.

14. The pharmaceutical composition according to claim 1, wherein the volume of the pharmaceutical composition is between 10% and 150% of the tumor volume.

15. The pharmaceutical composition according to claim 1, comprising 10 mM to 150 mM of the OGFR antagonist.

16. The pharmaceutical composition according to claim 1, comprising 20 mM to 140 mM of the OGFR antagonist.

17. The pharmaceutical composition according to claim 1, comprising 30 mM to 130 mM of the OGFR antagonist.

18. The pharmaceutical composition according to claim 1, comprising 40 mM to 120 mM of the OGFR antagonist.

19. The pharmaceutical composition according to claim 1, comprising 50 mM to 110 mM of the OGFR antagonist.

20. The pharmaceutical composition according to claim 1, comprising the OGFR antagonist in a concentration exceeding 100 mM.

21. The pharmaceutical composition according to claim 1, wherein the OGFR antagonist is naloxone.

22. A pharmaceutical composition for treating triple-negative breast cancer in a subject requiring such treatment, comprising a hydrogel carrier and an effective amount of an opioid growth factor receptor (OGFR) antagonist, wherein the OGFR antagonist is selected from naloxone, naltrexone, and combinations thereof, and the pharmaceutical composition is administered topically to a tumor site in a subject containing triple-negative breast cancer cells, and a single topical administration of the pharmaceutical composition results in a reduction of tumor volume compared to an untreated subject.

23. The pharmaceutical composition according to claim 22, wherein the concentration of the OGFR antagonist in the pharmaceutical composition is 1 mM to 10 mM.

24. The pharmaceutical composition according to claim 22, wherein the pharmaceutical composition is administered locally by intratumor injection.

25. The pharmaceutical composition according to claim 22, wherein the hydrogel carrier comprises a collagen-based gelatin hydrogel.

26. The pharmaceutical composition according to claim 22, wherein the hydrogel carrier comprises a poly-(butyric acid) / poly-(ethylene glycol) / poly-(butyric acid) hydrogel, a poly-(ethylene glycol) / poly-(butyric acid) / poly-(ethylene glycol) hydrogel, or a combination thereof.

27. The pharmaceutical composition according to claim 26, wherein the hydrogel carrier is poly-(ethylene glycol) / poly-(butyric acid) / poly-(ethylene glycol).

28. The pharmaceutical composition according to claim 22, wherein the pharmaceutical composition comprises a diluent, wherein the diluent enhances the solubility of the OGFR antagonist.

29. The pharmaceutical composition according to claim 28, wherein the diluent comprises a dimethyl sulfoxide (DMSO)-based saline solution or an acidified saline solution.

30. The pharmaceutical composition according to claim 29, wherein the diluent is a dimethyl sulfoxide (DMSO)-based physiological saline solution.

31. The pharmaceutical composition according to claim 30, wherein the DMSO-based saline solution comprises 1% volume / volume (v / v) DMSO to 80% v / v DMSO.

32. The pharmaceutical composition according to claim 30, wherein the DMSO-based physiological saline contains 50% v / v DMSO.

33. The pharmaceutical composition according to claim 29, wherein the acidified saline-based solution has a pH of 4.5 to 7.4.

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