Use of (S)-3-Amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid in the treatment of cancer

Abstract

Methods and compositions are provided for use in treating cancer and inhibiting the growth and metastasis of cancer, which comprise (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof.

Description

Technical Field

[0001] (Cross - reference to related applications) This application claims the priority of U.S. Provisional Patent Application 63 / 358,677, filed on July 6, 2022, the entire content of which is incorporated herein by reference.

[0002] (Field of the Invention) (Treatment of cancer with (S)-3 - amino - 4-(difluoromethylene)cyclopent - 1 - en - 1 - carboxylic acid or a pharmaceutically acceptable salt thereof)

Background Art

[0003] According to the World Health Organization (https: / / www.who.int / news - room / fact - sheets / detail / cancer), cancer is a general term for a large group of diseases that can affect any part of the body. The terms malignant tumor and neoplasm are also used. Cancer is a leading cause of death worldwide, with approximately 10 million deaths in 2020, accounting for nearly one in six deaths. The defining feature of cancer is the rapid formation of abnormal cells that grow beyond normal boundaries and invade adjacent parts of the body and spread to other organs, and the latter process is called metastasis. Widespread metastasis is the main cause of death from cancer.

[0004] Medulloblastoma (MB) is the most common pediatric malignant brain tumor. According to OMIM#155255, MB accounts for 16% of all pediatric brain tumors, and 40% of all cerebellar tumors in childhood are MB. Approximately 10 - 15% of medulloblastomas are diagnosed in infancy. MB occurs in a bimodal pattern, with peaks in incidence at 3 - 4 years and 8 - 9 years. MB is less than 1% of central nervous system (CNS) tumors in adults, and has the highest incidence in adults aged 20 - 34 years.

[0005] MB is defined by four molecular subgroups (Wnt, Shh, group 3, group 4) based on transcriptional and epigenetic profiles. See Manoranjan, et al. Nat Commun 11, 4323 (2020). Wnt (wingless) MB and Shh (sonic hedgehog) MB are named after signaling pathways thought to play important roles in the etiology of their subgroups. The Wnt signaling pathway is a group of signaling pathways that begin with proteins that transmit signals into cells via cell surface receptors. NT-activated medulloblastoma is defined by activating mutations in the WNT / β-catenin signaling pathway (e.g., CTNNB1 or germline APC), is often associated with loss of chromosome 6, and shows the best prognosis. Manoranjan, et al., supra. Wnt MB accounts for 10% of cases. Ibid.

[0006] The Shh signaling pathway is one of the major trafficking networks that control important events during development, namely the growth and patterning of the multicellular embryo. Abnormalities in the control and transmission of the Shh signaling pathway are involved in congenital defects, tissue regeneration, stem cell renewal, and cancer growth. Choudhry et al., Ann Neurosci. 2014;21(1):28-31. SHH-activated medulloblastoma (SHH-MB) accounts for 25-30% of all MB, occurs with a bimodal age distribution, includes many cases in infants and adults, but few pediatric cases. Menyhart and Gyorffy, Annals of Clinical and Translational Neurology 2019; 6(5): 990-1005. Different age groups are characterized by different survival outcomes and age-specific changes in regulatory pathways. Ibid.

[0007] The MBs in group 3 have a photoreceptor / retinal expression signature, and the MBs in group 4 express neuronal genes. Tamayo-Orrego, Lukas, and Frederic Charron. 1000Research vol. 8 F1000 Faculty Rev-1823. 29 Oct. 2019. Currently, it is thought that each different MB group is derived from specific progenitor cells and, therefore, determines the clinical and molecular behavior of the disease. Ibid.

[0008] Glioma is a type of brain tumor that develops from glial cells, which are special cells that surround and support the brain's nerve cells. According to the Genetic and Rare Diseases Information Center (GARD) of the National Institutes of Health (NIH) in the United States, gliomas are generally classified by the type of glial cell involved in the tumor: astrocytoma - a tumor that develops from star-shaped glial cells called astrocytes; ependymoma - a tumor that develops from ependymal cells lining the ventricles of the brain and the center of the spinal cord; oligodendroglioma - a tumor that affects oligodendrocytes, the myelin-forming glia of the central nervous system. Gliomas are classified on a scale of 1 to 4. Grade 1 gliomas usually grow slowly and often exhibit more benign behavior. Grade 2 and grade 3 gliomas grow more rapidly and often require more aggressive treatment. Grade 4 gliomas are the most aggressive type and are also known as glioblastomas. The symptoms of gliomas vary by type but may include headache, nausea and vomiting, confusion, personality changes, balance problems, vision problems, speech problems, and / or seizures. In research, 8 out of 19 members of the WNT, namely WNT1, 2, 2B, 3A, 5A, 6, 7A, and 7B, have been shown to be associated with the development of gliomas. Xu et al., Bioscience reports vol. 40 (3) (2020).

[0009] Breast cancer starts in breast tissue, and the cells usually form either in situ or invasive tumors. According to the NIH, breast cancer is the second most common cancer in women in the United States. Rarely, men can also get it. Certain gene markers such as BRCA1, BRCA2, and HER2 are associated with the risk of breast cancer, which may also increase the risk of ovarian cancer and other cancers. The types of breast cancer are determined by which breast cells become cancerous. Ductal carcinoma, which starts in the cells of the milk ducts and is the most common type; lobular carcinoma, which starts in the lobules and is more often seen in both breasts compared to other types of breast cancer; inflammatory breast cancer, where cancer cells block the lymphatic vessels in the skin of the breast; and Paget's disease of the breast, which is a cancer that involves the skin of the nipple. Examples of signs and symptoms of breast cancer include a new lump or thickening in or near the breast or in the armpit; changes in the size or shape of the breast; dimpling or puckering of the skin of the breast; nipple retraction inward; discharge from the nipple other than breast milk; scaling, redness, or swelling of the skin around the nipple or of the breast; and pain in any part of the breast. According to several studies approved by the National Library of Medicine, GABA has important prognostic value in breast cancer. The metastasis of breast cancer overexpresses many variables related to GABA, proliferates by metabolizing GABA as a biosynthetic energy source, and enables metastasis to the brain and cerebrospinal fluid.

[0010] Squamous cell carcinoma is the second most common form of skin cancer, characterized by abnormal and accelerated growth of the cells that make up the middle and outer layers of the skin. According to the Mayo Clinic and Healthline, squamous cell carcinoma of the skin can be aggressive in some cases but usually does not threaten life. Squamous cell carcinoma can also occur in other parts of the body such as the mouth and lungs, but this is much rarer. Many cases of squamous cell carcinoma of the skin are caused by long-term exposure to ultraviolet (UV) rays. Signs and symptoms of squamous cell carcinoma of the skin include hard, red nodules; flat sores with scaly crusts; new sores or raised areas that develop on old scars or ulcers; something that can progress to an open sore with scaly, flaky patches that develop on the lips; red sores or scaly patches that develop in the mouth; and red, raised patches or wart-like sores that develop on the anus or genitals.

[0011] Lung cancer occurs in the lungs and may spread to lymph nodes and other organs in the body. Approximately 40% of lung cancer patients develop brain metastases. According to the US Centers for Disease Control and Prevention (CDC), lung cancer is mainly classified into two types: small cell carcinoma and non-small cell carcinoma (including adenocarcinoma and squamous cell carcinoma). The growth and treatment methods vary for these types of lung cancer. Non-small cell lung cancer is more common than small cell lung cancer. Smokers have the highest risk of lung cancer. Lung cancer usually does not cause signs or symptoms in the early stages. The signs and symptoms of lung cancer typically appear when the disease progresses and may include a new, persistent cough that does not go away; coughing up blood; shortness of breath; chest pain; hoarseness; weight loss without trying; bone pain; and headaches.

[0012] Peritoneal cancer refers to cancer that has originated or spread within the peritoneal cavity. According to the NIH, even if cancer starts in the peritoneum or reaches the peritoneum from other sites (i.e., ovaries, colon, appendix, etc.), it is considered advanced at the time of reaching the peritoneum and may be referred to as peritoneal carcinomatosis. In patients in the early stages, there are often few symptoms until the disease progresses. When symptoms appear, they are often vague, non-specific symptoms such as fatigue; abdominal swelling; generalized abdominal pain; frequent urination; bowel changes; abnormal vaginal bleeding; abdominal tumors; unexpected weight loss; and a feeling of fullness during meals. As the disease progresses, fluid may accumulate in the abdomen, further increasing abdominal discomfort, nausea, vomiting, and shortness of breath. Complications of peritoneal cancer include intestinal obstruction and urinary tract obstruction, and stents or nephrostomy tubes may be required.

[0013] Hepatocellular carcinoma is the most common type of primary liver cancer. According to the Mayo Clinic and the American Cancer Society, hepatocellular carcinoma occurs most frequently in people with chronic liver diseases such as cirrhosis caused by hepatitis B or C infection, those who drink large amounts of alcohol, and those with fatty deposits in the liver. Most patients do not show symptoms in the early stages of primary liver cancer. When symptoms appear, they include unintentional weight loss; loss of appetite; upper abdominal pain; nausea and vomiting; general weakness and fatigue; abdominal swelling; jaundice; and the appearance of white, chalky stools. GABA may be a byproduct of liver disease and can contribute to hepatic encephalopathy in cirrhotic patients, which may in turn lead to hepatocellular carcinoma. A decrease in GABAergic activity may contribute to the etiology of hepatocellular carcinoma.

[0014] Gastric cancer can occur in any part of the stomach. In most parts of the world, gastric cancer forms in the body of the stomach. However, according to the Mayo Clinic, in the United States, gastric cancer is more likely to occur at the gastroesophageal junction. The cause of gastric cancer is not clear, but risk factors include gastroesophageal reflux disease; obesity; a diet high in salty and smoked foods; a diet low in fruits and vegetables; family history; Helicobacter pylori infection; gastritis; smoking; and gastric polyps. Symptoms of gastric cancer include difficulty swallowing; a feeling of fullness after eating small amounts; early satiety; heartburn; indigestion; nausea; stomach pain; unintentional weight loss; and vomiting. GABA has been associated with the growth of digestive tract cancers in some studies and functions as a tumor signaling molecule in gastric cancer.

[0015] Pancreatic cancer can originate from two types of cells in the pancreas: exocrine cells and neuroendocrine cells such as islet cells. According to the National Cancer Institute, the exocrine type is more common and is usually detected at an advanced stage. Pancreatic neuroendocrine tumors, also known as islet cell tumors, are less frequent but have a better prognosis. The cause of pancreatic cancer is not clear, but factors that may increase the risk include smoking and hereditary gene mutations. The symptoms of pancreatic cancer often do not appear until the disease progresses and include abdominal pain radiating to the back, loss of appetite or unexpected weight loss; jaundice; pale stools; dark urine; itchy skin; new diagnosis of diabetes or difficulty controlling existing diabetes; blood clots and fatigue. GABA has been associated with stimulating the growth of pancreatic cancer.

[0016] Glioblastoma, also known as grade IV astrocytoma, is an aggressive brain tumor that grows rapidly. It infiltrates the surrounding brain tissue but generally does not spread to distant organs. According to the American Association of Neurological Surgeons, glioblastoma can occur newly in the brain or progress from low-grade astrocytoma. The cause of glioblastoma is unknown, but glioblastoma tends to occur more frequently in the elderly and in men. The symptoms vary depending on the location of the tumor and include persistent headache; double vision or blurred vision; vomiting; loss of appetite; mood or personality changes; changes in thinking or learning ability; new onset of seizures; and progressive speech impairment.

[0017] Cervical cancer occurs in the lower part of the uterus that connects to the vagina. According to the CDC, various strains of the sexually transmitted human papillomavirus (HPV) are responsible for most cases of cervical cancer. In a very small number of people, the virus can survive for years and be involved in the process by which some cervical cells become cancerous. In early-stage cervical cancer, there are generally no symptoms. Symptoms of more advanced cervical cancer include vaginal bleeding after intercourse, between periods, or after menopause; heavy, foul-smelling, watery, bloody vaginal discharge; and pelvic pain or pain during intercourse.

[0018] Ovarian cancer occurs in the ovaries, or in related parts of the fallopian tubes or peritoneum. According to the CDC, ovarian cancer exhibits various different tumor types. The most common tumor type is high-grade serous carcinoma, which is found in approximately 70% of ovarian cancer cases. The cause of ovarian cancer is unknown, but factors such as family history, genetic mutations in BRCA1 or BRCA2, endometriosis, and middle to old age are cited as risk factors. Symptoms of ovarian cancer include abdominal fullness or swelling; weight loss; discomfort in the pelvic area; fatigue; back pain; changes in bowel habits; and frequent urination. Ovarian cancer is associated with GABA and may be used in detection.

[0019] Bladder cancer most often originates from the urothelial cells that line the bladder, ureters, and kidneys. According to the American Cancer Society, older adults are at higher risk of bladder cancer. Most bladder cancers are diagnosed at an early stage where treatment is highly possible, but the recurrence rate of bladder cancer is high. Symptoms of bladder cancer include hematuria; frequent urination; pain during urination; and back pain. GABA has been associated with the promotion and detection of bladder cancer in some studies.

[0020] Colorectal cancer begins in the colon or rectum and is referred to as colon cancer or rectal cancer depending on the origin. According to the American Cancer Society, most colorectal cancers begin as polyps that form inside the colon or rectum. Not all polyps become cancerous, but the risk varies depending on the type of polyp. Hyperplastic polyps and inflammatory polyps have a very low risk. Adenomatous polyps, sessile serrated polyps, and traditional serrated adenomas have a high risk and are referred to as "precancerous." Symptoms of colon cancer include persistent changes in bowel habits such as diarrhea or constipation, changes in the consistency of stools; rectal bleeding or bloody stools; persistent abdominal discomfort such as cramps, gas, or pain; weakness or fatigue; and unexplained weight loss. GABA has been associated with the growth and detection of colorectal cancer in some studies sponsored by the NIH.

[0021] Endometrial cancer or uterine cancer begins inside the uterus or in the glands. Most cases occur in women over 55 years old. Risk factors for endometrial cancer or uterine cancer include obesity, family history, age, and hormonal effectors (e.g., estrogen, contraception, pregnancy, and polycystic ovary syndrome). According to the Mayo Clinic, the main symptom is abnormal vaginal bleeding such as postmenopausal bleeding or bleeding between periods. Other symptoms include pelvic pain and pain during sexual intercourse, although some women have no symptoms at all. Endometrial cancer and uterine cancer are associated with GABA, have GABA receptors that promote cancer growth, and can be used for the detection of abnormalities.

[0022] Salivary gland cancer begins in one of the salivary glands, including the parotid gland, submandibular gland, and sublingual gland. According to the Mayo Clinic, most salivary gland tumors occur in the parotid gland. Most salivary gland tumors are benign, but sometimes they can become cancerous. Symptoms of salivary gland tumors include a lump or swelling in the jaw or near it, or in the neck or mouth; facial numbness; weakness of the muscles on one side of the face; persistent pain in the area of the salivary gland; difficulty swallowing; and difficulty opening the mouth wide.

[0023] Kidney cancer / renal cancer begins inside the renal tubules of the kidneys. According to the Mayo Clinic, the incidence of kidney cancer seems to be increasing, which is likely due to improved techniques for testing and imaging. Risk factors for kidney cancer include smoking, abuse of painkillers, obesity, family history, and high blood pressure. Kidney cancer can occur at any age, but young children are more likely to develop a type of kidney cancer called Wilms tumor. Kidney cancer often has the cancer confined to a small size within the kidney and is usually detected at an early stage when there are usually no symptoms. Later symptoms can include blood in the urine; persistent pain in the back or side; loss of appetite; unexplained weight loss; fatigue; and fever. GABA has been noted as a tumor signaling molecule that controls the growth of kidney cancer and can be used to detect the presence of kidney cancer.

[0024] Prostate cancer is cancer that occurs in a man's prostate gland and is the most common cancer among men in the United States. According to the American Cancer Society, prostate cancer usually grows very slowly, so finding and treating it before symptoms appear does not improve a patient's health or extend their life. Prostate cancer is rare in men under 40, but the risk of developing it increases rapidly after age 50. Additionally, genetic mutations in the BRCA1 or BRCA2 gene may increase the risk. Prostate cancer may not show signs or symptoms in its early stages, but as it progresses, it can cause symptoms such as urinary problems; a weak stream of urine; blood in the urine; blood in the semen; bone pain; weight loss without trying; and erectile dysfunction. GABA has been reported to increase cell proliferation associated with prostate cancer.

[0025] Vulvar cancer is a rare cancer that occurs on the outer surface of the female genital organs, including the labia majora and clitoris. It can occur at any age but is most common in older women. According to the NIH, persistent infection with the human papillomavirus can increase the risk of vulvar cancer, similar to smoking. Symptoms of vulvar cancer can include persistent itching; local pain or tenderness; bleeding other than during menstruation; skin changes, such as changes in color or thickening; and sores, wart-like bumps, open ulcers.

[0026] Thyroid cancer starts in the thyroid gland. The two most common types of thyroid cancer are papillary cancer and follicular cancer. Papillary thyroid cancer is the most common type and can occur at any age. Papillary thyroid cancer grows slowly and often spreads to the lymph nodes in the neck. Follicular cancer is less common and can spread through the blood to distant organs, particularly the lungs and bones. According to the American Thyroid Association, factors that may increase the risk of thyroid cancer can include a history of high-dose radiation exposure, family history, and being over 40. Most thyroid cancers do not cause any symptoms in their early stages, but as the cancer grows, it can cause a small nodule that can be felt through the skin of the neck; changes in the voice, such as an increased hoarseness; difficulty swallowing; swelling of the lymph nodes in the neck; pain in the neck or throat. Changes in GABA have also been shown to affect the growth of thyroid cancer.

[0027] Head and neck cancer usually begins in the squamous cells that line the mucosal surfaces of the head and neck, such as the mouth, pharynx, and larynx. Less common head and neck cancers include nasal cavity and paranasal sinus cancers and salivary gland cancers. Alcohol and tobacco are major risk factors for head and neck cancer. Hypopharyngeal cancer is strongly associated with the human papillomavirus. UV rays may contribute to the risk of lip cancer. Infection with the Epstein-Barr virus increases the risk of cancers of the nose and nasopharynx and salivary gland cancers. Exposure to chemicals such as wood dust, formaldehyde, asbestos, nickel, and radiation can increase the risk of oropharyngeal cancer. Symptoms vary depending on the type of cancer but include persistent open sores; unusual pain or bleeding; difficulty chewing, swallowing, or speaking; and local swelling. GABA has shown a proliferative effect on some head and neck cancers, including head and neck squamous cell carcinoma.

[0028] B-cell lymphoma is a type of non-Hodgkin lymphoma that begins in B cells. There are many subtypes within this category. According to the American Cancer Society, diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma are the most common subtypes. DLBCL is the most common type and occurs mainly in older adults. DLBCL is aggressive and tends to grow rapidly. Follicular lymphoma is less common, less aggressive, and very rare in younger people. Symptoms of non-Hodgkin lymphoma include swelling of lymph nodes in the neck, armpits, or groin; abdominal pain or swelling; chest pain, cough, or shortness of breath; persistent fatigue; fever; night sweats; and unexplained weight loss.

[0029] Chronic lymphocytic leukemia (CLL) is another subtype of B-cell non-Hodgkin lymphoma, in which most of the cancer cells are present in the blood and bone marrow. CLL progresses slowly and usually affects older people. The exact mechanism that causes CLL is unknown, but age and exposure to chemicals are thought to be risk factors. According to the American Cancer Society, CLL may not cause symptoms in the early stages, but when symptoms occur, they include swelling of lymph nodes; fatigue; pain due to an enlarged spleen; night sweats; weight loss; frequent infections; and easy bruising.

[0030] Acute lymphoblastic leukemia (ALL) is a type of blood and bone marrow cancer that affects white blood cells. According to the American Cancer Society, ALL is the most common childhood cancer and is very aggressive and responsive. Possible risk factors for ALL include prenatal X-ray exposure; radiation exposure; certain changes in chromosomes or genes; and certain genetic conditions, such as Down syndrome; neurofibromatosis type 1; Bloom syndrome; and constitutional mismatch repair deficiency. Symptoms include swollen lymph nodes; bruising; fever; bone and joint pain; bleeding from the gums; shortness of breath; fatigue and weakness; loss of appetite; and frequent infections. GABA has been observed in several studies to be involved in the growth and spread of ALL.

[0031] Hairy cell leukemia (HCL) is another very rare subtype of B-cell non-Hodgkin lymphoma. The affected cells are B lymphocytes found in the bone marrow, spleen, and blood, and have "hairy" projections. According to the Mayo Clinic, HCL occurs more often in men than women and is most common in adults over middle age. HCL is considered a chronic disease, but remission can sometimes be achieved with treatment for several years. Hairy cell leukemia may be discovered incidentally by a blood test for another disease or condition in people who do not have the signs and symptoms of hairy cell leukemia. When symptoms do appear, there is a feeling of fullness in the abdomen, discomfort when eating more than a little at a time; fatigue; easy bruising; recurrent infections; weakness; and unintentional weight loss.

[0032] Chronic myeloid leukemia (CML) is a rare type of cancer that starts in certain blood - forming cells of the bone marrow. CML is a slowly growing leukemia, but it can change into rapidly growing acute leukemia, which is difficult to treat. CML mainly occurs in adults, but very rarely in children. According to the Mayo Clinic, CML often does not cause symptoms and may be detected by a blood test. When symptoms do occur, they can include bone pain; easy bruising; fullness after a small meal; fatigue; fever; unintentional weight loss; loss of appetite; pain or a feeling of fullness under the left rib cage; night sweats.

[0033] Acute myeloid leukemia (AML) is a cancer of the blood and bone marrow. It is the most common type of acute leukemia in adults, is very aggressive, and progresses rapidly. According to the Mayo Clinic, men and adults over 65 are at higher risk of AML, as are smokers, people who have been exposed to radiation or dangerous chemicals, and those with other blood disorders. Symptoms of early - stage AML can sometimes resemble those of influenza or other common illnesses. In later stages, there can be fever; bone pain; weakness and fatigue; shortness of breath; pale skin; frequent infections; easy bruising; abnormal bleeding such as frequent nosebleeds or bleeding from the gums.

[0034] Melanoma is the most dangerous type of skin cancer. It grows rapidly and can spread to all organs. Melanoma starts from melanocytes that produce melanin. According to the National Cancer Institute of the United States, although the exact cause of melanoma is not clear, exposure to ultraviolet rays from sunlight or tanning lamps / beds increases the risk of developing melanoma. The risk of melanoma seems to be increasing in people under 40 years old, especially in women. The first symptoms of melanoma are often changes in existing moles, or the occurrence of new pigmentation or growths with abnormal appearance on the skin. Malignant moles look very different. Signs indicating the possibility of malignant pigmentation include when the shape of the growth is asymmetric, the border is irregular, the color changes, the diameter increases, or the color and shape seem to be evolving. There is a direct relationship between melanoma and GABA, and melanoma has GABA receptors that promote the growth of cancer tumors.

[0035] Testicular cancer occurs in the testicles and is the most common cancer in American men aged 15 - 35. Testicular cancer mainly has two pathological types: seminoma and nonseminoma. Seminoma grows slowly and mainly affects people aged 40 - 60. Nonseminoma grows more rapidly and mainly affects people in their teens, twenties, and thirties. Symptoms of testicular cancer include a lump or swelling in either testicle; a heavy feeling in the scrotum; dull pain in the abdomen or groin; sudden fluid accumulation in the scrotum; pain or discomfort in the testicle or scrotum; breast swelling or tenderness; and back pain. Testicular cancer usually affects only one testicle. There is a direct relationship between testicular cancer and GABA, and it has been shown that GABA promotes the growth of cancerous cells and changes sperm motility.

[0036] Brain metastases (metastatic brain tumors) and leptomeningeal metastases occur when cancer spreads from its site of origin or primary site. Brain metastases occur when cancer cells move into the brain itself. Leptomeningeal metastases (also known as leptomeningeal cancer, leptomeningeal carcinomatosis, leptomeningeal disease (LMD), neoplastic meningitis, meningeal metastases, and meningeal carcinomatosis) are a rare complication in which cancer spreads from the original tumor site to the meninges that surround the brain and spinal cord. The leptomeninges are the two innermost layers of tissue that cover the brain and spinal cord. Leptomeningeal metastases occur when cancer cells infiltrate the cerebrospinal fluid and spread throughout the central nervous system. The prognosis is generally poor, and the survival period is usually several months. Because it can metastasize to the subarachnoid space, the most common cancers that can involve the leptomeninges are breast cancer, lung cancer, and melanoma. There are also cases that arise from primary brain tumors such as medulloblastoma or glioma. The most common symptoms of leptomeningeal cancer are pain and seizures. Other symptoms include headache (usually accompanied by nausea, vomiting, and altered mental status), difficulty walking due to weakness or ataxia, memory impairment, incontinence, and sensory disturbances. In some cases, there may be symptoms of double vision, jaw numbness, back pain, limb weakness, sphincter-related problems, hydrocephalus, loss of urine control, difficulty walking, limb weakness and paresthesia, bowel or bladder dysfunction, double vision, sensory disturbances or loss of movement of the trigeminal nerve, cochlear dysfunction, confusion, and cognitive impairment.

[0037] γ-Aminobutyric acid (GABA) is metabolized by the transfer of an amino group by γ-aminobutyric acid aminotransferase (GABA-AT), also known as GABA-transaminase or ABAT. Inhibition of this enzymatic process reduces the breakdown of GABA, which leads to an increase in extracellular GABA and neuronal GABA concentrations. Vigabatrin is a GABA-AT inhibitor that has been used in the treatment of treatment-resistant epilepsy and childhood seizures. It has been suggested that the inhibitory effect of vigabatrin on IKCa channels is an important underlying mechanism of the vigabatrin-induced anti-neoplastic effect associated with gliomas. Hung et al., BMC Pharmacol Toxicol 22, 6 (2021). Vigabatrin has the potential for serious side effects. Its use is restricted because of the possibility of retinal toxicity and subsequent visual field defects. (1S,3S)-3-Amino + difluoromethylene-1-cyclopentanoic acid (also known as CPP-115) is a GABA-AT inhibitor that inactivates GABA-AT 186 times more efficiently than vigabatrin. Preclinical data for CPP-115 have reported that, when compared with vigabatrin, equivalent pharmacokinetics, improved tolerability, and a better toxicity profile are obtained at significantly lower drug doses. See Prescot et al., Neuropsychopharmacology (2018) 43, 646-654. See also U.S. Patent No. 9,993,449, which is incorporated herein by reference. (S)-3-Amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid (also known as OV329) is a GABA-AT inhibitor that has been shown to be 9.8 times more efficient than CPP-115 as an inactivator of GABA-AT. Ibid. U.S. Patent Nos. 9,603,820 and 9,993,449 describe the use of CPP-115 and OV329, respectively, for the treatment of hepatocellular carcinoma.

[0038] Many cancers are treated by surgery, chemotherapy, radiation, or combinations thereof. Chemotherapeutic agents used in the treatment of cancer are known to cause severe and unpleasant side effects in patients. For example, some chemotherapeutic agents can cause neuropathy, nephrotoxicity (e.g., hyperlipidemia, proteinuria, hypoproteinemia, combinations thereof, etc.), stomatitis, mucositisemesis, alopecia, eating disorders, esophagitis, amenorrhea, immunosuppression, anemia, high tone hearing loss, cardiotoxicity, fatigue, neuropathy, myelosuppression, etc. In some cases, chemotherapy is not effective or loses its effectiveness after any period of effectiveness during or immediately after the treatment regimen (i.e., the treatment regimen does not lead to a cure). Improved methods for the treatment of cancer, and compositions capable of delivering bioactive agents to assist in the treatment of cancer are still desirable. Furthermore, considering the high mortality rate due to cancer, there is a need for additional treatments to fight cancer. SUMMARY OF THE INVENTION

[0039] Methods and compositions for treating cancer are provided. In an embodiment, a method for treating cancer comprises administering to a subject in need thereof an effective amount of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof. In an embodiment, a composition for treating cancer comprising (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is administered to a subject in need thereof. In an embodiment, the cancer is medulloblastoma, glioma, breast cancer, squamous cell carcinoma, melanoma, lung cancer, peritoneal cancer, hepatocellular carcinoma, gastric cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, testicular cancer, bladder cancer, colorectal cancer, endometrial cancer or uterine cancer, salivary gland cancer, kidney cancer or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, head and neck cancer, B-cell lymphoma, chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), hairy cell leukemia, leptomeningeal carcinomatosis or chronic myelogenous leukemia, and subtypes thereof. In an embodiment, (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof reduces metastasis of the cancer. In an embodiment, (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof prevents metastasis of the cancer. In an embodiment, an effective amount of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is from about 0.0001 mg / kg to about 30 mg / kg. In an embodiment, (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof at 0.5 mg / kg to 6.5 mg / kg is administered to a subject in need thereof. In an embodiment, an effective amount of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is administered to the subject once a day, twice a day, three times a day, or four times a day.

[0040] Detailed Description Methods and compositions for treating cancer are provided. In embodiments, methods and compositions for treating cancer are provided that reduce or prevent metastasis without the undesirable side effects associated with commonly used chemotherapeutic agents. In an embodiment, a method of treating cancer comprises administering to a subject in need thereof an effective amount of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof (referred to interchangeably herein as OV329). In an embodiment, a method of inhibiting cancer growth and metastasis comprises administering to a subject in need thereof an effective amount of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof.

[0041] In leptomeningeal metastasis, a series of sequential steps must be achieved by tumor cells for metastasis to occur. Tumor cells proliferate at the primary site, invade the surrounding environment, enter the cerebrospinal fluid, and spread throughout the central nervous system. Metastatic tumor cells need to survive in the microenvironment of the meninges and cerebrospinal fluid. The meninges and cerebrospinal fluid provide a relatively nutrient-poor environment. Metastatic brain tumor cells, such as metastatic MB cells, maintain viability in the metabolite-poor cerebrospinal fluid by using GABA-AT as an alternative energy source, thereby promoting leptomeningeal metastasis formation. Martirosian et al., 2021, Cell Reports 35, 109302.

[0042] In the present disclosure, however, without wishing to be bound by any theory, administration of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid, a GABA-AT inhibitor, or a pharmaceutically acceptable salt thereof blocks or reduces GABA metabolism, such that metastatic MB cells cannot appropriately utilize GABA as an energy source alternative, thereby creating a less favorable malnourished environment for metastatic MB cells in cerebrospinal fluid and throughout the central nervous system. In this way, metastasis is prevented or reduced. Indeed, any cancer that utilizes GABA degradation via GABA-AT as an energy source, as described herein, is susceptible to the effect of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof to reduce or prevent the utilization of GABA as an energy source, thereby preventing or reducing tumor growth and / or metastasis.

[0043] In an embodiment, (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is administered to a subject at a low dose that inhibits GABA-AT but does not dramatically affect circulating GABA, which, as described above, may have a tumor and / or metastasis promoting effect.

[0044] Importantly, many anti-cancer compounds cannot cross the blood-brain barrier and are not available for the treatment of brain cancer. (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof can cross the blood-brain barrier and can treat brain cancer and leptomeningeal metastases.

[0045] As used herein, the term "cancer" means a growth disorder caused by or characterized by the proliferation of cells that have lost their sensitivity to normal growth control. Cancers of the same histotype usually begin in the same tissue and are classified into different subtypes based on their biological characteristics. The four general categories of cancer are carcinoma (derived from epithelial cells), sarcoma (derived from connective tissue or mesoderm), leukemia (derived from hematopoietic tissue), and lymphoma (derived from lymphoid tissue). Non-limiting examples of cancers that are susceptible to treatment with (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof include medulloblastoma, glioma, breast cancer, squamous cell carcinoma, melanoma, lung cancer, peritoneal cancer, hepatocellular carcinoma, gastric cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, testicular cancer, bladder cancer, colorectal cancer, endometrial cancer or uterine cancer, salivary gland cancer, kidney cancer or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, head and neck cancer, B-cell lymphoma, chronic lymphocytic leukemia, acute lymphoblastic leukemia, hairy cell leukemia or chronic myelogenous leukemia and subtypes thereof, including refractory versions of any of the foregoing cancers, or combinations of one or more of the foregoing cancers. The cancers described above can utilize GABA-AT to metabolize GABA as an energy source.

[0046] As used herein, the terms "cancer," "neoplasm," and "tumor" are used interchangeably, in either the singular or plural, and mean cells that have undergone malignant transformation rendering them pathological to a host organism. Primary cancer cells (i.e., cells obtained from near the site of malignant transformation) can be readily distinguished from non-cancer cells by established techniques, particularly histological examination. The definition of cancer cells used herein includes not only primary cancer cells, but also any cells derived from cancer cell progenitors. This includes metastatic cancer cells, in vitro cultures, and cell lines derived from cancer cells. When referring to types of cancer that typically manifest as solid tumors, a "clinically detectable" tumor is, for example, detectable based on a tumor mass by procedures such as CAT scan, MR imaging, X-ray, ultrasound, or palpation, and / or detectable by the expression of one or more cancer-specific antigens in a sample obtained from a patient. Leukemia is clinically detectable using one or more of a complete blood count, pallor, blood smear specimens, and bone marrow smear specimens. Progressive leukemia in a particular subject can manifest as a solid tumor.

[0047] (S)-3-Amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid can be represented by the following structure: [Chemical formula]

[0048] In an embodiment, (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid can be provided as an acid addition salt, zwitterionic hydrate, zwitterionic anhydrate, hydrochloride or hydrobromide, or in the form of a zwitterionic monohydrate. Examples of acid addition salts include, but are not limited to, maleic acid, fumaric acid, benzoic acid, ascorbic acid, succinic acid, oxalic acid, bis-methylenesalicylic acid, methanesulfonic acid, ethane-disulfonic acid, acetic acid, propionic acid, tartaric acid, salicylic acid, citric acid, gluconic acid, lactic acid, malic acid, mandelic acid, cinnamic acid, citraconic acid, aspartic acid, stearic acid, palmitic acid, itaconic acid, glycolic acid, pantothenic acid, p-aminobenzoic acid, glutamic acid, benzenesulfonic acid or the addition salts of theophylline acetic acid, and 8-halotheophyllines, such as 8-bromo-theophylline. In an embodiment, inorganic acid addition salts including, but not limited to, addition salts of hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfamic acid, phosphoric acid or nitric acid can be used.

[0049] As can be seen from the examples in this specification, an effective amount of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof for the treatment of cancer in this specification is surprisingly lacking in, or exhibits fewer, undesirable side effects normally associated with GABA-AT inhibitors such as vigabatrin, such as retinal toxicity and decreased motor coordination. The term "effective amount" as applied to (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof means an amount effective for preventing or treating cancer. The "effective amount" of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof can be determined by various factors such as, for example, the method of administration, the target site, the condition of the patient, etc. Therefore, when used in the human body, it is necessary to set an appropriate amount in consideration of safety and effectiveness. It is also possible to estimate the amount for human use from the effective amount obtained in animal experiments. See, for example, Reagan-Shaw et al., FASEB J. 2008 Mar;22(3):659-61.

[0050] The term "prevent" as used herein, including its grammatical variations such as "preventing" and "prevention" used herein, means the action of inhibiting or delaying the occurrence, progression, and recurrence of cancer by administration of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof. The term "treat" as used herein, including its grammatical variations such as "treating" and "treatment" used herein, means all types of actions of reducing the growth of cancer cells, promoting the death of cancer cells, and / or reducing the symptoms of cancer by administration of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof disclosed herein. "Treating cancer" herein includes inhibiting cancer growth and metastasis of cancer cells. For example, in embodiments, "treating" may include reducing seeding of metastatic cells or reducing seeding of tumor cells.

[0051] "Effective amount" as used herein is (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof in the range of about 0.0001 mg / kg to about 30 mg / kg. In embodiments, a relatively low dose of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof that inhibits GABA-AT without dramatically affecting circulating GABA is in the range of about 0.2 mg / kg to 10 mg / kg. In embodiments, a relatively low dose of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof that inhibits GABA-AT without dramatically affecting circulating GABA is in the range of about 0.5 mg / kg to about 6.5 mg / kg. In embodiments, about 20 mg to about 400 mg of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is administered once a day, twice a day, three times a day, or four times a day. For example, a pharmaceutical composition comprising an effective amount of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is about 20 mg to about 25 mg, about 25 mg to about 30 mg, about 30 mg to about 35 mg, about 35 mg to about 40 mg, about 40 mg to about 45 mg, about 45 mg to about 50 mg, about 50 mg to about 55 mg, about 55 mg to about 60 mg, about 60 mg to about 65 mg, about 65 mg to about 70 mg, about 70 mg to about 75 mg, about 75 mg to about 80 mg, about 80 mg to about 85 mg, about 85 mg to about 90 mg, about 90 mg to about 95 mg, about 95 mg to about 100 mg, about 100 mg to about 105 mg, about 105 mg to about 110 mg, about 110 mg to about 115 mg, about 115 mg to about 120 mg, about 120 mg to about 125 mg, about 130 mg to about 135 mg, about 140 mg to about 145 mg, about 150 mg to about 155 mg, about 160 mg to about 165 mg, about 170 mg to about 175 mg, about 180 mg to about 185 mg, about 190 mg to about 195 mg, about 200 mg to about 205 mg, about 210 mg to about 210 mg, about 215 mg to about 220 mg, about 225 mg to about 230 mg, about 235 mg to about 240 mg, about 245 mg to about 250 mg,It may contain (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof in an amount of about 255 mg to about 260 mg, about 265 mg to about 270 mg, about 275 mg to about 280 mg, about 285 mg to about 290 mg, about 295 mg to about 300 mg, about 305 mg to about 310 mg, about 315 mg to about 320 mg, about 325 mg to about 330 mg, about 335 mg to about 340 mg, about 345 mg to about 350 mg, about 355 mg to about 360 mg, about 365 mg to about 370 mg, about 375 mg to about 380 mg, about 385 mg to about 390 mg, about 395 mg to about 400 mg. In an embodiment, a pharmaceutical composition containing an effective amount of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is 20 mg, 21 mg, 22 mg, 23 mg, 24 mg, 25 mg, 26 mg, 27 mg, 28 mg, 29 mg, 30 mg, 31 mg, 32 mg, 33 mg, 34 mg, 35 mg, 36 mg, 37 mg, 38 mg, 39 mg, 40 mg, 41 mg, 42 mg, 43 mg, 44 mg, 45 mg, 46 mg, 47 mg, 48 mg, 49 mg, 50 mg, 51 mg, 52 mg, 53 mg, 54 mg, 55 mg, 56 mg, 57 mg, 58 mg, 59 mg, 60 mg, 61 mg, 62 mg, 63 mg, 64 mg, 65 mg, 66 mg, 67 mg, 68 mg, 69 mg, 70 mg, 71 mg, 72 mg, 73 mg, 74 mg, 75 mg, 76 mg, 77 mg, 78 mg, 79 mg, 80 mg, 81 mg, 82 mg, 83 mg, 84 mg, 85 mg, 86 mg, 87 mg, 88 mg, 89 mg, 90 mg, 91 mg, 92 mg, 93 mg, 94 mg, 95 mg, 96 mg, 97 mg, 98 mg, 99 mg, 100 mg, 101 mg, 102 mg, 103 mg, 104 mg, 105 mg, 106 mg, 107 mg, 108 mg, 109 mg, 110 mg, 111 mg, 112 mg, 113 mg, 114 mg, 115 mg, 116 mg, 117 mg, 118 mg, 119 mg, 120 mg, 121 mg, 122 mg, 123 mg, 124 mg, 125 mg, 126 mg, 127 mg, 128 mg, 129 mg, 130 mg, 135 mg, 140 mg, 145 mg, 150 mg, 155 mg, 160 mg, 165 mg, 170 mg,It contains (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof in an amount of 175 mg, 180 mg, 185 mg, 190 mg, 195 mg, 200 mg, 205 mg, 210 mg, 215 mg, 220 mg, 225 mg, 230 mg, 235 mg, 240 mg, 245 mg, 250 mg, 255 mg, 260 mg, 265 mg, 270 mg, 275 mg, 280 mg, 285 mg, 290 mg, 295 mg, 300 mg, 305 mg, 315 mg, 320 mg, 325 mg, 330 mg, 335 mg, 340 mg, 345 mg, 350 mg, 355 mg, 360 mg, 365 mg, 370 mg, 375 mg, 380 mg, 385 mg, 390 mg, or 400 mg.

[0052] In an embodiment, (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is administered to a subject at about 25 mg / day, 30 mg / day, 35 mg / day, 40 mg / day, 45 mg / day, 50 mg / day, 60 mg / day, 65 mg / day, 70 mg / day, 75 mg / day, 80 mg / day, 85 mg / day, 90 mg / day, 95 mg / day, 100 mg / day, 105 mg / day, 110 mg / day, 115 mg / day, 120 mg / day, 125 mg / day, 130 mg / day, 135 mg / day, 140 mg / day, 145 mg / day, 150 mg / day, 155 mg / day, 160 mg / day, 165 mg / day, 170 mg / day, 175 mg / day, 180 mg / day, 185 mg / day, 190 mg / day, 195 mg / day, 200 mg / day, 205 mg / day, 210 mg / day, 215 mg / day, 220 mg / day, 225 mg / day, 230 mg / day, 235 mg / day, 240 mg / day, 245 mg / day, 250 mg / day, 255 mg / day, 260 mg / day, 265 mg / day, 270 mg / day, 275 mg / day, 280 mg / day, 285 mg / day, 290 mg / day, 295 mg / day, 300 mg / day, 305 mg / day, 310 mg / day, 315 mg / day, 320 mg / day, 325 mg / day, 330 mg / day, 335 mg / day, 340 mg / day, 345 mg / day, 350 mg / day, 355 mg / day, 360 mg / day, 365 mg / day, 370 mg / day, 375 mg / day, 380 mg / day, 385 mg / day, 390 mg / day, 395 mg / day, or 400 mg / day one or more times. In an embodiment, the subject can start at a low dose and the dose can be increased over time.

[0053] In an embodiment, the administration step results in at least one of a 10% decrease in tumor size, at least a 10% decrease in the number of cancer metastases, or at least a 10% improvement in clinical signs and symptoms associated with cancer. As used herein, the term "administration" and its grammatical variations such as "administer" and "administering" mean providing a given substance to an individual or patient by any suitable method. For example, (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof can be administered enterally, e.g., orally or parenterally (e.g., by intravenous, subcutaneous, intraperitoneal, or topical application as an injection). The dosage varies depending on the patient's weight, age, gender, health status, diet, administration time, administration method, excretion rate, disease severity, etc. Liquid formulations for oral administration of (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof include suspensions, oral liquids, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweetening agents, flavoring agents, preservatives, etc. may be included. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, suppositories, etc.

[0054] In an embodiment, (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is administered to a subject having cancer via a pharmaceutical composition. The pharmaceutical composition herein can be administered by any device capable of moving the active substance to the target cells. The pharmaceutical composition herein encompasses dosage forms. The dosage forms herein encompass unit dosages. In embodiments, as discussed below, various dosage forms, including conventional formulations and modified release formulations, can be administered one or more times a day. In embodiments, (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is administered to the subject once or twice a day (such as in the morning or at night), or three times a day (morning, noon, and evening) or four times a day (morning, noon, evening). Any suitable route of administration can be utilized, for example, oral, rectal, nasal, pulmonary, vaginal, sublingual, transdermal, subcutaneous, intravenous, intraarterial, intramuscular, intraperitoneal, intratumoral, intracranial, and intraventricular routes.

[0055] Suitable dosage forms include tablets, capsules, oral liquids, powders, aerosols, topical liquids, transdermal modalities such as patches, creams, and ointments, parenteral formulations, and suppositories. In an embodiment, a medicament for treating cancer or inhibiting cancer growth and metastasis is manufactured using (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof. Those skilled in the art are proficient in pharmaceutical compounding techniques for formulating and manufacturing pharmaceutical compositions. See, for example, E. W. Martin ed., Remington's Pharmaceutical Sciences, 18th ed. (1990), Mack Publishing Co.

[0056] In an embodiment, a method of treating cancer, comprising inhibiting cancer growth and metastasis, the method comprising administering to a subject in need thereof a pharmaceutical composition comprising (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof, wherein the composition provides improvement of cancer symptoms for more than 1 hour after administration to the subject. In an embodiment, a method of treating cancer, comprising inhibiting cancer growth and metastasis, the method comprising administering to a subject in need thereof a composition comprising (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof, wherein the composition provides improvement of one or more cancer symptoms for more than 2 hours after administration to the subject. In an embodiment, a method of treating cancer, comprising inhibiting cancer growth and metastasis, the method comprising administering to a subject in need thereof a pharmaceutical composition comprising (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof, wherein the composition provides improvement of one or more cancer symptoms for more than 3 hours after administration to the subject. In an embodiment, a method of treating cancer, comprising inhibiting cancer growth and metastasis, the method comprising administering to a subject in need thereof a pharmaceutical composition comprising (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof, wherein the composition provides improvement of one or more cancer symptoms for more than 4 hours after administration to the subject. In an embodiment, a method of treating cancer, comprising inhibiting cancer growth and metastasis, the method comprising administering to a subject in need thereof a pharmaceutical composition comprising (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof, wherein the composition provides improvement of one or more cancer symptoms for more than 6 hours after administration to the subject.In an embodiment, a method of treating cancer, comprising inhibiting the growth and metastasis of cancer, the method comprising administering to a subject in need thereof a pharmaceutical composition comprising (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof, wherein the composition provides improvement of one or more cancer symptoms beyond 8 hours, 10 hours, 12 hours, 14 hours, 16 hours, 18 hours, 20 hours, 22 hours or 24 hours after administration to the subject. In an embodiment, the pharmaceutical composition provides improvement of the subject's next-day function. For example, the pharmaceutical composition may provide improvement of one or more cancer symptoms beyond, for example, about 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 14 hours, 16 hours, 18 hours, 20 hours, 22 hours or 24 hours after administration and waking from a night's sleep.

[0057] In an embodiment, the pharmaceutical composition herein may be provided with a conventional release profile or a modified release profile. The pharmaceutical composition may be prepared using a pharmaceutically acceptable "carrier" composed of materials considered to be safe and effective. The "carrier" includes all components other than the active ingredient present in the formulation. The term "carrier" includes, but is not limited to, diluents, binders, lubricants, disintegrants, fillers, and coating compositions. Those skilled in the art are familiar with such pharmaceutical carriers and methods of formulating pharmaceutical compositions using such carriers. In an embodiment, the pharmaceutical composition herein may include pharmaceutically acceptable additives such as, for example, starch, pregelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose, mannitol, gum arabic, alpha starch, corn starch, cellulose powder, hydroxypropyl cellulose, Opadry, sodium starch glycolate, carnauba wax, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, sucrose, dextrose, sorbitol, talc, etc. In an embodiment, the pharmaceutically acceptable additives can be added to the composition in an amount of 0.1 to 90 parts by weight, but is not limited thereto.

[0058] In an embodiment, the pharmaceutical composition herein is a modified release dosage form that provides a modified release profile. The modified release profile can exhibit an immediate release, delayed release, or sustained release profile. Conventional (or unmodified) release oral dosage forms such as tablets, capsules, suppositories, syrups, solutions, and suspensions typically release the medicament into the oral cavity, stomach, or intestine when the tablet, capsule shell, or suppository dissolves, or in the case of syrups, solutions, and suspensions, when they are swallowed. The pattern of drug release from a modified release (MR) dosage form is intentionally altered from that of conventional dosage forms to achieve the desired therapeutic objective and / or better patient compliance. MR formulations include orally disintegrating dosage forms (ODDFs) that provide immediate release, sustained release dosage forms, delayed release dosage forms (e.g., enteric-coated), and pulsatile release dosage forms.

[0059] An ODDF is a solid dosage form containing a pharmaceutical or active ingredient that rapidly disintegrates, usually within a few seconds, when placed on the tongue. The disintegration time of an ODDF generally ranges from 1 to 2 seconds to about 1 minute. ODDFs are designed to rapidly disintegrate or dissolve upon contact with saliva. This dosage method can be beneficial for people who may have problems swallowing tablets, whether due to physical weakness or mental nature. Subjects in pain may exhibit such behavior. ODDFs can rapidly deliver the drug to the bloodstream via the mucosa, resulting in a rapid onset of action. Examples of ODDFs include orally disintegrating tablets, capsules, and rapidly dissolving films and wafers.

[0060] Extended release dosage forms (ERDFs) have an extended release profile and enable a reduction in the dosing frequency compared to conventional dosage forms, such as solutions or unmodified release dosage forms. ERDFs provide a prolonged duration of action of the drug. Suitable formulations for providing an extended release profile are well known in the art. For example, coated sustained release beads or granules (the terms "beads" and "granules" are used interchangeably herein) in which (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is applied to beads, such as confectionery nonpareil beads, and then coated with a conventional release retardant material such as wax, enteric coating, are well known in the art. In embodiments, the beads can be formed by mixing (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof with a material to provide a mass from which the drug leaches. In embodiments, the beads can be processed to provide different release rates by varying the properties of the coating or mass, such as thickness, porosity, use of different materials, etc. Beads having different release rates can be combined in a single dosage form to provide variable or continuous release. The beads may be placed in capsules or compressed into tablets.

[0061] In an embodiment, the modified dosage forms herein incorporate delayed release dosage forms having a delayed release profile. The delayed release dosage forms can include delayed release tablets or delayed release capsules. A delayed release tablet is a solid dosage form that releases a drug (or drugs), such as (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof, at a time other than promptly after administration. A delayed release capsule is a solid dosage form in which a drug is enclosed within a hard or soft soluble container made of a suitable form of gelatin and releases the drug at a time other than immediately after administration. For example, enteric-coated tablets, capsules, particles, and beads are well-known examples of delayed release dosage forms. Enteric-coated tablets, capsules, particles, and beads pass through the stomach and release the drug in the intestine. In an embodiment, a delayed release tablet is a solid dosage form containing an aggregate of pharmaceutical particles that releases a drug (or drugs) at a time other than immediately after administration. In an embodiment, the aggregate of pharmaceutical particles is covered with a coating that delays the release of the drug. In an embodiment, a delayed release capsule is a solid dosage form containing an aggregate of pharmaceutical particles that releases a drug (or drugs) at a time other than immediately after administration. In an embodiment, the aggregate of pharmaceutical particles is covered with a coating that delays the release of the drug.

[0062] Sustained release dosage forms are known to those skilled in the art. For example, (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is applied to beads, such as confectionery nonpareil beads, and then coated with a conventional release retardant material such as wax, enteric coating, etc., resulting in coated sustained release beads or granules. In an embodiment, the beads can be formed by mixing (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof with a material to provide a mass from which the drug leaches. In an embodiment, the beads can be processed to provide different release rates by varying the properties of the coating or mass, such as thickness, porosity, use of different materials, etc. In an embodiment, enteric granules of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof can be contained in an enteric capsule or tablet that releases the granules in the small intestine. In an embodiment, the granules have a coating such that the coated granules remain intact until at least the ileum is reached, and then provide a delayed release of the drug in the colon. Suitable enteric coating materials, such as Eudragit® coatings such as methacrylic acid and methyl methacrylate polymers, are well known in the art. The granules can be placed in capsules or compressed into tablets.

[0063] In an embodiment, (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is incorporated into a porous inert carrier that provides a delayed release profile. In an embodiment, the porous inert carrier incorporates channels or passages through which the drug diffuses into the surrounding fluid. In an embodiment, (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is incorporated into an ion exchange resin to provide a delayed release profile. The delayed action can be attributed to a predetermined rate of drug release from the resin when the drug-resin complex contacts gastrointestinal fluid and ionic components dissolved therein. In an embodiment, a membrane is utilized to control the rate of release from a drug-containing reservoir. In an embodiment, a liquid formulation can also be utilized to provide a delayed release profile. For example, a liquid formulation consisting of solid particles dispersed throughout a liquid phase in which the particles do not dissolve. The suspensions of the present invention are formulated to enable at least a reduction in the dosing frequency as compared to drugs presented as conventional dosage forms (e.g., solutions or rapid drug release, as conventional solid dosage forms). For example, suspensions of ion exchange resin components or microbeads.

[0064] In an embodiment, (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is incorporated into a polymeric delivery vehicle made from cellulose, agarose, polymethacrylate, polystyrene, and polyacrylamide, and composites thereof.

[0065] In an embodiment, (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is incorporated into a hydrogel-based delivery vehicle. Examples of suitable hydrogels include polyvinyl alcohol, sodium alginate, chitosan, polyvinyl pyrrolidone, derivatives of polyacrylic acid and / or polymethacrylic acid, and composites thereof.

[0066] In an embodiment, the pharmaceutical composition described herein is suitable for parenteral administration, including, for example, intramuscular (i.m.), intravenous (i.v.), subcutaneous (s.c.), intraperitoneal (i.p.), or intrathecal (i.t.) administration. Parenteral compositions must be sterile for administration by injection, infusion, or implantation into the body and may be packaged in either single-dose or multi-dose containers. In an embodiment, a liquid pharmaceutical composition for parenteral administration to a subject contains an active substance, for example, (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof, in any of the respective amounts described above. In an embodiment, a pharmaceutical composition for parenteral administration is formulated as a total volume of, for example, 10 ml, 20 ml, 25 ml, 50 ml, 100 ml, 200 ml, 250 ml, or 500 ml. In an embodiment, the composition is contained in a bag, glass vial, plastic vial, or bottle.

[0067] The pharmaceutical composition for parenteral administration provided herein may contain one or more excipients, for example, a solvent, solubility enhancer, suspending agent, buffer, isotonic agent, stabilizer, or antibacterial preservative. When used, the excipients of the parenteral composition do not adversely affect the stability, bioavailability, safety, and / or efficacy of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof used in the composition. Thus, a parenteral composition is provided in which there is no incompatibility between any of the components of the dosage form.

[0068] In an embodiment, the parenteral composition (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof contains a stabilizing amount of at least one excipient. For example, the excipient may be selected from the group consisting of buffers, solubilizing agents, isotonic agents, antioxidants, chelating agents, antibacterial agents, and preservatives. One of ordinary skill in the art will understand that an excipient may have one or more functions and may be classified into one or more defined groups.

[0069] In an embodiment, in a parenteral composition ((S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof and an additive), the excipient is present, for example, at a weight % (w / v) of less than about 10%, less than about 5%, less than about 2.5%, less than about 1% or less than about 0.5%. In an embodiment, the excipient is present, for example, at a weight % of about 1.0% to about 10%, about 10% to about 25%, about 15% to about 35%, about 0.5% to about 5%, about 0.001% to about 1%, about 0.01% to about 1%, about 0.1% to about 1%, or about 0.5% to about 1%. In an embodiment, the excipient is present, for example, at a weight % of about 0.001% to about 1%, about 0.01% to about 1%, about 1.0% to about 5%, about 10% to about 15%, or about 1% to about 15%.

[0070] In an embodiment, there is provided a parenteral composition of an active substance, for example (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof, wherein the pH of the composition is from about 4.0 to about 8.0. In an embodiment, the pH of the composition is, for example, about 5.0 to about 8.0, about 6.0 to about 8.0, about 6.5 to about 8.0. In an embodiment, the pH of the composition is, for example, about 6.5 to about 7.5, about 7.0 to about 7.8, about 7.2 to about 7.8, or about 7.3 to about 7.6. In an embodiment, the pH of the aqueous solution is, for example, about 6.8, about 7.0, about 7.2, about 7.4, about 7.6, about 7.7, about 7.8, about 8.0, about 8.2, about 8.4 or about 8.6.

[0071] It should be understood that the dosages of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof provided herein are applicable to all dosage forms described herein, including conventional dosage forms, modified dosage forms, and the parenteral formulations described herein. One of ordinary skill in the art will determine the appropriate amount according to criteria such as dosage form, route of administration, tolerance of the subject, effectiveness, treatment goal and treatment effect, among other pharmaceutically acceptable criteria.

[0072] In an embodiment, (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof can be administered alone or in combination therapy, where the subject is also undergoing one or more cancer therapies such as surgery, chemotherapy, radiation therapy, hyperthermia therapy, immunotherapy, hormone therapy, and / or laser therapy.

[0073] In an embodiment, the combination therapy can include, for example, one or more chemotherapeutic agents, targeting agents such as antibody kinase inhibitors; hormonal agents, etc. The combination therapy can also include conventional therapies including, but not limited to, antibody administration, vaccine administration, cytotoxic agents, natural amino acid polypeptides, nucleic acids, nucleotide analogs, and biological response modifiers. Two or more compounds can be used together or sequentially. For example, anti-cancer agents that are well-known in the art and can be used therapeutically in combination with (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof. Examples of chemotherapeutic agents include alkylating agents, antimetabolites, natural products, hormones and antagonists, and various agents. Examples of alkylating agents include nitrogen mustards, such as mechlorethamine, cyclophosphamide, ifosfamide, melphalan (L-sarcolysin), and chlorambucil; ethyleneimines and methylmelamines, such as hexamethylmelamine and thiotepa; alkyl sulfonates, such as busulfan; nitrosoureas, such as carmustine (BCNU), semustine (methyl-CCNU), lomustine (CCNU), and streptozotocin; DNA synthesis antagonists, such as estramustine phosphate; and triazines, such as dacarbazine (DTIC, dimethyl-triazenoimidazole carboxamide) and temozolomide. Examples of antimetabolites include folic acid analogs, such as methotrexate (amethopterin); pyrimidine analogs, such as fluorouracin (5-fluorouracil, 5-FU, 5FU), floxuridine (fluorodeoxyuridine, FUdR), cytarabine (cytosine arabinoside), and gemcitabine; purine analogs, such as mercaptopurine (6-mercaptopurine, 6-MP), thioguanine (6-thioguanine, TG), and pentostatin (2'-deoxycoformycin, deoxycoformycin), cladribine, and fludarabine; and topoisomerase inhibitors, such as amsacrine.Examples of natural products include vinca alkaloids such as vinblastine (VLB) and vincristine; taxanes such as paclitaxel (Abraxane) and docetaxel (Taxotere); epipodophyllotoxins such as etoposide and teniposide; camptothecin such as topotecan and irinotecan; antibiotics such as dactinomycin (actinomycin D), daunorubicin (daunomycin, rubidomycin), doxorubicin, bleomycin, mitomycin (mitomycin C), idarubicin, epirubicin; enzymes such as L-asparaginase; and biological response modifiers such as interferon alpha and interleukin 2. Examples of hormones and antagonists include luteinising releasing hormone agonists such as buserelin; corticosteroids such as prednisone and related preparations; progestins such as hydroxyprogesterone caproate, medroxyprogesterone acetate and megestrol acetate; estrogens such as diethylstilbestrol and ethinylestradiol and related preparations; estrogen antagonists such as tamoxifen and anastrozole; androgens such as testosterone propionate and fluoxymesterone and related preparations; androgen antagonists such as flutamide and bicalutamide; and gonadotropin releasing hormone analogs such as leuprolide. Examples of various agents include thalidomide; platinum coordination complexes such as cisplatin, oxaliplatin and carboplatin; anthracenediones such as mitoxantrone; substituted ureas such as hydroxyurea; methylhydrazine derivatives such as procarbazine (N-methylhydrazine, MIH); adrenocortical suppressants such as mitotane and aminoglutethimide; RXR agonists such as bexarotene; and tyrosine kinase inhibitors such as imatinib. In some embodiments, the additional cancer therapy is bortezomib administration.

[0074] Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0075] As used herein, the terms "about" or "approximately" mean within an acceptable error range of a particular value as determined by one of ordinary skill in the art, which depends in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, "about" can mean within or exceeding three standard deviations, according to the practice in the art. Alternatively, "about" can mean within a range of up to 20%, 10%, 5%, and / or 1% of a given value.

[0076] "Improvement" refers to the treatment of cancer, including all types of cancer, and means a change in a better direction. "Improvement" can be either subjective or objective.

[0077] "Improvement in the next-day function" or "if there is an improvement in the next-day function" means an improvement after waking up from a sleep period of one night, where the beneficial effect of the administration of (S)-3-amino-4-(difluoromethylenyl)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is applied to cancer symptoms and is distinguishable, either subjectively by the subject or objectively by an observer, during a period of time after waking up, such as 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 24 hours, etc.

[0078] "PK" means the pharmacokinetic profile. C max is defined as the maximum plasma drug concentration (ng / ml) estimated during the experiment. T max is, C max is defined as the time (minutes) at which C 0-∞It is the total area under the plasma drug concentration-time curve (ng·hr / ml or μg·hr / ml) from drug administration until the drug disappears. The area under the curve is controlled by clearance. Clearance is defined as the volume of blood or plasma (ml / min) from which the drug content is completely removed per unit time.

[0079] "Treating", "treatment", or "treat" can mean reducing, improving, alleviating, remitting, calming, inhibiting, reversing, and / or reducing cancer or cancer symptoms in a subject. In embodiments, "treating", "treat", or "treatment" can mean preventing the appearance of clinical symptoms of a disease or condition in a subject who has or may have a predisposition to the disease or condition but has not yet experienced or exhibited clinical or subclinical symptoms of the disease or condition. The benefit to the subject being treated can be statistically significant, mathematically significant, or at least perceptible to the subject and / or healthcare provider. Nevertheless, prophylactic (preventive) treatment and therapeutic (curative) treatment are two distinct embodiments of the disclosure herein.

[0080] "Pharmaceutically acceptable" means "generally considered safe", e.g., physiologically tolerable, and typically does not produce allergic or similar unfavorable reactions such as gastric upset when administered to humans, for molecular entities and compositions. In embodiments, the term means molecular entities and compositions that are approved by federal or state regulatory agencies as a GRAS list under Sections 204(s) and 409 of the Federal Food, Drug, and Cosmetic Act, and are the subject of premarket review and approval by the FDA or similar lists, the United States Pharmacopeia, or other generally recognized pharmacopeias for use in animals, and more particularly in humans.

[0081] "Administered concomitantly", "administered in combination", "combination of", "administered together" are used interchangeably and mean that two or more agents are administered during the course of treatment. The agents can be administered simultaneously or separately at intervals. The agents can be administered in a single dosage form or in separate dosage forms.

[0082] "Subjects in need thereof" includes individuals diagnosed with cancer or at risk of developing cancer with reasonable certainty. Methods and compositions comprising (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof can be provided to any individual, including, for example, when the subject is a neonate, infant, pediatric subject (6 months to 12 years), adolescent subject (12 years to 18 years) or adult (18 years or older). The subjects are mammals such as humans. "Patient" and "subject" can be used interchangeably herein.

[0083] As used herein, the term "pharmaceutically acceptable salt" means a derivative of a compound as defined herein, where the parent compound is modified by making a salt of the acid or base thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines; and alkali or organic salts of acidic residues such as carboxylic acids. Pharmaceutically acceptable salts include conventional non-toxic salts or quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. Such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid and nitric acid; and organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, naphthalenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid and isethionic acid salts, but are not limited thereto. Pharmaceutically acceptable salts can be synthesized from the parent compound containing basic or acidic moieties by conventional chemical methods.

Example

[0084] The examples provided herein are included only to reinforce the disclosure herein and should not be construed as limiting in any way.

[0085] Example 1 (S)-3-Amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid safety evaluation

[0086] The non-GLP safety evaluation regarding the changes in retinal function and structure was evaluated in rats. In the initial results, (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid was shown to have good tolerance at a dose up to 3 mg / kg / day for 45 days for all eye endpoints evaluated (fundus examination, ERG, OCT, and histological evaluation). Furthermore, the toxicity of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid was evaluated in non-GLP single-dose and repeated-dose toxicity tests in rats and dogs. In the 10-day administration in rats, good tolerance was shown at a dose of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid up to 15 mg / kg in males and up to 10 mg / kg in females. When administered to male and female dogs for 10 days, (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid showed good tolerance at a dose up to 1.0 mg / kg. (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid did not show genotoxicity in the Ames assay. The Ames assay is a well-known method that uses several strains of the bacterium Salmonella typhimurium to test whether a specific chemical causes mutations in the DNA of the test organism. In the Irwin test in mice, no impairment of central nervous system or cardiovascular system safety was confirmed. The Irwin observation test is commonly used to evaluate the effects of new substances on behavior and physiological functions. Roux et al., Curr Protoc Pharmacol. 2005 Jan 1;Chapter 10:Unit 10.

[0087] Evidence of genotoxicity or hERG channel activity attributable to (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid was not confirmed in vitro. In adult mice, an acute no-observed-adverse-effect level (NOAEL) of 6 mg / kg was determined by evaluating the behavioral neurotoxicity of untreated animals following oral administration. In another cohort, when (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid was administered orally at 1 mg / kg / day for 45 days, the compound was well tolerated, with no decrease in body weight gain, no significant behavioral impairment, and no effect on retinal function as measured by electroretinogram (i.e., retinal B-wave amplitude) compared to the control group. These results are in contrast to those of a direct comparison with vigabatrin, where significant decreases in body weight and retinal B-wave amplitude were observed at doses with much lower GABA-AT inhibitory activity. Based on these results, it is clear that inhibition of GABA-AT by (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid does not necessarily involve retinal toxicity such as that of vigabatrin.

[0088] Converting the above doses to human doses according to Reagan-Shaw et al., FASEB J. 2008 Mar;22(3):659-61 gives the following:

[0089] 15 mg / kg rat = 2.4 mg / kg human

[0090] 10 mg / kg rat = 1.62 mg / kg human

[0091] 1 mg / kg / day rat = 0.48 mg / kg / day human

[0092] 1 mg / kg dog = 1.54 mg / kg human

[0093] 6 mg / kg mouse = 0.49 mg / kg human

[0094] 1 mg / kg / day mouse = 0.08 mg / kg / day human

[0095] Example 2 (S)-3-Amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid pharmacokinetics and metabolism profile

[0096] After oral administration of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid to both rats and dogs, the time to maximum concentration (Tmax) was 15 - 30 minutes and the elimination half-life (t1 / 2) was approximately 75 - 90 minutes. Dogs showed higher exposure (AUC basis) than rats for an equivalent oral dose. In vitro studies in hepatocytes have shown that the clearance of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid was significantly higher in rats than in dogs. Intrinsic clearance (Cl int ) = 24.3 (rat), 13.1 (human) and 8.97 mL / min / kg (dog). (S)-3-Amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid was minimally (4 - 20%) bound to plasma proteins in rats, dogs and humans at 37°C.

[0097] Example 3 Prospective evaluation of the efficacy of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid in a mouse model of medulloblastoma and leptomeningeal metastases The xenograft model is established according to Martirosian et al., 2021, Cell Reports 35, 109302. Therefore, animal experiments are performed on 9- to 10-week-old female athymic nude mice housed in ventilated cages (3-5 animals per cage) (n = 20). Animals are housed in an animal room designated for immunodeficient mice with all cages, bedding, and feed sterilized to ensure a pathogen-free environment. Animals are monitored before and after the experiment and are determined to be healthy and pain-free. Animals are not used for other procedures prior to transplantation surgery. All animals are humanely euthanized when they reach a specific time point or when pathological signs, including the appearance of tumor symptoms (paralysis, hydrocephalus, weight loss, head tilt), appear.

[0098] Using the D283 cell line (commercially available from the American Type Culture Collection (ATCC)), a human medulloblastoma cell line that divides rapidly, and Med-2112FH (MD cells commercially available from the Brain Tumor Resource Center of the Fred Hutchinson Cancer Research Institute), three medulloblastoma xenograft models (cohorts) were established: (1) a primary tumor model in which 2x105 D283 scrambled, GABA-AT knockdown (KD), and GABA-AT overexpression (OE) medulloblastoma cells were transplanted into the cerebellum, (2) a competitive leptomeningeal model in which 1x105 D283 GABA-AT KD and 1x105 GABA-AT OE medulloblastoma cells were transplanted into the leptomeningeal space, and (3) a clinically important double injection model that depicts primary and metastatic medulloblastoma by injecting 1x105 Med-2112FH into both the cerebellum and the leptomeningeal space. Mice were first anesthetized with 5% isoflurane (Vetone, Cat#502017) and maintained with 2% - 2.5% isoflurane. To model primary disease, a stereotactic frame was used to inject tumor cells into the cerebellum. Cells were transplanted 1 mm into the cerebellum, 6.47 Bregma, 1 mm lateral to the sagittal suture. To model leptomeningeal disease, the surgical setup mirrors cerebellar injection, but cells are injected into the leptomeningeal space as previously described (Xavier et al., 2018 J. Vis. Exp. 135, 57378.). All transplanted lines express luciferase for in vivo bioluminescence imaging (BLI) after transplantation. BLI was performed on days 3 and 7 after injection in the single injection model and then weekly thereafter. Mice were monitored for the development of symptoms related to tumor burden and humanely euthanized as needed. In the double injection model, BLI was performed on day 3 after injection and then every 3 - 4 days thereafter. Mice were euthanized on day 21 after transplantation to observe the immediate effect of the leptomeningeal microenvironment on medulloblastoma cells.

[0099] In this study, three treatment regimens were utilized with a randomized crossover design, including 1 mg / kg, 3 mg / kg, 5 mg / kg, 6 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, and 40 mg / kg doses of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid in saline and a 3 ml / kg body weight dose of saline as a solvent control, with once-daily, twice-daily, or three-times-daily IP administration. Each animal group was subjected to at least three cycles of chemotherapy treatment and continued for the purpose of result determination. Each cohort was evaluated to compare the increase in survival time compared to untreated animals. Qualitative symptoms observed in the mouse model include irregular stride, impairment of cranial nerve function, and decreased motor coordination and performance. Some of the quantitative behavioral tests include the mouse rotarod, forced air challenge, screen reversal test, horizontal wire test, and stride analysis. Magnetic resonance imaging (MRI) is used to confirm and monitor tumor growth and as an anatomical biomarker of treatment response.

[0100] It should be understood that the examples and embodiments provided herein are illustrative examples and embodiments. Those skilled in the art will envision various changes to the examples and embodiments that are consistent with the scope of the disclosure herein. Such changes are intended to be encompassed by the claims.

Claims

**Claim 1** A method for inhibiting cancer growth and metastasis, comprising administering to a subject in need thereof a composition comprising (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof in an amount of from about 0.0001 mg / kg to about 30 mg / kg. **Claim 2** The method for inhibiting cancer growth and metastasis according to claim 1, wherein the composition comprises (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof in an amount of from about 0.1 mg / kg to about 15 mg / kg. **Claim 3** The method for inhibiting cancer growth and metastasis according to claim 2, wherein the composition comprises (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof in an amount of from about 0.2 mg / kg to about 10 mg / kg. **Claim 4** The method for inhibiting cancer growth and metastasis according to claim 3, wherein the composition comprises (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof in an amount of from about 0.5 mg / kg to about 6.5 mg / kg. **Claim 5** The method for inhibiting cancer growth and metastasis according to claim 1, wherein the cancer is medulloblastoma, glioma, breast cancer, squamous cell carcinoma, melanoma, lung cancer, peritoneal cancer, hepatocellular carcinoma, gastric cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, testicular cancer, bladder cancer, colorectal cancer, endometrial cancer or uterine cancer, salivary gland cancer, kidney cancer or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, head and neck cancer, B-cell lymphoma, chronic lymphocytic leukemia, acute lymphoblastic leukemia, hairy cell leukemia, leptomeningeal carcinomatosis, or chronic myelogenous leukemia and subtypes thereof. **Claim 6** The method for inhibiting cancer growth and metastasis according to claim 1, wherein the subject is a human. **Claim 7** The method for inhibiting cancer growth and metastasis according to claim 1, wherein the subject is also receiving one or more cancer therapies selected from the group consisting of surgery, chemotherapy, radiation therapy, hyperthermia therapy, immunotherapy, hormone therapy or laser therapy. **Claim 8** The method for inhibiting cancer growth and metastasis according to claim 1, wherein the composition is administered enterally or parenterally. **Claim 9** The method for inhibiting cancer growth and metastasis according to claim 8, wherein the composition is administered orally, rectally, transdermally, intramuscularly, intravenously, subcutaneously, intraperitoneally, or intrathecally.

10. A method for treating cancer, comprising administering to a subject in need thereof a composition comprising (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof in an amount of about 0.0001 mg / kg to about 30 mg / kg, thereby treating the cancer.

11. The method for treating cancer according to claim 10, wherein the composition comprises (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof in an amount of about 0.1 mg / kg to about 15 mg / kg.

12. The method for treating cancer according to claim 11, wherein the composition comprises (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof in an amount of about 0.2 mg / kg to about 10 mg / kg.

13. The method for treating cancer according to claim 12, wherein the composition comprises (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof in an amount of about 0.5 mg / kg to about 6.5 mg / kg.

14. The method for treating cancer according to claim 10, wherein the cancer is medulloblastoma, glioma, breast cancer, squamous cell carcinoma, melanoma, lung cancer, peritoneal cancer, hepatocellular carcinoma, gastric cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, testicular cancer, bladder cancer, colorectal cancer, endometrial cancer or uterine cancer, salivary gland cancer, kidney cancer (renal cancer), prostate cancer, vulvar cancer, thyroid cancer, head and neck cancer, B-cell lymphoma, chronic lymphocytic leukemia, acute lymphoblastic leukemia, hairy cell leukemia, leptomeningeal carcinomatosis, or chronic myelogenous leukemia and subtypes thereof.

15. The method for treating cancer according to claim 10, wherein the subject is a human.

16. The method for treating cancer according to claim 10, wherein the subject is also receiving one or more cancer therapies selected from the group consisting of surgery, chemotherapy, radiation therapy, hyperthermia therapy, immunotherapy, hormone therapy, or laser therapy.

17. The method for inhibiting cancer growth and metastasis according to claim 10, wherein the composition is administered enterally or parenterally.

18. The method for inhibiting cancer growth and metastasis according to claim 17, wherein the composition is administered orally, rectally, transdermally, intramuscularly, intravenously, subcutaneously, intraperitoneally or intrathecally.

19. (S)-3-Amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof for use in inhibiting cancer growth and metastasis.

20. The use according to claim 19, wherein about 0.1 mg / kg to about 15 mg / kg of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is used.

21. The use according to claim 19, wherein the cancer is medulloblastoma, glioma, breast cancer, squamous cell carcinoma, melanoma, lung cancer, peritoneal cancer, hepatocellular carcinoma, gastric cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, testicular cancer, bladder cancer, colorectal cancer, endometrial cancer or uterine cancer, salivary gland cancer, kidney cancer or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, head and neck cancer, B-cell lymphoma, chronic lymphocytic leukemia, acute lymphoblastic leukemia, hairy cell leukemia, leptomeningeal carcinomatosis, or chronic myelogenous leukemia and subtypes thereof.

22. (S)-3-Amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof for use in treating cancer.

23. The use according to claim 22, wherein about 0.1 mg / kg to about 15 mg / kg of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is used.

24. The use according to claim 22, wherein the cancer is medulloblastoma, glioma, breast cancer, squamous cell carcinoma, melanoma, lung cancer, peritoneal cancer, hepatocellular carcinoma, gastric cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, testicular cancer, bladder cancer, colorectal cancer, endometrial cancer or uterine cancer, salivary gland cancer, kidney cancer or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, head and neck cancer, B-cell lymphoma, chronic lymphocytic leukemia, acute lymphoblastic leukemia, hairy cell leukemia, leptomeningeal carcinomatosis, or chronic myelogenous leukemia and subtypes thereof.

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