Use of (s)-3-amino-4-(difluoromethylene) cyclopent-1-en-1-carboxylic acid in treatment of cancer

Abstract

Provided are methods for treating cancer and inhibiting proliferation and metastasis of cancer and compositions comprising (S)-3-amino-4-(difluoromethylene) cyclopent-1-en-1-carboxylic acid, or a pharmaceutically acceptable salt thereof, for treating cancer and inhibiting proliferation and metastasis of cancer.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of and priority to U.S. Provisional Application No. 63 / 358,677, filed on July 6, 2022, and is incorporated herein by reference in its entirety. Technical Field

[0002] Treating cancer with (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof. background

[0003] According to the World Health Organization (https: / / www.who.int / news-room / fact-sheets / detail / cancer), cancer is an umbrella term for a large group of diseases that can affect any part of the body. Other terms used are malignant neoplasm and neoplasm. Cancer is the leading cause of death worldwide, with a death count of nearly 10 million in 2020, or nearly one in six deaths. A defining characteristic of cancer is the rapid production of abnormal cells that grow beyond their usual boundaries and can then invade adjacent parts of the body and spread to other organs; the latter process is known as metastasis. Extensive metastasis is the leading cause of cancer death.

[0004] Medulloblastoma (MB) is the most common malignant pediatric 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% to 15% of medulloblastomas are diagnosed during infancy. MB occurs in a bimodal pattern, with peak incidence between 3 and 4 years of age and between 8 and 9 years of age. MB accounts for less than 1% of central nervous system (CNS) tumors in adults, with the highest incidence in adults aged 20 to 34 years.

[0005] MB are defined by four molecular subgroups (Wnt, Shh, Group 3, Group 4) based on transcriptional and epigenetic profiles. See, Manoranjan et al. Nat Commun11, 4323 (2020). Wnt (wingless) and Shh (Sonic Hedgehog) MBs are named after the signaling pathways that are thought to play a prominent role in the pathogenesis of this subgroup. The Wnt signaling pathway is a group of signal transduction pathways that initiate with proteins that transmit signals into the cell via cell surface receptors. NT-activated medulloblastomas are defined by activating mutations in the WNT / β-catenin signaling pathway (e.g., CTNNB1 or germline APC), are often associated with loss of chromosome 6, and have the most favorable prognosis. Manoranjan et al., supra. Wnt MBs account for 10% of cases. Ibid.

[0006] The Shh signaling pathway is one of the major trafficking networks that regulates key events during development, namely, growth and patterning of the multicellular embryo. Abnormalities in the regulation and transduction of the Shh signaling pathway have been associated with birth 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 MBs and occurs with a bimodal age distribution, including many infants and adults but fewer 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 alterations in regulatory pathways. Ibid.

[0007] Group 3 MBs have a photoreceptor / retinal expression signature, whereas group 4 MBs express neuronal genes. Tamayo-Orrego, Lukas, and Frédéric Charron. 1000Research Volume 8, F1000 Faculty Rev-1823. October 29, 2019. Current thinking is that each distinct group of MBs derives from a specific cell of origin, thus dictating the clinical and molecular behavior of the disease. Ibid.

[0008] Gliomas are brain tumors that develop from glial cells, which are specialized cells that surround and support neurons in the brain. According to the National Institutes of Health (NIH), Genetic and Rare Diseases Information Center (GARD), gliomas are often classified based on the type of glial cells involved in the tumor: Astrocytomas — tumors that develop from star-shaped glial cells called astrocytes; Ependymomas — tumors that arise from ependymal cells that line the center of the brain ventricles and spinal cord; Oligodendrogliomas — tumors that affect oligodendrocytes (myelinating glia of the central nervous system). Gliomas are graded from grade I to grade IV. Grade I gliomas are usually slow growing and often behave in a more benign way. Grade II and III gliomas can grow faster, and they often require more aggressive treatment. Grade IV gliomas are the most aggressive type and are also known as glioblastomas. Symptoms of gliomas vary by type, but can include headaches, nausea and vomiting, confusion, personality changes, balance problems, vision problems, speech difficulties, and / or seizures. In the study, eight of the 19 WNTs, namely WNT1, 2, 2B, 3A, 5A, 6, 7A, and 7B, were shown to be associated with glioma development. Xu et al., Bioscience reports Volume 40(3)(2020).

[0009] Breast cancer begins in breast tissue, and the cells typically form tumors that can be in situ or invasive. According to the NIH, breast cancer is the second most common type of cancer in women in the United States. Rarely, it may also affect men. Certain genetic markers are associated with the risk of breast cancer, such as BRCA1, BRCA2, and HER2, which may increase the risk of ovarian cancer and other cancers. Different types of breast cancer are generally based on which breast cells turn into cancer. These types include: ductal carcinoma, which starts in the cells of the ducts and is the most common type; lobular carcinoma, which starts in the lobules and is more common in both breasts than other types of breast cancer; inflammatory breast cancer, in which cancer cells block lymph vessels in the skin of the breast; Paget's disease of the breast, which is a cancer involving the skin of the nipple. Signs and symptoms of breast cancer include new lumps or thickenings in or near the breast or in the armpit; changes in the size or shape of the breast; shallow dimples or wrinkles in the skin of the breast; nipple turning inward into the breast; nipple discharge other than breast milk; scaly, red, or swollen skin in the nipple area or breast; and pain in any area of ​​the breast. According to several studies accepted by the U.S. National Library of Medicine, GABA has a significant prognostic value in breast cancer. Breast cancer metastases overexpress many GABA-related variables and are able to proliferate by metabolizing GABA as a biosynthetic energy source, enabling metastatic spread to the brain and cerebrospinal fluid.

[0010] Squamous cell carcinoma is the second most common form of skin cancer and is characterized by abnormal, 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 is not usually life-threatening, although it can be aggressive. Squamous cell carcinoma can also be found in other areas of the body, including the mouth and lungs, but is much rarer. Most squamous cell carcinomas of the skin are caused by long-term exposure to ultraviolet (UV) radiation. Signs and symptoms of squamous cell carcinoma of the skin include firm, red nodules; flat sores with a scaly crust; new sores or raised areas over old scars or ulcers; rough, scaly patches on the lips that may turn into open sores; red sores or rough patches inside the mouth; and red, raised patches or wart-like sores on or in the anus or genitals.

[0011] Lung cancer starts in the lungs and may spread to the lymph nodes or other organs of the body. Up to 40 percent of people with lung cancer develop brain metastases. According to the Centers for Disease Control and Prevention (CDC), there are two main types of lung cancer, called small cell lung cancer and non-small cell lung cancer (which includes adenocarcinoma and squamous cell carcinoma). These types of lung cancer grow differently and are treated differently. Non-small cell lung cancer is more common than small cell lung cancer. People who smoke are at greatest risk for lung cancer. Lung cancer usually does not cause signs and symptoms in its earliest stages. Signs and symptoms of lung cancer usually occur in the later stages of the disease and can include: a new cough that doesn't go away; coughing up blood; shortness of breath; chest pain; hoarseness; losing weight without trying; bone pain; and headaches.

[0012] Cancer of the peritoneum is a cancer that begins in the abdominal cavity or has spread to the abdominal cavity. According to the NIH, whether the cancer starts from the peritoneum or spreads from other places (i.e., ovaries, colon, appendix, etc.), it is considered to be advanced once it enters the peritoneum, and can be referred to as peritoneal cancer spread. Patients in the early stages will usually have very few symptoms until the disease develops. When symptoms occur, they are usually vague and non-specific, such as fatigue; abdominal swelling; diffuse abdominal pain; frequent urination; intestinal changes; abnormal vaginal bleeding; abdominal mass; unexpected weight loss; and a feeling of fullness when eating. As the disease progresses, fluid may accumulate in the abdomen, causing additional abdominal discomfort, nausea and vomiting, and shortness of breath. Complications of peritoneal cancer can include intestinal and urinary tract obstruction, sometimes requiring stents or nephrostomy tubes.

[0013] Hepatocellular carcinoma is the most common type of primary liver cancer. According to the Mayo Clinic and the American Cancer Society, hepatocellular carcinoma most often occurs in people with chronic liver disease (such as cirrhosis caused by hepatitis B or hepatitis C infection), as well as in people who drink a lot of alcohol and have liver fat accumulation. Most patients do not show symptoms in the early stages of primary liver cancer. When symptoms do occur, they can include: weight loss without trying to lose weight; loss of appetite; upper abdominal pain; nausea and vomiting; general weakness and fatigue; abdominal swelling; jaundice; and white chalky stools. GABA may be a byproduct of liver disease and causes hepatic encephalopathy in patients with cirrhosis, which may further lead to hepatocellular carcinoma. A reduction in GABAergic activity may contribute to the pathogenesis of hepatocellular carcinoma.

[0014] Stomach cancer can affect any part of the stomach. In most parts of the world, stomach cancer forms in the body of the stomach. However, in the United States, stomach cancer is more likely to affect the gastroesophageal junction, according to the Mayo Clinic. While it's not clear what causes stomach cancer, factors that increase your risk include: gastroesophageal reflux disease; obesity; a diet high in salt and rich in smoked foods; a diet low in fruits and vegetables; a family history of Helicobacter pylori infection; gastritis; smoking; and stomach polyps. Symptoms of stomach cancer can include: difficulty swallowing; feeling bloated after eating; feeling full after eating a small amount; heartburn; indigestion; nausea; stomach pain; unintentional weight loss; and vomiting. In several studies, GABA has been linked to the proliferation of gastrointestinal cancer types and as a tumor signaling molecule in gastric cancer.

[0015] Pancreatic cancer can develop from two types of cells in the pancreas: exocrine cells and neuroendocrine cells, such as islet cells. The exocrine type is more common and is usually discovered in later stages, according to the National Cancer Institute. Pancreatic neuroendocrine tumors, also called islet cell tumors, are less common but have a better prognosis. It's not clear what causes pancreatic cancer, but some factors that may increase your risk include smoking and inherited gene mutations. Symptoms of pancreatic cancer usually don't appear until the disease is advanced and can include: abdominal pain that radiates to the back; loss of appetite or unintentional weight loss; jaundice; light-colored stools; dark urine; itchy skin; a new diagnosis of diabetes or existing diabetes that becomes more difficult to control; blood clots; and fatigue. GABA has been linked to stimulating pancreatic cancer growth.

[0016] Glioblastoma, also known as grade IV astrocytoma, is a fast-growing and aggressive brain tumor. It invades nearby brain tissue but generally does not spread to distant organs. According to the American Association of Neurological Surgeons, glioblastoma can arise de novo in the brain or evolve from a low-grade astrocytoma. The cause of glioblastoma is unknown, but glioblastoma tends to be more common in older people and men. Symptoms can vary based on the location of the tumor but can include persistent headaches; double vision or blurred vision; vomiting; loss of appetite; changes in mood and personality; changes in thinking and learning ability; new onset of seizures; and progressive speech difficulties.

[0017] Cervical cancer develops in the lower part of the uterus that connects to the vagina. According to the CDC, various strains of the human papillomavirus (a sexually transmitted infection) play a role in causing most cervical cancers. In a small percentage of people, the virus can survive for years, leading to a process that causes some cervical cells to become cancerous. Early cervical cancer usually produces no symptoms. Symptoms of more advanced cervical cancer include: vaginal bleeding after intercourse, between periods, or after menopause; watery, bloody vaginal discharge that may be thick and have a foul odor; and pelvic pain or pain during intercourse.

[0018] Ovarian cancer starts in the ovaries, or in the associated areas of the fallopian tubes or peritoneum. According to the CDC, ovarian cancer presents in a variety of different tumor types. The most common tumor type is high-grade serous carcinoma, which occurs in about 70 percent of ovarian cancer cases. The cause of ovarian cancer is unknown, but risk factors include family history, genetic mutations in BRCA1 or BRCA2, endometriosis, and being middle-aged or older. Signs and symptoms of ovarian cancer can include: abdominal bloating or swelling; weight loss; discomfort in the pelvic area; fatigue; back pain; changes in bowel habits; and frequent urination. Ovarian cancer has been linked to GABA, and GABA can be tested for.

[0019] Bladder cancer most often begins in the urothelial cells that line the bladder, ureters, and kidneys. Older people are at a higher risk for bladder cancer, according to the American Cancer Society. Most bladder cancers are diagnosed in the early stages when the cancer is highly treatable, but bladder cancer has a high recurrence rate. Symptoms of bladder cancer include: blood in the urine; frequent urination; painful urination; and back pain. In several studies, GABA has been linked to the promotion and detection of bladder cancer.

[0020] Colorectal cancer originates in the colon or rectum and, depending on the source, may be called colon cancer or rectal cancer. According to the American Cancer Society, most colorectal cancers begin as polyps in the lining of the colon or rectum. Not all polyps become cancerous, but the risk may be higher or lower 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 are high risk and are called "precancerous." Symptoms of colon cancer include: persistent changes in bowel habits, including diarrhea or constipation, or changes in stool consistency; rectal bleeding or blood in the stool; persistent abdominal discomfort, such as cramping, gas, or pain; weakness or fatigue; and unexplained weight loss. In several NIH-sponsored studies, GABA has been associated with the proliferation and detection of colorectal cancer.

[0021] Endometrial cancer, or uterine cancer, begins in the lining or glands of the uterus. Most cases occur in women over the age of 55. Risk factors for endometrial or uterine cancer include: obesity, family history, age, and hormonal effectors (e.g., estrogen, birth control, pregnancy, and polycystic ovary syndrome). According to the Mayo Clinic, the key symptom is abnormal vaginal bleeding, such as postmenopausal bleeding or bleeding between periods. Other symptoms can include pelvic pain and pain during sexual intercourse, but some women experience no symptoms at all. Endometrial and uterine cancers are associated with GABA and have GABA receptors, which promote cancer proliferation and can be used to detect abnormalities.

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

[0023] Kidney / renal cancer begins in the lining of the kidney tubules. According to the Mayo Clinic, the incidence of kidney cancer appears to be increasing, most likely due to technological improvements in testing and imaging. Risk factors for kidney cancer include smoking, misuse 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 is usually discovered in its early stages, when the cancer is small, confined to the kidney, and often without symptoms. Later stage symptoms may develop, including blood in the urine; persistent back or side pain; loss of appetite; unexplained weight loss; tiredness; and fever. GABA has emerged as a tumor signaling molecule that controls the proliferation of kidney cancer and can be used to detect its presence.

[0024] Prostate cancer develops 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, and finding and treating it before symptoms develop may not improve a man's health or help them live longer. Prostate cancer is rare in men under the age of 40, but the risk of developing prostate cancer rises rapidly after age 50. In addition, inherited mutations in the BRCA1 or BRCA2 genes may increase the risk. Prostate cancer may not cause signs or symptoms in its early stages, but more advanced stages may cause symptoms such as: difficulty urinating; decreased force of urine stream; blood in the urine; blood in the semen; bone pain; weight loss without trying to lose weight; and erectile dysfunction. GABA has been reported to increase cell proliferation associated with prostate cancer.

[0025] Vulvar cancer is a rare cancer that develops on the outer surface of the female genitalia, which includes the labia and clitoris. Although it can occur at any age, it is most common in older adults. Persistent infection with the human papillomavirus may increase the risk of vulvar cancer, as does smoking, according to the NIH. Symptoms of vulvar cancer can include: persistent itching; localized pain and tenderness; bleeding that is not from menstruation; skin changes, such as color changes or thickening; and lumps, wart-like bumps, or open sores.

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

[0027] Head and neck cancer usually starts in the squamous cells that line the mucosal surfaces of the head and neck, such as the inside of the mouth, throat, and larynx. Less common head and neck cancers include cancers of the paranasal sinuses and nasal cavity, as well as salivary gland cancers. Alcohol and tobacco are major risk factors for head and neck cancer. Oropharyngeal cancer is also closely associated with human papillomavirus. UV light may increase the risk of cancer of the lips. Epstein-Barr virus infection may increase the risk of nasal cancer, postnasal cancer, and salivary gland cancer. Chemical and radiation exposure, including wood dust, formaldehyde, asbestos, nickel, etc., may increase the risk of nasopharyngeal cancer. Symptoms can vary based on the type of cancer, but can include: persistent open sores; abnormal pain or bleeding; difficulty chewing, swallowing, and speaking; and localized swelling. GABA has been shown to have 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 originates in B cells. There are many subtypes that fall into 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 people. DLBCL tends to be aggressive and fast-growing. Follicular lymphoma is less common and less aggressive and is very rare in younger people. Symptoms of non-Hodgkin lymphoma can include swollen lymph nodes in the neck, armpits, or groin; abdominal pain or swelling in the abdomen; chest pain, cough, or difficulty breathing; 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 found 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 chemical exposure appear to be risk factors. According to the American Cancer Society, CLL may not cause any symptoms in the early stages, but when symptoms do appear, they can include: swollen lymph nodes; fatigue; fever; pain due to an enlarged spleen; night sweats; weight loss; frequent infections; and easy bruising.

[0030] Acute lymphoblastic leukemia (ALL) is a cancer of the blood and bone marrow that affects white blood cells. According to the American Cancer Society, ALL is the most common childhood cancer, and it is very aggressive and fast-acting. Possible risk factors for ALL include: exposure to x-rays before birth; exposure to radiation; 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 can include: enlarged lymph nodes; bruises; fever; bone and joint pain; bleeding gums; shortness of breath; fatigue and weakness; loss of appetite; and frequent infections. GABA is involved in the proliferation and spread of ALL and has been observed in several studies.

[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, with projections that give the "hairy" appearance. According to the Mayo Clinic, HCL affects more men than women, and it most often develops in middle-aged or older adults. HCL is considered a chronic disease because it may never completely go away, although treatment can lead to years of remission. Some people have no signs or symptoms of hairy cell leukemia, but a blood test for another disease or condition may inadvertently reveal hairy cell leukemia. When symptoms do occur, they can include: a feeling of fullness in the abdomen that makes eating more than a little uncomfortable; fatigue; easy bruising; recurring infections; weakness; and unintentional weight loss.

[0032] Chronic myeloid leukemia (CML) is a rare type of cancer that begins in certain blood-forming cells in the bone marrow. CML is a slow-growing leukemia, but it can turn into a fast-growing acute leukemia that is difficult to treat. CML occurs mainly in adults but very rarely in children. According to the Mayo Clinic, CML usually does not cause symptoms and may not be detected during a blood test. When symptoms do occur, they can include: bone pain; easy bleeding; feeling full after eating small amounts of food; fatigue; fever; unintentional weight loss; loss of appetite; pain or fullness on the left side below the ribs; and 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 and is very aggressive and progresses quickly. According to the Mayo Clinic, men and adults over 65 are at higher risk for AML, as are smokers, people previously exposed to radiation and hazardous chemicals, and people with other blood disorders. Symptoms of early-stage AML may be similar to those of the flu or other common illnesses. Later stages may have symptoms that include: fever; bone pain; lethargy and fatigue; shortness of breath; pale skin; frequent infections; easy bruising; and unusual bleeding, such as frequent nosebleeds and bleeding gums.

[0034] Melanoma is the most dangerous type of skin cancer. It grows quickly and can spread to any organ. Melanoma originates in melanocytes, cells that produce melanin. According to the National Cancer Institute, the exact cause of melanoma is unknown, but exposure to ultraviolet radiation from the sun or tanning lamps / beds increases the risk of developing melanoma. The risk of developing melanoma appears to increase in people under the age of 40, especially women. The first melanoma symptoms are usually changes in existing moles or the development of new pigmentation or abnormal-looking growths on the skin. Malignant moles vary widely in appearance. Some signs that the pigmentation may be malignant are if the growth is asymmetrical in shape; has irregular borders; changes in color; has growth in diameter; or appears to be evolving in color or shape. Melanoma and GABA have demonstrated a direct relationship, and melanoma has GABA receptors that proliferate cancerous growths.

[0035] Testicular cancer develops in the testicles and is the most common cancer in American men between the ages of 15 and 35. There are two main forms: seminomas and nonseminomas. Seminomas grow slowly and mainly affect people between the ages of 40 and 60. Nonseminomas grow faster and mainly affect teenagers, people in their 20s and 30s. Symptoms of testicular cancer include a lump or enlargement in either testicle; a feeling of heaviness in the scrotum; dull pain in the abdomen or groin; a sudden accumulation of fluid in the scrotum; pain or discomfort in the testicle or scrotum; enlargement or tenderness in the breast; and back pain. Testicular cancer usually affects only one testicle. Testicular cancer and GABA have demonstrated a direct relationship, with GABA promoting cancer cell growth and changes in sperm motility.

[0036] Brain metastasis (metastatic brain tumor) and leptomeningeal metastasis occur when cancer spreads from the origin or primary cancer site. Brain metastasis occurs when cancer cells move to the brain itself. Leptomeningeal metastasis (also known as leptomeningeal cancer, leptomeningeal cancer spread, leptomeningeal disease (LMD), tumor meningitis, meningeal metastasis, and meningeal cancer spread) is a rare complication of cancer in which the disease spreads from the original tumor site to the meninges surrounding the brain and spinal cord. The pia mater is the innermost two layers of tissue covering the brain and spinal cord. Leptomeningeal metastasis occurs when cancer cells invade the cerebrospinal fluid and spread throughout the central nervous system. Prognosis is usually poor, and survival is usually measured in months. The most common cancers that involve the leptomeninges are breast cancer, lung cancer, and melanoma, because they can metastasize to the subarachnoid space. This condition may also be caused by primary brain tumors such as medulloblastoma or glioma. The most common symptoms of leptomeningeal cancer are pain and seizures. Other symptoms may include headaches (often associated with nausea, vomiting, dizziness), gait difficulties due to weakness or ataxia, memory problems, incontinence, paresthesias. In some cases, symptoms may include double vision, jaw numbness, back pain, leg weakness, sphincter-related problems, hydrocephalus, loss of urination, difficulty walking, limb weakness and paresthesias, bowel and bladder dysfunction, double vision, loss of trigeminal nerve sensation or movement, cochlear dysfunction, confusion, and cognitive impairment.

[0037] γ-Aminobutyric acid (GABA) is metabolized by transamination by γ-aminobutyric acid transaminase (GABA-AT), also known as GABA-transaminase or ABAT. Inhibition of this enzymatic process reduces GABA degradation, leading to increased extracellular and neuronal GABA concentrations. Vigabatrin is a GABA-AT inhibitor that has been used to treat refractory epilepsy and infantile spasms. It has been suggested that inhibition of IKCa channels by vigabatrin may be an important potential mechanism for vigabatrin-induced antitumor effects associated with gliomas. Hung et al., BMC Pharmacol Toxicol 22, 6 (2021). Vigabatrin is associated with several potentially serious side effects. Its use is limited due to potential 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 is 186 times more efficient than vigabatrin in inactivating GABA-AT. It is reported that preclinical data of CPP-115 show that significantly lower drug doses provide comparable pharmacokinetics, improved tolerability, and a more favorable toxicity profile when compared to vigabatrin. 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, which is 9.8 times more efficient than CPP-115 as an inactivator of GABA-AT. Same as above. U.S. Patent Nos. 9,603,820 and 9,993,449, respectively, describe the use of CPP-115 and OV329 for the treatment of hepatocellular carcinoma.

[0038] Many cancers are treated with surgery, chemotherapy, radiation or a combination thereof. Chemotherapeutic agents for the treatment of cancer are known to produce serious and unpleasant side effects in patients. For example, some chemotherapeutic agents can cause neuropathy, nephrotoxicity (for example, hyperlipidemia, proteinuria, hypoproteinemia, combinations thereof, etc.), stomatitis, mucositis, alopecia, anorexia, esophagitis, amenorrhea, decreased immunity, anemia, high-pitched hearing loss, cardiotoxicity, fatigue, neuropathy, bone marrow suppression, to name a few. In some cases, during treatment or soon after the treatment regimen draws a conclusion (that is, the treatment regimen does not result in cure), chemotherapy is invalid or loses effectiveness after a period of time. It is still desirable to be used for the improved method for the treatment of cancer and to be able to deliver a bioactive agent to help treat the composition of cancer. In addition, in view of the high mortality level attributable to cancer, there is a demand for the other treatment of the disease. Overview

[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, the compound comprising ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof for treating cancer 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 lymphocytic leukemia (ALL), hairy cell leukemia, leptomeningeal carcinoma spread or chronic myeloblastic 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 aforementioned cancers. In an embodiment, ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof prevents metastasis of the aforementioned cancers. In an embodiment, ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is between about 0.0001 mg / kg and about 30 mg / kg. In an embodiment, 0.5 mg / kg to 6.5 mg / kg of ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof. In embodiments, an effective amount of ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof. Details

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

[0041] In leptomeningeal metastasis, in order for metastasis to occur, tumor cells must complete a series of sequential steps. At the primary site, tumor cells proliferate, invade the surrounding environment, enter the cerebrospinal fluid and spread throughout the central nervous system. Metastatic tumor cells will then 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 cancer cells such as metastatic MB cells use GABA-AT to maintain viability in metabolite-deficient cerebrospinal fluid by using GABA as an energy source substitute, thereby promoting the formation of leptomeningeal metastasis. Martirosian et al., 2021, Cell Reports 35, 109302.

[0042] According to the present disclosure, but not wishing to be bound by any theory, by administering (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof as a GABA-AT inhibitor, GABA metabolism is blocked or reduced, and metastatic MB cells are unable to fully utilize GABA as an energy source alternative, thereby creating an even more hostile nutrient-poor environment for metastatic MB cells in the cerebrospinal fluid and throughout the central nervous system. In this way, metastasis is prevented or reduced. In fact, any cancer described herein that utilizes GABA catabolism via GABA-AT as an energy source is susceptible to the effects of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof in terms of reducing or preventing 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 significantly affect circulating GABA, which, as discussed above, can have pro-tumor and / or metastatic effects.

[0044] It is noteworthy that many anticancer compounds cannot cross the blood-brain barrier and cannot be used to treat 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 treat brain cancer and leptomeningeal metastasis.

[0045] As used herein, the term "cancer" refers to a proliferative disorder caused by or characterized by the proliferation of cells that lose sensitivity to normal growth controls. Cancers of the same tissue type usually originate from the same tissue and can be divided into different subtypes based on their biological characteristics. The four major categories of cancer are carcinomas (epithelial cell origin), sarcomas (connective tissue or mesodermal origin), leukemias (hematopoietic tissue origin), and lymphomas (lymphoid tissue origin). Non-limiting examples of cancers 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 lymphocytic leukemia, hairy cell leukemia or chronic myeloblastic leukemia and subtypes thereof, including refractory forms of any of the above cancers, or a combination of one or more of the above cancers. The above cancers can utilize GABA-AT to metabolize GABA as an energy source.

[0046] As used herein, the terms "cancer", "neoplasm" and "tumor" are used interchangeably and, in the singular or plural, refer to cells that have undergone malignant transformation that makes them pathological to the host organism. Primary cancer cells (i.e., cells obtained near the site of malignant transformation) can be easily distinguished from non-cancerous cells by well-established techniques, particularly histological examination. The definition of cancer cells as used herein includes not only primary cancer cells, but also any cells derived from cancer cell ancestors. This includes metastatic cancer cells, and in vitro cultures and cell lines derived from cancer cells. When referring to a cancer that is usually manifested as a solid tumor, a "clinically detectable" tumor is a tumor that is detectable based on tumor mass, for example, by procedures such as CAT scans, MR imaging, X-rays, ultrasound or palpation, and / or due to the expression of one or more cancer-specific antigens in a sample that can be obtained from a patient. Leukemia can be detected clinically using one or more of a complete blood count, pallor, blood smear, and bone marrow smear. Advanced leukemia in some subjects can be manifested as a solid tumor.

[0047] ( SThe structure of )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid can be represented as follows:

[0048] In an embodiment, ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid can be provided as acid addition salt, zwitterion hydrate, zwitterion anhydrate, hydrochloride or hydrobromide or in the form of zwitterion monohydrate. Acid addition salt includes but is not limited to maleic acid, fumaric acid, benzoic acid, ascorbic acid, succinic acid, oxalic acid, dimethoxy salicylic acid, methanesulfonic acid, ethanedisulfonic 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 theophylline acetic acid addition salt and 8-halogenated theophylline such as 8-bromotheophylline. In embodiments, inorganic acid addition salt can be used, including but not limited to hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfamic acid, phosphoric acid or nitric acid addition salt.

[0049] As can be seen from the embodiments herein, the (S)-3-amino-4-(difluoromethylene) cyclopent-1-ene-1-carboxylic acid or its pharmaceutically acceptable salt for the treatment of cancer herein surprisingly has no or shows less undesirable side effects commonly associated with GABA-AT inhibitors such as vigabatrin, such as retinal toxicity and reduced motor coordination. When applied to (S)-3-amino-4-(difluoromethylene) cyclopent-1-ene-1-carboxylic acid or its pharmaceutically acceptable salt, the term "effective amount" refers to the amount for effectively preventing or treating cancer. The "effective amount" of (S)-3-amino-4-(difluoromethylene) cyclopent-1-ene-1-carboxylic acid or its pharmaceutically acceptable salt can be determined by a variety of factors, for example, method of administration, target site, patient condition, etc. Therefore, in view of safety and efficacy, the dosage when used in human body should be determined with an appropriate amount. The amount used for humans can also be estimated according to the effective amount determined by animal experiments. See, e.g., Reagan-Shaw et al., FASEBJ. 2008 Mar;22(3):659-61.

[0050] As used herein, the term "prevent", including its grammatical variants such as "preventing" and "prevention", refers to the effect of inhibiting or delaying the development, spread and recurrence of cancer by administering (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or its pharmaceutically acceptable salt. As used herein, the term "treat", including its grammatical variants such as "treating" and "treatment", refers to all types of effects of reducing the proliferation of cancer cells, promoting the death of cancer cells and / or reducing the symptoms of cancer by administering (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or its pharmaceutically acceptable salt according to the disclosure herein. "Treatment of cancer" herein includes inhibiting the proliferation of cancer and the metastasis of cancer cells. For example, in an embodiment, "treatment" can include reducing the inoculation of metastatic cells or reducing the inoculation of tumor cells.

[0051] An "effective amount" herein ranges from about 0.0001 mg / kg to about 30 mg / kg ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof. In embodiments, inhibition of GABA-AT without significantly affecting circulating GABA ( S A relatively low dose of )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof ranges from about 0.2 mg / kg to 10 mg / kg. In embodiments, inhibition of GABA-AT without significantly affecting circulating GABA ( SRelatively low doses of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof range from about 0.5 mg / kg to about 6.5 mg / kg. In embodiments, from 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 can contain from 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 240 mg to about 250 mg, 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 350 mg to about 360 mg, about 375 mg to about 380 mg, about 385 mg to about 390 mg, about 395 mg to about 400 mg, about 410 mg to about 420 mg, about 430 mg to about 440 mg 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.mg of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof. 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 comprises 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 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, 88mg, 89 mg, 90 mg, 91 mg, 92mg, 93 mg, 94 mg, 95 mg, 96 mg, 97 mg, 98 mg, 99 100 mg 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, 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, 280390 mg or 400 mg of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof.

[0052] In an embodiment, ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is administered 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, 185 190 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, 360 365 mg / day, 370 mg / day, 375 mg / day, 380 mg / day, 385 mg / day, 390 mg / day, 395 mg / day or 400 mg / day is administered to a subject. In an embodiment, a subject can start with a low dose and the dose can be increased over time.

[0053] In embodiments, the administering step results in at least one of: a 10% decrease in tumor size, a 10% decrease in the number of cancer metastases, or an at least 10% improvement in clinical signs and symptoms associated with cancer. As used herein, the term "administration" and its grammatical variants such as "administer" and "administering" means providing a predetermined substance to an individual or patient by any suitable method. For example, S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof can be administered enterally, such as orally, or parenterally (e.g., by intravenous, subcutaneous, intraperitoneal or topical application of injectable preparations). The dosage can vary depending on the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate, severity of the disease, etc. For oral administration ( S Liquid preparations of 4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof include suspensions, oral solutions, emulsions, syrups, etc. In addition to water and liquid paraffin as commonly used simple diluents, various excipients such as wetting agents, sweeteners, flavoring agents, preservatives, etc. may also be included. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, 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 suffering from cancer via a pharmaceutical composition. The pharmaceutical compositions herein may be administered by any device capable of moving the active substance to the target cells. The pharmaceutical compositions herein encompass dosage forms. The dosage forms herein encompass unit doses. In embodiments, as discussed below, a variety of dosage forms including conventional formulations and modified release formulations may be administered once or more per 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 (e.g., morning and / or night), or three times a day (morning, afternoon and evening), or four times a day (morning, afternoon, evening and night). Any suitable route of administration can be used, 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, transdermal forms such as topical liquids, patches, creams and ointments, parenteral formulations, and suppositories. In an embodiment, ( S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is used to manufacture a medicament for treating cancer or inhibiting cancer proliferation and metastasis. Those skilled in the art are familiar with drug formulation techniques for preparing and manufacturing pharmaceutical compositions. See, for example, EW Martin, ed., Remington's Pharmaceutical Sciences, 18th edition (1990), Mack Publishing Co.

[0056] In an embodiment, a method for treating cancer (including inhibiting cancer proliferation and metastasis) is provided, the method comprising administering a pharmaceutical composition comprising (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof to a subject in need thereof, wherein the composition provides an improvement in cancer symptoms for more than 1 hour after administration to the subject. In an embodiment, a method for treating cancer (including inhibiting cancer proliferation and metastasis) is provided, the method comprising administering a pharmaceutical composition comprising (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof to a subject in need thereof, wherein the composition provides an improvement in one or more cancer symptoms for more than 2 hours after administration to the subject. In an embodiment, a method for treating cancer (including inhibiting cancer proliferation and metastasis) is provided, the method comprising administering a pharmaceutical composition comprising (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof to a subject in need thereof, wherein the composition provides improvement in one or more cancer symptoms for more than 3 hours after administration to the subject. In an embodiment, a method for treating cancer (including inhibiting cancer proliferation and metastasis) is provided, the method comprising administering a pharmaceutical composition comprising (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof to a subject in need thereof, wherein the composition provides improvement in one or more cancer symptoms for more than 4 hours after administration to the subject. In an embodiment, a method for treating cancer (including inhibiting cancer proliferation and metastasis) is provided, the method comprising administering a pharmaceutical composition comprising (S)-3-amino-4-(difluoromethylene) cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof to a subject in need thereof, wherein the composition provides an improvement in one or more cancer symptoms for more than 6 hours after administration to the subject. In an embodiment, a method for treating cancer (including inhibiting cancer proliferation and metastasis) is provided, the method comprising administering a pharmaceutical composition comprising (S)-3-amino-4-(difluoromethylene) cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof to a subject in need thereof, wherein the composition provides an improvement in one or more cancer symptoms for more than 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 an improvement in the next day function of the subject. For example, the pharmaceutical composition can provide improvement in one or more cancer symptoms that persists for more than about, e.g., 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 embodiments, the pharmaceutical compositions herein may be provided with conventional release or modified release profiles. Pharmaceutical compositions may be prepared using pharmaceutically acceptable "carriers" comprising materials that are considered safe and effective. "Carriers" include all components present in a pharmaceutical formulation except for one or more active ingredients or ingredients. 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 compounding pharmaceutical compositions using such carriers. In an embodiment, the pharmaceutical composition herein may include pharmaceutically acceptable additives, which are exemplified by starch, gelatinized starch, microcrystalline cellulose, milk sugar, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose, mannitol, gum arabic, pregelatinized starch, corn starch, cellulose powder, hydroxypropyl cellulose, Opadry, sodium starch glycolate, carnauba wax, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, white sugar, dextrose, sorbitol, talc, etc. In an embodiment, pharmaceutically acceptable additives may be added to the composition in an amount of 0.1 to 90 parts by weight, but are not limited thereto.

[0058] In an embodiment, the pharmaceutical composition herein is a modified release dosage form providing a modified release profile. The modified release profile can present an immediate release profile, a delayed release profile or an extended release profile. Conventional (or unmodified) release oral dosage forms such as tablets, capsules, suppositories, syrups, solutions and suspensions typically release the drug into the mouth, stomach or intestine as the tablet, capsule shell or suppository dissolves, or in the case of syrups, solutions and suspensions, the drug is released into the mouth, stomach or intestine when they are swallowed. In order to achieve the desired therapeutic purpose and / or better patient compliance, the mode of drug release from a modified release (MR) dosage form is intentionally changed from the mode of drug release of a conventional dosage form. The types of MR drug products include orally disintegrating dosage forms (ODDF) providing immediate release, extended release dosage forms, delayed release dosage forms (e.g., enteric coated) and pulsed release dosage forms.

[0059] ODDF is a solid dosage form containing a pharmaceutical substance or active ingredient, which usually disintegrates rapidly within a few seconds when placed on the tongue. The disintegration time of ODDF is usually in the range of from one or two seconds to about one minute. ODDF is designed to disintegrate or dissolve rapidly after contact with saliva. This mode of administration can be beneficial to people who may have problems swallowing tablets, whether the problem is essentially from physical weakness or mental illness. Subjects in pain may show such behavior. ODDF can provide rapid delivery of drugs to blood flow through mucous membranes, resulting in rapid onset. The example of ODDF includes orally disintegrating tablets, capsules, and rapidly dissolving films and wafers.

[0060] Extended release dosage forms (ERDF) have an extended release profile and are those dosage forms that allow a reduction in dosing frequency compared to the dosing frequency presented by conventional dosage forms such as solutions or unmodified release dosage forms. ERDF provides a sustained duration of drug action. Suitable formulations that provide an extended release profile are well known in the art. For example, coated slow-release beads or granules ("beads" and "granules" are used interchangeably herein), wherein ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is applied to beads, such as confectioners nonpareil beads, and then coated with conventional release-delaying materials such as wax, enteric coatings, and the like. In an embodiment, beads may be formed wherein ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is mixed with a material to provide a mass from which the drug is leached. In an embodiment, the beads can be engineered to provide different release rates by changing the properties of the coating or mass, such as thickness, porosity, using different materials, etc. Beads with different release rates can be combined into a single dosage form to provide variable or continuous release. The beads can be contained in a capsule or compressed into a tablet.

[0061] In an embodiment, the modified dosage forms herein include delayed release dosage forms having a delayed release profile. The delayed release dosage form may include a delayed release tablet or a delayed release capsule. A delayed release tablet is a solid dosage form that releases a drug (or drugs) at a time different from immediate release after administration, such as ( S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof. Delayed release capsules are solid dosage forms in which the drug is encapsulated in a hard or soft soluble container made of a suitable form of gelatin, and the solid dosage form releases a drug (or more drugs) at a time different from immediate release after administration. For example, enteric-coated tablets, capsules, granules and beads are well-known examples of delayed release dosage forms. Enteric-coated tablets, capsules, granules 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 comprising a conglomerate of pharmaceutical particles that releases a drug (or more drugs) at a time different from immediate release after administration. In an embodiment, the agglomerate of the 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 that comprises a conglomerate of pharmaceutical particles that releases a drug (or more drugs) at a time different from immediate release after administration. In an embodiment, the agglomerates of pharmaceutically acceptable particles are covered with a coating that delays the release of the drug.

[0062] Delayed release dosage forms are known to those skilled in the art. For example, coated delayed release beads or granules, wherein ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is applied to beads, such as sugar granule beads, and then coated with conventional release delaying materials such as wax, enteric coatings, and the like. In an embodiment, beads may be formed wherein ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is mixed with a material to provide a mass from which the drug is leached. In embodiments, the beads can be engineered to provide different release rates by varying the properties of the coating or mass, such as thickness, porosity, use of different materials, etc. In embodiments, ( S Enteric coated granules of 1-(2-((4-((2-( ...))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))))

[0063] In the implementation scheme, ( 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 embodiments, the porous inert carrier incorporates channels or pathways from which the drug diffuses into the surrounding fluid. In embodiments, ( 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. When the drug-resin complex contacts the gastrointestinal fluid and the ionic components dissolved therein, the drug is released from the resin at a predetermined rate, which can result in a delayed effect. In an embodiment, a membrane is used to control the rate of release from a reservoir containing the drug. In an embodiment, a liquid product can also be used to provide a delayed release profile. For example, a liquid product composed of solid particles dispersed throughout a liquid phase, the particles being insoluble in the liquid phase. The suspension is formulated to at least allow a reduction in the frequency of administration of the drug as compared to the frequency of administration of the drug presented as a conventional dosage form (e.g., as a conventional solid dosage form of a solution or rapid release drug). For example, a suspension of an ion exchange resin component or microbeads.

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

[0065] In the implementation scheme, ( 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 composite materials thereof.

[0066] In embodiments, the pharmaceutical compositions described herein are suitable for parenteral administration, including, for example, intramuscular (im), intravenous (iv), subcutaneous (sc), intraperitoneal (ip), or intrathecal (it). Parenteral compositions must be sterile for administration by injection, infusion, or implantation into the body, and may be packaged in single-dose or multi-dose containers. In embodiments, a liquid pharmaceutical composition for parenteral administration to a subject comprises any corresponding amount of the active substance described above, for example, ( S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof. In an embodiment, the pharmaceutical composition for parenteral administration is formulated to a total volume of about, 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, a glass vial, a plastic vial, or a bottle.

[0067] The pharmaceutical compositions provided herein for parenteral administration may contain one or more excipients, such as solvents, solubility enhancers, suspending agents, buffers, isotonic agents, stabilizers, or antimicrobial preservatives. When used, the excipients of the parenteral compositions will not affect the ( S The invention relates to a parenteral composition wherein the stability, bioavailability, safety and / or efficacy of the parenteral composition is adversely affected by the presence of any incompatibility between any of the components of the dosage form.

[0068] In an embodiment, comprising ( S The parenteral composition of 2-(4-(2-fluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof comprises a stabilizing amount of at least one excipient. For example, the excipient may be selected from the group consisting of a buffer, a solubilizer, a tonicity agent, an antioxidant, a chelating agent, an antimicrobial agent, and a preservative. It will be appreciated by those skilled in the art that an excipient may have more than one function and be classified into one or more defined groups.

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

[0070] In an embodiment, a parenteral composition of an active substance is provided, the 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 between about 4.0 and about 8.0. In embodiments, the pH of the composition is, for example, between about 5.0 and about 8.0, about 6.0 and about 8.0, about 6.5 and about 8.0. In embodiments, the pH of the composition is, for example, between about 6.5 and about 7.5, about 7.0 and about 7.8, about 7.2 and about 7.8, or about 7.3 and about 7.6. In embodiments, 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 ( S The dosage amount of )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is applicable to all dosage forms described herein, including conventional dosage forms, modified dosage forms, and parenteral formulations described herein. Those skilled in the art will determine the appropriate amount depending on criteria such as dosage form, route of administration, subject tolerance, efficacy, therapeutic goals and therapeutic benefits, and 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, thermal therapy, immunotherapy, hormone therapy and / or laser therapy.

[0073] In embodiments, combination therapy may include, for example, one or more of a chemotherapeutic agent; a targeting agent such as an antibody; a kinase inhibitor; a hormonal agent, etc. Combination therapy may also include conventional therapy, including but not limited to administration of antibodies, administration of vaccines, administration of cytotoxic agents, natural amino acid polypeptides, nucleic acids, nucleotide analogs, and biological response modifiers. Two or more combined compounds may be used together or sequentially. For example, an anticancer agent well known in the art may be used with ( S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof is used as a treatment. Examples of chemotherapeutic agents include alkylating agents, antimetabolites, natural products, hormones and antagonists, and other 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 streptozocin (streptozotocin); DNA synthesis antagonists, such as estramustine phosphate; and triazines, such as dacarbazine (DTIC, dimethyl-triazainylimidazolecarboxamide) and temozolomide. Examples of antimetabolites include folic acid analogs such as methotrexate (methotrexate / amethopterin); pyrimidine analogs such as fluorouracil (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; camptothecins, such as topotecan and irinotecan; antibiotics, such as dactinomycin (actinomycin D), daunorubicin (daunorubicin), 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 luteinizing releasing hormone agonists such as buserelin; adrenocortical hormones such as prednisone and related products; progestins such as hydroxyprogesterone caproate, medroxyprogesterone acetate, and megestrol acetate; estrogens such as diethylstilbestrol and ethinyl estradiol and related products; estrogen antagonists such as tamoxifen and anastrozole; androgens such as testosterone propionate and fluoxymesterone and related products; androgen antagonists such as flutamide and bicalutamide; and gonadotropin-releasing hormone analogs such as leuprolide.Examples of other 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 defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present disclosure belongs.

[0075] As used herein, the term "about" or "approximately" means within an acceptable error range for a particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, according to the practice in the art, "about" can mean within 3 or more than 3 standard deviations. Alternatively, "about" can mean a range of up to 20%, up to 10%, up to 5%, and / or up to 1% of a given value.

[0076] "Improvement" refers to treatment of cancer, including all forms of cancer, and means getting better. "Improvement" can be subjective or objective.

[0077] "Improvement in next-day function" or "wherein there is an improvement in next-day function" refers to an improvement after awakening from an overnight sleep period, wherein administration of ( S The beneficial effects of )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof are applicable to cancer symptoms and persist for a period of time after waking up, such as 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 24 hours, etc., which is subjectively discernible by the subject or objectively discernible by an observer.

[0078] "PK" refers to pharmacokinetic profile. 最大 T is defined as the highest estimated plasma drug concentration (ng / ml) during the experimental period. 最大 is defined as when C 最大 The time (min) when AUC is estimated. 0-∞ It is the total area under the plasma drug concentration-time curve from the time the drug is administered until it is eliminated (ng·hr / ml or µg·hr / ml). The area under the curve is determined by the clearance rate. The clearance rate is defined as the volume of blood or plasma that completely clears its drug content per unit time (ml / min).

[0079] "Treating," "treatment," or "treat" may refer to the following: reducing, improving, alleviating, ameliorating, mitigating, inhibiting, reversing, and / or alleviating a cancer or cancer symptom in a subject. In embodiments, "treating," "treatment," or "treatment" may refer to preventing the appearance of clinical symptoms of a disease or condition in a subject who may be suffering from or susceptible to a disease or condition but who has not yet experienced or displayed clinical or subclinical symptoms of the disease or condition. The benefit to the subject being treated may be statistically significant, mathematically significant, or at least perceptible to the subject and / or to the physician. Nevertheless, prophylactic (preventive) treatment and therapeutic (curative) treatment are two separate embodiments of the disclosure herein.

[0080] "Pharmaceutically acceptable" refers to molecular entities and compositions that are "generally regarded as safe," e.g., they are physiologically tolerable and typically do not produce allergic reactions or similar adverse reactions such as stomach upset and the like when administered to humans. In embodiments, the term refers to molecular entities and compositions that are approved by a regulatory agency of a federal or state government, as a GRAS list or similar list under sections 204(s) and 409 of the Federal Food, Drug and Cosmetic Act that has undergone premarket review and is approved by the FDA, the U.S. Pharmacopeia, or another generally recognized pharmacopoeia for use in animals and more particularly in humans.

[0081] "Co-administered with," "administered in combination with," "combination of," or "administered together with" may be used interchangeably and mean the administration of two or more agents in the course of therapy. The agents may be administered together at the same time or separately at spaced-apart intervals. The agents may be administered in a single dosage form or in separate dosage forms.

[0082] "Subjects in need thereof" include individuals who have been diagnosed with cancer or are reasonably certain to be at risk of developing cancer. SThe methods and compositions of producing 4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof can be provided to any individual, including, for example, where the subject is a neonate, infant, pediatric subject (6 months to 12 years old), adolescent subject (age 12 years to 18 years old), or adult (over 18 years old). Subjects include mammals, such as humans. "Patient" and "subject" can be used interchangeably herein.

[0083] As used herein, the term "pharmaceutically acceptable salt" refers to a derivative of a compound as defined herein, wherein the parent compound is modified by preparing an acid salt or a basic salt thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; and alkali metal 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, for example, formed from non-toxic inorganic or organic acids. Such conventional non-toxic salts include, but are not limited to, those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid and nitric acid; and salts prepared from following 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 hydroxyethanesulfonic acid. Pharmaceutically acceptable salts can be synthesized by conventional chemical methods from parent compounds comprising a basic or acidic moiety. Example

[0084] The examples provided herein are included merely to enhance the disclosure herein and should not be considered limiting in any respect.

[0085] Example 1 ( S Safety Assessment of 3-Amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic Acid Non-GLP safety assessments of changes in retinal function and structure were evaluated in rats. Preliminary results indicate that for all evaluated ocular endpoints (funduscopy, ERG, OCT, and histology), ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid was well tolerated at doses up to 3 mg / kg / day for 45 days. In addition, ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid toxicity. In rats, ( S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid was well tolerated at doses up to 15 mg / kg in males and 10 mg / kg in females. In 10-day dosing in male and female dogs, ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid was well tolerated at doses up to 1.0 mg / kg. S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid was not genotoxic in the Ames assay. The Ames assay is a well-known method that uses several strains of the bacterium Salmonella typhimurium ( Salmonella typhimurium ) strains to test whether a given chemical can cause mutations in the DNA of the test organism. No CNS or cardiovascular safety concerns were found in the mouse Irwin study. The Irwin observation test is often used to evaluate the effects of new substances on behavior and physiological function. Roux et al., Curr Protoc Pharmacol. 2005 Jan 1; Chapter 10: Unit 10.

[0086] No protein was identified in vitro ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid-induced genotoxicity or hERG Evidence of channel activity. An acute no-observed adverse effect level (NOAEL) of 6 mg / kg orally was established in adult mice by evaluating behavioral neurotoxicity in naive animals. In a separate group, 1 mg / kg / day was administered orally ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid for 45 days showed that the compound was well tolerated, with no reduction in weight gain relative to controls, no significant behavioral deficits, and no effects on retinal function as measured by electroretinogram (i.e., retinal B-wave amplitude). These findings are in contrast to the findings of head-to-head comparisons with vigabatrin, in which doses with considerably less GABA-AT inhibitory activity resulted in significant weight loss and reduced retinal B-wave amplitude. Based on these findings, it is clear that ( S The inhibition of GABA-AT by 3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid is not necessarily associated with vigabatrin-like retinal toxicity.

[0087] According to Reagan-Shaw et al., FASEB J. 2008 Mar;22(3):659-61, converting the above doses to human doses yields the following: 15 mg / kg rat = 2.4 mg / kg human 10 mg / kg rat = 1.62 mg / kg human 1 mg / kg / day rat = 0.48 mg / kg / day human 1 mg / kg dog = 1.54 mg / kg human 6 mg / kg mouse = 0.49 mg / kg human 1 mg / kg / day mouse = 0.08 mg / kg / day human Example 2 ( S Pharmacokinetics and metabolism of 3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid Oral administration in rats and dogs ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid, the maximum concentration time (T 最大 ) is 15-30 minutes, and the elimination half-life (t1 / 2) is about 75-90 minutes. For equivalent oral doses, the exposure (based on AUC) is higher in dogs than in rats. In vitro studies in hepatocytes showed ( S The clearance of 3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid in rats was significantly greater than that in dogs; the intrinsic clearance (Cl int ) = 24.3 mL / min / kg (rat), 13.1 mL / min / kg (human) and 8.97 mL / min / kg (dog). ( S )-3-Amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid is minimally bound to proteins in rat, dog, and human plasma at 37°C (4% to 20%).

[0088] Example 3 Prospective evaluation of the efficacy of (S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid in mouse models of medulloblastoma and leptomeningeal metastasis A xenograft model will be established according to Martirosian et al., 2021, Cell Reports 35, 109302. Therefore, animal experiments will be performed on 9-10 week old female athymic nude mice housed in ventilated cages (3-5 animals per cage) (n=20). The animals will be housed in an animal room designated for immunodeficient mice, and all cages, bedding, and feed will be sterilized to ensure a pathogen-free environment. Before and after the experiment, the animals will be monitored and determined to be healthy and pain-free. Animals will not be used for other procedures prior to the transplantation procedure. All animals will be humanely euthanized when a specific time point is reached or signs of morbidity appear, including the development of tumor symptoms (paralysis, hydrocephalus, weight loss, head tilt).

[0089] Three medulloblastoma xenograft models (groups) were established using the rapidly dividing human medulloblastoma cell line D283 cells (commercially available from the American Type Culture Collection (ATCC)) and Med-2112FH (MD cells commercially available from the Brain Tumor Resource Laboratory at Fred Hutchinson Cancer Research): (1) 2 × 10 5 D283 disrupted, GABA-AT knockout (KD), and GABA-AT overexpressing (OE) medulloblastoma cells were transplanted into the cerebellum as primary tumor models. (2) 1×10 5 D283 GABA-AT KD and 1 × 10 5 A competitive leptomeningeal model in which 1×10 GABA-AT OE medulloblastoma cells were transplanted into the cisterna magna, and (3) 1×10 5 Med-2112FH is injected into both the cerebellum and the occipital cistern to depict a clinically relevant double injection model of primary and metastatic medulloblastoma. The mice will initially be anesthetized under 5% isoflurane (Vetone, catalog number 502017) and maintained at 2%-2.5% isoflurane. In order to simulate primary disease, tumor cells will be injected into the cerebellum using a stereotaxic frame. The cells will be transplanted to the -6.47 bregma, 1 mm lateral to the sagittal suture, and 1 mm in the cerebellum. In order to simulate leptomeningeal disease, the surgical setting is similar to the cerebellar injection, however, as previously shown, the cells will be injected into the occipital cistern (Xavier et al., 2018 J. Vis. Exp.135, 57378.). All transplanted strains will express luciferase for in vivo bioluminescent imaging (BLI) after transplantation. For the single injection model, BLI will be performed 3 days and 7 days after injection, and once a week thereafter. Mice will be monitored for the development of symptoms related to tumor burden and humanely euthanized when indicated. For the double injection model, BLI will be performed 3 days after injection and every 3-4 days thereafter. Mice will be euthanized 21 days after transplantation to observe the immediate effects of the leptomeningeal microenvironment on medulloblastoma cells.

[0090] Three treatment regimens will be used in this study and will include 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 of leuprolide in saline in a randomized crossover design. S)-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid and saline as vehicle control in a volume of 3 ml / kg body weight once a day, twice a day or three times a day IP administration. Each group of animals will receive at least three cycles of chemotherapy and maintained to determine the results. Each group will be evaluated to compare the increase in survival time compared to untreated animals. Qualitative symptoms observed in the mouse model include irregular stride length, impaired cranial nerve function, and decreased motor coordination and performance. Several quantitative behavioral assays will include mouse rotating rod, forced air challenge, screen flip test, horizontal wire test and stride length analysis. Magnetic resonance imaging (MRI) will be used to confirm and monitor tumor growth and as an anatomical biomarker of treatment response.

[0091] It should be understood that the embodiments and embodiments provided herein are exemplary embodiments and embodiments. Those skilled in the art will envision various modifications of the embodiments and embodiments consistent with the scope of the disclosure herein. Such modifications are intended to be covered by the claims.

Claims

1. A method for inhibiting cancer proliferation and metastasis, comprising administering to a subject in need thereof a drug comprising from about 0.0001 mg / kg to about 30 mg / kg ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof.

2. The method for inhibiting cancer proliferation and metastasis according to claim 1, wherein the composition comprises from 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.

3. The method for inhibiting cancer proliferation and metastasis according to claim 2, wherein the composition comprises from about 0.2 mg / kg to about 10 mg / kg of ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof.

4. The method for inhibiting cancer proliferation and metastasis according to claim 3, wherein the composition comprises from about 0.5 mg / kg to about 6.5 mg / kg of ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof.

5. The method for inhibiting cancer proliferation 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 lymphocytic leukemia, hairy cell leukemia, leptomeningeal carcinoma spread or chronic myeloblastic leukemia and subtypes thereof. The method for inhibiting cancer proliferation and metastasis according to claim 1 , wherein the subject is a human.

7. The method for inhibiting cancer proliferation and metastasis according to claim 1, wherein the subject is also undergoing one or more cancer therapies selected from the group consisting of surgery, chemotherapy, radiotherapy, thermal therapy, immunotherapy, hormone therapy or laser therapy.

8. The method for inhibiting cancer proliferation and metastasis according to claim 1, wherein the composition is administered enterally or parenterally.

9. The method for inhibiting cancer proliferation 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 drug comprising from about 0.0001 mg / kg to about 30 mg / kg ( S )-3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof, thereby treating cancer.

11. The method for treating cancer according to claim 10, wherein the composition comprises from 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.

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

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

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 or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, head and neck cancer, B-cell lymphoma, chronic lymphocytic leukemia, acute lymphocytic leukemia, hairy cell leukemia, leptomeningeal carcinoma or chronic myeloblastic 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 undergoing one or more cancer therapies selected from the group consisting of surgery, chemotherapy, radiation therapy, thermal therapy, immunotherapy, hormone therapy, or laser therapy.

17. The method for treating cancer according to claim 10, wherein the composition is administered enterally or parenterally.

18. The method for treating cancer according to claim 17, wherein the composition is administered orally, rectally, transdermally, intramuscularly, intravenously, subcutaneously, intraperitoneally or intrathecally. 19.( S Use of 3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof in inhibiting cancer proliferation 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.

21. The method of 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 lymphocytic leukemia, hairy cell leukemia, leptomeningeal carcinoma or chronic myeloblastic leukemia and subtypes thereof. twenty two.( S Use of 3-amino-4-(difluoromethylene)cyclopent-1-ene-1-carboxylic acid or a pharmaceutically acceptable salt thereof in the treatment of 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.

24. The method of 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 lymphocytic leukemia, hairy cell leukemia, leptomeningeal carcinoma or chronic myeloblastic leukemia and subtypes thereof.

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