Novel selective inhibitors of lysyl oxidase

Abstract

The present invention relates to novel fluoroarylamine sulfone derivatives capable of inhibiting certain amine oxidase enzymes. These compounds are useful for various indications, such as the treatment of fibrosis, cancer, and / or scarring in human subjects and pets and livestock. Furthermore, the present invention relates to pharmaceutical compositions containing these compounds and their uses.

Description

[Technical Field]

[0001] The present invention relates to novel fluoroarylamine sulfone derivatives capable of inhibiting certain amine oxidase enzymes. These compounds are useful for treating various indications, such as fibrosis, cancer, and / or scarring, in human subjects and pets and livestock. In addition, the present invention relates to pharmaceutical compositions containing these compounds and their use. [Background technology]

[0002] A family of five closely related enzymes is associated with fibrotic diseases and metastatic cancer. The enzymes are lysyl oxidase (LOX), the first described family member, as well as LOX-like 1 (LOXL1), LOXL2, LOXL3, and LOXL4. Lysyl oxidase isozymes are copper-dependent amine oxidases that initiate covalent crosslinking of collagen and elastin. Their primary function is oxidative deamination of lysine and hydroxylysine amino acid side chains to aldehydes, which spontaneously react with adjacent residues to promote crosslinking of collagen and elastin. The resulting crosslinked chains contribute to extracellular matrix (ECM) stability and make them less susceptible to proteolysis by enzymes such as matrix metalloproteinases (MMPs). Lysyl oxidase enzyme activity is crucial for maintaining normal tone and elasticity of connective tissues in many organ systems of the body.

[0003] Lysyl oxidase isozymes belong to the larger group of amine oxidases, which includes flavin-dependent and copper-dependent oxidases, characterized by the nature of their catalytic cofactor. Flavin-dependent enzymes include monoamine oxidase-A (MAO-A), monoamine oxidase-B (MAO-B), polyamine oxidase, and lysyl demethylase (LSD1). Copper-dependent enzymes include semicarbazide-sensitive amine oxidase (vascular adhesion protein-1, SSAO / VAP-1), retinal amine oxidase, diamine oxidase, and lysyl oxidase isozymes. Copper-dependent amine oxidases have a second cofactor that differs slightly from enzyme to enzyme. In SSAO / VAP-1, this is an oxidized tyrosine residue (TPQ, which is oxidized to a quinone), whereas in lysyl oxidase isozymes, TPQ is further processed by the addition of an adjacent lysine residue (forming LTQ).

[0004] Lysyl oxidase isozymes exhibit distinct expression patterns in vivo, suggesting that specific isozymes have specific biological roles. Catalytically active forms of LOX have been identified in the cytoplasmic and nuclear compartments, and research is ongoing to clarify their roles in these compartments. LOX itself plays a major role in, for example, epithelial-mesenchymal transition (EMT), cell migration, adhesion, transformation, and gene regulation. Different patterns of LOX expression / activity are associated with distinct pathological processes, such as fibrotic disorders, Alzheimer's disease and other neurodegenerative processes, and tumor progression and metastasis.

[0005] The directed replacement of dead or damaged cells with connective tissue after injury is an evolutionarily conserved survival mechanism, most prominent in humans, where it plays a beneficial role after trauma, infection, or disease. Progressive scarring can occur after more chronic and / or repeated injury, leading to partial or total dysfunction of the affected organ. Fibrosis can be caused by a variety of factors, including chronic infection, chronic exposure to alcohol and other toxins, autoimmune and allergic reactions, surgery, radiation therapy, and chemotherapy. This pathological process can therefore occur in almost any organ or tissue in the body, typically resulting from weeks or months of simultaneous inflammation, tissue destruction, and repair. In this context, fibrosis most commonly affects the lungs, liver, skin, kidneys, and cardiovascular system.

[0006] Liver fibrosis can occur as a complication of hemochromatosis, Wilson's disease, alcoholism, schistosomiasis, viral hepatitis, bile duct obstruction, exposure to toxins, and metabolic disorders. Liver fibrosis is characterized by the accumulation of extracellular matrix, which is qualitatively distinct from the extracellular matrix in normal liver. This fibrosis can progress to cirrhosis, liver failure, cancer, and ultimately death.

[0007] Fibrous tissue can accumulate in the heart and blood vessels as a result of high blood pressure, hypertensive heart disease, atherosclerosis and myocardial infarction, with the accumulation of extracellular matrix or fibrous deposition causing the vasculature to stiffen and the cardiac tissue itself to stiffen.

[0008] Pulmonary arterial hypertension (PAH) is a rare and rapidly fatal condition characterized by elevated pulmonary arterial pressure and caused by increased pulmonary vascular resistance. PAH is a heterogeneous condition with diverse causes, but there is increasing recognition that it is associated with other diseases, such as connective tissue disorders and scleroderma. Pathological hallmarks of PAH include vessel wall remodeling with excessive extracellular matrix (ECM) deposition and cross-linking. Lysyl oxidase is dysregulated in the pulmonary vasculature of patients with idiopathic pulmonary arterial hypertension (IPAH), contributing to the persistence of ECM components through cross-linking and inappropriate collagen and elastin remodeling. PAH patients have a poor prognosis. Pharmacological targeting of lysyl oxidase offers therapeutic interventions for which few or no currently existent therapeutic options exist.

[0009] A strong association between fibrosis and increased lysyl oxidase activity has been demonstrated, for example, in experimental liver fibrosis in rats, models of pulmonary fibrosis, arterial fibrosis, dermal fibrosis, and adriamycin-induced renal fibrosis in rats. Among these experimental models of human disease, the most significant increase in enzyme activity was seen in a rat model of CCl4-induced liver fibrosis. In these studies, low levels of enzyme activity in healthy livers were increased 15- to 30-fold in fibrotic livers.

[0010] In humans, there is also a significant correlation between lysyl oxidase activity measured in plasma and the progression of liver fibrosis. Lysyl oxidase activity levels are normally low in the serum of healthy individuals, but are significantly increased in chronic active hepatitis and even more so in cirrhosis. Thus, lysyl oxidase may serve as a marker of internal fibrosis.

[0011] Lysyl oxidase isozymes are highly regulated by hypoxia-inducible factor 1α (HIF-1α) and TGF-β, the two most important growth factors that cause fibrosis. Because collagen cross-linking occurs in all types of fibrosis, lysyl oxidase isozyme inhibitors can be used in idiopathic pulmonary fibrosis, scleroderma, renal fibrosis, or hepatic fibrosis.

[0012] In normal wound healing, granulation tissue formation is a transient process that provides a scaffold for re-epithelialization and repair. Subsequently, tissue is remodeled, resulting in the formation of a normotrophic scar. However, after injury, humans are unable to regenerate normal skin. Instead, scar formation (cicatrization) occurs during the repair (or healing) process. Scars are aesthetically and functionally inferior to skin. Scarring is a chronic problem, and excessive or hypertrophic scars, along with their associated aesthetic, functional, and psychological sequelae, remain significant challenges in the treatment of deep skin injuries and burns. A key factor contributing to the poor appearance and reduced flexibility of scars, especially hypertrophic scars, is changes to collagen in the dermal layer. In scar tissue, collagen (mainly collagen I) is more densely packed and tightly aligned in parallel bundles. In normal skin, collagen is less densely packed and has a more "basketweave"-like structure. These changes in both collagen structure and quantity are the main cause of the disfiguring appearance of scars, leading to loss of flexibility, discomfort and functional problems.

[0013] Dermal fibrosis, or excessive scarring of the skin, is the result of an excessive healing response and is characterized by disproportionate fibroblast proliferation and extracellular matrix (ECM) production in the dermis. Clinically, dermal fibrosis manifests as thickened, firm, and hardened areas of skin. The spectrum of fibrotic skin disorders is broad and includes, but is not limited to, hypertrophic scars, keloids, scleroderma (diffuse and focal subtypes), edematous sclerosis (Buschke's disease), systemic amyloidosis, lipodermatosis, progeria, scleroderma syndrome, Dupuytren's contracture, nephrogenic fibrosing dermatosis (NFD), mixed connective tissue disease, scleromyxedema, graft-versus-host disease (GVHD), and eosinophilic fasciitis. While each of these disorders has its own unique etiology and clinical characteristics, they all involve collagen overproduction and altered collagen remodeling. One potential mechanism for altered ECM remodeling is covalent crosslinking. This suggests that LOX family enzymes are directly involved in the development of skin fibrosis. Expression of LOX and LOXL1-4 is increased in scar fibroblasts compared with normal skin fibroblasts, and LOX and LOXL1 are the predominant isoforms found in skin tissue.

[0014] Keloids or keloidal scars are a type of scar that occurs at the site of a healed skin injury due to the abnormal deposition of granulation tissue (type 3 collagen), which is then gradually replaced by type 1 collagen. This abnormal collagen deposition then results from an imbalance between net collagen synthesis and deposition and collagen degradation. Keloids are firm, rubbery lesions or shiny fibrous nodules that can vary in color from pink to the color of human skin, or from red to dark brown.

[0015] Histologically, keloids are fibrous tumors characterized by atypical fibroblastic aggregates with excessive deposition of extracellular matrix components, particularly collagen, fibronectin, elastin, and proteoglycans. In most cases, the lesions have a relatively acellular center with thick, abundant collagen bundles, forming nodules in the deeper dermal layers of the lesion. Keloids are difficult to treat because these lesions can cause significant pain, pruritus, and physical disfigurement. Furthermore, their appearance may not improve over time, and if located over a joint, they may limit mobility.

[0016] Physiological manipulation of collagen deposition / crosslinking and collagen degradation (via collagenase activity) offers, at least theoretically, an opportunity to reduce keloid severity and induce scars with improved physical properties.

[0017] Studies in patients with large pedunculated keloids have shown that treatment with β-aminopropionitrile (BAPN) or penicillamine (a nonselective pan-LOX inhibitor) and colchicine (a collagenase activity stimulator) after excision of the keloid and grafting of the defect has a measurable beneficial effect on superficial scarring.

[0018] Studies using two complementary in vitro skin-like models, human skin equivalent (hSE) and self-organizing stromal tissue, identified LOXL4 as the key isoform mediating the TGF-β-induced fibrotic phenotype.

[0019] The scarring process is a major problem and challenge in the eye and surrounding structures. Ocular scarring plays a major role in either primary disease (e.g., corneal and conjunctival scarring) or treatment failure (e.g., post-operative trabeculectomy).

[0020] Glaucoma is a disease in which the optic nerve is damaged, resulting in progressive, irreversible vision loss. Elevated intraocular pressure (IOP) is one of the major risk factors for the development and progression of glaucoma. Most glaucoma treatments aim to lower intraocular pressure by either reducing aqueous humor formation within the eye or increasing aqueous humor outflow from the eye, as in the case of glaucoma filtration surgery. Trabeculectomy (the current gold standard for IOP management) is a filtration procedure that creates an opening in the anterior chamber under a partial-thickness scleral flap, allowing aqueous humor to outflow from the eye. Postoperative scarring is a major cause of treatment failure. The antimetabolites mitomycin C (MMC) and 5-fluorouracil (5-FU) are currently used in clinical practice to limit postoperative ocular scar tissue formation. Although these agents have been shown to improve IOP following filtration surgery, they are nonselective and associated with significant side effects. Safer, more targeted antifibrotic agents are needed.

[0021] Gingival fibromatosis is a rare and heterogeneous group of disorders that manifest as slowly progressive, localized, or diffuse fibrous hypertrophy (gingival overgrowth or hypertrophy) of keratinized gingiva. In severe cases, excess tissue can cover the tooth crowns, causing masticatory, aesthetic, speech, functional, and periodontal problems. Gingival overgrowth can be hereditary or idiopathic, and can be associated with oral inflammatory diseases or other systemic disorders. However, the majority of cases are due to side effects of systemic medications, such as the antiseizure drug phenytoin, the immunosuppressant cyclosporine A, and certain antihypertensive dihydropyridine calcium channel blockers (particularly nifedipine). The pathological manifestations of gingival overgrowth include the excessive accumulation of extracellular matrix proteins, of which collagen I is the most prevalent. One recognized mechanism of drug-induced gingival overgrowth is epithelial motility (EMT). EMT is a process in which epithelial cells transdifferentiate into fibrogenic fibroblast-like cells, resulting in weakened interactions between gingival cells and the extracellular matrix. Damaged epithelium, basement membrane, and underlying stroma result in TGF-β stimulation of lysyl oxidase enzyme activity, contributing to connective tissue fibrosis.

[0022] The consistent and potent inhibition of fibrosis by lysyl oxidase isoenzyme blockers is due to the lack of cross-linking activity, which renders collagen susceptible to degradation by proteolytic enzymes such as MMPs. Therefore, any type of fibrosis should be ameliorated by treatment with a lysyl oxidase isoenzyme inhibitor. Given the diverse contributions of all lysyl oxidase isoenzymes in fibrosis, inhibitors that consistently and potently inhibit all lysyl oxidase isoenzymes, i.e., pan-LOX inhibitors, should be the most effective.

[0023] Rheumatoid arthritis (RA) is a systemic autoimmune disorder characterized by chronic, painful inflammation of the joint lining. However, in some individuals, the condition progresses, causing painful swelling and inflammation in surrounding tissues and other body systems, such as the skin, eyes, lungs, heart, and blood vessels. Therefore, RA is a painful and debilitating disease that can result in significant loss of function and mobility in the hands, wrists, and feet. Active RA begins in a few joints but can progress to affect multiple joints. Synovial hyperplasia, involving infiltrating immune cells and resident synovial fibroblasts (SFs), is a typical feature of RA. RA synovial fibroblasts (RASFs) are the most common cell type at the site of infiltration and are the primary cause of joint destruction. Activated RASFs can migrate and are therefore involved in the spread of arthritis between joints. Cytokines from infiltrating immune cells induce the activation and proliferation of synovial fibroblasts. These activated SFs then generate a pathogenic stroma, perpetuating chronic inflammation and ultimately leading to cartilage and bone destruction. Transplantation of RASFs together with human cartilage into severe combined immunodeficient mice demonstrated in vivo migration of activated RASFs and spread of disease to the site of transplanted human cartilage. Furthermore, RASFs actively degrade cartilage, whereas control transplants of synovial fibroblasts from osteoarthritis (OA) patients and skin fibroblasts from healthy donors did not. RASFs differ from non-activated healthy fibroblasts by their morphology and gene expression. RASFs are characterized by the expression of anti-apoptotic proto-oncogenes and the absence of tumor suppressor genes. The production of pro-inflammatory cytokines and chemokines by RASFs further attracts immune cells to the synovium. Furthermore, the production of matrix metalloproteinase (MMP) enzymes promotes cartilage invasion and destruction.

[0024] The type II collagen-induced arthritis (CIA) model is frequently used as an animal model of RA because it adequately recapitulates the characteristic immunological, pathological, and arthritic symptoms observed in humans. High levels of LOX expression have been demonstrated in CIA rats in the synovium, synovial fluid, and serum. Inhibition of LOX with β-aminopropionitrile (BAPN, a pan-LOX inhibitor) was found to attenuate inflammation, synovial hyperplasia, angiogenesis, and the expression of MMP-2 and MMP-9, indicating that LOX promotes synovial hyperplasia and angiogenesis in CIA rats. Furthermore, knockdown of LOXL2 and antibodies against LOXL2 attenuated collagen deposition, proliferation, and invasion of RASFs.

[0025] Although there is no cure for RA, many treatments are available that can alleviate symptoms and slow disease progression. However, these treatments are associated with significant side effects, in part related to suppression of the immune system. Selective drugs that target RASFs could offer a more useful therapy for RA.

[0026] Osteoarthritis (OA) is a disease characterized by the degeneration of articular cartilage and the underlying bone. Due primarily to "wear and tear," OA results in joint pain and stiffness. The joints most commonly affected are those of the fingers, knees, back, and hips. Unlike other forms of arthritis (such as RA), osteoarthritis only affects the joints. Often, joints on one side of the body are affected more than the other. OA is a progressive, debilitating disease that can significantly impact work and normal daily activities.

[0027] Synovial fibrosis is a major cause of OA and is a manifestation of an imbalance between fibroblast proliferation and collagen synthesis and degradation, which leads to excessive deposition of collagen in the extracellular matrix (ECM), causing synovial thickening and stiffening.

[0028] Genes encoding several lysyl oxidase family enzymes, including LOX, LOXL2, LOXL3, and LOXL4, have been shown to be highly expressed in mice with experimental OA and in humans with end-stage OA.

[0029] Given that many members of the lysyl oxidase family of enzymes contribute in various ways to the pathogenesis of both rheumatoid arthritis and osteoarthritis, pan-LOX inhibitors could potentially offer more efficacious therapies.

[0030] BAPN is a widely used, irreversible lysyl oxidase inhibitor based on a nonselective mechanism. Since the 1960s, BAPN has been used in animal studies (primarily rats, mice, and hamsters) and has been shown to be effective in reducing collagen content in various models (e.g., CCl4, bleomycin, quartz, cancer) and tissues (e.g., liver, lung, and dermis). However, studies in human patients with scleroderma have shown that BAPN is poorly tolerated, highlighting the need for safer alternatives.

[0031] Lysyl oxidase-catalyzed collagen crosslinks can proceed via two pathways: the allysine pathway and the hydroxyallysine pathway. In the hydroxyallysine pathway, premature divalent crosslinks such as dehydro-dihydroxylysinonorleucine (deH-DHLNL) and dehydro-hydroxylysinonorleucine (deH-HLNL) are initially formed, which then further progress (through a lysyl oxidase-independent reaction) to mature trivalent crosslinks between three collagen molecules, resulting in the formation of deoxypyridinoline (DPD) and pyridinoline (PYD). These mature and premature crosslinks can be measured by LC-MS / MS.

[0032] Lysyl oxidase isozymes are not only involved in the cross-linking of elastin and collagen during wound healing and fibrosis, but also regulate cell motility and signal transduction. Their intracellular and nuclear functions are associated with gene regulation, potentially leading to tumorigenesis and tumor progression. Both down- and up-regulation of lysyl oxidase isozymes in tumor tissues and cancer cell lines has been described, suggesting a dual role for lysyl oxidase isozymes and LOX propeptides as both metastasis-promoting and tumor-suppressing genes.

[0033] In addition to its role in tissue remodeling, LOX isozymes also play important roles in primary cancer and metastasis. Tumor growth is primarily associated with a reactive stroma composed of fibroblasts called cancer-associated fibroblasts (CAFs). Mice subcutaneously inoculated with an equal mixture of tumor and CAF cells exhibit accelerated growth rates and a higher incidence of metastasis. CAF knockout models have been shown to promote tumorigenesis, but this is a rather abstract scenario compared to the tumor microenvironment in patients. CAFs have been shown to have increased LOX expression compared to normal fibroblasts. Utilizing LOX inhibitors in the context of cancer may affect both the tumor and stromal compartments, helping to reduce tumor growth and metastasis.

[0034] Emerging evidence suggests a link between idiopathic pulmonary fibrosis and lung cancer, but further testing is needed. In mouse models of both lung and liver in which chemical or radiation-induced fibrosis is present, alpha-smooth muscle actin (a marker of fibroblasts), LOX expression, and metastatic tumor growth are increased, and this is reversed by an LOX antibody.

[0035] To date, increased lysyl oxidase isozyme mRNA and / or protein has been observed in patient samples from The Cancer Genome Atlas (TCGA) for breast cancer, CNS cancer cell lines, head and neck squamous cell carcinoma, esophageal cancer, kidney cancer, lung cancer, prostate cancer, clear cell renal cell carcinoma and lung carcinoma, ovarian cancer, uterine cancer, melanoma, and osteosarcoma. TCGA patient gene expression data for the LOX family are shown in Table 1. A plus sign indicates higher than average gene expression within this dataset.

[0036] [Table 1]

[0037] Statistically significant clinical correlations between lysyl oxidase isoenzyme expression and tumor progression have been observed in breast cancer, head and neck squamous cell carcinoma, myelofibrosis, prostate cancer, pancreatic cancer, ovarian cancer, and clear cell renal cell carcinoma. The role of lysyl oxidase isoenzymes in tumor progression has been most extensively examined in breast cancer, using in vitro migration / invasion models and in vivo mouse models of tumorigenesis and metastasis. Increased lysyl oxidase isoenzyme expression was observed in hypoxic patients and was associated with estrogen receptor-negative status (ER-), decreased overall survival in ER and lymph node-negative patients who did not receive adjuvant systemic treatment, and decreased bone metastasis-free survival in ER and lymph node-negative patients. In vivo models demonstrated that LOX inhibitors may benefit breast cancer patients with bone metastases by regulating bone homeostasis independently of receptor activator of nuclear factor-κB ligand (RANKL). Lysyl oxidase isozyme mRNA was demonstrated to be upregulated in invasive and metastatic cell lines (MDA-MB-231 and Hs578T) and in more aggressive breast cancer cell lines and distant metastatic tissues compared with primary cancer tissues.

[0038] The pathogenic process in primary myelofibrosis involves primary megakaryocyte-heavy clonal myeloproliferation and a paraneoplastic stromal reaction, including myelofibrosis, osteosclerosis, angiogenesis, and extramedullary hematopoiesis. The bone marrow reaction includes excessive deposition of extracellular matrix proteins such as fibrillar collagen, hypocellularity, activation and recruitment of bone marrow fibroblasts, excessive production of cytokines and growth factors, and other changes that result in impaired hematopoietic capacity. Secondary myelofibrosis can result from polycythemia vera or essential thrombocytosis. In myelofibrosis, disease progression correlates with an increase in the number of megakaryocytes overexpressing LOX. In a GATA1-low mouse model of myelofibrosis, disease progression (including increases in megakaryocyte numbers, fibrosis, and spleen size) was significantly attenuated by a pan-LOX inhibitor.

[0039] For most tumor types, surgical resection is the first choice of treatment. A wound healing response is initiated by surgery and can correlate with increased metastatic spread. In breast cancer models, abdominal surgery has been shown to increase lung metastasis. Furthermore, this was shown to be driven by systemic LOX. Injection of plasma (containing LOX) collected from abdominally operated mice into tumor-bearing mice increased lung metastasis. Blocking systemic LOX induced by surgery with BAPN suppressed metastasis and improved survival.

[0040] In models of colon cancer, breast cancer, and melanoma, LOX expression has been shown to be increased in tumor-associated endothelial cells and stimulate angiogenesis and tumor growth.

[0041] In patients with pancreatic, breast, lung, ovarian, and colon cancer, high collagen content correlates with elevated LOX gene expression, chemotherapy resistance, and significantly reduced survival rates. The combination of LOX inhibitors (both BAPN and LOX antibodies) and standard-of-care chemotherapy in desmoplastic tumor mouse models reduced tumor interstitial pressure and caused vasodilation. Increased vascular flow increases the concentration of chemotherapy agents at the primary tumor site, reducing metastatic burden and improving survival rates.

[0042] In head and neck squamous cell carcinoma, increased expression of lysyl oxidase isozymes was observed in association with CA-IX, a marker of hypoxia, and was associated with decreased cancer-specific survival, overall survival, and metastasis-free survival.In oral squamous cell carcinoma, lysyl oxidase isozyme mRNA expression was upregulated compared with normal mucosa.

[0043] Gene expression profiling of gliomas identified lysyl oxidase isozymes that were overexpressed as part of a molecular signature indicative of invasiveness and associated with aggressive tumors that strongly correlated with poor patient survival. Lysyl oxidase isozyme protein expression was increased in glioblastoma and astrocytoma tissues, as well as in invasive U343 and U251 cultured astrocytoma cells.

[0044] In tissues, lysyl oxidase isozyme mRNA was upregulated in prostate cancer compared with benign prostatic hyperplasia, correlated with Gleason score, and was associated with both high grade and short time to recurrence.

[0045] In renal clear cell carcinoma (RCC), smoking is associated with allelic imbalance at chromosome 5q23.1, where the LOX gene is located, possibly involving gene duplication.

[0046] SiHa cervical cancer cells showed increased invasion in vitro under hypoxic / anoxic conditions, which was suppressed by inhibition of extracellular catalytic lysyl oxidase activity by treatment with BAPN as well as LOX antisense oligos, LOX antibodies, LOX shRNA, or extracellular copper chelators.

[0047] In a genetically engineered mouse model of ovarian cancer (ApoE knockout), desmoplastic tumors with increased LOX gene expression were formed. Treatment with BAPN significantly improved survival and reduced lung metastasis. Certain tumors in ovarian cancer patients harbor a single nucleotide polymorphism, G473A, in the LOX gene. Two independent studies have shown that individuals with the G473A polymorphism are more likely to develop ovarian cancer.

[0048] In human primary oral squamous cell carcinoma (OSCC), the expression levels of lysyl oxidase enzymes (particularly LOX and LOXL2) and lysyl hydroxylases are significantly elevated, especially in tumors with advanced regional lymph node metastasis (RLNM). Both reducible or immature crosslinks (deH-DHLNL and deH-HLNL) and nonreducible or mature crosslinks (DPD and PYD) are significantly elevated in OSCC compared with normal tissue.

[0049] The findings described herein provide a strong rationale for combination therapy involving LOX isoenzyme inhibitors and anti-tumor therapy in patients.

[0050] Recently, CCT365623, a reversible pan-LOX inhibitor, has been utilized in a breast cancer model (MMTV-PyMT) to suppress metastasis and improve survival.

[0051] Those skilled in the art will recognize that drug-like properties are essential to enable drug development. Successful drug development requires finding compounds that combine multiple drug-like properties, including but not limited to: Physicochemical properties of the drug, including solubility, permeability, lipophilicity, and stability Pharmacological properties, including absorption, distribution, metabolism, and excretion (ADME); transporter activity; efficacy at the intended biological target; duration of action; and minimal nonspecific and off-target pharmacological effects. Activity at desired location Drug safety; minimal toxicity

[0052] Considering the complex role of lysyl oxidase family enzymes in the aforementioned diseases, pan-LOX inhibitors with favorable drug-like properties could offer clear and important therapeutic benefits.

[0053] Scientific and patent literature describes the therapeutic efficacy of small molecule inhibitors of lysyl oxidase isoenzymes and antibodies to LOX and LOXL2 in animal models of fibrosis and cancer metastasis. Some known MAO inhibitors have also been reported to inhibit lysyl oxidase isoenzymes (e.g., the MAO-B inhibitor mofegiline shown below). This inhibitor is a member of the haloarylamine family of MAO inhibitors, and the halogen in mofegiline is fluorine. Fluoroarylamine inhibitors are described in U.S. Pat. No. 4,454,158. Patents have been issued claiming fluoroarylamines and chloroarylamines, such as MDL72274 (shown below), as inhibitors of lysyl oxidase (U.S. Patent Nos. 4,943,593; 4,965,288; 5,021,456; 5,059,714; 5,182,297; and 5,252,608). Many of the compounds claimed in these patents have also been reported to be potent MAO-B and SSAO / VAP-1 inhibitors. [ka]

[0054] Further fluoroarylamine inhibitors are described in US Patent No. 4,699,928. Other examples structurally related to mofegiline can be found in WO 2007 / 120528.

[0055] WO 2009 / 066152 discloses a family of 3-substituted 3-haloarylamines that are inhibitors of SSAO / VAP-1 useful as treatments for a variety of indications, including inflammatory diseases. None of these documents specifically discloses the fluoroarylamine compounds of the present invention.

[0056] Antibodies to LOX and LOXL2 are disclosed in U.S. Patent Application Publication No. 2009 / 0053224, along with methods for diagnostic and therapeutic use. Anti-LOX and anti-LOXL2 antibodies can be used to identify and treat conditions such as fibrotic conditions, angiogenesis, or to prevent the transition from an epithelial to a mesenchymal cell state: U.S. Patent Application Publication No. 2011 / 0044907.

[0057] WO 2017 / 136871 and WO 2017 / 136870 disclose haloallylamine indole and azaindole derivative inhibitors of lysyl oxidase and their uses.

[0058] WO 2018 / 157190 discloses haloarylamine pyrazole derivative inhibitors of lysyl oxidase and uses thereof.

[0059] WO 2020 / 024017 discloses haloarylamine sulfone derivative inhibitors of lysyl oxidase and uses thereof. WO 2021 / 012014 discloses difluorohaloarylamine sulfone derivative inhibitors of lysyl oxidase.

[0060] WO 2017 / 141049 and WO 2019 / 073251 disclose a family of methylamine and bridged homopiperazine derivatives as lysyl oxidase inhibitors and their use in the treatment of cancer and fibrosis-related diseases, respectively.

[0061] Another class of lysyl oxidase inhibitors is disclosed in WO 2003 / 097612, WO 2006 / 053555, and US Patent Application Publication No. 2008 / 0293936.

[0062] Further LOXL2 inhibitors are disclosed in WO 2020 / 099886, WO 2018 / 048930, WO 2017 / 015221, WO 2017 / 003862, WO 2016 / 144702 and WO 2016 / 144703. Summary of the Invention

[0063] The present invention provides novel substituted fluoroallylamine compounds that inhibit lysyl oxidase (LOX), lysyl oxidase-like 2 (LOXL2), and other lysyl oxidase isozymes. Surprisingly, these novel 2-sulfonylmethyl-3-fluoroallylamine compounds are potent inhibitors of human LOX and LOXL isozymes. These compounds have been found to have favorable properties for topical or oral application. Furthermore, these novel compounds also selectively inhibit specific LOX and LOXL isozymes relative to other enzymes in the amine oxidase family and have favorable stability and safety profiles.

[0064] A first aspect of the present invention is a compound of formula I: [ka] [In the formula, A is aryl; Each R 1 is XR 4 , halogens, deuterium, C 1~6 Alkyl, OC 1~6 Alkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, -CN, -NO2, -C(O)OR 5 , -C(O)NR 6 R 7 , -S(O)NR 6 R 7 , -S(O)2R 8 , -NR 9 C(O)R 10 , and -NR 9 S(O)2R 10 each C is independently selected from the group consisting of 1~6Alkyl, aryl, heteroaryl, cycloalkyl and heterocycloalkyl are substituted with halogen, -OH, and -C 1~4 optionally substituted with one or more substituents selected from the group consisting of alkyl; R 2 is H or F; R 3 is H or F; provided that R 2 or R 3 Only one of the is F; X is selected from the group consisting of O, CH2, and S(O)2; R 4 is selected from the group consisting of aryl and heterocycloalkyl; each aryl is selected from the group consisting of -S(O)R 8 each heterocycloalkyl optionally substituted by ═O; R 5 is hydrogen and C 1~6 selected from the group consisting of alkyl; R 6 and R 7 is hydrogen and C 1~6 independently selected from the group consisting of alkyl; R 8 is C 1~6 is alkyl; R 9 is hydrogen or C 1~6 is alkyl; R 10 is C 1~6 is alkyl; and n is 0, 1, 2, 3, 4, or 5, or a pharmaceutically acceptable salt thereof.

[0065] A second aspect of the present invention provides a pharmaceutical composition comprising a compound according to the first aspect of the invention, or a pharmaceutically acceptable salt or solvate thereof, and at least one pharmaceutically acceptable excipient, carrier or diluent.

[0066] A third aspect of the present invention provides a method for inhibiting the amine oxidase activity of any one of LOX, LOXL1, LOXL2, LOXL3 and LOXL4 in a subject in need thereof, the method comprising administering to the subject an effective amount of a compound according to the first aspect of the invention, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition according to the second aspect of the invention.

[0067] A fourth aspect of the present invention provides a method for treating a condition by inhibiting the activity of any one of LOX, LOXL1, LOXL2, LOXL3 and LOXL4 proteins, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to the first aspect of the invention, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition according to the second aspect of the invention.

[0068] A fifth aspect of the present invention provides the use of a compound according to the first aspect of the invention, or a pharmaceutically acceptable salt or solvate thereof, for the manufacture of a medicament for treating a condition by inhibiting the activity of any one of LOX, LOXL1, LOXL2, LOXL3 and LOXL4 proteins.

[0069] A sixth aspect of the invention provides a compound according to the first aspect of the invention, or a pharmaceutically acceptable salt or solvate thereof, for use in treating a condition by inhibiting the activity of any one of LOX, LOXL1, LOXL2, LOXL3 and LOXL4 proteins.

[0070] In one embodiment of the methods and uses of the present invention, the condition is selected from fibrosis, cancer, and myeloid malignancies.

[0071] In one embodiment of the methods and uses of the present invention, the condition is scarring.

[0072] A further aspect of the present invention provides a method for treating or preventing keloid disease or scarring, which method comprises administering to a subject in need thereof a therapeutically effective amount of a compound according to the present invention, or a pharmaceutically acceptable salt thereof.

[0073] In one embodiment of the composition of the invention, the pharmaceutical composition is a topical composition. In one embodiment of the method of the invention, the pharmaceutical composition is administered topically.

[0074] In one embodiment of the composition of the invention, the pharmaceutical composition is an oral composition. In one embodiment of the method of the invention, the pharmaceutical composition is administered orally.

[0075] Contemplated herein are combination therapies, wherein the method further comprises co-administering an additional therapeutic agent used for the treatment of cancer, fibrosis, inflammation, immunosuppression, angiogenesis, fungal infection, bacterial infection, metabolic conditions, pain, and pruritus.

[0076] definition Below are some definitions that may be helpful in understanding the description of the present invention. These are intended as general definitions and in no way limit the scope of the present invention to only these terms, but are presented for a better understanding of the description that follows.

[0077] Unless the context requires otherwise or unless specifically stated to the contrary, any integer, step or element of the invention described herein as a singular integer, step or element specifically encompasses both the singular and plural forms of the described integer, step or element.

[0078] Throughout this specification, unless the context requires otherwise, the word "comprise" or variations such as "comprises" or "comprising" will be understood to imply the inclusion of a stated step or element or integer or group of steps or elements or integers, but not the exclusion of any other step or element or integer or group of elements or integers. Thus, in the context of this specification, the term "comprising" means "including primarily, but not necessarily solely."

[0079] Those skilled in the art will appreciate that the invention described herein is susceptible to variations and modifications other than those specifically described. The invention is to be understood to include all such variations and modifications. The invention also includes all steps, features, compositions, and compounds referred to or shown in this specification, individually or collectively, and any and all combinations of such steps or features, or any two or more thereof.

[0080] As used herein, the term "alkyl" includes within its meaning monovalent ("alkyl") and divalent ("alkylene") straight- or branched-chain saturated hydrocarbon groups having 1 to 6 carbon atoms, e.g., 1, 2, 3, 4, 5, or 6 carbon atoms. A straight- or branched-chain alkyl group may be attached at any available point to produce a stable compound. For example, the term alkyl includes, but is not limited to, methyl, ethyl, 1-propyl, isopropyl, 1-butyl, 2-butyl, isobutyl, tert-butyl, amyl, 1,2-dimethylpropyl, 1,1-dimethylpropyl, pentyl, isopentyl, hexyl, 4-methylpentyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 1,2,2-trimethylpropyl, 1,1,2-trimethylpropyl, and the like.

[0081] As used herein, the terms "aryl" or "arylene" and variations thereof refer to monovalent ("aryl") and divalent ("arylene") mono-, polynuclear, conjugated, and fused analogs of aromatic hydrocarbons having 6 to 10 carbon atoms. A fused analog of aryl refers to an aryl group fused to a monocyclic cycloalkyl or monocyclic heterocyclyl group where the point of attachment is on the aromatic moiety. Examples of aryl and its fused analogs include phenyl, naphthyl, indanyl, indenyl, tetrahydronaphthyl, 2,3-dihydrobenzofuranyl, tetrahydrobenzopyranyl, 1,4-benzodioxanyl, and the like. A "substituted aryl" is an aryl independently substituted with one or more, preferably one, two, or three, substituents attached to any available atom to produce a stable compound.

[0082] As used herein, the terms "heteroaryl" and variations such as "heteroaromatic group" or "heteroarylene" include within their meaning monovalent ("heteroaryl") and divalent ("heteroarylene"), mono-, polynuclear, conjugated, and fused heteroaromatic groups having 5 to 10 atoms, in which 1 to 4 ring atoms or 1 to 2 ring atoms are heteroatoms independently selected from O, N, NH, and S. Heteroaryl is also intended to include oxidized S or N, such as sulfinyl, sulfonyl, and N-oxide of a tertiary ring nitrogen. A carbon or nitrogen atom serves as the point of attachment of the heteroaryl ring structure, providing a stable compound is produced. The heteroaromatic group can be a C1-9 heteroaromatic. A fused analog of heteroaryl refers to a heteroaryl group fused to a monocyclic cycloalkyl or monocyclic heterocyclyl group in which the point of attachment is on the aromatic moiety. Examples of heteroaryl groups and their fused analogs include pyrazolyl, pyridyl, oxazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, triazinyl, thienyl, benzoxazolyl, benzothiazolyl, benzimidazolyl, benzofuranyl, benzothiophenyl, furo(2,3-b)pyridyl, indolyl, isoquinolyl, imidazopyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, pyridonyl, phenanthrolinyl, quinolyl, isoquinolinyl, imidazolinyl, thiazolinyl, pyrrolyl, furanyl, thiophenyl, oxazolyl, isoxazolyl, isothiazolyl, triazolyl, and the like. "Nitrogen-containing heteroaryl" refers to a heteroaryl in which any heteroatom is N. "Substituted heteroaryl" refers to a heteroaryl that is independently substituted with one or more, preferably one, two, or three, substituents attached to any available atom to produce a stable compound.

[0083] As used herein, the terms "heterocyclyl" and variations such as "heterocycloalkyl" include within their meaning monovalent ("heterocyclyl") and divalent ("heterocyclylene"), saturated or partially saturated (non-aromatic), monocyclic, bicyclic, polycyclic, or fused hydrocarbon groups having 3 to 10 ring atoms, in which 1 to 4 or 1 to 2 ring atoms are heteroatoms independently selected from O, N, NH, or S, SO, or SO2, and the point of attachment can be carbon or nitrogen. A fused analog of heterocyclyl refers to a monocyclic heterocycle fused to an aryl or heteroaryl group in which the point of attachment is on the non-aromatic portion. A heterocyclyl group is a C 3~8 The heterocycloalkyl group can be C 3~6 The heterocyclyl group can be C 3~5 It can be heterocyclyl. Examples of heterocyclyl groups and their fused analogs include pyrrolidinyl, thiazolidinyl, piperidinyl, piperazinyl, imidazolidinyl, 2,3-dihydrofuro(2,3-b)pyridyl, benzoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, dihydroindolyl, quinuclidinyl, azetidinyl, morpholinyl, tetrahydrothiophenyl, tetrahydrofuranyl, tetrahydropyranyl, and the like. The term also includes partially unsaturated monocyclic rings that are not aromatic, such as 2- or 4-pyridones or N-substituted uracils attached through nitrogen. Heterocycloalkyls can be substituted, for example, heterocycloalkyls can be substituted by =0.

[0084] As used herein, the term "halogen" or variations such as "halide" or "halo" refers to fluorine, chlorine, bromine and iodine.

[0085] As used herein, the term "heteroatom" or variations such as "hetero-" or "heterogroup" refers to O, N, NH, and S.

[0086] Generally, "substituted" refers to an organic group (e.g., an alkyl group) as defined herein in which one or more bonds to a hydrogen atom are replaced with a bond to a non-hydrogen atom or a non-carbon atom. Substituted groups also include groups in which one or more bonds to a carbon or hydrogen atom are replaced with one or more bonds, including double or triple bonds, to a heteroatom. Thus, unless otherwise specified, a substituent is substituted with one or more substituents. In some embodiments, a substituent is substituted with 1, 2, 3, 4, 5, or 6 substituents.

[0087] As used herein, the term "optionally substituted" means that the group to which this term refers can be unsubstituted or substituted with one or more groups independently selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, halo, haloalkyl, hydroxyl, hydroxyalkyl, alkoxy, thioalkoxy, alkenyloxy, haloalkoxy, NO, NH(alkyl), N(alkyl), alkylamino, dialkylamino, acyl, alkenoyl, alkynoyl, acylamino, diacylamino, acyloxy, alkylsulfonyl, alkylsulfonyloxy, sulfonamido, heterocyclooxy, heterocycloamino, haloheterocycloalkyl, alkylsulfenyl, alkylcarbonyloxy, phosphorus-containing groups (e.g., phosphono and phosphinyl), aryl, heteroaryl, alkylaryl, aralkyl, alkylheteroaryl, cyano, ═O, COH, COalkyl, C(O)NH, —C(O)NH(alkyl), and —C(O)N(alkyl). Preferred substituents include halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, hydroxy(C1-C6)alkyl, C3-C6 cycloalkyl, C(O)OH, NHC(O)C1-C4 alkyl, C(O)C1-C4 alkyl, NH2, NHC1-C4 alkyl, N(C1-C4 alkyl)2, SO2(C1-C4 alkyl), OH, and CN. Particularly preferred substituents include C 1~4 Alkyl, C 1~4Alkoxy, SO2 (C1-C4 alkyl), =O, halogen, OH, hydroxy (C 1~3 ) alkyl (e.g., C(CH3)2OH), and C 1~3 Haloalkyl (e.g., CF3, CH2CF3) is included.

[0088] The present invention includes within its scope all stereoisomers and isomeric forms of the compounds disclosed herein, including all diastereomeric isomers, racemates, enantiomers, and mixtures thereof. It is also understood that compounds described by Formula I can exist as E and Z isomers, also known as cis and trans isomers. Thus, the present disclosure should be understood to include, for example, E, Z, cis, trans, (R), (S), (L), (D), (+), and / or (-) forms of the compounds, as appropriate in each case. Where no specific stereoisomerism is shown in a structure, it should be understood that any and all possible isomers are encompassed. The compounds of the present invention encompass all conformational isomers. The compounds of the present invention may also exist in one or more tautomeric forms, including both single tautomers and mixtures of tautomers. Also included within the scope of the present invention are all polymorphs and crystalline forms of the compounds disclosed herein.

[0089] The present invention includes within its scope different isotopes of atoms. Any atom not specifically designated as a particular isotope is meant to represent any stable isotope of that atom. Thus, the present disclosure should be understood to include deuterium isotopes of hydrogen.

[0090] All references cited in this application are specifically incorporated by cross-reference in their entirety. The mention of such a document should not be construed as an admission that the document forms part of the common general knowledge or is prior art.

[0091] In the context of this specification, the term "administering" and its variations such as "administer" and "administration" include contacting, applying, delivering, or providing a compound or composition of the invention to an organism or surface by any suitable means. In the context of this specification, the term "treatment" refers to any and all uses that cure a disease state or symptom, prevent the establishment of a disease, or in any way prevent, hinder, slow, or reverse the progression of a disease or other undesirable condition.

[0092] In the context of this specification, the term "topical administration" or variations of that term (including "topical application") is meant to include applying, contacting, delivering or providing a compound or composition of the present invention to the skin or a localized area of ​​the body.

[0093] In the context of this specification, the term "topical administration" or variations of that term (including "topical application") is meant to include applying, contacting, delivering or providing a compound or composition of the present invention to the skin or a localized area of ​​the body.

[0094] In the context of this specification, the term "effective amount" is intended to include a non-toxic amount of a compound or composition of the present invention that is sufficient to provide the desired effect. Thus, the term "therapeutically effective amount" is intended to include a non-toxic amount of a compound or composition of the present invention that is sufficient to provide the desired therapeutic effect. The exact amount required will vary from subject to subject, depending on factors such as the species being treated, the subject's sex, age, and general health, the severity of the condition being treated, the specific agent being administered, and the mode of administration. Therefore, it is not possible to specify an exact "effective amount." However, for any given case, one of ordinary skill in the art can determine an appropriate "effective amount" using only routine experimentation. [Brief explanation of the drawings]

[0095] [Figure 1a] FIG. 1 shows the time-dependent inhibitory effect of Compound 1 on LOXL1. [Figure 1b]FIG. 1 shows the time-dependent inhibitory effect of Compound 1 on LOXL2. [Figure 2] FIG. 1 shows the diffusion of Compound 1 through human skin. [Figure 3] FIG. 1 shows the efficacy of Compound 1 in a mouse model of sclerosis (bleo=bleomycin; ns=not significant; ****=P≦0.0001; Cmpd1=Compound 1). [Figure 4] FIG. 1 shows reduction of collagen cross-links after treatment with Compound 1 in a rodent injury model (*=P≦0.05). DETAILED DESCRIPTION OF THE INVENTION

[0096] The present invention relates to novel fluoroallylamine sulfone derivatives capable of inhibiting lysyl oxidase (LOX), lysyl oxidase-like 2 (LOXL2) and other lysyl oxidase isozymes.

[0097] In particular, the present invention provides compounds of formula I: [ka] [In the formula, A is aryl; Each R 1 is XR 4 , halogens, deuterium, C 1~6 Alkyl, OC 1~6 Alkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, -CN, -NO2, -C(O)OR 5 , -C(O)NR 6 R 7 , -S(O)NR 6 R 7 , -S(O)2R 8 , -NR 9 C(O)R 10 , and -NR 9 S(O)2R 10 each C is independently selected from the group consisting of 1~6 Alkyl, aryl, heteroaryl, cycloalkyl and heterocycloalkyl are substituted with halogen, -OH, and -C1~4 optionally substituted with one or more substituents selected from the group consisting of alkyl; R 2 is H or F; R 3 is H or F; provided that R 2 or R 3 Only one of the is F; X is selected from the group consisting of O, CH2, and S(O)2; R 4 is selected from the group consisting of aryl and heterocycloalkyl; each aryl is selected from the group consisting of -S(O)R 8 each heterocycloalkyl optionally substituted by ═O; R 5 is hydrogen and C 1~6 selected from the group consisting of alkyl; R 6 and R 7 is hydrogen and C 1~6 independently selected from the group consisting of alkyl; R 8 is C 1~6 is alkyl; R 9 is hydrogen or C 1~6 is alkyl; R 10 is C 1~6 is alkyl; n is 0, 1, 2, 3, 4 or 5, or a pharmaceutically acceptable salt thereof.

[0098] In some embodiments, the invention comprises a polymorphic form, solvate, hydrate, or tautomeric form of a compound of Formula I. In some embodiments, the invention comprises a polymorphic form of a compound of Formula I. In other embodiments, the invention comprises a solvate or hydrate of a compound of Formula I. In further embodiments, the invention comprises a tautomeric form of a compound of Formula I.

[0099] In one embodiment of the compounds of the present invention, A is selected from the group consisting of phenyl, naphthyl, and 1,4-benzodioxanyl. In another embodiment of the compounds of the present invention, A is phenyl.

[0100] In one embodiment of the compounds of the present invention, R 1 is XR 4 , halogens, deuterium, C 1~6 Alkyl, OC 1~6 Alkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, -CN, -NO2, -C(O)OR 5 , -C(O)NR 6 R 7 , -S(O)NR 6 R 7 , -S(O)2R 8 , -NR 9 C(O)R 10 , and -NR 9 S(O)2R 10 each C is independently selected from the group consisting of 1~6 Alkyl, aryl, heteroaryl, cycloalkyl and heterocycloalkyl are substituted with halogen, -OH, and -C 1~4 In another embodiment of the compounds of the present invention, each R 1 is XR 4 , halogens, deuterium, C 1~6 Alkyl, OC 1~6 Alkyl, heteroaryl, -CN, -NO2, -C(O)OR 5 , -C(O)NR 6 R 7 and -S(O)2R 8 each C is independently selected from the group consisting of 1~6 Alkyl and heteroaryl are substituted with halogen and -C 1~6 In a further embodiment of the compounds of the present invention, each R 1 is XR 4 , halogen, deuterium, -C 1~4independently selected from the group consisting of alkyl, -OCH3, heteroaryl, -CN, -NO2, -C(O)OH, -C(O)NH2, and -S(O)2CH3; 1~4 Alkyl and heteroaryl may contain fluorine and -C 1~4 In another embodiment of the compounds of the present invention, each R 1 is XR 4 , F, Cl, Br, deuterium, -methyl, -CF3, isopropyl, -OCH3, thiazolyl, pyrazolyl, -CN, -NO2, -C(O)OH, -C(O)NH2, -S(O)2CH3, wherein each thiazolyl or pyrazolyl is optionally substituted with methyl.

[0101] In one embodiment of the compounds of the present invention, R 2 is H or F, and R 3 is H or F, provided that R 2 or R 3 and only one of R is F. In another embodiment of the compounds of the invention, 2 is F and R 3 is H. In a further embodiment of the compounds of the invention, R 2 is H and R 3 is F.

[0102] In one embodiment of a compound of the invention, X is selected from the group consisting of O, CH2, and S(O)2. In another embodiment of a compound of the invention, X is selected from the group consisting of O and S(O)2. In another embodiment of a compound of the invention, X is O. In a further embodiment of a compound of the invention, X is CH2. In another embodiment of a compound of the invention, X is S(O)2.

[0103] In one embodiment of the compounds of the present invention, R 4 is selected from the group consisting of aryl and heterocycloalkyl; each aryl is selected from the group consisting of -S(O)R 8and each heterocycloalkyl is optionally substituted by ═O. In another embodiment of the compounds of the invention, R 4 is phenyl and [ka] wherein each phenyl is optionally substituted with -S(O)2CH3. In a further embodiment, R 4 is phenyl. In another embodiment, R 4 is phenyl substituted with -S(O)2CH3. In a further embodiment, R 4 teeth, [ka] is.

[0104] In one embodiment of the compounds of the present invention, R 5 is hydrogen and C 1~6 In another embodiment of the compounds of the present invention, R 5 is selected from the group consisting of hydrogen and methyl. In a further embodiment of the compounds of the invention, R 5 is hydrogen. In another embodiment of the compounds of the present invention, R 5 is methyl.

[0105] In one embodiment of the compounds of the present invention, R 6 and R 7 is hydrogen and C 1~6 In another embodiment of the compound of the present invention, R 6 and R 7 are both hydrogen. In a further embodiment of the compounds of the invention, R 6 and R 7 are both methyl.

[0106] In one embodiment of the compounds of the present invention, R 8 is C 1~6 In another embodiment of the compounds of the present invention, R8 is methyl.

[0107] In one embodiment of the compounds of the present invention, R 9 is hydrogen or C 1~6 In another embodiment of the compounds of the present invention, R 9 is hydrogen. In a further embodiment of the compounds of the invention, R 9 is C 1~6 In another embodiment of the compounds of the present invention, R 9 is methyl.

[0108] In one embodiment of the compounds of the present invention, R 10 is C 1~6 In another embodiment of the compounds of the present invention, R 10 is methyl.

[0109] In one embodiment of a compound of the invention, n is 0, 1, 2, 3, 4, or 5. In another embodiment of a compound of the invention, n is 0. In a further embodiment of a compound of the invention, n is 0, 1, or 2. In another embodiment of a compound of the invention, n is 1, 2, or 3. In another embodiment of a compound of the invention, n is 1 or 2. In a further embodiment of a compound of the invention, n is 1. In another embodiment of a compound of the invention, n is 2. In a further embodiment of a compound of the invention, n is 3. In another embodiment of a compound of the invention, n is 4. In a further embodiment of a compound of the invention, n is 5.

[0110] In one embodiment, the present invention provides a compound of formula Ia: [ka] [In the formula, A, R 1 and n is as defined for compounds of formula I, or a pharmaceutically acceptable salt thereof.

[0111] In one embodiment, the present invention provides a compound of formula 1b: [ka] [In the formula, Each R 1a is XR 4 , halogens, deuterium, C 1~4 independently selected from the group consisting of alkyl and -CN; Each R 1b is XR 4 , halogen, deuterium, -C 1~4 Alkyl, -OC 1~4 Alkyl, heteroaryl, -CN, -NO2, -C(O)OR 5 , -C(O)NR 6 R 7 , -S(O)2R 8 each -C independently selected from the group consisting of 1~4 Alkyl and heteroaryl are substituted with halogen and -C 1~4 optionally substituted with one or more substituents selected from the group consisting of alkyl; R 1c is deuterium, -C 1~4 Alkyl, -CN, -C(O)OR 5 and -S(O)2R 8 selected from the group consisting of: X is selected from the group consisting of O, CH2, and S(O)2; R 4 is selected from the group consisting of aryl and heterocycloalkyl; 4 is -S(O)2R 8 is optionally substituted by; R 5 is hydrogen and C 1~4 selected from the group consisting of alkyl; R 6 and R 7 is hydrogen and C 1~4 independently selected from the group consisting of alkyl; R 8 is C 1~4 or a pharmaceutically acceptable salt thereof.

[0112] In one embodiment of the compound of Formula 1b, each R 1ais XR 4 each R is independently selected from the group consisting of chlorine, deuterium, methyl, and —CN; 1b is XR 4 , halogen, deuterium, -C 1~4 independently selected from the group consisting of alkyl, -OCH3, heteroaryl, -CN, -NO2, -C(O)OH, -C(O)NH2, and -S(O)2CH3; 1~4 Alkyl and heteroaryl may contain fluorine and -C 1~4 optionally substituted with one or more substituents selected from the group consisting of alkyl; R 1c is selected from the group consisting of deuterium, methyl, -CN, -C(O)OH, and -S(O)2CH3; X is selected from the group consisting of O, CH2, and S(O)2; R 4 teeth, Phenyl and [ka] wherein each phenyl is optionally substituted with -S(O)2CH3.

[0113] In a further embodiment of the compound of Formula 1b, each R 1a is XR 4 each R is independently selected from the group consisting of chlorine, deuterium, methyl, and —CN; 1b is XR 4 , F, Cl, Br, deuterium, -methyl, -CF3, isopropyl, -OCH3, thiazolyl, pyrazolyl, triazolyl-CN, -NO2, -C(O)OH, -C(O)NH2, -S(O)2CH3, wherein each thiazolyl or pyrazolyl is optionally substituted with methyl; R 1c is selected from the group consisting of deuterium, methyl, -CN, -C(O)OH, and -S(O)2CH3; X is selected from the group consisting of O, CH2, and S(O)2; R 4 is phenyl and [ka] wherein each phenyl is optionally substituted with -S(O)2CH3.

[0114] Exemplary compounds according to the present invention include those set forth in Table 2, or a pharmaceutically acceptable salt thereof:

[0115] [Table 2-1]

[0116] [Table 2-2]

[0117] [Table 2-3]

[0118] [Table 2-4]

[0119] [Table 2-5]

[0120] In particular, the present invention provides the following compounds: [ka] or a pharmaceutically acceptable salt, polymorphic form, solvate, hydrate or tautomeric form thereof.

[0121] In one embodiment, the compound is a pharmaceutically acceptable salt, polymorphic form, solvate, hydrate, or tautomeric form of the compound.

[0122] In one embodiment, the compound is a pharmaceutically acceptable salt of the compound.

[0123] In another embodiment, the compound is the hydrochloride salt of: [ka]

[0124] In one embodiment, the compound of the invention has the chemical name (E)-3-fluoro-2-((phenylsulfonyl)methyl)prop-2-en-1-amine.

[0125] Preparation of Compounds of Formula I The compounds described herein are synthesized using standard synthetic techniques using methods known in the art.

[0126] The compounds are prepared using standard organic techniques, including but not limited to those described in Advanced Organic Chemistry, 6th Edition, March, John Wiley and Sons Inc. Standard procedures for the use of protecting groups to temporarily protect functional groups such as alcohols, amines, and carboxylic acids are described, for example, in Protecting Groups in Organic Synthesis, 4th Edition, John Wiley and Sons Inc.

[0127] Alternative reaction conditions for the chemical transformations described herein may include, for example, variations in solvents, reaction temperatures, reaction times, as well as different chemical reagents.

[0128] [ka]

[0129] P is a functional group used to protect the nitrogen functionality. Examples of P include carbonates such as tert-butyloxycarbonyl (BOC), 9-fluorenylmethyloxycarbonyl (FMOC), and benzyloxycarbonyl (CBZ) groups. In some embodiments, allyl bromide A-1 (the synthesis of A-1 is described in WO 2021 / 258159 and WO 2018 / 158140) is treated with an arylsulfinic acid sodium salt A-3 in a suitable solvent, such as DMF, to provide allyl sulfone A-4. In some embodiments, the solvent is acetone or acetonitrile. In some embodiments, A-3 can be prepared from the corresponding arylsulfonyl chloride A-2 by treatment with sodium sulfite and sodium carbonate in a suitable solvent, such as water. In other embodiments, the reaction is carried out for 1 to 5 hours. In some embodiments, standard amine deprotection conditions provide A-5. In some embodiments, the deprotection conditions involve treatment of A-4 with HCl solution. In other embodiments, HCl in 1,4-dioxane is used. In some embodiments, HCl in diethyl ether is used. In some embodiments, HCl in ethyl acetate is used. In other embodiments, the deprotection conditions involve treatment of A-4 with TFA.

[0130] [ka]

[0131] In some embodiments, thiol derivative B-2 is treated with allyl bromide A-1 in the presence of a suitable base using a suitable polar solvent to give thioether derivative B-5. In some embodiments, the base is Cs2CO3. In other embodiments, the base is K2CO3. In some embodiments, the solvent is DMF. In other embodiments, the solvent is acetone or acetonitrile. In some embodiments, sulfone derivative B-6 is obtained by treating thioether B-5 with a suitable oxidizing agent in a suitable solvent. In some embodiments, the oxidizing agent is hydrogen peroxide. In other embodiments, sodium tungstate is added. In still other embodiments, the oxidizing agent is Oxone® (potassium peroxymonosulfate). In some embodiments, the solvent is methanol. In some embodiments, standard amine deprotection conditions give B-7. In some embodiments, the deprotection conditions involve treatment of B-6 with HCl solution. In other embodiments, HCl in 1,4-dioxane is used. In other embodiments, the deprotection conditions involve treatment of B-6 with TFA.

[0132] [ka]

[0133] In some embodiments, the allyl sulfide derivative C-2 can be obtained from A-1 by treatment with thiourea, followed by base-promoted hydrolysis in a suitable solvent at a suitable temperature.

[0134] In some embodiments, the solvent is methanol. In some embodiments, the temperature is reflux. In other embodiments, the temperature is room temperature. In some embodiments, allyl sulfide C-2 can be coupled with aryl iodide C-3 in the presence of a suitable catalyst and base to provide allyl sulfide C-4. In some embodiments, the catalyst is CuO. In other embodiments, the catalyst is CuI or a palladium-containing catalyst. In some embodiments, the base is potassium hydroxide. In other embodiments, the base is Cs2CO3. In some embodiments, the solvent used in the coupling reaction is DMSO. In other embodiments, the solvent is water or DMF. In some embodiments, the temperature at which the coupling reaction is carried out is 120°C. In some embodiments, the sulfone derivative C-5 can be obtained by treating thioether C-4 with a suitable oxidizing agent in a suitable solvent. In some embodiments, the oxidizing agent is hydrogen peroxide. In other embodiments, sodium tungstate is added. In still other embodiments, the oxidizing agent is Oxone®. In some embodiments, the solvent is methanol. In some embodiments, standard amine deprotection conditions provide C-6. In some embodiments, the deprotection conditions involve treatment of C-5 with HCl solution. In other embodiments, HCl in 1,4-dioxane is used. In other embodiments, the deprotection conditions involve treatment of C-5 with TFA.

[0135] [ka]

[0136] In some embodiments, the acid derivative D-1 is converted to the corresponding amide D-3 using standard peptide coupling techniques known to those skilled in the art. In some embodiments, the carboxylic acid derivative D-1 and an appropriately substituted amine HNR 1 R 2Standard peptide coupling conditions between D-1 and D-2 provide amide D-3. In some embodiments, standard amine deprotection conditions provide D-4 from D-3 and D-2 from D-1. In some embodiments, deprotection conditions involve treatment of D-1 or D-3 with HCl solution. In other embodiments, HCl in 1,4-dioxane is used. In other embodiments, deprotection conditions involve treatment of D-1 or D-3 with TFA.

[0137] [ka]

[0138] In some embodiments, ester E-1 is converted to the corresponding acid derivative E-2 by treatment with a suitable base in water and a suitable co-solvent. In some embodiments, the base is sodium hydroxide. In other embodiments, the base is potassium hydroxide. In some embodiments, the co-solvent used is methanol. In other embodiments, the co-solvent is methanol and THF. In some embodiments, standard amine deprotection conditions provide E-3. In some embodiments, the deprotection conditions involve treatment of E-2 with HCl solution. In other embodiments, HCl in 1,4-dioxane is used. In other embodiments, the deprotection conditions involve treatment of E-2 with TFA.

[0139] [ka]

[0140] In some embodiments, boronate ester F-2 can be obtained from an aryl halide by treatment with a suitable diboron reagent, a palladium-based catalyst, and a base in a suitable solvent. In some embodiments, the diboron reagent is bis(pinacolato)diboron. In some embodiments, the palladium catalyst is bis(diphenylphosphino)ferrocene-palladium(II) dichloride dichloromethane complex. In some embodiments, the base is potassium acetate. In some embodiments, the solvent is 1,4-dioxane. In some embodiments, biaryl derivative F-3 can be obtained by applying standard Suzuki-type coupling conditions between boronate ester F-2 and a suitably substituted aryl or heteroaryl. In some embodiments, the palladium catalyst used in the coupling reaction is tetrakis(triphenylphosphine)palladium(0). In some embodiments, the base used is tribasic potassium phosphate. In other embodiments, the solvent used is 1,4-dioxane. In still other embodiments, the temperature at which the coupling reaction is carried out is 90°C. In some embodiments, biaryl derivative F-3 can be obtained from boronic ester F-2 and a suitably substituted aryl or heteroaryl by treatment with copper acetate and a suitable base. In some embodiments, the base is pyridine. In other embodiments, the reaction is carried out in a solvent such as acetonitrile.

[0141] Cis / trans (E / Z) isomers can be separated by conventional techniques well known to those skilled in the art, such as chromatography and fractional crystallization.

[0142] Therapeutic Uses and Formulations Another aspect of the present invention relates to a pharmaceutical composition comprising a compound of the present invention, or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable diluent, excipient or adjuvant.

[0143] The present invention also relates to the use of compounds of the present invention in therapy, particularly for inhibiting the lysyl oxidase family members LOX, LOXL1, LOXL2, LOXL3, and LOXL4. In one embodiment, the present invention provides for the selective inhibition of a specific lysyl oxidase isozyme. In another embodiment, the present invention provides for the simultaneous inhibition of two, three, four, or five lysyl oxidase isozymes. The relative inhibitory potency of a compound can be determined by the amount required to inhibit the amine oxidase activity of LOX, LOXL1, LOXL2, LOXL3, and LOXL4 in a variety of ways, including in vitro assays using recombinant or purified human proteins or recombinant or purified non-human enzymes, cell assays expressing normal rodent enzymes, cell assays transfected with human proteins, and in vivo studies in rodents and other mammalian species.

[0144] In one embodiment, the compounds of the invention are long-lasting inhibitors of the lysyl oxidase family members LOX, LOXL1, LOXL2, LOXL3, and LOXL4. In one embodiment, the compounds of the invention exhibit activity at compound concentrations greater than IC 50 A compound of the present invention is a long-lasting inhibitor of a LOX or LOXL1-4 enzyme if the inhibition of LOX or LOXL1-4 enzyme activity continues by more than 50% after the activity of the LOX or LOXL1-4 enzyme drops to less than 50%. In one embodiment, a compound of the present invention exhibits sustained inhibition of a LOX or LOXL1-4 enzyme over a 30 minute period. In one embodiment, a compound of the present invention exhibits sustained inhibition of a LOX or LOXL1-4 enzyme over a 24 hour period. In one embodiment, a compound of the present invention is an irreversible inhibitor of the lysyl oxidase family members LOX, LOXL1, LOXL2, LOXL3, and LOXL4.

[0145] Accordingly, a further aspect of the present invention relates to a method for inhibiting the amine oxidase activity of any one of LOX, LOXL1, LOXL2, LOXL3, or LOXL4 in a subject in need thereof, comprising administering to the subject an effective amount of a compound of the present invention, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition thereof.

[0146] In one embodiment, the invention relates to a method of inhibiting the amine oxidase activity of LOXL2. In another embodiment, the invention relates to inhibiting the amine oxidase activity of LOX and LOXL2. In a further embodiment, the invention relates to a method of inhibiting the amine oxidase activity of LOX.

[0147] As previously mentioned, LOX and LOXL1-4 enzymes are members of a large family of flavin- and copper-dependent amine oxidases, which includes SSAO / VAP-1, monoamine oxidase-B (MAO-B), and diamine oxidase (DAO). In one embodiment, compounds of the invention selectively inhibit members of the lysyl oxidase isozyme family with respect to SSAO / VAP-1, MAO-B, or DAO. In one embodiment, compounds of the invention selectively inhibit members of the lysyl oxidase isozyme family with respect to other members of the amine oxidase family.

[0148] Compounds containing an amine moiety can be metabolized as substrates by amine oxidase, which can reduce therapeutically effective compound concentrations and generate harmful products. The compounds of the present invention are stable in the presence of SSAO / VAP-1, MAO-A, MAO-B, DAO, and dog serum.

[0149] The present invention also discloses methods of using the compounds of the present invention to inhibit one or more lysyl oxidase isozymes (LOX, LOXL1, LOXL2, LOXL3, and LOXL4) in patients suffering from fibrotic diseases, and methods of treating fibrotic diseases. Furthermore, the present invention discloses methods of using the compounds of the present invention to inhibit one or more lysyl oxidase isozymes (LOX, LOXL1, LOXL2, LOXL3, and LOXL4) in patients suffering from cancer, including metastatic cancer, and methods of treating cancer and metastatic cancer.

[0150] In a further aspect of the present invention, there is provided a method for treating a condition by inhibiting the activity of any one of LOX, LOXL1, LOXL2, LOXL3 and LOXL4 proteins, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition thereof.

[0151] In another aspect, a method for treating a condition modulated by any one of LOX, LOXL1, LOXL2, LOXL3 and LOXL4 is provided, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition thereof.

[0152] In one embodiment of the method of the present invention, the condition is selected from the group consisting of fibrosis, cancer, and arthritis.

[0153] In another aspect, the present invention provides methods for reducing extracellular matrix formation by treating human subjects, companion animals, and livestock with the fluoroarylamine inhibitors of the lysyl oxidase isozyme family of the invention described herein.

[0154] The above methods are applicable where the condition is fibrosis. As used herein, "fibrosis" includes diseases such as cystic fibrosis, pulmonary fibrosis, liver fibrosis, renal fibrosis, scleroderma, radiation-induced fibrosis, Dupuytren's contracture, Peyronie's disease, scarring, and other diseases in which excessive fibrosis contributes to the pathology.

[0155] In one embodiment, the fibrosis is selected from the group consisting of mediastinal fibrosis, myelofibrosis, retroperitoneal fibrosis, progressive giant fibrosis, nephrogenic systemic fibrosis, Crohn's disease, keloids, scleroderma / systemic sclerosis, arthrofibrosis, Dupuytren's contracture, adhesive capsulitis, pancreatic fibrosis, intestinal fibrosis, hepatic fibrosis, pulmonary fibrosis, renal fibrosis, cardiac fibrosis, fibrostenosis, cystic fibrosis, idiopathic pulmonary fibrosis, radiation-induced fibrosis, Peyronie's disease, and scleroderma, or associated with respiratory disease, abnormal wound healing and repair, scarring, hypertrophic scarring / keloids, post-surgical scarring, cardiac arrest, and all conditions in which excessive or abnormal deposition of fibrous material is associated with disease, injury, transplantation, or surgery. In another embodiment, the fibrosis is selected from the group consisting of hepatic fibrosis, pulmonary fibrosis, renal fibrosis, cardiac fibrosis, scarring, and scleroderma. In a further embodiment, the fibrosis is selected from the group consisting of keloids, scars, hypertrophic scars, scleroderma and Dupuytren's contracture.

[0156] In one embodiment, renal fibrosis includes, but is not limited to, diabetic nephropathy, vesicoureteral reflux, tubulointerstitial renal fibrosis, glomerulonephritis or glomerular nephritis (including focal segmental glomerulosclerosis and membranous glomerulonephritis, IgA nephropathy, and membranoproliferative glomerulonephritis). In one embodiment, liver fibrosis leads to liver cirrhosis and includes related conditions such as chronic viral hepatitis, nonalcoholic fatty liver disease (NAFLD), alcoholic steatohepatitis (ASH), nonalcoholic steatohepatitis (NASH), primary biliary cirrhosis (PBC), biliary cirrhosis, and autoimmune hepatitis.

[0157] In one embodiment, the fibrosis is selected from keloids, scars, ocular scars, hypertrophic scars, scleroderma, Dupuytren's contracture, and Peyronie's disease. In one embodiment, the hypertrophic scar results from a burn. In one embodiment, the hypertrophic scar is caused by trauma. In another embodiment, the hypertrophic scar is caused by a surgical procedure. In one embodiment, the keloid is caused by trauma. In another embodiment, the keloid is caused by a surgical procedure. In a further embodiment, the keloid is the result of skin damage caused by acne, burns, chickenpox, perforations, abrasions, surgical incisions, or vaccination sites.

[0158] A further aspect of the present invention provides a method for treating or preventing keloid disease or scarring, comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to the present invention, or a pharmaceutically acceptable salt thereof. In one embodiment, the compound is administered topically. In another embodiment, the compound is administered orally.

[0159] In one embodiment, the keloid disease or scar is a keloid, hypertrophic scar, post-surgical scar, burn scar, post-traumatic scar, Dupuytren's contracture, or is the result of skin damage caused by trauma or a surgical procedure, or caused by acne, burns, chickenpox, infection, perforation, abrasion, surgical incision, or vaccination site. In one embodiment, the treatment aids in wound healing and improves the subject's skin compliance or skin appearance.

[0160] The above methods are also applicable when the condition is a proliferative disease, such as cancer. In one embodiment, the cancer is selected from the group consisting of lung cancer, breast cancer, colon cancer, anal cancer, pancreatic cancer, prostate cancer, ovarian cancer, liver and bile duct cancer, esophageal cancer, non-Hodgkin's lymphoma, bladder cancer, uterine cancer, glioma, glioblastoma, medulloblastoma, and other tumors of the brain, myelofibrosis, kidney cancer, head and neck cancer, gastric cancer, multiple myeloma, testicular cancer, germ cell tumors, neuroendocrine tumors, cervical cancer, oral cancer, carcinoids of the gastrointestinal tract, breast, and other organs, signet ring cell carcinoma, sarcoma, fibrosarcoma, hemangioma, hemangiomatosis, hemangiopericytoma, pseudoangiomatous stromal hyperplasia of the breast, and other organs. hyperplasia, myofibroblastoma, fibromatosis, inflammatory myofibroblastic tumor, lipoma, angiolipoma, granular cell tumor, neurofibroma, neurilemmoma, angiosarcoma, liposarcoma, rhabdomyosarcoma, osteosarcoma, mesenchymal tumors including leiomyoma or leiomyosarcoma.

[0161] In one embodiment, the proliferative disorder is a myeloid malignancy selected from the group consisting of myelodysplastic syndromes (MDS), myelodysplastic neoplasms (MPN), MDS / MPS overlap syndromes, and acute myeloid leukemia (AML), including acute promyelocytic leukemia (APL).

[0162] In one embodiment, the cancer is selected from the group consisting of breast cancer, head and neck squamous cell carcinoma, brain cancer, prostate cancer, renal cell carcinoma, liver cancer, lung cancer, oral cancer, cervical cancer, and tumor metastasis.

[0163] In one embodiment, lung cancer includes lung adenocarcinoma, squamous cell carcinoma, large cell carcinoma, bronchioloalveolar carcinoma, non-small cell carcinoma, small cell carcinoma, and mesothelioma. In one embodiment, breast cancer includes ductal carcinoma, lobular carcinoma, inflammatory breast cancer, clear cell carcinoma, and mucinous carcinoma. In one embodiment, colorectal cancer includes colon cancer and rectal cancer. In one embodiment, pancreatic cancer includes pancreatic adenocarcinoma, pancreatic islet cell carcinoma, and neuroendocrine tumors.

[0164] In one embodiment, ovarian cancer includes ovarian epithelial or surface epithelial-stromal tumors, including serous tumors, endometrioid tumors, and mucinous cystadenocarcinomas, as well as sex cord-stromal tumors. In one embodiment, liver cancer and bile duct cancer include hepatocellular carcinoma, cholangiocarcinoma, and hemangioma. In one embodiment, esophageal cancer includes esophageal adenocarcinoma and squamous cell carcinoma. In one embodiment, uterine cancer includes endometrial adenocarcinoma, uterine papillary serous carcinoma, uterine clear cell carcinoma, uterine sarcoma and leiomyosarcoma, and mixed Müllerian tumor. In one embodiment, kidney cancer includes renal cell carcinoma, clear cell carcinoma, and Wilms' tumor. In one embodiment, head and neck cancer includes squamous cell carcinoma. In one embodiment, gastric cancer includes gastric adenocarcinoma and gastrointestinal stromal tumor.

[0165] In one embodiment, the cancer is selected from the group consisting of pancreatic cancer, liver cancer, breast cancer, myelofibrosis, and mesothelioma.

[0166] In one embodiment, the compounds of the present invention may be for use in the treatment of non-metastatic cancer. In another embodiment, the compounds of the present invention may be for use in the treatment of metastatic cancer. In a further embodiment, the compounds of the present invention may be for use in the prevention or treatment of tumor metastasis.

[0167] The above methods are applicable where the condition is arthritis, which as used herein includes rheumatoid arthritis and osteoarthritis.

[0168] In one embodiment of the method of the present invention, the subject is selected from the group consisting of humans, pets and livestock. In another embodiment of the method of the present invention, the subject is a human.

[0169] A further aspect of the present invention provides the use of a compound of the present invention, or a pharmaceutically acceptable salt or solvate thereof, for the manufacture of a medicament for treating a condition by inhibiting the activity of any one of the LOX, LOXL1, LOXL2, LOXL3 and LOXL4 proteins.

[0170] Another aspect of the present invention provides the use of a compound of the present invention, or a pharmaceutically acceptable salt or solvate thereof, for the manufacture of a medicament for treating a condition modulated by any one of LOX, LOXL1, LOXL2, LOXL3 and LOXL4.

[0171] Pharmaceutical and / or therapeutic preparations In another embodiment of the present invention, there is provided a composition comprising a compound of the present invention and at least one pharmaceutically acceptable excipient, carrier, or diluent. The compound of the present invention may also be present as a suitable salt, including a pharmaceutically acceptable salt.

[0172] The phrase "pharmaceutically acceptable carrier" refers to any carrier known to those skilled in the art to be suitable for a particular mode of administration. Further, the compounds may be formulated as the sole pharmaceutically active ingredient in the composition or may be combined with other active ingredients.

[0173] The phrase "pharmaceutically acceptable salt" refers to any salt preparation suitable for use in pharmaceutical applications. A pharmaceutically acceptable salt means a salt that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, etc., and that is commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art and include acid addition salts and base salts. Hemisalts of acids and bases can also be formed. Pharmaceutically acceptable salts include amine salts of mineral acids (e.g., hydrochloride, hydrobromide, sulfate, etc.); and amine salts of organic acids (e.g., formate, acetate, lactate, malate, tartrate, citrate, ascorbate, succinate, maleate, butyrate, valerate, fumarate, sulfonate, etc.).

[0174] For compounds having a basic moiety, a suitable pharmaceutically acceptable salt may be an acid addition salt. For example, a suitable pharmaceutically acceptable salt of such a compound can be prepared by mixing a pharmaceutically acceptable acid (e.g., hydrochloric acid, sulfuric acid, methanesulfonic acid, succinic acid, fumaric acid, maleic acid, benzoic acid, phosphoric acid, acetic acid, oxalic acid, carbonic acid, tartaric acid, or citric acid) with the compound of the present invention.

[0175] S.M. Berge et al. provide a detailed description of pharmaceutically acceptable salts in J. Pharmaceutical Sciences, 1977, 66:1-19. Salts can be prepared in situ during the final isolation and purification of the compounds of the invention, or separately by reacting the free base function with a suitable organic acid. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, digluconate, cyclopentanepropionate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, 2-hydroxybenzoate, and the like. Examples of suitable base salts include dimethylsulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oleate, palmitate, pamoate, pectinate, 3-phenylpropionate, phosphate, pivalate, propionate, stearate, succinate, sulfate, tartrate, toluenesulfonate, undecanoate, valerate, and the like. Suitable base salts are formed from bases which form non-toxic salts. Examples include arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine, and zinc salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like, as well as non-toxic ammonium, quaternary ammonium, and amine cations, including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, dimethylamine, trimethylamine, triethylamine, triethanolamine, and the like.

[0176] Pharmaceutically acceptable salts of compounds can be prepared by methods known to those skilled in the art, such as, for example: (i) reacting the compound with the desired acid or base; (ii) removing an acid- or base-labile protecting group from a suitable precursor of the compound, or ring-opening a suitable cyclic precursor, e.g., a lactone or lactam, using a desired acid or base; or (iii) Converting one salt of the compound to another by reaction with an appropriate acid or base or using a suitable ion exchange column.

[0177] Reactions (i) to (iii) above are typically carried out in solution. The resulting salt can be precipitated and recovered by filtration or by evaporation of the solvent. The degree of ionization in the resulting salt can vary from completely ionized to almost non-ionized.

[0178] Thus, for example, suitable pharmaceutically acceptable salts of compounds according to the invention can be prepared by mixing a pharmaceutically acceptable acid such as hydrochloric acid, sulfuric acid, methanesulfonic acid, succinic acid, fumaric acid, maleic acid, benzoic acid, phosphoric acid, acetic acid, carbonic acid, tartaric acid, or citric acid with a compound of the invention. Accordingly, suitable pharmaceutically acceptable salts of compounds of the invention include acid addition salts.

[0179] The compounds of the present invention can exist in both unsolvated and solvated forms.The term "solvate" is used herein to describe a molecular complex comprising a compound of the present invention and a stoichiometric amount of one or more pharmaceutically acceptable solvent molecules (e.g., ethanol).The term "hydrate" is used when the solvent is water.

[0180] The compositions herein comprise the compounds provided herein.In one embodiment, the compounds of the present invention are formulated into suitable pharmaceutical preparations, such as solutions, suspensions, tablets, creams, gels, dispersible tablets, pills, capsules, powders, sustained-release preparations or elixirs for oral administration, or sterile solutions or suspensions for parenteral administration, as well as transdermal patch preparations and dry powder inhalers.In one embodiment, the compounds are formulated into pharmaceutical compositions using techniques and procedures well known in the art.

[0181] In the composition, an effective concentration of the compound or its pharmaceutically acceptable derivative is mixed with a suitable pharmaceutical carrier. The compound can be derivatized as the corresponding salt, base, solvate, or hydrate before formulation, as described above. The concentration of the compound in the composition is effective to deliver an amount that, upon administration, treats, prevents, or ameliorates one or more symptoms of the disease or disorder being treated.

[0182] In one embodiment, the composition is formulated for single administration. To formulate the composition, a weight fraction of the compound is dissolved, suspended, dispersed, or otherwise mixed in a selected carrier at an effective concentration such that the condition being treated is alleviated, prevented, or one or more symptoms are ameliorated.

[0183] The active compound is included in a pharmaceutically acceptable carrier in an amount sufficient to exert a therapeutically useful effect without undesirable side effects in the treated patient. The therapeutically effective concentration can be empirically determined by testing the compound in the in vitro and in vivo systems described herein, and then extrapolated therefrom to dosages for humans.

[0184] The concentration of active compound in the pharmaceutical composition will vary depending on absorption, distribution, inactivation, and excretion rates of the active compound, the physicochemical properties of the compound, the administration schedule and dosage, and other factors known to those of skill in the art.

[0185] Administration may occur at intervals of minutes, hours, days, weeks, months, or years, or may be continuous over any one of these time periods. A suitable dosage is in the range of about 0.1 ng / kg body weight to 0.1 g / kg body weight per dose. The dosage is preferably in the range of 1 μg to 0.1 g / kg body weight per dose, for example, 1 mg to 0.1 g / kg body weight per dose. Suitably, the dosage is in the range of 1 μg to 50 mg / kg body weight per dose, for example, 1 μg to 20 mg / kg body weight per dose, or 1 μg to 10 mg / kg body weight per dose. Other suitable dosages may be in the range of 1 mg to 25 mg / kg body weight per dose, for example, 1 mg to 10, 20, 50, or 100 mg / kg body weight per dose, or 10 μg to 100 mg / kg body weight per dose. In one embodiment, the dosage is in the range of 1 mg to 10 mg / kg body weight per dose.

[0186] Alternatively, the effective dose is up to about 10 mg / cm 2 or up to about 1 mg / cm 2 , about 0.5mg / cm 2 , about 0.2mg / cm 2 , about 0.1mg / cm 2 , about 0.05mg / cm 2 , approximately 0.02 mg / cm 2 , or about 0.01 mg / cm 2 For example, about 0.1 μg / cm 2 ~About 1mg / cm 2 , or about 1 μg / cm 2 ~Approx. 1mg / cm 2 , about 10μg / cm 2 ~About 1mg / cm 2 , about 10μg / cm 2 ~about 0.1mg / cm 2 , about 10μg / cm 2 ~about 0.01mg / cm 2 , about 10μg / cm 2 ~Approx. 500μg / cm 2 , about 10μg / cm 2 ~Approx. 200μg / cm 2 , about 10μg / cm 2 ~Approx. 100μg / cm 2 , about 10μg / cm2 ~Approx. 50μg / cm 2 , about 20μg / cm 2 ~About 1mg / cm 2 , about 50μg / cm 2 ~About 1mg / cm 2 , about 100μg / cm 2 ~About 1mg / cm 2 , about 200μg / cm 2 ~About 1mg / cm 2 , about 500pg / cm 2 ~About 1mg / cm 2 , about 50μg / cm 2 ~Approx. 500μg / cm 2 , about 50μg / cm 2 ~Approx. 200μg / cm 2 , about 100μg / cm 2 ~Approx. 500μg / cm 2 or approximately 200 μg / cm 2 ~Approx. 500μg / cm 2 The range may be:

[0187] Suitable dosages and administration regimens will be determined by the attending physician and may vary depending on the particular condition being treated, the severity of the condition, and the general health, age, and weight of the subject.

[0188] When a compound exhibits insufficient solubility, methods for solubilizing the compound can be used. Such methods are known to those skilled in the art and include, but are not limited to, the use of cosolvents such as dimethyl sulfoxide (DMSO), the use of surfactants such as TWEEN®, dissolving in aqueous sodium bicarbonate, formulating the compound of interest as nanoparticles, etc. Compound derivatives (e.g., prodrugs of the compound) can also be used to formulate effective pharmaceutical compositions.

[0189] Upon mixing or addition of the compounds, the resulting mixture may be a solution, suspension, emulsion, etc. The form of the resulting mixture will vary depending upon many factors, including the intended mode of administration and the solubility of the compound in the selected carrier or vehicle. The effective concentration is sufficient for ameliorating the symptoms of the disease, disorder, or condition being treated and may be empirically determined.

[0190] Pharmaceutical compositions are provided in unit dosage forms such as tablets, capsules, pills, powders, granules, sterile parenteral solutions or suspensions, and oral solutions or suspensions, and oil-water emulsions, containing a suitable amount of the compound or a pharmaceutically acceptable derivative thereof for administration to humans and animals.

[0191] In one embodiment, pharmaceutically therapeutically active compounds and derivatives thereof are formulated and administered in unit dosage forms or multiple dosage forms. The active ingredient can be administered at once or in several small doses at intervals. As used herein, a unit dosage form refers to a physically discrete unit suitable for human and animal subjects, individually packaged as known in the art. Each unit dosage contains a predetermined amount of a therapeutically active compound sufficient to produce a desired therapeutic effect, together with the necessary pharmaceutical carrier, vehicle, or diluent. Examples of unit dosage forms include ampoules and syringes, and individually packaged tablets or capsules. A unit dosage form can be administered in small amounts or repetitions thereof. A multiple dosage form is a plurality of identical unit dosage forms packaged in a single container for administration in separate unit dosage forms. Examples of multiple dosage forms include vials, bottles of tablets, or capsules. Thus, a multiple dosage form is a plurality of unit dosages that are not separated in packaging.

[0192] Actual methods for preparing such dosage forms are known, or will be apparent, to those skilled in the art.

[0193] Dosage forms or compositions can be prepared containing active ingredient in the range of 0.005% to 100% (by weight), with the remainder consisting of non-toxic carriers. Methods for preparing these compositions are known to those skilled in the art. Contemplated compositions contain 0.001% to 100% (by weight) active ingredient, and in one embodiment 0.1 to 95% (by weight), in another embodiment 75 to 85% (by weight), and in another embodiment 0.1 to 25% (by weight) active ingredient. The amount of active substance in such therapeutically useful compositions is such that an effective dosage can be achieved.

[0194] Mode of administration Convenient modes of administration include injection (subcutaneous, intravenous, etc.), oral administration, inhalation, transdermal application, topical administration to the skin, eye, ear, or oral surfaces, vaginal, or rectal administration. Depending on the route of administration, the formulation and / or compound may be coated with a material to protect the compound from the action of enzymes, acids, and other natural conditions that may inactivate the compound's therapeutic activity. The compound may also be administered parenterally or intraperitoneally.

[0195] Orally Administered Composition Oral pharmaceutical dosage forms can be solid, gel, or liquid. Solid dosage forms include tablets, capsules, granules, and bulk powders. Types of oral tablets include compressed chewable lozenges and tablets that may be enteric-coated, sugar-coated, or film-coated. Capsules can be hard or soft gelatin capsules, and granules and powders can be provided in non-effervescent or effervescent form in combination with other ingredients known to those skilled in the art.

[0196] Solid Composition for Oral Administration In certain embodiments, the formulation is a solid dosage form, in one embodiment, a capsule or tablet. Tablets, pills, capsules, lozenges, etc. may contain one or more of the following ingredients: binders; lubricants; diluents; glidants; disintegrants; colorants; sweeteners; flavoring agents; wetting agents; emetic coatings; and film coatings, or compounds of similar nature. Examples of binders include microcrystalline cellulose, tragacanth gum, glucose solution, acacia mucilage, gelatin solution, molasses, polyvinylpyrrolidine, povidone, crospovidone, sucrose, and starch paste. Lubricants include talc, starch, magnesium or calcium stearate, lycopodium, and stearic acid. Diluents include, for example, lactose, sucrose, starch, kaolin, salt, mannitol, and dicalcium phosphate. Glidants include, but are not limited to, colloidal silicon dioxide. Disintegrants include croscarmellose sodium, sodium starch glycolate, alginic acid, corn starch, potato starch, bentonite, methylcellulose, agar, and carboxymethylcellulose. Coloring agents include, for example, any of the approved certified water-soluble FD and C dyes, mixtures thereof, and water-insoluble FD and C dyes suspended on alumina hydrate. Sweetening agents include sucrose, lactose, mannitol, and artificial sweeteners such as saccharin, as well as various spray-dried flavors. Flavoring agents include, but are not limited to, natural flavors extracted from plants such as fruits, and synthetic blends of compounds that produce a pleasant sensation, such as peppermint and methyl salicylate. Wetting agents include propylene glycol monostearate, sorbitan monooleate, diethylene glycol monolaurate, and polyoxyethylene laural ether. Emetic coatings include fatty acids, fats, waxes, shellac, ammoniated shellac, and cellulose acetate phthalate.Film coatings include hydroxyethylcellulose, sodium carboxymethylcellulose, polyethylene glycol 4000 and cellulose acetate phthalate.

[0197] The compound or its pharmaceutically acceptable derivative can be provided in a composition that protects it from the acidic environment of the stomach. For example, the composition can be formulated with an enteric coating that maintains its integrity in the stomach and releases the active compound in the intestine. The composition can also be formulated in combination with an antacid or other similar ingredient.

[0198] When the dosage unit is a capsule, it may contain, in addition to the above-mentioned materials, a liquid carrier such as fatty oil. Furthermore, the dosage unit may contain various other materials that modify the physical form of the dosage unit (for example, sugar coating and other enteric agents). The compound may also be administered as a component of an elixir, suspension, syrup, wafer, sprinkle, chewing gum, etc. The syrup may contain, in addition to the active compound, sucrose as a sweetener and certain preservatives, dyes and colorings, and flavorings.

[0199] The active materials can also be mixed with other active materials that do not impair the desired action, or with materials that supplement the desired action, such as antacids, H2 blockers, and diuretics. The active ingredient is a compound described herein or a pharmaceutically acceptable derivative thereof. Higher concentrations of the active ingredient, up to about 98% by weight, may be included.

[0200] In all embodiments, tablet and capsule formulations may be coated as known to those skilled in the art to modify or sustain dissolution of the active ingredient, and thus may be coated with conventional enteric digestible coatings such as phenylsalicylate, waxes, and cellulose acetate phthalate.

[0201] Oral liquid composition Liquid oral dosage forms include aqueous solutions, emulsions, suspensions, solutions and / or suspensions reconstituted from non-effervescent granules, and effervescent preparations reconstituted from effervescent granules. Aqueous solutions include, for example, elixirs and syrups. Emulsions are either oil-in-water or water-in-oil.

[0202] Liquid pharmaceutically administrable compositions can be prepared, for example, by dissolving, dispersing, or otherwise mixing the active compound(s) defined above and optional pharmaceutical adjuvants in a carrier, for example, water, saline, aqueous dextrose, glycerol, glycol, ethanol, etc., thereby forming a solution or suspension. If desired, the pharmaceutical composition to be administered can also contain minor amounts of non-toxic auxiliary substances, such as wetting agents, emulsifiers, solubilizing agents, pH buffering agents, and the like, such as acetate salts, sodium citrate, cyclodextrin derivatives, sorbitan monolaurate, triethanolamine sodium acetate, triethanolamine oleate, and other similar agents.

[0203] Elixirs are clear, sweetened, hydroalcoholic preparations. Pharmaceutically acceptable carriers used in elixirs include solvents. Syrups are concentrated aqueous solutions of sugars, such as sucrose, and may contain preservatives. Emulsions are two-phase systems in which one liquid is dispersed in the form of small droplets throughout another liquid. Pharmaceutically acceptable carriers used in emulsions are non-aqueous liquids, emulsifiers, and preservatives. Suspensions use pharmaceutically acceptable suspending agents and preservatives. Pharmaceutically acceptable substances used in non-effervescent granules to be reconstituted into liquid oral dosage forms include diluents, sweeteners, and wetting agents. Pharmaceutically acceptable substances used in effervescent granules to be reconstituted into liquid oral dosage forms include organic acids and carbon dioxide sources. Coloring agents and flavoring agents are used in all of the above dosage forms.

[0204] Solvents include glycerin, sorbitol, ethyl alcohol, and syrup. Examples of preservatives include glycerin, methyl and propylparaben, benzoic acid, sodium benzoate, and ethanol. Examples of non-aqueous liquids used in emulsions include mineral oil and cottonseed oil. Examples of emulsifying agents include gelatin, acacia, tragacanth, bentonite, and surfactants such as polyoxyethylene sorbitan monooleate. Suspending agents include sodium carboxymethylcellulose, pectin, tragacanth, Veegum, and acacia. Sweetening agents include sucrose, syrup, glycerin, and artificial sweeteners such as saccharin. Wetting agents include propylene glycol monostearate, sorbitan monooleate, diethylene glycol monolaurate, and polyoxyethylene lauryl ether. Organic acids include citric acid and tartaric acid. Carbon dioxide sources include sodium bicarbonate and sodium carbonate. Coloring agents include any of the approved certified water soluble FD and C dyes, and mixtures thereof.Flavoring agents include natural flavors extracted from fruits and other plants, and synthetic blends of compounds which produce a pleasant taste sensation.

[0205] For a solid dosage form, the solution or suspension, for example, in propylene carbonate, vegetable oils or triglycerides, is in one embodiment encapsulated in a gelatin capsule. For a liquid dosage form, the solution, for example, in a polyethylene glycol, can be diluted with a sufficient quantity of a pharmaceutically acceptable liquid carrier, for example, water, to be easily measured for administration.

[0206] Alternatively, liquid or semisolid oral formulations can be prepared by dissolving or dispersing the active compound or salt in vegetable oils, glycols, triglycerides, propylene glycol esters (e.g., propylene carbonate) and other similar carriers, and encapsulating these solutions or suspensions in hard or soft gelatin capsule shells. Briefly, such formulations include, but are not limited to, those containing a compound provided herein, a dialkylated mono- or polyalkylene glycol (including, but not limited to, 1,2-dimethoxymethane, diglyme, triglyme, tetraglyme, polyethylene glycol-350-dimethyl ether, polyethylene glycol-550-dimethyl ether, polyethylene glycol-750-dimethyl ether (350, 550, and 750 refer to the approximate average molecular weight of the polyethylene glycol)), and one or more antioxidants (e.g., butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate, vitamin E, hydroquinone, hydroxycoumarin, ethanolamine, lecithin, cephalin, ascorbic acid, malic acid, sorbitol, phosphoric acid, thiodipropionic acid and its esters, and dithiocarbamates).

[0207] Other formulations include, but are not limited to, aqueous alcohol solutions containing pharmaceutically acceptable acetals. The alcohols used in these formulations are pharmaceutically acceptable water-miscible solvents having one or more hydroxyl groups, including, but not limited to, propylene glycol and ethanol. Acetals include, but are not limited to, di(lower alkyl) acetals of lower alkyl aldehydes, such as acetaldehyde diethyl acetal.

[0208] Injections, solutions and emulsions Parenteral administration, characterized in one embodiment by injection, either subcutaneously, intramuscularly, or intravenously, is also contemplated herein. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid prior to injection, or as emulsions. Injectables, solutions, and emulsions also contain one or more excipients. Suitable excipients are, for example, water, saline, dextrose, glycerol, or ethanol. In addition, if desired, the administered pharmaceutical composition may also contain small amounts of nontoxic auxiliary substances, such as wetting or emulsifying agents, pH buffers, stabilizers, solubility enhancers, and other similar agents, such as sodium acetate, sorbitan monolaurate, triethanolamine oleate, and cyclodextrins.

[0209] Implantation of a slow-release or sustained-release system, such that a constant level of dosage is maintained, is also contemplated herein. Briefly, the compounds provided herein are dispersed in a solid internal matrix, such as polymethyl methacrylate, polybutyl methacrylate, plasticized or unplasticized polyvinyl chloride, plasticized nylon, plasticized polyethylene terephthalate, natural rubber, polyisoprene, polyisobutylene, polybutadiene, polyethylene, ethylene-vinyl acetate copolymer, silicone rubber, polydimethylsiloxane, silicone carbonate copolymer, hydrophilic polymers such as hydrogels of esters of acrylic and methacrylic acid, collagen, cross-linked polyvinyl alcohol, and cross-linked partially hydrolyzed polyvinyl acetate; The outer polymer membrane is insoluble in body fluids, and is surrounded by such outer polymer membranes as polyethylene, polypropylene, ethylene / propylene copolymer, ethylene / ethyl acrylate copolymer, ethylene / vinyl acetate copolymer, silicone rubber, polydimethylsiloxane, neoprene rubber, chlorinated polyethylene, polyvinyl chloride, vinyl acetate, vinylidene chloride, ethylene, and vinyl chloride copolymer of propylene, ionomer polyethylene terephthalate, butyl rubber, epichlorohydrin rubber, ethylene / vinyl alcohol copolymer, ethylene / vinyl acetate / vinyl alcohol terpolymer, and ethylene / vinyloxyethanol copolymer.The compound diffuses through the outer polymer membrane in a release rate-controlling step.The percentage of active compound contained in such parenteral compositions varies widely depending on its specific nature, the activity of the compound, and the needs of the patient.

[0210] Parenteral administration of the composition includes intravenous, subcutaneous, and intramuscular administration. Preparations for parenteral administration include sterile solutions ready for injection, sterile dry soluble products (including hypodermic tablets) such as lyophilized powders ready to be combined with a solvent immediately before use, sterile suspensions ready for injection, sterile dry insoluble products ready to be combined with a vehicle immediately before use, and sterile emulsions. The solutions may be aqueous or non-aqueous.

[0211] If administered intravenously, suitable carriers include saline or phosphate buffered saline (PBS), and solutions containing viscosity enhancing and solubilizing agents such as glucose, polyethylene glycol, and polypropylene glycol, and mixtures thereof.

[0212] Pharmaceutically acceptable carriers used in parenteral preparations include aqueous vehicles, non-aqueous vehicles, antibacterial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, emulsifying agents, sequestering or chelating agents, and other pharmaceutically acceptable substances.

[0213] Examples of aqueous vehicles include sodium chloride injection, Ringer's injection, isotonic dextrose injection, sterile water injection, dextrose and lactated Ringer's injection. Non-aqueous parenteral vehicles include fixed oils of vegetable origin, olive oil, cottonseed oil, corn oil, sesame oil, and peanut oil. Bacteriostatic or fungistatic concentrations of antibacterial agents, including phenols or cresols, mercurials, benzyl alcohol, chlorobutanol, methyl and propyl p-hydroxybenzoic acid esters, thimerosal, benzalkonium chloride, and benzethonium chloride, must be added to parenteral preparations packaged in multidose containers. Isotonic agents include sodium chloride and dextrose. Buffers include phosphates and citrates. Antioxidants include sodium bisulfate. Local anesthetics include procaine hydrochloride. Suspending and dispersing agents include sodium carboxymethylcellulose, hydroxypropyl methylcellulose, and polyvinylpyrrolidone. Emulsifying agents include polysorbate 80 (TWEEN® 80). Sequestering or chelating agents of metal ions include EDTA. Pharmaceutical carriers also include ethyl alcohol, polyethylene glycol, and propylene glycol for water-miscible vehicles; and sodium hydroxide, hydrochloric acid, citric acid, or lactic acid for pH adjustment.

[0214] The concentration of the pharmaceutically active compound is adjusted so that an effective amount is obtained upon injection to produce the desired pharmacological effect. The exact dose will vary depending on the age, weight and condition of the patient or animal, as is known in the art.

[0215] Unit-dose parenteral preparations are packaged in ampoules, vials, or syringes with needles. All preparations for parenteral administration must be sterile, as is known and practiced in the art.

[0216] Illustratively, intravenous or intraarterial infusion of a sterile aqueous solution containing an active compound is an effective mode of administration. Another embodiment is a sterile aqueous or oily solution or suspension containing an active material injected as necessary to produce the desired pharmacological effect.

[0217] Injectables are designed for local and systemic administration. In one embodiment, a therapeutically effective dose is formulated to contain a concentration of the active compound relative to the tissue to be treated of at least about 0.1% w / w up to about 90% w / w or more, and in certain embodiments, greater than 1% w / w.

[0218] The compound may be suspended in micronized or other suitable form, or may be derivatized to produce a more soluble active product, or may be derivatized to produce a prodrug. The form of the resulting mixture will vary depending on many factors, such as the intended mode of administration and the solubility of the compound in the selected carrier or vehicle. The effective concentration is sufficient to ameliorate the symptoms of the condition and can be empirically determined.

[0219] freeze-dried powder Also of interest herein are lyophilized powders, which can be reconstituted for administration as solutions, emulsions, and other mixtures, which can also be reconstituted and formulated as solids or gels.

[0220] Sterile lyophilized powders are prepared by dissolving a compound provided herein or a pharmaceutically acceptable derivative thereof in a suitable solvent. The solvent may contain excipients that improve the stability or other pharmacological components of the powder or a reconstituted solution prepared from the powder. Excipients that can be used include, but are not limited to, dextrose, sorbitol, fructose, corn syrup, xylitol, glycerin, glucose, sucrose, or other suitable agents. The solvent may also contain a buffer (e.g., citric acid, sodium phosphate, potassium phosphate, etc.) or other similar buffers known to those of skill in the art (in one embodiment, near neutral pH). Subsequent sterile filtration of the solution followed by lyophilization under standard conditions known to those of skill in the art provides the desired formulation. In one embodiment, the resulting solution is dispensed into vials for lyophilization. Each vial contains a single dose or multiple doses of the compound. The lyophilized powder can be stored under appropriate conditions, for example, at about 4°C to room temperature.

[0221] This lyophilized powder is reconstituted with water for injection to provide a formulation for use in parenteral administration. To reconstitute, the lyophilized powder is added to sterile water or other suitable carrier. The exact amount will vary depending on the compound selected. Such amounts can be empirically determined.

[0222] Topical administration Topical compositions typically comprise an active substance, a pharmaceutically acceptable carrier, and optionally one or more additional components, such as, for example, to help form a desired delivery vehicle for the active substance.Topical mixtures are prepared as described for local and systemic administration.The resulting mixtures can be solutions, suspensions, emulsions, etc., and are formulated as creams, gels, jelly, waxes, ointments, emulsions, solutions, elixirs, lotions, suspensions, tinctures, pastes, foams, aerosols, irrigation, sprays, suppositories, bandages, skin patches, and / or combinations thereof, or any other formulation suitable for topical administration.

[0223] Carrier The active agent can be combined with a pharmaceutically acceptable carrier or diluent to form a pharmaceutical composition or formulation. In such pharmaceutical formulations, the active agent or therapeutic composition can be combined with a "carrier" that is physiologically compatible with the skin or mucosal tissue of humans or animals to which it is topically administered. Typically, the carrier is substantially inert, except for its inherent surfactant properties, which can aid in the formation of a solution or suspension of the active ingredient. The composition may contain other physiologically active ingredients that do not interfere with the efficacy of the active agent in the composition. In some embodiments, the carrier can be a liquid or gel-based material for use in liquid or gel formulations. The particular formulation will depend, in part, on the desired route or mode of administration.

[0224] Suitable carrier materials include any carrier or vehicle commonly used as a base for solutions, dispersions, emulsions, gels, creams, ointments, lotions, pastes, or foams for topical administration. Examples include emulsifiers, inert carriers including hydrocarbon bases, emulsifying bases, non-toxic solvents, or water-soluble bases.

[0225] Many suitable liquid- or gel-based carriers are known in the art. The carrier should be capable of dissolving or dispersing the active agent at effective levels, optionally with the aid of a non-toxic surfactant. Examples include water, saline, alcohol (e.g., methanol, ethanol, propanol, or butanol), glycerol, glycols (e.g., ethylene glycol, propylene glycol, or ethoxydiglycol), polyethylene glycols (e.g., MW 400-20,000), water-alcohol / glycol blends, and the like. Suitable carriers and diluents for certain embodiments include, for example, water, saline, isotonic saline (e.g., phosphate-buffered saline), aqueous dextrose, glycerol, ethoxydiglycol, dimethyl sulfoxide (DMSO), and the like, or combinations thereof.

[0226] Suitable carriers further include aqueous and oily carriers, such as white petrolatum, isopropyl myristate, lanolin or lanolin alcohol, mineral oil, aromatic or essential oils, nasturtium oil extract, sorbitan monooleate, cetostearyl alcohol (together or in various combinations), and detergents (e.g., polysorbates (Tween), e.g., polysorbate 20, 40, 60, or 80; polyoxyl stearate; or sodium lauryl sulfate), etc. One or more carrier materials can be mixed with water to form lotions, gels, creams, semi-solid compositions, etc. Other suitable carriers include water-in-oil or oil-in-water emulsions, as well as mixtures of emulsifiers and emollients with solvents (e.g., sucrose stearate, sucrose cocoate, sucrose distearate, mineral oil, propylene glycol, 2-ethyl-1,3-hexanediol, polyoxypropylene-15-stearyl ether, water, or combinations thereof). For example, emulsions containing water, glycerol stearate, glycerin, mineral oil, synthetic spermaceti, cetyl alcohol, or combinations thereof can be used. Preservatives, such as one or more of butylparaben, methylparaben, propylparaben, benzyl alcohol, and ethylenediaminetetraacetic acid salts, can also be included in the carrier. The composition of the carrier can be varied as long as it does not significantly inhibit the pharmacological activity of the active ingredients of the therapeutic composition.

[0227] Suitable pharmaceutically acceptable carriers include, but are not limited to, creams such as Cetaphil Moisturizing Cream (Galderma Laboratories, LP), QV Cream (Lision Hong), Sorbolene, etc. In some embodiments, the pharmaceutically acceptable carrier includes a lotion such as Alpha Keri Moisturizing Lotion (Mentholatum), DermaVeen Moisturizing Lotion (Dermatech Laboratories), QV Skin Lotion (Lision Hong), Cetaphil Moisturizing Lotion (Galderma Laboratories, LP).

[0228] Gelling and Thickening Agents The compositions described herein may contain one or more gelling agents to increase the viscosity of the composition. Examples of gelling agents and thickening agents include, but are not limited to, fatty acids, fatty acid salts and esters, fatty alcohols, synthetic polymers, modified cellulose, xanthan gum, or combinations thereof. Examples of suitable synthetic polymers include polyethylene glycol (PEG), polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), various Pluronics (poloxamers), or carbomers (e.g., Carbomer 940 or Carbomer 934). Examples of suitable modified celluloses include methylcellulose, carboxymethylcellulose (CMC), hydroxyethylcellulose (HEC), hydroxymethylcellulose (HMC), hydroxypropylcellulose (HPC), hydroxypropyl-methylcellulose (HPMC), or other cellulose-based gelling agents.

[0229] Various gelling agents are commercially available and can be obtained in many suitable molecular weights and ranges. For example, the molecular weight of the gelling agent can be about 1 kDa to about 1,000 kDa, about 10 kDa to about 1,000 kDa, about 100 kDa to about 1,000 kDa, or about 50 kDa to about 500 kDa. Examples of thickening agents include lanolin, hard paraffin, liquid paraffin, white petrolatum, soft yellow paraffin or soft white paraffin, white beeswax, yellow beeswax, propolis (propionium), cetostearyl alcohol, cetyl alcohol, dimethicone, emulsifying wax, microcrystalline wax, oleyl alcohol, and stearyl alcohol. The gelling or thickening agent may be present in the formulation at about 0.05% to about 20% by weight, typically about 0.1% to about 10% by weight, about 0.1% to about 5% by weight, about 0.5% to about 2% by weight, about 0.8% to about 2% by weight, or about 1 to 1.5% by weight. In one embodiment, the composition comprises 0.5% to 15% by weight, 1% to 10% by weight, or 2% to 10% by weight of one or more thickening or gelling agents.

[0230] One or more gelling or thickening agents can be included in a single formulation. Such agents can be used with a liquid carrier to form a spreadable gel, paste, ointment, soap, etc., which can be applied directly to the user's skin.

[0231] pH adjuster The topical formulation of the present invention may also contain a pH adjusting agent. In one embodiment, the pH adjusting agent is a base. Suitable pH adjusting bases include bicarbonates, carbonates, and hydroxides, such as alkali metal or alkaline earth metal hydroxides, and transition metal hydroxides. In another embodiment, the pH adjusting agent is an acid, an acid salt, or a mixture thereof. In a further embodiment, the pH adjusting agent is a buffering agent. Suitable buffering agents include citric acid / citric acid buffer, acetic acid / acetic acid buffer, phosphoric acid / phosphate buffer, formic acid / formic acid buffer, propionic acid / propionic acid buffer, lactic acid / lactic acid buffer, carbonate / carbonate buffer, ammonium / ammonia buffer, etc.

[0232] Solutions and Dispersions The solution of the active substance or its salt can be prepared in water, optionally mixed with a non-toxic surfactant.Dispersions can be prepared in glycerol, liquid polyethylene glycol, triacetin, or pharmaceutically acceptable oils, or their mixtures.Under normal conditions of storage and use, the preparations can contain preservatives to prevent the growth of microorganisms.

[0233] Pharmaceutical dosage forms include sterile aqueous solutions or dispersions containing the active ingredient suitable for the extemporaneous preparation of sterile solutions or dispersions, optionally encapsulated in liposomes. The final dosage form must be fluid and stable under the conditions of manufacture and storage. The liquid carrier or vehicle can be a solvent or liquid dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, liquid polyethylene glycol, etc.), vegetable oils, non-toxic glyceryl esters, and suitable mixtures thereof. The proper fluidity of the composition can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions, or by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents (e.g., parabens, chlorobutanol, phenol, sorbic acid, thiomersal, etc.). In many cases, it is preferable to include isotonic agents, such as sugars, buffers, or sodium chloride. Absorption of the composition can be prolonged by agents delaying absorption, such as aluminum monostearate and / or gelatin.

[0234] Solution can be prepared by incorporating desired amount of active substance in suitable solvent or oil together with various other components listed herein, and then optionally sterilize by filtration as needed.When powder is used to prepare solution, preparation method can include vacuum drying and freeze-drying technology, thereby obtaining powder of active substance and any additional desired components present in the prepared solution.

[0235] gel Gels are clear, sticky, jelly-like semi-solids or solids prepared from high molecular weight polymers in an aqueous or alcoholic base. Alcoholic gels often dry and cool. Non-alcoholic gels are more lubricious. Gels or jellies can be produced using suitable gelling agents, including but not limited to gelatin, tragacanth, carbomer, or cellulose derivatives, and may contain glycerol as a humectant, emollient, and / or preservative. In some embodiments, the gel formulation contains the same or similar ingredients as the solution or dispersion plus the gelling agent.

[0236] The gel may include a non-ionic copolymer gelling agent. In one embodiment, the gelling agent is a non-ionic polyoxyethylene-polyoxypropylene copolymer gel, e.g., a Pluronic gel such as Pluronic F-127 (BASF Corp.), providing a Pluronic gel-based formulation. This gel may be advantageous because it is liquid at low temperatures but rapidly hardens at physiological temperatures, confining drug release to or near the application site. Other formulations may include carboxymethylcellulose (CMC)-based formulations, hydroxymethylcellulose (HMC)-based formulations, hydroxypropylcellulose (HPC)-based formulations, or hydroxypropylmethylcellulose (HPMC)-based formulations.

[0237] cream Creams are viscous liquids or semisolid emulsions of the oil-in-water or water-in-oil type. Cream bases are water-washable and contain an oil phase, an emulsifier, and an aqueous phase. Water-in-oil creams can be formulated by using a suitable emulsifier with similar properties to, but not limited to, a fatty alcohol such as cetyl alcohol or cetostearyl alcohol and an emulsifying wax. Oil-in-water creams can be formulated using an emulsifier such as cetomacrogol emulsifying wax. Suitable properties include the ability to modify the viscosity of the emulsion and both physical and chemical stability over a wide range of pH. Water-soluble or water-miscible cream bases can contain a preservative system and can be buffered to maintain an acceptable physiological pH.

[0238] The oil phase, also called the "internal" phase, is generally composed of petrolatum and a fatty alcohol such as cetyl alcohol or stearyl alcohol. The aqueous phase usually, although not necessarily, exceeds the oil phase in volume and generally contains a humectant (a substance that absorbs water or helps another substance retain water, such as glycerin, sorbitol, or urea).

[0239] The emulsifier in cream formulations is generally a nonionic, anionic, cationic, or amphoteric surfactant. Examples of emulsifiers include, but are not limited to, fatty alcohol polyoxyethylene ether (Peregal A-20), stearates such as polyoxyl stearates (Softener SG), glyceryl stearate and PEG-5 glyceryl stearate, PEG-2 glyceryl stearate, cetyl alcohol, dithranol, or a combination thereof. Oil phase components include, but are not limited to, dimethicone, dimethiconol, cyclomethicone, diisopropyl adipate, cetyl alcohol, stearyl alcohol, paraffin, petrolatum, almond oil, stearic acid, or a combination thereof. In certain embodiments, aqueous components include, but are not limited to, purified water, glycerol (glycerin), propylene glycol, ethylparaben, humectants, or a combination thereof.

[0240] ointment Ointments are semi-solid preparations containing active substances incorporated into a fatty, waxy, or synthetic base. Ointments are typically based on petrolatum or other petroleum derivatives. The specific ointment base used will provide suitable drug delivery and other desired properties (e.g., emollient properties, etc.), as will be understood by those skilled in the art. Like other carriers or vehicles, ointment bases are typically inert, stable, non-irritating, and non-sensitizing.

[0241] Ointment bases can be broadly classified into four classes: oleaginous bases; emulsifiable bases; emulsion bases; and water-soluble bases. Oleaginous ointment bases can include, for example, vegetable oils, animal fats, and semi-solid hydrocarbons obtained from petroleum. Emulsifiable ointment bases, also known as absorbent ointment bases, contain little or no water and can include, for example, hydroxystearin sulfate, anhydrous lanolin, and hydrophilic petrolatum. Emulsion ointment bases are either water-in-oil (W / O) emulsions or oil-in-water (O / W) emulsions, and the oil component can include, for example, cetyl alcohol, glyceryl monostearate, lanolin, and stearic acid. Water-soluble ointment bases can be prepared from polyethylene glycols of various molecular weights.

[0242] lotion Lotion is a liquid or semi-liquid preparation in which solid particles containing active agents are present in a water or alcohol base.Lotion is usually a suspension of solids, and may include a liquid oily emulsion of the oil-in-water type.Lotion is often a desirable formulation because it is a more fluid composition that is easy to apply.In general, lotions are advantageous for finely dividing insoluble materials in lotions.Lotions typically contain suspending agents to obtain better dispersion, and compounds that are useful for localizing and retaining active agents in contact with the skin, such as methylcellulose, sodium carboxymethylcellulose, etc.

[0243] paste Paste is a semi-solid dosage form in which the active agent is suspended in a suitable base.Depending on the nature of the base, paste is divided into fatty paste or one made from a single-phase aqueous gel.The base in fatty paste is generally petrolatum, hydrophilic petrolatum, etc.Paste made from a single-phase aqueous gel generally incorporates carboxymethylcellulose, etc. as a base.

[0244] Form Foam preparations can be formulated using inert propellants to be delivered from pressurized aerosol cans via suitable applicators.Suitable excipients for the formulation of foam base include but are not limited to propylene glycol, emulsifying wax, cetyl alcohol, and glyceryl stearate.Possible preservatives include methylparaben and propylparaben.

[0245] Therefore, the compositions described herein can be formulated for topical or transdermal administration in any desired form, including sustained-release or delayed-release preparations. The formulations can include known antioxidants (e.g., vitamin E); buffering agents; lubricants (e.g., synthetic or natural beeswax); sunscreens (e.g., para-aminobenzoic acid); and cosmetic agents (e.g., colorants, fragrances, essential oils, moisturizers, or drying agents).

[0246] Auxiliaries such as casein, gelatin, albumin, or sodium alginate may also be included in various formulations. Adjuvants such as fragrances and additional antibacterial agents can be added to optimize the properties for a given use. Examples of fragrances include ylang-ylang oil, lavender oil, powdered fragrance, jasmine, gardenia oil, or green tea oil. In addition, substances such as wetting agents or emulsifiers, stabilizers, or pH buffers may also be included. When an aqueous carrier is used, the composition is typically near neutral pH (± about 1 or about 2 pH units).

[0247] Further examples of dermatological ingredients and compositions for delivering active agents to the skin are known in the art, and such dermatological compositions can be used in place of other active agents or in combination with the active agents described herein.

[0248] The above compositions can be prepared using standard formulation techniques. For example, compositions containing an active substance or a salt of an active substance can be milled to reduce particle size. A second active substance can then be added with a small amount of a carrier, such as polysorbate 80 and / or ethoxydiglycol, to moisten the active substance. This mixture can then be incorporated into a desired amount of oil using the principle of geometric dilution until a smooth, uniform suspension is formed. This suspension can then be combined with other ingredients, such as fragrance, to provide a therapeutic composition. The suspension can also be combined with other ingredients to form various formulations, such as gels, jellies, creams, ointments, waxes, lotions, pastes, foams, or aerosols. The suspension, or gels, jellies, creams, ointments, waxes, lotions, or pastes, can also be incorporated into patches, such as occlusive patches, to further improve transdermal penetration.

[0249] nanoparticles Nanoparticles can be composed of materials such as polymers, lipids (e.g., liposomes), or dendrimers (e.g., PAMAM, PEI, PPI, or polylysine-containing dendrimers). For example, nanoemulsions containing nanoparticles containing active agents can be used to provide different physical and chemical properties. For example, nanoparticles can allow deeper penetration into the skin, delivering treatment to more layers of the skin and beyond. Nanoparticles can be coated and / or embedded (encapsulated) in a composition. Nanoparticles can be formed using techniques such as electrospraying. Nanoparticles can be formed by milling particles containing an active agent to a desired size. Nanoparticles can include a surface coating.

[0250] The diameter of the nanoparticles can be from about 1 nm to about 100 nm, from about 100 nm to about 2500 nm, or from about 2500 nm to about 10,000 nm. The nanoparticles can be formed from polymers or surfactants. Polymers and surfactants include, but are not limited to, nonionic surfactants, anionic surfactants, such as carboxylates, such as alkyl carboxylate-fatty acid salts, or carboxylate fluorosurfactants; sulfates, such as alkyl sulfates (e.g., sodium lauryl sulfate) or alkyl ether sulfates (e.g., sodium laureth sulfate); sulfonates, such as docusate (e.g., sodium dioctyl sulfosuccinate) or alkyl benzene sulfonates; and phosphate esters, such as alkyl aryl ether phosphates or alkyl ether phosphates; cationic surfactants, including fatty amine salts and quaternary ammonium salts; zwitterionic surfactants, such as those with quaternary amine groups and sulfonic acid or carboxyl groups, natural polymers, and synthetic polymers, including, but not limited to, alginates, hyaluronates, chitosan, gelatin, celluloses, such as ethyl cellulose, PVP, PEG, or acylates. In one embodiment, the surfactant comprises polyoxyethylene, poloxamer, poloxamine, or polysorbate.

[0251] The compound or its pharmaceutically acceptable derivative can be formulated as an aerosol for topical application, such as by inhalation. These formulations for administration to the respiratory tract can be in the form of an aerosol or solution for a nebulizer, or as a very fine particle powder for inhalation, alone or in combination with an inert carrier such as lactose. In such cases, the particles of the formulation have a diameter of less than 50 microns in one embodiment, and less than 10 microns in one embodiment.

[0252] The compounds can be formulated for topical or local application, for example, topical application to the skin and mucous membranes, for example, in the eye or mouth, application in the form of gels, creams, and lotions, as well as application to the eye, or intracisternally or intrathecally. Topical administration is contemplated for dermal or transdermal delivery, and for administration to the eye or mucous membranes (including oral mucosa), or for inhalation therapy. Nasal, buccal, lingual, and sublingual formulations of the active compound alone or in combination with other pharmaceutically acceptable excipients listed herein can also be administered.

[0253] These solutions, particularly those intended for ophthalmic use, can be formulated as 0.01% to 10% (volume %) isotonic solutions at a pH of about 5 to 7 containing appropriate salts.

[0254] Compositions for other routes of administration Other routes of administration (eg, transdermal patches, including iontophoretic and electrophoretic devices, vaginal administration, and rectal administration) are also contemplated herein.

[0255] Transdermal patches, including iontophoretic and electrophoretic devices, are well known to those skilled in the art. For example, pharmaceutical dosage forms for rectal administration include rectal suppositories, capsules, and tablets for systemic administration. As used herein, a rectal suppository refers to a solid body for insertion into the rectum that melts or softens at body temperature and releases one or more pharmacologically or therapeutically active ingredients. Pharmaceutically acceptable substances utilized in rectal suppositories are a base or vehicle and an agent to raise the melting point. Examples of bases include cocoa butter (theobroma oil), glycerin-gelatin, carbowax (polyoxyethylene glycol), and appropriate mixtures of mono-, di-, and triglycerides of fatty acids. Combinations of various bases can be used. Agents to raise the melting point of the suppository include spermaceti and wax. Rectal suppositories can be prepared by either compression or molding. In one embodiment, the weight of a rectal suppository is approximately 2 to 3 gm.

[0256] Tablets and capsules for rectal administration are manufactured using the same pharmaceutically acceptable substance and by the same methods as for formulations for oral administration.

[0257] Other transdermal delivery methods known in the art or described herein include, for example, 1) the use of chemical penetration enhancers or skin enhancers; 2) liposome-mediated delivery; 3) electroporation; 5) sonophoresis (ultrasound); 6) mechanical (e.g., microporation) devices, and / or 7) pneumatically directed methods.

[0258] Suitable methods for transdermal delivery of drugs described herein can include methods directed to enhancing transport of substances through skin pores, for example, by increasing the transport rate through existing pores or by increasing the number of available skin pores by creating artificial pores.

[0259] Transdermal delivery can be achieved by the use of chemical enhancers or penetration enhancers, including, for example, pharmaceutically acceptable oils such as vegetable oils, nut oils, ethoxylated oils, PEG, linoleic acid, ethanol, methanol, and / or agents that degrease the stratum corneum. Suitable oils include meadowfoam oil, castor oil, jojoba oil, corn oil, sunflower oil, and sesame oil, all of which may be optionally ethoxylated. Furthermore, transdermal patches can be used for topical or transdermal delivery of the compositions described herein. Patches can also be adapted for delivery of dried or lyophilized forms of the compositions described herein. Other patch technologies can be used in conjunction with the compositions described herein, for example, in patch reservoirs. Transdermal delivery can also be achieved by liposome-mediated delivery methods (e.g., delivery facilitated by the application of a lipophilic membrane composition). Transdermal delivery systems can also be used in combination with a wide range of iontophoretic or electrotransport systems. The sonophoresis technique can be used to apply one ultrasonic frequency to the skin, or two or more different ultrasonic frequencies (e.g., one low and one high ultrasonic frequency) to the skin. As with the other techniques described above, this technique can be used in combination with other techniques, such as prior to topical application of the compositions described herein, including the application of a transdermal patch.

[0260] Another transdermal drug delivery technique that can be used in conjunction with the compositions described herein includes using a device that uses air pressure to inject a small stream of the composition through the top layer of the skin without using a needle. The air pressure gun can be the same or similar to the device used to vaccinate children. For diabetic patients who take insulin once a day, small disposable pen-like devices are also suitable.

[0261] Targeted formulations The compounds provided herein, or their pharmaceutically acceptable derivatives, can also be formulated to target specific tissues, receptors, or other areas of the body of the subject to be treated.Many of these targeting methods are well known to those skilled in the art.All of these targeting methods are contemplated herein for use in the compositions of the present invention.

[0262] Liposomal suspensions, including tissue-targeted liposomes such as tumor-targeted liposomes, may also be suitable as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art. For example, liposome formulations can be prepared. Briefly, egg phosphatidylcholine and brain phosphatidylserine (molar ratio 7:3) can be dried in a flask to form liposomes such as multilamellar vesicles (MLVs). A solution of a compound provided herein in phosphate-buffered saline (PBS) lacking divalent cations is added, and the flask is shaken until the lipid membrane is dispersed. The resulting vesicles are washed to remove unencapsulated compound, pelleted by centrifugation, and then resuspended in PBS.

[0263] Coadministration with other drugs According to another aspect of the present invention, it is contemplated that the compounds described herein can be administered to a subject in need in combination with a pharmaceutical agent that is considered by those skilled in the art to be the current standard treatment for the subject's condition.This combination can provide one or more benefits to the subject, such as reducing the dosage required to achieve the same benefit, achieving the desired therapeutic effect in a shorter time, etc.

[0264] The compound of the present invention can be administered together with other drugs as part of a treatment regimen.For example, it may be desirable to administer a combination of active compounds to treat a specific disease or condition.Therefore, within the scope of the present invention, two or more pharmaceutical compositions, at least one of which contains the compound of the present invention, can be combined in the form of a kit suitable for co-administration of the compositions.

[0265] In one embodiment of the method of the present invention, the compound of the present invention can be administered in combination with a second therapeutic agent. In one embodiment, the second therapeutic agent can be selected from one or more of the following categories: (i) Anticancer drugs, such as cisplatin, oxaliplatin, carboplatin, cyclophosphamide, nitrogen mustard, uracil mustard, bendamustine, melphalan, chlorambucil, chlormethine, busulfan, temozolamide, nitrosoureas, ifosamide, melphalan, pipobroman, triethylene-melamine, triethylenethiophoporamine, carmustine, lomustine, streptozocin, and dacarbazine, gemcitabine, 5-fluorouracil, Uracil, tegafur, raltitrexed, methotrexate, pemetrexed, leucovorin, cytosine arabinoside, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, pentostatine, hydroxyurea, trifluridine, trifluracil, adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin, mithramycin, vincristine, vinblastine, vindesine and vinorelbine, Taxol, Taxotere, eribulin, carfilzomib, bortezomib, etoposide, teniposide, amsacrine, topotecan, irinotecan, mitoxantrone, camptothecin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, ara-C, paclitaxel (Taxol™), nab-paclitaxel, docetaxel, mithramycin, deoxycoformycin, mitomycin-C, L-asparaginase, IFN-α, azacitidine, decitabine or its orally bioavailable combination products with cedazuridine, bortezomi ... bortezomib, bortezomib, bortezomib, bortezomib, bortezomib, bortezomib, bortezomib, bortezomib, bortezomib, bortezomib, bortezomib, bortezomib, bortezomib, bortezomib, bortezomib, bortezomib, bortezomib, bortezomib, bortezomib, bortezomib, bortezomib Rinostat, MS-275, panobinostat, romidepsin, valproic acid, mocetinostat (MGCD0103), pracinostat SB939, belinostat, panobinostat, irabectedin, tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene, iodoxifene, bicalutamide, flutamide, nilutamide, cyproterone acetate, goserelin, leuprorelin, buserelin, progestogens, megestrol acetate, anastrozole, letrozole, vorazole, exemestane, finasteride,Navelbene, CPT-II, anastrazole, letrazole, capecitabine, cyclophosphamide, ifosfamide, and droloxafine; and abiraterone, enzalutamide, lanreotide, dasatinib, bosutinib (SKI-606), trastuzumab, pertuzumab, panitumumab, cetuximab, gefitinib, erlotinib, 6-acrylamide- / -V-(3-chloro-4-fluorophenyl)-7-(3-morpholinopropoxy)-quinazolin-4-amine (CI 1033), afatinib, vandetanib, osimertinib, rociletinib, lapatinib, CTLA-4, 4-IBB, PD-I, imatinib, nilotinib (AMN107), sorafenib, tipifarnib and lonafarnib, vemurafenib, dabrafenib, trametinib, cobimetinib, ponatinib, palbociclib, Verolimus, ruxolitinib, ibrutinib, ceritinib, crizotinib, ectinib, cabozantirsib, vismodegib, soniiegib, BAL3833, regorafenib, vandetanib, vatalanib, sunitinib, axitinib, pazopanib, lenvatinib, talimogene laherparepvec laherparepvec, denosumab, obinulumab, biinatumumab, dinutuximab, idarucizumab, daratumumab, necitumumab, elotuzumab, olaratumumab, alemtuzumab, rituximab, ibritumomab tiuxetan tiuxetan), ofatumumab, peginterferon α-2b, aldesleukin, Gardasil, Cervarix, Oncophage, sipressel-T (Provenge) gp100, Ad.p53DC, pembrolizumab, atezolizumab, indoximod, MK-3475, nivolumab, MEDI-4736, RG-7446, ipilumumab, brentuximab vedotin, ilastuzumab emtansine, fludarivine (Fludara), cladribine, pentostatin, idelalisib, perifosine, birinapant, LCL161,AEG40730, SM-164, LBW242, ML101, AT-406, GDC-0917, AEG35156, HGS1029, bortezomib, ixazomib, carfilzomib, marizomib (NPI-0052), MLN9708, olaparib, rucaparib; anti-apoptotic, venetoclax, navitoclax, chimeric antigen receptor, (ii) anti-inflammatory agents, such as meloxicam, feoprofen, oxaprozin, salsalate, etoricoxib, tenoxicam, aspirin, nabumetone, flurbiprofen, mefenamic acid, phenylbutazone, lornoxicam, indomethacin, etodolac, diflunisal, ketoprofen, valdecoxib, tolfenamic acid, piroxicam, sulindac, tolmetin, ketorolac, loxoprofen, acetaminophen, bromfenac, diclofenac, ibuprofen, meclofenamate, nabumetone, naproxen, nepafenac, celecoxib, triamcinolone acetonide, colchicine, hydrocortisone, hydrocortisone acetate, methylprednisolone, aclomethasone dipropionate, emricasan, PXS-4728 (BI 1467335), namodenoson, roflumilast, crisaborole, PF-07038124, etrasimod, litorcitinib, ruxolitinib, pacritinib, delgocitinib, tofacitinib, eldulatinib, brepoxitinib, momelotinib, PF-07295324, PF-07259955, (iii) antihypertensives, such as hydrochlorothiazide, chlorthalidone, furosemide, spironolactone, triamterene, amiloride, benazepril, captopril, lisinopril, enalapril, ramipril, fosinopril, moexipril, perindopril, quinapril, trandolapril, losartan, candesartan, valsartan, telmisartan, clonidine, methyldopa, propranolol, Nolol, nadolol, timolol, pindolol, labetolol, metoprolol, atenolol, esmolol, betaxolol, carvedilol, prazosin, terazosin, doxazosin, phenoxybenzamine, phentolamine, verapamil, diltiazem, nifedipine, felodipine, amlodipine, nimodipine, diazoxide, minoxidil, pinacidil, nicorandil, hydralazine, sodium nitroprusside, (iv) antifibrotic agents, such as pirfenidone, nintedanib, cenicriviroc, selonsertib, lanifibranor, nimasimab, nitrazoxanide, NGM282, aparalenone, tipelukast, Actimmune, ponatinib, lenvatinib, dovitinib, lucitanib, danusertinib, brivatinib, erdafitinib, bexotegrast, PD173074, PD166 866, AZD4547, BGJ398, LY2874455, TAS-120, ARQ087, BLU9931, FGF401, BAY-1163877, ENMD-2076, IMCA1, FGF401, DEBIO1347, FIIN-2, G P-369, PRO-001, H3B-6527, BAY1187982, MFGR1877S, FP-1039, BLU554, PRN1371, S49076, SU6668, SU5416, TERN-501, MGL-3196, VK2809. (v) antiangiogenic agents, such as axitinib, bevacizumab, cabozantinib, everolimus, lenalidomide, lenvatinib, pazopanib, ramucirumab, regorafenib, vandetanib, vatalanib, sunitinib, aflibercept (ziv-aflibercept), thalidomide, pomalidomide, (vi) Antidiabetic agents, such as gliclazide, glimepiride, repaglinide, metformin, pioglitazone, rosiglitazone, acarbose, ringliptin, saxagliptin, sitagliptin, alogliptin, exenatide, liraglutide, dulaglutide, lixisenatide, semaglutide, canagliflozin, dapagliflozin, empagliflozin, and ertugliflozin. (vii) immunosuppressants, such as prednisone, budesonide, prednisolone, tofacitinib, cyclosporine, tacrolimus, sirolimus, everolimus, azathioprine, leflunomide, mycophenolate, abatacept, adalimumab, anakinra, certolizumab, etanercept, golimumab, infliximab, ixekizumab, natalizumab, rituximab, secukinumab, tocilizumab, ustekinumab, vedolizumab, basiliximab, and daclizumab; (viii) antibacterial agents, such as daptomycin, delafloxacin, telavancin, ceftaroline, fidaxomicin, amoxicillin, ampicillin, becampicillin, carbenicillin, cioxacillin, dicloxacillin, flucioxacillin, mezlocillin, nafcillin, oxacillin, penicillin G, penicillin V, piperacillin, pivampicillin, pivmecillinam, ticarcillin, cefacetrile, cefadroxil, cephalexin, cephalosporin, Glycine, cephalonium, cephaloridine, cephalothin, cephapirin, cefatrizine, cefazaflour, cefazedone, cefazolin, cephradine, cefroxadine, ceftezole, cefaclor, cefamandole, cefmetazole, cefonicid, cefotetan, cefoxitin, cefprozil, cefuroxime, cefuzonam, cefcapene, cefdaloxime, cefdinir, cefditoren, cefetamet, cefixime, cefmenoxime, cefodizime, cefotaxime, cefpimizole, cefpodoxime, cefteram, ceftibuten, cefti Ofur, ceftiolene, ceftizoxime, ceftriaxone, cefoperazone, ceftazidime, cefclidin, cefepime, cefluprenam, cefoselis, cefozopran, cefpirome, cefquinome, ceftobiprole, ceftaroline, cefaclomezine, cephaloram, cefaparol, cefcanel, cefedrolol, cefenpidone, cefetorzole, cefibitril, cefmatilen, cefmepidium, cefovecin, cefoxazole, cefrotil, cefsumidium, cefractatime, cef Tioxide, ceftazidime, avibactam, ceftolozane, tazobactam, aztreonam, vaborbactam, imipenem, doripenem, ertapenem, meropenem, azithromycin, erythromycin, clarithromycin, dirithromycin, roxithromycin, telithromycin, clindamycin, lincomycin, pristinamycin, quinupristin, dalfopristin, amikacin, gentamicin, kanamycin, neomycin, netilmicin, paromomycin, streptomycin, tobramycin, flumequine, nalidixic acid,Oxolinic acid, piromidic acid, pipemidic acid, losoxacin, ciprofloxacin, enoxacin, lomefloxacin, nadifloxacin, norfloxacin, ofloxacin, pefloxacin, rufloxacin, balofloxacin, gatifloxacin, grepafloxacin, levofloxacin, moxifloxacin, pazufloxacin, sparfloxacin, temafloxacin, tosufloxacin, besifloxacin, delafloxacin, clinafloxacin, gemifloxacin, prulifloxacin, sitafloxacin, trovafloxacin cin, sulfamethizole, sulfamethoxazole, sulfisoxazole, trimethoprim, demeclocycline, doxycycline, minocycline, oxytetracycline, tetracycline, tigecycline, chloramphenicol, metronidazole, tinidazole, nitrofurantoin, vancomycin, teicoplanin, telavancin, linezolid, cycloserine, rifampin, rifabutin, rifapentine, rifalazil, bacitracin, polymyxin B, viomycin, capreomycin, (ix) antifungal agents, such as amorolfine, butenafine, naftifine, terbinafine, bifonazole, butoconazole, clotrimazole, econazole, fenticonazole, ketoconazole, isoconazole, luliconazole, miconazole, omoconazole, oxiconazole, sultaconazole, sulconazole, tioconazole, terconazole, albaconazole, efinaconazole, fluconazole, isavuconazole, itraconazole, posaconazole, ravuconazole, voriconazole, abafungin, amphotericin B, nystatin, natamycin, trichomycin, anidulafungin, caspofungin, micafungin, tolnaftate, flucytosine, butenafine, griseofulvin, ciclopirox, selenium sulfide, tavaborole, (x) antipruritic agents, such as doxepin, tacrolimus, pimecrolimus, menthol, capsaicin, salicylic acid, pramoxine, lidocaine, polidocanol, N-palmitoylethanolamine, prednisone, prednisolone, mirtazapine, paroxetine, fluvoxamine, sertraline, naltrexone, methylnaltrexone, butorphanol, nalfurafine, gabapentin, pregablin, aprepitant, loratadine, desloratadine, cetirizine, levocetirizine, NGX-4010, TS-022, (xi) metabolic agents, for example, obeticholic acid, elafibranor, aramchol, seladelpal, MGL-3196, tropifexor, MSDC-0602K, BMS-986036, semaglutide, EDP-305, gemcabene, PF-05221304, PF-06865571, PF-06835919, LIK066, LMB763, vitamin E, acarbose, miglitol, pramlintide, alogliptan, linagliptan, saxagliptin, sitagliptin, and albiglutide , dulaglutide, exenatide, liraglutide, lixisenatide, insulin, nateglinide, repaglinide, metformin, canagliflozin, dapagliflozin, empagliflozin, chlorpropamide, glimepiride, glipizide, glyburide, tolazamide, tolbutamide, rosiglitazone, pioglitazone, atorvastatin, amlodipine, simvastatin, ezetimibe, lovastatin, sitagliptin, cholestyramine, colesevelam, colestipol, fenofibrate, gemcitabine Fibrozil, fenofibric acid, niacin, icosapent, mipomersen, lomitapide, evolocumab, alirocumab, fluvastatin, pravastatin, rosuvastatin, pitavastatin, simvastatin, cerivastatin, allopurinol, lesinurad, pegloticase, febuxostat, rasburicase, ivacaftor, velaglucerase alfa, imiglucerase, alglucosidase alfa, laronidase, cerliponase alfa alglucerase, idursulfase, taliglucerase alfa, agalsidase beta, sebelipase alfa, bestronidase alfa, galsulfase, elosulfase alfa, eliglustat, burosumab, migalastat, sapropterin, metreleptin, nitisinone, pegvalase, asfotase alfa, inotersen, miglustat, orlistat, sodium phenylbutyrate, glycerol phenylbutyrate.

[0266] In one embodiment, the compounds of the present invention can be administered in combination with other therapeutic treatments. For example, the compounds of the present invention can be administered in combination with radiation therapy or chemotherapy. In one embodiment, the compounds of the present invention can be administered in combination with one or more additional anti-tumor agents and / or radiation therapy for the treatment of cancer. In one embodiment, the compounds of the present invention can be administered in combination with wound covers and wound dressings. In one embodiment, the compounds of the present invention can be administered in combination with scar covers and scar dressings.

[0267] When two or more active ingredients are co-administered, the active ingredients can be administered simultaneously, sequentially, or separately. In one embodiment, a compound of the invention is co-administered simultaneously with a second therapeutic agent. In another embodiment, a compound of the invention and a second therapeutic agent are administered sequentially. In a further embodiment, a compound of the invention and a second therapeutic agent are administered separately.

[0268] The present invention will now be described in more detail, by way of example only, with reference to the following non-limiting examples, which are intended to provide illustrations of the invention and should not be construed as limiting the generality of the disclosure herein throughout. [Example]

[0269] For purposes of this specification, the following abbreviations have the indicated meanings: rt=room temperature t=time min=minutes ℃=Celsius temperature h=time RT=retention time aq = aqueous sat=saturated NaHMDS = sodium hexamethyldisilazide THF = tetrahydrofuran DMF = dimethylformamide TFA = trifluoroacetic acid DIBAL = diisobutylaluminum hydride MTBE = methyl tert-butyl ether DIPEA = N,N-diisopropylethylamine HATU = 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate LC-MS or LCMS = Liquid Chromatography Mass Spectrometry rpm = revolutions per minute NMR=nuclear magnetic resonance DMSO = dimethyl sulfoxide Boc = tert-butyloxycarbonyl HPLC = High-Performance Liquid Chromatography (including High-Pressure Liquid Chromatography) TLC = Thin-Layer Chromatography V = volume cone=concentration RP = reversed phase (chromatography) NP = normal phase (chromatography)

[0270] Experiment: General Methods All commercially available solvents and reagents were used as received. Where appropriate, reactions were carried out under an argon atmosphere. Reactions were monitored by either analytical thin-layer chromatography (TLC) or analytical liquid chromatography-mass spectrometry (LC-MS) using reversed-phase conditions and recorded on either a Shimadzu LC-MS 2020 instrument or an Agilent LC / MSD 1200 instrument. Purification of intermediates and final compounds was carried out using column chromatography or preparative HPLC, as needed. Normal-phase column chromatography was carried out under medium pressure on either silica gel or prepacked silica gel cartridges using a flash chromatography system (CombiFlash Rf200, Teledyne Isco Systems, USA). Reverse-phase column chromatography was carried out under low pressure on prepacked C18 cartridges using a flash chromatography system (Reveleris® X2). The eluent was monitored by UV light (λ = 254 / 280 nm). 1H-NMR spectra were recorded using either a Bruker 300 MHz NMR spectrometer or a Bruker Fourier 400 / 300 MHz spectrometer. Chemical shifts (δ) are reported as parts per million (ppm) relative to tetramethylsilane (TMS; internal standard). The following abbreviations are used for multiplicities: s = singlet; br s = broad singlet; d = doublet; t = triplet; q = quartet; m = multiplet; br m = broad multiplet. Low-resolution mass spectra (MS) were obtained as electrospray atmospheric pressure ionization (ES-API) mass spectra and recorded on either a Shimadzu LCMS 2020 instrument or an Agilent LC / MSD 1200 instrument using reversed-phase conditions. All animal experiments performed were in accordance with institutional guidelines and approval from the local ethical committee.

[0271] Unless otherwise indicated, conventional methods of mass spectrometry, NMR, HPLC, and protein chemistry and biochemical techniques are used. Unless otherwise stated, starting materials for chemical syntheses and biological applications are available from commercial sources.

[0272] The LC-MS method used for the analysis is described below: Method A: Shimadzu LC-MS 2020 instrument; LC-MS column: Restek Raptor ARC-18 5 μm (4.6 × 150 mm); mobile phase A: 0.1% formic acid in Milli-Q water, mobile phase B: 0.1% formic acid in methanol (gradient grade); flow rate: 1.5 mL / min; column oven temperature: 40 °C; run time: 9.4 min. Injection volume: 5 µL; binary gradient flow program: start from 95% water (0.1% formic acid) and 5% methanol (0.1% formic acid), go to 50% water (0.1% formic acid) and 50% methanol (0.1% formic acid) at 1.40 min, change to 5% water (0.1% formic acid) and 95% methanol (0.1% formic acid) at 6.00 min, hold the conditions until 7.40 min, and return to 95% water (0.1% formic acid) and 5% methanol (0.1% formic acid) from 7.41 min to 9.40 min.

[0273] Method B: Agilent LC / MSD 1200 series quadrupole mass spectrometer (column: ODS 2000 (50 × 4.6 mm, 5 μm) operated in ES (+) or (−) ionization mode; T = 30 °C; flow rate = 1.5 mL / min; detection wavelength: 214 nm).

[0274] Experimental details for the synthesis of intermediate 1 (Int-1) are disclosed in WO 2021 / 258159 and WO 2018 / 158140, the contents of which are incorporated by reference.

[0275] Example 1 Preparation of (E)-3-Fluoro-2-((phenylsulfonyl)methyl)prop-2-en-1-amine Hydrochloride (Gram Scale Process; Compound 1) [ka]

[0276] Procedure A: Preparation of tert-butyl (E)-(3-fluoro-2-((phenylsulfonyl)methyl)allyl)carbamate [ka]

[0277] To a stirred solution of Int-1 (2.50 g, 9.32 mmol) in DMF (12 mL) was added sodium benzenesulfinate (2.30 g, 14.0 mmol) in one portion. The resulting mixture was stirred at room temperature for 2 h. After this time, TLC analysis showed no detectable Int-1. Water (100 mL) was added, and the reaction mixture was stirred at room temperature for 5 min. The precipitated solid was collected by filtration, and the solid was washed with water (2 × 25 mL). The solid was dried in an oven at 60 °C overnight to give tert-butyl (E)-(3-fluoro-2-((phenylsulfonyl)methyl)-allyl)carbamate (2.80 g, 91%). 1H NMR (300MHz,CDCl3)δ ppm:7.96-7.85(m,2H),7.72(td,J=7.2,1.5Hz,1H),7.62(ddd,J=8.3,6.4,1.4Hz,2H),6.39(d,J=80. 5Hz, 1H), 5.03 (s, 1H), 3.98 (ddd, J = 6.4, 2.8, 1.4Hz, 2H), 3.75 (d, J = 2.9Hz, 2H), 1.46 (d, J = 1.2Hz, 9H).

[0278] Procedure B: Preparation of (E)-3-fluoro-2-((phenylsulfonyl)methyl)prop-2-en-1-amine hydrochloride (Compound 1) [ka]

[0279] To a stirred suspension of tert-butyl (E)-(3-fluoro-2-((phenylsulfonyl)methyl)allyl)carbamate (2.80 g, 8.50 mmol) in MeOH (10 mL) was added HCl (4.0 M in 1,4-dioxane; 5.0 mL). The resulting mixture was stirred at room temperature for 2 hours. TLC analysis after this time showed complete consumption of the starting material. The reaction mixture was concentrated in vacuo to remove volatiles. Ethyl acetate (25 mL) was added and the mixture was stirred for 5 minutes. The precipitated solid was collected by filtration and the solid was washed with ethyl acetate (25 mL). The solid was dried at 60° C. for 1 hour to give compound 1 (2.09 g, 93%). 1 H NMR(300MHz, methanol-d4)δ ppm:8.03-7.93(m,2H),7.87-7.76(m,1H),7.75-7.65(m,2H),6.78(d,J=80.5Hz,1H),4.10(d,J=2.8Hz,2H),3.86(d,J=2.5Hz,2H).MS:230.1[M+H] + ;RT=2.85 minutes (Method A).

[0280] Preparation of (E)-3-Fluoro-2-((phenylsulfonyl)methyl)prop-2-en-1-amine Hydrochloride (Kilogram Scale Process; Compound 1) [ka]

[0281] Procedure C: Preparation of tert-butyl (E)-(3-fluoro-2-((phenylsulfonyl)methyl)allyl)-carbamate [ka]

[0282] A reaction vessel was charged with Int-1 (8.4 kg, 1.0 equiv.), sodium benzenesulfinate (7.7 kg, 1.5 equiv.), and DMF (33.6 L, 4 V). The resulting mixture was stirred at room temperature for 20 h. Water (67 L, 8 V) was added to the reactor, and stirring was continued for 1 h. The solid was collected by filtration, and the filter cake was washed with water (25.2 L, 3 V). The crude solid was dried under vacuum to give the crude product (8.9 kg, 95.8% purity; residual Int-1: 82 ppm). The crude product was dissolved in ethyl acetate (3 V) at 60 °C. To this was added n-heptane (30 V) dropwise. The mixture was then cooled to 20° C., and the resulting solid was collected by filtration and dried under vacuum to give tert-butyl (E)-(3-fluoro-2-((phenylsulfonyl)methyl)-allyl)carbamate (8.6 kg, residue Int-1: 32 ppm).

[0283] Procedure D: Preparation of (E)-3-fluoro-2-((phenylsulfonyl)methyl)prop-2-en-1-amine hydrochloride (Compound 1) [ka]

[0284] To a reaction vessel containing tert-butyl (E)-(3-fluoro-2-((phenylsulfonyl)methyl)-allyl)carbamate (8.6 kg, 1.0 equiv.) and ethyl acetate (51.6 L, 6 V) under nitrogen was added HCl (4 M in ethyl acetate; 25.8 L, 3 V) dropwise. The resulting mixture was stirred at 25-30°C for 16 h. The solid was collected by filtration, and the filter cake was washed with ethyl acetate (25.8 L, 3.0 V). The solid was then dried under vacuum at 45-50°C to give compound 1 (6.3 kg).

[0285] Preparation of (E)-3-((2-(aminomethyl)-3-fluoroallyl)sulfonyl)benzonitrile hydrochloride (compound 2) [ka]

[0286] Procedure E: Preparation of sodium 3-cyanobenzenesulfinate [ka]

[0287] To a stirred solution of sodium sulfite (945 mg, 7.50 mmol) and sodium carbonate (1.06 g, 10.0 mmol) in water (10 mL) was added 3-cyanobenzenesulfonyl chloride (1.01 g, 5.00 mmol) in portions over 5 min at room temperature. The mixture was stirred at room temperature for 4 h. The mixture was concentrated to approximately 5 mL. The reaction mixture was directly purified by RP chromatography (Reveleris® C18 40 g; eluent: Solvent A = (12 mM aqueous HCl); Solvent B = methanol; 100% A for 4.8 min, followed by a gradient of 0 to 50% B for 4.2 min. Final hold at 50% B for 1 min; flow rate 32 mL / min). The desired fractions were pooled, and saturated aqueous NaHCO3 (2.0 mL) was added. The mixture was concentrated in vacuo to give crude sodium 3-cyanobenzenesulfinate (700 mg, 74%), which was used further as is. 1 H NMR (300MHz, deuterium oxide) δ7.88(td,J=1.7,0.6Hz,1H),7.81(dt,J=7.8,1.4Hz,1H),7.78(td,J=7.7,1.5Hz,1H),7.58(td,J=7.8,0.6Hz,1H).

[0288] Procedure F: Preparation of tert-butyl (E)-(2-(((3-cyanophenyl)sulfonyl)methyl)-3-fluoroallyl)carbamate [ka]

[0289] To a stirred mixture of crude sodium 3-cyanobenzenesulfinate (284 mg, ca. 1.5 mmol) in DMF (2.5 mL) was added Int-1 (268 mg, 1.00 mmol). The resulting mixture was stirred at room temperature for 1 h. TLC analysis after this time showed that Int-1 was completely consumed. Water (10 mL) was added and the product was extracted with ethyl acetate (3 x 10 mL). The combined organics were washed with brine, dried over Na2SO4, and then concentrated in vacuo. The crude material was purified by NP chromatography (CombiFlash® Redisep column 12 g silica; eluent: Solvent A = cyclohexane; Solvent B = ethyl acetate; 10% B for 1 min, followed by a gradient from 10% to 60% B for 8 min. Final hold at 60% B for 2 min. Flow rate 30 mL / min) to give tert-butyl (E)-(2-(((3-cyanophenyl)sulfonyl)methyl)-3-fluoroallyl)carbamate (95 mg, 27%). 1 H NMR(300MHz,CDCl3)δ 8.24(d,J=1.8Hz,1H),8.16(dt,J=7.9,1.5Hz,1H),7.99(dt,J=7.8,1.4Hz,1H),7.82-7.73(m,1 H),6.49(d,J=80.2Hz,1H),4.97(s,1H),4.04-3.91(m,2H),3.81(d,J=2.7Hz,2H),1.46(s,9H).

[0290] Procedure G: Preparation of (E)-3-((2-(aminomethyl)-3-fluoroallyl)sulfonyl)benzonitrile hydrochloride (compound 2) [ka]

[0291] To a suspension of tert-butyl (E)-(2-(((3-cyanophenyl)sulfonyl)methyl)-3-fluoroallyl)carbamate (94.0 mg, 0.27 mmol) in 1,4-dioxane (2.0 mL) at room temperature was added HCl (4.0 M in 1,4-dioxane; 2.0 mL, 8.0 mmol). The resulting mixture was stirred at room temperature for 2 hours. TLC analysis after this time indicated complete consumption of the starting material. The reaction mixture was concentrated in vacuo. Ethyl acetate (5 mL) was added and the mixture was stirred for 5 minutes. The precipitated product was then collected by filtration, and the filter "cake" was washed with ethyl acetate (5 mL). The product was dried at 60° C. for 1 hour to give compound 2 (55.0 mg, 71%). 1 H NMR(300MHz,DMSO-d6)δ 8.41(brs,4H),8.29(d,J=7.8Hz,1H),8.22(d,J=8.0Hz,1H),7.91(t,J=7.9Hz ,1H),6.81(d,J=81.2Hz,1H),4.66-4.31(m,2H),3.70-3.48(m,2H).LC-MS:m / z 255[M+H] + ;RT=2.45 minutes (Method A).

[0292] Procedure H: Preparation of tert-butyl (E)-(3-fluoro-2-(((3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)sulfonyl)methyl)allyl)carbamate [ka]

[0293] A stirred solution of tert-butyl (E)-(2-(((3-bromophenyl)sulfonyl)methyl)-3-fluoroallyl)carbamate (0.41 g, 1.0 mmol), bis(pinacolato)diboron (0.28 g, 1.1 mmol), and potassium acetate (0.29 g, 3.00 mmol) in 1,4-dioxane (10.0 mL) was purged with nitrogen for 5 minutes. 1,1′-bis(diphenylphosphino)ferrocene-palladium(II) dichloride dichloromethane complex (41 mg, 0.05 mmol) was then added to the solution, and the resulting mixture was heated at 80° C. overnight. The reaction mixture was then cooled to room temperature and diluted with water (25 mL). The product was extracted with ethyl acetate (3×25 mL). The combined organics were washed with brine, dried over NaSO, and then concentrated in vacuo to give crude tert-butyl (E)-(3-fluoro-2-(((3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)sulfonyl)methyl)-allyl)carbamate (0.46 g, 99%). The crude material was carried on immediately to the next step without purification.

[0294] Procedure I: Preparation of tert-butyl (E)-(3-fluoro-2-(((3-(thiazol-2-yl)phenyl)sulfonyl)methyl)allyl)-carbamate [ka]

[0295] A solution of 2-bromothiazole (164 mg, 1.00 mmol), tert-butyl (E)-(3-fluoro-2-(((3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)sulfonyl)methyl)allyl)carbamate (228 mg, 0.50 mmol), and tribasic potassium phosphate (0.46 g, 2.00 mmol) in 1,4-dioxane (8.0 mL) and water (2.0 mL) was degassed by bubbling argon through it for 5 minutes. Tetrakis(triphenylphosphine)palladium(0) (58 mg, 0.05 mmol) was then added under argon, and the reaction mixture was heated at 90° C. for 5 hours. TLC analysis after this time indicated the reaction was complete. The reaction mixture was cooled to room temperature, diluted with ethyl acetate (50 mL), and then filtered through a plug of Celite™. The filtrate was then concentrated in vacuo. The crude material was purified by NP chromatography (CombiFlash® Redisep column 12 g silica; eluent: solvent A = cyclohexane; solvent B = ethyl acetate; gradient 0% to 40% B over 10 min. Final hold at 40% B for 5 min. Flow rate 30 mL / min) to give tert-butyl (E)-(3-fluoro-2-(((3-(thiazol-2-yl)phenyl)sulfonyl)methyl)allyl)carbamate (120 mg, 51%). 1 H NMR(300MHz,CDCl3)δ 8.48(t,J=1.8Hz,1H),8.28(ddd,J=7.8,1.8,1.1Hz,1H),7.99-7.90(m,2H),7.69(t,J=7.8Hz,1H),7.46(d,J=3. 3Hz,1H), 6.46(d,J=80.5Hz,1H),5.12(s,1H),4.00(ddd,J=6.4,2.7,1.3Hz,2H),3.85-3.76(m,2H),1.44(s,9H).

[0296] Procedure J: Preparation of tert-butyl (E)-(3-fluoro-2-(((3-(2-methylthiazol-5-yl)phenyl)sulfonyl)-methyl)allyl)carbamate [ka]

[0297] A solution of 5-bromo-2-methylthiazole (178 mg, 1.00 mmol), tert-butyl (E)-(3-fluoro-2-(((3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)sulfonyl)methyl)allyl)carbamate (227 mg, 0.50 mmol), tribasic potassium phosphate (0.46 g, 2.0 mmol) in 1,4-dioxane (8 mL) and water (2 mL) was degassed by bubbling argon through it for 5 minutes. Tetrakis(triphenylphosphine)palladium(0) (58 mg, 0.05 mmol) was then added under argon, and the reaction mixture was heated at 90° C. for 5 hours. TLC analysis after this time showed traces of unreacted starting material. The reaction mixture was cooled to room temperature, diluted with ethyl acetate (50 mL), and then filtered through a plug of Celite®. The filtrate was concentrated in vacuo. The crude material was adsorbed onto silica gel and purified by NP chromatography (CombiFlash® Redisep column 12 g silica; eluent: Solvent A = cyclohexane; Solvent B = ethyl acetate; 10% B for 1 min, followed by a gradient of 10% to 40% B for 10 min. Final hold at 40% B for 7 min. Flow rate 30 mL / min). LCMS analysis indicated approximately 10% of an unidentified impurity. Therefore, this material was further purified by RP chromatography (Reveleris® C18 24 g; eluent: Solvent A = 12 mM aqueous HCl; Solvent B = methanol; 20% B for 3 min, followed by a gradient of 20 to 80% B for 15 min. Flow rate 32 mL / min) to give tert-butyl (E)-(3-fluoro-2-(((3-(2-methylthiazol-5-yl)phenyl)sulfonyl)methyl)allyl)-carbamate (75 mg, 31%). 1 H NMR(300MHz,CDCl3)δ 8.03(d,J=1.9Hz,1H),7.92(s,1H),7.82(dd,J=7.9,1.8Hz,2H),7.66-7.59(m,1H),6.47(d,J= 80.5Hz, 1H), 5.05 (s, 1H), 4.03-3.93 (m, 2H), 3.79 (d, J=2.7Hz, 2H), 2.78 (s, 3H), 1.45 (s, 9H).

[0298] Procedure K: Preparation of tert-butyl (E)-(3-fluoro-2-(((3-(3-methyl-1H-pyrazol-1-yl)phenyl)sulfonyl)methyl)allyl)carbamate [ka]

[0299] A mixture of 3-methyl-1H-pyrazole (123 mg, 1.50 mmol), cupric acetate (272 mg, 1.50 mmol), pyridine (0.16 mL, 2.00 mmol) and tert-butyl (E)-(3-fluoro-2-(((3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)sulfonyl)methyl)allyl)carbamate (455 mg, 1.00 mmol) in acetonitrile (10 mL) was stirred at room temperature overnight. The reaction mixture was concentrated in vacuo and the crude material was purified by NP chromatography (CombiFlash® Redisep column 24 g silica; eluent: Solvent A = cyclohexane; Solvent B = ethyl acetate; 10% B in 1 min, followed by a gradient of 10% to 50% B in 9.5 min, flow rate 35 mL / min) to give tert-butyl (E)-(3-fluoro-2-(((3-(3-methyl-1H-pyrazol-1-yl)phenyl)sulfonyl)methyl)allyl)carbamate (25 mg, 6%).

[0300] Procedure L: Preparation of tert-butyl (E)-(3-fluoro-2-(((3-(phenylsulfonyl)phenyl)sulfonyl)methyl)-allyl)carbamate [ka]

[0301] A stirred mixture of tert-butyl (E)-(2-(((3-bromophenyl)sulfonyl)methyl)-3-fluoroallyl)carbamate (500 mg, 1.22 mmol), sodium carbonate (130 mg, 1.22 mmol), sodium benzenesulfinate (402 mg, 2.45 mmol), N,N'-dimethylethylenediamine (3.0 mg, 34 pmol), and cupric acetate (3.0 mg, 17 μmol) in DMSO (2.0 mL) was purged with argon. The mixture was then heated at 100°C for 4 h. LCMS analysis after this time showed approximately 20% of the desired product. The reaction mixture was cooled to room temperature and then diluted with water (20 mL). The product was extracted with ethyl acetate (3 x 20 mL). The combined organic extracts were washed with brine, dried over Na2SO4, and then concentrated in vacuo. The crude material was purified by NP chromatography (CombiFlash® Redisep column 12 g silica; eluent: Solvent A = cyclohexane; Solvent B = ethyl acetate; 20% B for 1 min, followed by a gradient of 20% to 40% B for 3.5 min, hold at 40% B for 1.5 min, and finally a gradient of 40% to 60% B for 4 min. Flow rate 30 mL / min) to give tert-butyl (E)-(3-fluoro-2-((3-(phenylsulfonyl)phenyl)sulfonyl)-methyl)allyl)carbamate (80 mg, 14%). 1 H NMR(300MHz,CDCl3)δ 8.48-8.45(m,1H),8.27(ddd,J=7.9,1.8,1.1Hz,1H),8.11(ddd,J=7.8,1.8,1.1Hz,1H),8.03-7.95(m,2H),7.78(td,J=7. 9,0.5Hz,1H),7.69-7.53(m,3H),6.40(d,J=80.2Hz,1H),4.98(s,1H),3.98-3.91(m,2H),3.81-3.76(m,2H),1.46(s,9H).

[0302] [Table 3-1]

[0303] [Table 3-2]

[0304] [Table 3-3]

[0305] [Table 3-4]

[0306] [Table 3-5]

[0307] [Table 3-6]

[0308] Preparation of (E)-2-(((3-chlorophenyl)sulfonyl)methyl)-3-fluoroprop-2-en-1-amine hydrochloride (compound 19) [ka]

[0309] Procedure M: Preparation of tert-butyl (E)-(2-(((3-chlorophenyl)thio)methyl)-3-fluoroallyl)carbamate [ka]

[0310] To a stirred mixture of cesium carbonate (456 mg, 1.40 mmol) and 3-chlorothiophenol (217 mg, 1.50 mmol) in DMF (2.5 mL) was added Int-1 (268 mg, 1.00 mmol). The resulting mixture was stirred at room temperature for 2 h. TLC analysis after this time showed that Int-1 was completely consumed. Water (10 mL) was added, and the product was extracted with ethyl acetate (3 × 10 mL). The combined organic extracts were washed with brine, dried over NaSO, and concentrated to give tert-butyl (E)-(2-(((3-chlorophenyl)thio)methyl)-3-fluoroallyl)carbamate (350 mg). This material was used directly in the next step.

[0311] Procedure N: Preparation of tert-butyl (E)-(2-(((3-chlorophenyl)sulfonyl)methyl)-3-fluoroallyl)-carbamate [ka]

[0312] To a stirred solution of tert-butyl E-(2-(((3-chlorophenyl)thio)methyl)-3-fluoroallyl)carbamate (350 mg) in methanol (5 mL) was added hydrogen peroxide (30% w / v, 4.0 mL), followed by sodium tungstate (100 mg). The mixture was stirred at room temperature for 2 hours. LCMS analysis after this time showed that the starting material had been completely consumed. Water (20 mL) was added and the product was extracted with ethyl acetate (3×10 mL). The combined organic extracts were washed with brine, dried over Na2SO4, and then concentrated in vacuo. The residue was purified on silica gel (CombiFlash® Redisep column 12 g silica; eluent: Solvent A = cyclohexane; Solvent B = ethyl acetate; 100% cyclohexane in 1 min, followed by a gradient of 0% to 40% ethyl acetate in cyclohexane in 7 min, flow rate 30 mL / min) to give tert-butyl (E)-(2-(((3-chlorophenyl)sulfonyl)methyl)-3-fluoroallyl)-carbamate (0.31 g, 91%). 1H NMR(300MHz,CDCl3)57.92(t,J=1.9Hz,1H),7.81(dt,J=7.8,1.4Hz,1H),7.69(ddd,J=8.1,2.1,1.1Hz,1H),7.56(t,J =7.9Hz,1H),6.46(d,J=80.4Hz,1H),5.02(s,1H),4.00(ddd,J=6.4,2.8,1.3Hz,2H),3.81-3.71(m,2H),1.47(s,9H).

[0313] Procedure O: Preparation of (E)-2-(((3-chlorophenyl)sulfonyl)methyl)-3-fluoroprop-2-en-1-amine hydrochloride (compound 19) [ka]

[0314] To a suspension of tert-butyl (E)-(2-(((3-chlorophenyl)sulfonyl)methyl)-3-fluoroallyl)-carbamate (0.33 g, 0.90 mmol) in methanol (5.0 mL) was added HCl (2.0 M in diethyl ether; 5.0 mL, 10.0 mmol). The resulting mixture was stirred at room temperature for 2 hours. TLC analysis after this time indicated that the reaction was complete. The reaction mixture was concentrated in vacuo. Ethyl acetate (5 mL) was added and the mixture was stirred for 5 minutes. The precipitated product was collected by filtration and the filter cake was washed with ethyl acetate (5 mL). The solid was dried at 60° C. for 1 hour to give (E)-2-(((3-chlorophenyl)sulfonyl)methyl)-3-fluoroprop-2-en-1-amine hydrochloride (190 mg, 70%). 1 H NMR(300MHz,DMSO-d6)δ 8.35(s,3H),7.98(t,J=1.9Hz,1H),7.90(ddt,J=8.3,4.2,1.3Hz,2H),7.73(t,J=7.9H z,1H),6.85(d,J=81.3Hz,1H),4.44(d,J=2.6Hz,2H),3.62(d,J=2.4Hz,2H).LC-MS:m / z 264[M+H] + ;RT=2.83 minutes (Method A).

[0315] Procedure P: Preparation of 2-(3-nitrophenyl)-2H-1,2,3-triazole [ka]

[0316] A stirred mixture of 1-bromo-3-nitrobenzene (1.01 g, 5.00 mmol), tribasic potassium phosphate (2.12 g, 10.0 mmol), 2H-triazole (691 mg, 10.0 mmol), N,N'-dimethylethylenediamine (44 mg, 0.50 mmol), cuprous iodide (17 μL, 0.50 mmol), cuprous oxide (72 mg, 0.50 mmol), and N,N'-dimethylethylenediamine (55 μL, 0.50 mmol) in DMF (25 mL) was purged with argon. The mixture was heated to reflux for 6 h. TLC analysis after this time indicated the formation of the desired product and traces of unreacted starting material. The reaction mixture was cooled to room temperature and diluted with water (70 mL). The mixture was stirred at room temperature for 5 min, and the resulting solid was collected by filtration. The crude solid was purified on silica gel (CombiFlash® Redisep column 24 g silica; eluent: Solvent A = cyclohexane; Solvent B = ethyl acetate; 10% B for 1 min, followed by a gradient of 10% to 16% B for 1 min. hold. Then 16% B for 3 min. Then a gradient of 16% to 50% B for 1 min. Finally, a gradient of 50% to 60% B for 9 min. Flow rate 30 mL / min) to give 2-(3-nitrophenyl)triazole (350 mg, 37%). 1 H NMR(300MHz, CDCl3)δ 9.00(t,J=2.2Hz,1H),8.47(ddd,J=8.2,2.1,1.0Hz,1H),8.24(ddd,J=8.2,2.2,1.0Hz,1H),7.91(s,2H),7.71(t,J=8.2Hz,1H).

[0317] Procedure Q: Preparation of 3-(2H-1,2,3-triazol-2-yl)aniline [ka]

[0318] A mixture of 2-(3-nitrophenyl)triazole (350 mg, 1.84 mmol) and 10% (w / w) Pd / C (40 mg) in methanol (25 mL) was hydrogenated at 50 psi for 1 h. TLC analysis indicated the absence of starting material and low product. The Pd / C was removed by filtration, and the filtrate was concentrated in vacuo to give crude 3-(triazol-2-yl)aniline (300 mg, 1.87 mmol, quantitative yield). This material was used in the next step without purification.

[0319] Procedure R: Preparation of 3-(2H-1,2,3-triazol-2-yl)benzenethiol [ka]

[0320] A stirred solution of crude 3-(2H-1,2,3-triazol-2-yl)aniline (300 mg, 1.87 mmol) and concentrated aqueous HCl (0.47 mL, 5.62 mmol), water (5.0 mL), and THF (5.0 mL) was cooled to 0 °C. A solution of sodium nitrite (142 mg, 2.06 mmol) in water (2.0 mL) was added in four portions. The resulting mixture was stirred at 0 °C for 30 min. A solution of potassium ethyl xanthate (453 mg, 2.81 mmol) in water (2.0 mL) was then added, and the mixture was stirred at 0 °C for 20 min. The mixture was gradually warmed to 60 °C over 15 min. The mixture was then cooled to room temperature, after which aqueous NaOH (2.0 M; 3.0 mL) was added. Stirring was then continued at room temperature for 1 h. The reaction mixture was concentrated in vacuo to approximately 2 mL. This solution was used directly in the next step without isolation or purification.

[0321] Procedure S: Preparation of 1-(3-(methylsulfonyl)phenoxy)-2-nitrobenzene [ka]

[0322] A mixture of 3-(methylsulfonyl)phenol (344 mg, 2.00 mmol) and potassium carbonate (300 mg, 2.17 mmol) in DMF (3.0 mL) was stirred for 5 minutes. To this, 1-fluoro-2-nitro-benzene (282 mg, 2.00 mmol) was added in one portion, and the resulting mixture was heated at 100° C. for 2 hours. TLC analysis after this time indicated that the reaction was complete. The reaction mixture was cooled to room temperature and then diluted with water (30 mL). The product was extracted with ethyl acetate (3×25 mL). The combined organic extracts were washed with brine, dried over NaSO, and then concentrated in vacuo to give 1-(3-(methylsulfonyl)phenoxy)-2-nitrobenzene (530 mg, 90%). 1 H NMR(300MHz,CDCl3)δ 8.04(dd,J=8.1,1.7Hz,1H),7.75(ddd,J=7.8,1.7,1.0Hz,1H),7.69-7.52(m,3H),7.42-7.30(m,2H),7.16(dd,J=8.3,1.2Hz,1H).

[0323] Procedure T: Preparation of 2-(3-(methylsulfonyl)phenoxy)aniline [ka]

[0324] To a stirred solution of 1-(3-(methylsulfonyl)phenoxy)-2-nitrobenzene (530 mg, 1.81 mmol), THF (20 mL), and saturated aqueous NH4Cl (10 mL) was added zinc powder (1.00 g). The resulting suspension was stirred at room temperature for 30 minutes. LCMS analysis after this time indicated complete consumption of the starting material. Saturated aqueous NaHCO3 (10 mL) and ethyl acetate (20 mL) were added, and the mixture was stirred for 5 minutes. The organic layer was separated, washed with brine, dried over Na2SO4, and then concentrated in vacuo to provide 2-(3-(methylsulfonyl)phenoxy)aniline (450 mg, 95%). This material was used directly in the next step without further purification.

[0325] Preparation of (E)-3-((2-(aminomethyl)-3-fluoroallyl)sulfonyl)benzamide hydrochloride (compound 27) [ka]

[0326] Procedure U: Preparation of (E)-3-((2-(aminomethyl)-3-fluoroallyl)sulfonyl)benzamide hydrochloride (compound 27) [ka]

[0327] A mixture of (E)-3-((2-(((tert-butoxycarbonyl)amino)methyl)-3-fluoroallyl)sulfonyl)benzoic acid (150 mg, 0.40 mmol), ammonium chloride (43.0 mg, 0.80 mmol), and N,N-diisopropylethylamine (0.28 mL, 1.61 mmol) in DMF (2.0 mL) was stirred for 10 min. HATU (229 mg, 0.60 mmol) was added, and stirring was continued at room temperature for 1 h. LCMS analysis after this time indicated the reaction was complete. The reaction mixture was loaded directly onto a 40 g RP column and purified by chromatography (Reveleris® C18 40 g; eluent: Solvent A = 12 mM aqueous HCl; Solvent B = methanol; 5% B for 3 min, followed by a gradient of 5-65% B for 15 min. Flow rate 40 mL / min). Fractions containing the desired product were combined and concentrated in vacuo. During concentration, deprotection of -NHBoc occurred, yielding the hydrochloride salt. After complete removal of water under vacuum, acetonitrile (4 mL) was added, and the mixture was stirred for 5 minutes. The mixture containing the precipitated product was transferred to a vial, and the solid was spun down in a centrifuge (4000 rpm, 4 minutes). The liquid was carefully decanted, and the solid residue was dried at 60 °C to give compound 27 (65 mg, 52%). 1H NMR(300MHz,DMSO-d6)δ 8.44(d,J=1.7Hz,1H),8.35(d,J=12.5Hz,4H),8.29-8.20(m,1H),8.10-7.96(m,1H),7.79(t, LC-MS:m / z 273[M+H] + ;RT=2.82 minutes (Method A).

[0328] Preparation of (E)-4-((2-(aminomethyl)-3-fluoroallyl)sulfonyl)benzoic acid hydrochloride (compound 31) [ka]

[0329] Procedure V: Preparation of (E)-4-((2-(((tert-butoxycarbonyl)amino)methyl)-3-fluoroallyl)sulfonyl)benzoic acid [ka]

[0330] To a stirred solution of methyl (E)-4-((2-(((tert-butoxycarbonyl)amino)methyl)-3-fluoroallyl)sulfonyl)benzoate (340 mg, 0.88 mmol) in methanol (10 mL) was added aqueous NaOH (2.0 M; 2.50 mL, 5.00 mmol). The resulting mixture was stirred at room temperature for 1 h. LCMS analysis after this time showed complete hydrolysis of the methyl ester. The reaction mixture was then concentrated in vacuo. Water (10 mL) was added to dissolve the residue. The mixture was acidified with 2 M HCl, causing a white solid to precipitate. The solid product was collected by filtration, and the solid was washed with water and then dried at 60 °C to afford (E)-4-((2-(((tert-butoxycarbonyl)amino)methyl)-3-fluoroallyl)sulfonyl)benzoic acid (328 mg, 100%). This material was used directly in the next step without further purification.

[0331] Procedure W: Preparation of (E)-4-((2-(aminomethyl)-3-fluoroallyl)sulfonyl)benzoic acid hydrochloride (compound 31) [ka]

[0332] To a stirred suspension of (E)-4-((2-(((tert-butoxycarbonyl)amino)methyl)-3-fluoroallyl)sulfonyl)benzoic acid (328 mg, 0.88 mmol) in dichloromethane (5 mL) was added TFA (5.0 mL). The resulting mixture was stirred at room temperature for 1 hour. LCMS analysis after this time showed complete consumption of the starting material. The reaction mixture was then concentrated in vacuo. Ethyl acetate (10 mL) was added and the mixture was again concentrated in vacuo. This process was repeated two more times to ensure complete removal of residual TFA. The residue was then dissolved in water (2 mL). To this was added aqueous HCl (2.0 M, 2.0 mL), resulting in the formation of a white precipitate. The resulting suspension was stirred for 5 minutes. The solid was collected by filtration and washed with water (1 mL). The product was dried at 60° C. for 2 hours to give compound 31 (0.22 g, 81%). 1 H NMR(300MHz,DMSO-d6)δ 8.34(s,3H),8.20(d,J=8.4Hz,2H),8.04(d,J=8.4Hz,2H),6.82(d,J=81.2Hz,1H),4.42(d,J=2.6Hz,2H),3.60(d,J=2.3Hz,2H).LC-MS:m / z 274[M+H] + (Method A).

[0333] [Table 4-1]

[0334] [Table 4-2]

[0335] [Table 4-3]

[0336] [Table 4-4]

[0337] [Table 4-5]

[0338] Preparation of (E)-1-(3-((2-(aminomethyl)-3-fluoroallyl)sulfonyl)benzyl)azepan-2-one hydrochloride (compound 36) [ka]

[0339] Procedure X: Preparation of tert-butyl 6-((3-iodobenzyl)amino)hexanoate [ka]

[0340] A mixture of tert-butyl 5-bromopentanoate (300 mg, 1.27 mmol), (3-iodophenyl)methanamine hydrochloride (341 mg, 1.27 mmol), and potassium carbonate (525 mg, 3.80 mmol) in DMF (3.0 mL) was stirred at room temperature overnight. LCMS analysis indicated that the reaction was complete. Water (15 mL) was then added, and the product was extracted with ethyl acetate. The combined organic extracts were washed with brine, dried over Na2SO4, and then concentrated in vacuo. The crude product was purified by RP chromatography (Reveleris® C18 40 g; eluent: solvent A = 12 mM aqueous HCl; solvent B = methanol; 20% B for 3 min, followed by a gradient from 20 to 44% B for 5 min. Final hold at 44% B for 3.5 min.; flow rate 40 mL / min) to give tert-butyl 6-((3-iodobenzyl)amino)hexanoate (0.30 g, 59%).

[0341] Procedure Y: Preparation of 6-((3-iodobenzyl)amino)hexanoic acid [ka]

[0342] To a solution of tert-butyl 6-((3-iodobenzyl)amino)hexanoate (30 mg, 0.74 mmol) in CHCl (3.0 mL) was added TFA (3.0 mL). The resulting mixture was stirred at room temperature for 2 h. LCMS analysis after this time indicated the reaction was complete. The volatiles were then removed in vacuo. Ethyl acetate (5 mL) was added and the mixture was concentrated to remove residual TFA to provide crude 6-((3-iodobenzyl)amino)hexanoic acid (300 mg, quantitative yield). This material was used in the next step without further purification.

[0343] Procedure Z: Preparation of 1-(3-iodobenzyl)azepan-2-one [ka]

[0344] To a stirred solution of 6-((3-iodobenzyl)amino)hexanoic acid (300 mg, 0.74 mmol) in DMF (3.0 mL) was added N,N-diisopropylethylamine (0.65 mL, 3.72 mmol) and HATU (424 mg, 1.12 mmol). The resulting mixture was stirred at room temperature for 2 h. LCMS analysis after this time indicated the reaction was complete. The reaction mixture was diluted with water (25 mL). The product was extracted with ethyl acetate, and the combined organic extracts were washed with brine, dried over Na2SO4, and concentrated in vacuo. The crude product was purified by NP chromatography (CombiFlash® Redisep column 24 g silica; eluent: Solvent A = cyclohexane; Solvent B = ethyl acetate; 0% B for 1 min, followed by a gradient of 0% to 50% B for 13 min, then hold at 50% B for 3 min. Flow rate 30 mL / min) to give 1-(3-iodobenzyl)azepan-2-one (0.15 g, 60%). 1H NMR(300MHz,CDC13)δ 7.63-7.57(m,2H),7.24(dt,J=7.6,1.4Hz,1H),7.10-7.02(m,1H),4.53(s,2H),3.34-3.25(m, 2H), 2.61(tt,J=4.2,1.9Hz,2H),1.72(dt,J=7.8,2.7Hz,4H),1.52(qd,J=5.8,3.9,3.1Hz,2H).

[0345] Procedure AA: Preparation of tert-butyl (E)-(3-fluoro-2-(mercaptomethyl)allyl)carbamate [ka]

[0346] A stirred mixture of Int-1 (540 mg, 2.01 mmol) and thiourea (169 mg, 2.22 mmol) in methanol (10 mL) was heated to reflux for 1.5 h. LCMS analysis after this time indicated complete formation of the intermediate S-alkylisothiourea. Aqueous NaOH (2.0 M; 1.20 mL, 2.40 mmol) was then added, and the reaction mixture was heated to reflux for 1 h. LCMS analysis indicated the absence of the isothiourea intermediate and the formation of the desired product. The reaction mixture was concentrated in vacuo, and the residue was adsorbed onto silica gel. Normal-phase chromatography (CombiFlash® Redisep column 24 g silica; eluent: Solvent A = cyclohexane; Solvent B = ethyl acetate; 10% B for 1 min, followed by a gradient from 10% to 50% B for 7 min, then a 2 min hold at 50% B; flow rate 30 mL / min) afforded the impure title compound (170 mg, 19%). LC-MS analysis of the purified material showed some impurities and a purity of approximately 50%. This material was carried on to the next step without further purification.

[0347] Procedure AB: Preparation of tert-butyl (E)-(3-fluoro-2-(((3-((2-oxoazepan-1-yl)methyl)phenyl)thio)methyl)allyl)carbamate [ka]

[0348] A stirred mixture of 1-(3-iodobenzyl)azepan-2-one (150 mg, 0.46 mmol), tert-butyl (E)-(3-fluoro-2-(mercaptomethyl)allyl)carbamate (168 mg, 0.46 mmol), potassium hydroxide (45 mg, 0.68 mmol), and finely powdered cupric oxide (3.63 mg, 46 μmol) in DMSO (1.0 mL) was heated to 80° C. under nitrogen for 6 h. LCMS analysis after this time indicated the desired product had formed. The reaction mixture was cooled to room temperature and then diluted with water (10 mL). The product was extracted with ethyl acetate (3×10 mL). The combined organic layers were washed with brine, dried over NaSO, and then concentrated in vacuo. The crude material was purified by NP chromatography (CombiFlash® Redisep column 24 g silica; eluent: Solvent A = cyclohexane; Solvent B = ethyl acetate; 10% B for 1 min, followed by a gradient of 10% to 75% B over 12 min. Flow rate 30 mL / min) to give tert-butyl (E)-(3-fluoro-2-(((3-((2-oxoazepan-1-yl)methyl)phenyl)thio)methyl)allyl)carbamate (40 mg, 21%).

[0349] Procedure AC: Preparation of tert-butyl (E)-(3-fluoro-2-(((3-((2-oxoazepan-1-yl)methyl)phenyl)sulfonyl)methyl)allyl)carbamate [ka]

[0350] To a stirred solution of tert-butyl (E)-(3-fluoro-2-(((3-((2-oxoazepan-1-yl)methyl)phenyl)thio)-methyl)allyl)carbamate (40 mg, 0.09 mmol) in methanol (2.0 mL) was added hydrogen peroxide (30% w / v, 1.0 mL), followed by a solution of sodium tungstate dihydrate (31 mg, 0.09 mmol) in water (0.5 mL). The resulting mixture was then stirred at room temperature for 2 hours. LCMS analysis after this time showed the complete absence of starting material. Water (10 mL) was added and the product was extracted with ethyl acetate. The combined organic extracts were washed with brine, dried over Na2SO4, and concentrated in vacuo to give tert-butyl (E)-(3-fluoro-2-(((3-((2-oxoazepan-1-yl)methyl)phenyl)sulfonyl)methyl)allyl)carbamate (20 mg, 46%). This material was carried on to the final step without purification.

[0351] Procedure AD: Preparation of (E)-1-(3-((2-(aminomethyl)-3-fluoroallyl)sulfonyl)benzyl)azepan-2-one hydrochloride (compound 36) [ka]

[0352] (E)-(3-Fluoro-2-(((3-((2-oxoazepan-1-yl)methyl)phenyl)sulfonyl)methyl)allyl)carbamate (20 mg, 0.04 mmol) was added to a solution of HCl (4.0 M in dioxane; 1.0 mL). The resulting mixture was stirred at room temperature for 2 hours. LC-MS analysis after this time showed that the starting material was completely consumed. The reaction mixture was then concentrated in vacuo. Ethyl acetate (2 mL) was added, and the mixture was stirred for 5 minutes. The mixture containing the precipitated product was transferred to a vial, and the solid was spun down in a centrifuge (4000 rpm, 4 minutes). The liquid was carefully decanted, and the solid residue was dried at 60° C. for 1 hour to give compound 36 (2.5 mg, 15%). 1H NMR(300MHz, methanol-d4)δ 7.88(d,J=8.3Hz,2H),7.69(dt,J=14.9,7.5Hz,2H),6.77(d,J=80.4Hz,1H),4.71(s,2H),4.10(d,J=2.9Hz,2H ),3.85(d,J=2.3Hz,2H),3.54-3.44(m,2H),2.72-2.59(m,2H),1.77(d,J=10.9Hz,4H),1.58(s,2H).LC-MS:m / z 377[M+Na] + ;RT=3.00 minutes (Method A).

[0353] Preparation of (Z)-3-fluoro-2-((phenylsulfonyl)methyl)prop-2-en-1-amine hydrochloride (compound 37) [ka]

[0354] Compound 37 was prepared by a two-step process starting from the E / Z mixture of intermediate 1 (E / Z)Int-1) using chemistry similar to that described herein for the synthesis of compound 1. The final compound (37) was obtained in pure isomeric form by application of HPLC separation. The detailed synthesis of E / Z)Int-1 is disclosed in WO 2013 / 163675.

[0355] Compound 37: 1 H NMR(400MHz, methanol-d4)δ 7.96(dd,J=8.0,0.8Hz,2H),7.75(dt,J=7.2,0.8Hz,1H),7.65(dd,J=8.4,7.6 Hz,2H),7.06(d,J=79.6Hz,1H),4.22(d,J=2.0Hz,2H),3.76(d,J=2.8Hz,2H).

[0356] Example 2 Methods for determining the ability of Compound 1 of the present invention to inhibit LOX and LOXL1-4 from different sources LOX and LOXL protein family members can be obtained as recombinant active proteins from commercial sources, extracted from animal tissues such as bovine aorta, tendons, and pig skin, or prepared from cell cultures. The inhibitory effects of compounds of the present invention were tested against a given LOX or LOXL preparation using a method based on the detection of hydrogen peroxide by the Amplex Red oxidation assay [Zhou et al., 1997]. Assays were developed using either a 384- or 96-well format. Briefly, in a standard black, clear-bottom 384-well plate assay, 25 μL of a dilution of either the isozyme or ortholog in 1.2 M urea, 50 mM sodium borate buffer (pH 8.2) was added to each well in the presence of 1 μM mofegiline and 0.5 mM pargyline (to inhibit SSAO, MAO-B, and MAOA, respectively; not required if the enzymes are derived from recombinant or purified forms). Test compounds were dissolved in DMSO and tested in concentration-response curves (CRCs) with typically 11 data points in the micromolar or nanomolar range after 30 min of incubation at 37 °C. Then, 25 μL of reaction mixture containing 2x KM concentrations of putrescine (Sigma-Aldrich, e.g., 20 mM for LOX and LOXL1, or 10 mM for LOXL2 and 4 mM for LOXL3 and LOXL4) prepared in 1.2 M urea, 50 mM sodium borate buffer (pH 8.2), 120 μM Amplex Red (Thermo Fisher Scientific), and 1.5 U / mL horseradish peroxidase (Sigma-Aldrich) was added to the corresponding wells. For 96-well plates, the above volumes were doubled. Fluorescence (RFU) was read every 2.5 min for 30 min at 37 °C using an excitation 544 nm and emission 590 nm (Optima; BMG Labtech). The kinetic slope / minute for each well was calculated using MARS data analysis software (BMG labtech), and this value was used to estimate IC50 values ​​(Dotmatics). The ability of compounds of the invention to inhibit the amine oxidase activity of LOX and other family members is shown in Table 5.Bovine LOX was used as an alternative to human recombinant LOX due to its similar pharmacological properties and the often poor and unreliable behavior of commercially available or otherwise obtained human recombinant LOX. Table 6 shows the corresponding results obtained after incubation with the enzyme for 2 hours at 37°C.

[0357] [Table 5]

[0358] [Table 6] I C 50 :A=<2μM, B=<5μM, C=<10μM, D=>10μM

[0359] Example 3 The compounds of the present invention exhibit sustained inhibition of LOXL1 and LOXL2. To achieve meaningful pharmacological effects in the presence of high substrate concentrations, compounds that exhibit sustained and prolonged inhibition of LOX and LOXL1-4 are advantageous over competitive inhibitors, as the pharmacological effects may outlast the presence of unbound inhibitors. In preferred embodiments, compounds of the invention exhibit sustained inhibition of LOX and LOXL1-4.

[0360] Method for measuring sustained inhibition of LOX and LOXL1-4 by Compound 1 Jump dilution experiments: Assays were developed using a 96-well format, with the starting enzyme concentration set 100-fold higher than the inhibition test. The enzyme was added at 10× IC 50 The mixture was incubated for 40 minutes at 37°C in the presence of a test inhibitor at a concentration of 0.01%. After incubation, the mixture was diluted 50-fold in assay buffer and then further diluted 2-fold in the Amplex Red-horseradish peroxidase-putrescine reaction mixture (as in Example 2) before fluorescence measurement. Results were expressed as the percent signal recovery at 21-30 minutes compared to the uninhibited control. LOXL1 is used as a surrogate for LOX due to its similar pharmacological behavior.

[0361] The reversible standards used as controls showed nearly complete activity recovery after 30 minutes for both LOXL1 and LOXL2 (88.1% and 89.7%, respectively). In contrast, the known pan-LOX irreversible inhibitors BAPN and Compound 1 showed sustained inhibition of LOXL1 and LOXL2, with the LOXL1 / LOXL2 enzymes recovering only 0.3% / 11.2% for BAPN and 2.5% / 2.5% for Compound 1 between 21 and 30 minutes.

[0362] Example 4 Method for measuring the ability of Compound 1 to inhibit human recombinant SSAO / VAP-1 Human recombinant SSAO / VAP-1 amine oxidase activity was measured using a coupled colorimetric method as described for monoamine oxidases, copper-containing amine oxidases, and related enzymes [Holt A. and Palcic M., 2006]. Briefly, a cloned cDNA template corresponding to residues 34–763 of human SSAO / VAP-1, incorporating a mouse Ig kappa (κ) signal sequence, an N-terminal Flag epitope tag, and a tobacco etch virus (TEV) cleavage site, was assembled into a mammalian expression vector (pLO-CMV) by Geneart AG. This vector containing the human SSAO / VAP-1 residues was transfected into the CHO-K1 glycosylation mutant cell line Lec 8. A clone stably expressing human SSAO / VAP-1 was isolated and cultured on a large scale. Active human SSAO / VAP-1 was purified and recovered using immunoaffinity chromatography and used as a source of SSAO / VAP-1 activity. A high-throughput fluorescent assay was developed using either a 96- or 384-well format. Briefly, in a standard 384-well plate assay, 25 μL of purified human SSAO / VAP-1 (0.25 μg / mL) in 0.1 M sodium phosphate buffer (pH 7.4) was added to each well. Test compounds were dissolved in DMSO and incubated with human SSAO / VAP-1 for 30 min at 37°C before being tested in a concentration-response curve (CRC), typically with 4 to 11 data points in the micromolar or nanomolar range. After the 30-min incubation, 25 μL of a reaction mixture containing 600 μM benzylamine (Sigma-Aldrich), 120 μM Amplex Red (Thermo Fisher Scientific), and 1.5 U / mL horseradish peroxidase (Sigma-Aldrich) prepared in 0.1 M sodium phosphate buffer (pH 7.4) was added to the corresponding well. Fluorescence units (RFU) were read every 2.5 min for 30 min at 37°C, with excitation at 544 nm and emission at 590 nm (Optima; BMG labtech). MARS data analysis software (BMG labtech) was used to calculate the kinetic slope / min for each well, and this value was used to calculate the IC50 Values ​​were estimated (Dotmatics). The ability of Compound 1 to inhibit SSAO / VAP-1 is shown in Table 7.

[0363] Example 5 Method for determining the ability of Compound 1 to inhibit human recombinant MAO-B The specificity of Compound 1 of the present invention was tested by measuring its ability to inhibit MAO-B activity in vitro using recombinant human MAO-B (0.02 mg / mL; Sigma Aldrich). The assay was performed similarly to that for human SSAO / VAP-1 (Example 4), except that the substrate benzylamine was used at 100 μM. The ability of Compound 1 to inhibit MAO-B is shown in Table 7.

[0364] Example 6 Methods for Determining the Ability of Compound 1 to Inhibit Human Recombinant MAO-A The specificity of compound 1 of the present invention was tested by measuring its ability to inhibit MAO-A activity in vitro using recombinant human MAO-A (0.003 mg / mL; Sigma Aldrich). The assay was performed similarly to human SSAO / VAP-1 (Example 4), except that the incubation with the test compound was extended to 2 hours and the substrate tyramine was used at 200 μM instead of benzylamine. The MAO-A inhibitory potency of compound 1 is shown in Table 7.

[0365] Example 7 Method for measuring the inhibitory activity of compound 1 on human recombinant DAO The specificity of compound 1 of the present invention was tested by measuring its ability to inhibit DAO activity in vitro using recombinant human DAO (100 ng / mL; kindly provided by Prof. Boehm, Department of Clinical Pharmacology, Medical University, Vienna). The assay was performed in a similar manner to that for human SSAO / VAP-1 (Example 4), except that the incubation with the test compound was extended to 2 hours and the substrate putrescine was used at 200 μM instead of benzylamine. The ability of compound 1 to inhibit DAO is shown in Table 7.

[0366] LOX and LOXL1-4 enzymes are members of a large family of flavin- and copper-dependent amine oxidases, including SSAO / VAP-1, monoamine oxidase-B (MAO-B), monoamine oxidase-A (MAO-A), and diamine oxidase (DAO). Compound 1 of the present invention selectively inhibits members of the LOX family of enzymes with respect to SSAO / VAP-1, MAO-B, MAO-A, and DAO. Examples of high selectivity are shown in Table 7.

[0367] [Table 7]

[0368] Example 8 Measurement of time-dependent inhibition of LOXL1 and LOXL2 by compound 1 The time-dependent inhibitory effect of Compound 1 of the present invention was tested against given LOXL1 and LOXL2 preparations using a method based on the detection of hydrogen peroxide by the Amplex Red oxidation assay [Zhou et al. 1997]. The assay was developed for use in either 384- or 96-well formats. Briefly, in a standard black, clear-bottom 384-well plate assay, 25 μL of a dilution of either the isozyme or ortholog in 1.2 M urea, 50 mM sodium borate buffer (pH 8.2) was added to each well in the presence of 1 μM mofegiline and 0.5 mM pargyline (to inhibit SSAO, MAO-B, and MAO-A, respectively; not required if the enzyme was recombinant or purified). The test compound (Compound 1) was dissolved in DMSO and incubated with the enzyme at 37°C for different times, followed by a concentration-response curve (CRC) of typically 11 data points in the micromolar or nanomolar range. Then, 25 μL of reaction mixture containing 2x KM concentration of putrescine (Sigma Aldrich, e.g., 20 mM for LOXL1 or 10 mM for LOXL2) prepared in 1.2 M urea, 50 mM sodium borate buffer (pH 8.2), 120 μM Amplex Red (Thermo Fisher Scientific), and 1.5 U / mL horseradish peroxidase (Sigma Aldrich) was added to the corresponding wells. Fluorescence (RFU) was read every 2.5 min for 30 min at 37 °C, with excitation at 544 nm and emission at 590 nm (Optima; BMG labtech). The MARS data analysis software (BMG labtech) was used to calculate the kinetic slope / min for each well, and this value was used to determine the IC. 50 Values ​​were estimated (Dotmatics). Figure 1a / b and Table 8a / b show that Compound 1 exhibits time-dependent LOXL1 and LOXL2 inhibition, with apparent potency increasing over time.

[0369] [Table 8]

[0370] [Table 9]

[0371] Example 9 The following examples in Table 9 are intended to be provided to illustrate topical formulations and should not be construed as limiting the generality of the disclosure herein throughout, It will be understood that numerous variations and modifications are possible while remaining within the scope of the present invention.

[0372] [Table 10]

[0373] Preparation of topical formulations To a 1 L glass beaker containing 120 g of petrolatum, 75 g of cetostearyl alcohol was added. The mixture was then heated to 70°C in a heat bath. The mixture was thoroughly stirred until all ingredients were dissolved to obtain an oil phase. 5 g of monobasic sodium phosphate (NaH2PO4) was placed in a 1 L glass bottle. 600 g of Millipore water was added. The mixture was thoroughly stirred until all solids were dissolved to obtain a sodium phosphate buffer solution. 383.0 g of sodium phosphate buffer solution was placed in a 500 mL beaker, to which the test compound was added, and the pH of the formulation was adjusted. The mixture was heated to 70°C in a heat bath. 150 g of propylene glycol was added to a 500 mL glass bottle, and 25 g of cetomacrogol 1000 BPC flakes were added. The mixture was heated to 70°C in a heat bath with stirring until a homogeneous solution was obtained. This solution was combined with the sodium phosphate buffer solution. The mixture was thoroughly stirred until all ingredients were dissolved to obtain an aqueous phase. The oil and aqueous phases were removed from the heat bath. The aqueous phase was transferred to an aluminum mixer bowl, and the oil phase was added while mixing. The mixture was gradually cooled to 25-30°C and homogenized to obtain a cream formulation. To prepare cream formulations containing 0.3%, 1.0%, 1.5%, 2.0%, and 4.0% Compound 1, an appropriate amount of Compound 1 was dissolved in sodium phosphate buffer before being combined with the oil phase.

[0374] Example 10 The stability of a drug product formulation can significantly impact the duration and cost of drug development, the nature of the testing required to support a regulatory submission, and ultimately its safety and approval. It is important to minimize the amount of impurities or degradation products formed over time due to interactions between the various components in the formulation. This can be particularly important in compositions designed to increase skin permeability.

[0375] Compound 1 exhibited good stability in topical formulation 1, with peak percentage of compound exceeding 99.5% at all time points tested, as shown in Table 10.

[0376] [Table 11]

[0377] Example 11 Genotoxicity is a term that describes the property of chemicals related to their ability to cause damage to genetic material, leading to mutations and possibly the development of various types of cancer. Evaluating the genotoxic effects of various substances used as ingredients in drugs is extremely important, as it can directly affect the developability and usefulness of drugs.

[0378] Bacterial reverse mutation test The test article, Compound 1, was tested for its mutagenic potential in a bacterial reverse mutation assay. Salmonella typhimurium strains TA98, TA100, TA1535, and TA1537 and Escherichia coli strain WP2uvrA(pKM101) were used in three stages: a preliminary toxicity test, an initial mutation assay, and a confirmatory mutation assay. The bacterial test strains were exposed to the test article in the presence and absence of a metabolic activation system (S9 fraction prepared from Aroclor 1254-induced rat liver).

[0379] The test article was soluble in DMSO at 50 mg / mL and was used as the selection vehicle in the mutation assay. The test article was found to be stable at 20 μg / mL and 50,000 μg / mL in the vehicle DMSO for 6 hours at room temperature and for 4 days at refrigerated temperatures.

[0380] In preliminary toxicity studies, Salmonella typhimurium TA100 was directly plated and exposed to the test article at 39.0625, 78.125, 156.25, 312.5, 625, 1250, 2500, and 5000 μg / plate, along with a DMSO control, to select test doses for the mutation assay.

[0381] The results of this preliminary toxicity study showed that the test article did not precipitate on basal agar plates up to 5000 μg / plate, regardless of whether metabolic activation was present or absent.

[0382] The test article did not exhibit toxicity to the test strain at any of the doses tested, as the intensity of background flora and mean revertant colony numbers were comparable to the DMSO control, with or without metabolic activation. Based on these observations, the highest OECD 471 recommended dose of 5000 μg / plate was tested in the mutation assay.

[0383] Bacterial test strains were exposed to the test articles in triplicate at 50, 158, 500, 1581, and 5000 μg / plate. Initial mutation assays were performed using the direct plate uptake exposure format, and confirmatory mutation assays were performed using the preincubation exposure format. A vehicle control (DMSO) and appropriate positive controls were run simultaneously. For all test strains, the mean and standard deviation of the number of revertant colonies were calculated for each test dose and control.

[0384] The results of both the initial and confirmatory mutation assays showed that the test article did not exhibit any positive increase in mutagenicity at any dose tested, with or without metabolic activation, in all test strains when compared to the respective vehicle control plates.

[0385] Results of concentration analysis of dose formulation samples from the initial and confirmatory mutation assays confirmed that the highest dose level of 5000 μg / plate was achieved, supporting the validity of the study conclusions.

[0386] Under identical test conditions, the mean number of revertant colonies in the positive control increased more than three-fold, demonstrating the sensitivity of the assay procedure used.

[0387] This study demonstrated that Compound 1 was not mutagenic in the bacterial reverse mutation assay up to a dose of 5000 μg / plate under the test conditions used.

[0388] Example 12 Compound 1 exhibits good diffusion through human skin. Skin permeability is essential when considering topical applications to treat skin conditions. One method for assessing skin permeability is to use a Franz diffusion cell system with human skin as the membrane (Particle Sciences; Technical Brief 2009; Vol. 10). A topically formulated donor compound (Compound 1) is applied to the upper chamber, and diffusion through the skin is monitored at relevant time points by sampling the solution in the receptor chamber through a sampling port. The concentration of Compound 1 was determined by quantitative HPLC analysis.

[0389] Compound 1 exhibits good permeability and diffusion across the membrane (from a topical formulation containing 2% Compound 1) in a time-dependent manner, reaching high levels in the receptor chamber after 24 hours (Figure 2).

[0390] Example 13 Mouse model of sclerosis Subcutaneous bleomycin (0.1 U / kg) was administered every 2 days (for a total of 21 days) to male C57BL / 6 mice to induce skin fibrosis as a model of sclerosis. Starting on day 3, lesions were treated with vehicle or various concentrations of Compound 1 in a topical formulation. Histological examination was completed after 21 days. The histological analysis is shown in Figure 3 and demonstrates a significant improvement in the mean collagen score.

[0391] Example 14 Rodent injury models Mice were inoculated with full-thickness skin (3 cm of the flank of the mouse). 2 ) were injured by excision. In the treatment group, a 4% Compound 1 solution was applied topically once daily from 24 hours after injury until 1 week after injury. The wounds were then left to heal for an additional week. Mice were euthanized 4–6 weeks after injury, and tissues were analyzed for collagen content, hydroxyproline as a surrogate marker of collagen content, extracted cross-linking biomarkers: HLNL, DHLNL (reduced forms of deH-HLNL and deH-DHLNL, respectively), PYD, and DPD, elastin content, and overall morphology and histological changes (using polarized light microscopy, immunohistochemistry, standard staining markers, and LCMSMS).

[0392] Immature and mature crosslinks (measured using LCMS and normalized to protein content) were reduced in treated tissues compared to controls (Figure 4).

[0393] References Holt, A., Palcic, MM, (2006). A peroxidase-coupled continuous absorbance plate-reader assay for flavin monoamine oxidases, copper-containing amine oxidases and related enzymes. Nat Protoc, 1(5), 2498-2505 Zhou et al.(1997).A stable nonfluorescent derivative of resorufin for the fluorometric determination of trace hydrogen peroxide:applications in detecting the activity of phagocyte NADPH oxidase and other oxidases.Anal.Biochem,253,162-168。

Claims

1. Formula I: 【Chemistry 1】 [In the formula, A is an aryl; Each R １ is X-R ４ , halogen, deuterium, C １～６ alkyl, O-C １～６ alkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, -CN, -NO ２ , -C(O)OR ５ , -C(O)NR ６ R ７ , -S(O) ２ NR ６ R ７ , -S(O) ２ R ８ , -NR ９ C(O)R １０ , and -NR ９ S(O) ２ R １０ is independently selected from the group consisting of; each C １～６ alkyl, aryl, heteroaryl, cycloalkyl and heterocycloalkyl is optionally substituted by one or more substituents selected from the group consisting of halogen, -OH, and -C １～４ alkyl; R ２ is either H or F; R ３ is H or F; however, R ２ or R ３ Only one of them is F; X is O, CH ２ , and S(O) ２ Selected from the group consisting of; R ４ It is selected from the group consisting of aryls and heterocycloalkyls; each aryl is -S(O) ２ R ８ Each heterocycloalkyl group is optionally substituted by =O; R ５ is hydrogen and C １～６ Selected from the group consisting of alkyl groups; R ６ and R ７ is hydrogen and C １～６ Independently selected from the group consisting of alkyls; R ８ C １～６ It is alkyl; R ９ is hydrogen or C １～６ It is alkyl; R １０ C １～６ It is alkyl; n is 0, 1, 2, 3, 4, or 5. A compound thereof, or a pharmaceutically acceptable salt thereof.

2. R ２ F is R ３ The compound according to claim 1, wherein H is present, or a pharmaceutically acceptable salt thereof.

3. A is a compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein A is selected from the group consisting of phenyl, naphthyl, and 1,4-benzodioxanyl.

4. Each R １ However, X-R ４ , halogen, deuterium, C １～６ Alkyl, O-C １～６ Alkyl, heteroaryl, -CN, -NO ２ , -C(O)OR ５ , -C(O)NR ６ R ７ and -S(O) ２ R ８ Independently selected from the group consisting of; each C １～６ Alkyl and heteroaryl compounds, halogens and -C １～４ The compound according to claim 1, or a pharmaceutically acceptable salt thereof, which is optionally substituted with one or more substituents selected from the group consisting of alkyl groups.

5. Formula Ib: 【Chemistry 2】 [In the formula, Each R １ａ X-R ４ , halogen, deuterium, C １～４ Independently selected from the group consisting of alkyl and -CN; Each R １ｂ X-R ４ , halogen, deuterium, -C １～４ Alkyl, -O-C １～４ Alkyl, heteroaryl, -CN, -NO ２ , -C(O)OR ５ , -C(O)NR ６ R ７ , -S(O) ２ R ８ Independently selected from the group consisting of, each -C １～４ Alkyl and heteroaryl compounds are halogens and -C １～４ It is optionally substituted with one or more substituents selected from the group consisting of alkyl groups; R １ｃ is deuterium, -C １～４ Alkyl, -CN, -C(O)OR ５ and -S(O) ２ R ８ Selected from the group consisting of; X is O, CH ２ , and S(O) ２ Selected from the group consisting of; R ４ It is selected from the group consisting of aryls and heterocycloalkyls; each aryl is -S(O) ２ R ８ Each heterocycloalkyl group is optionally substituted by =O; R ５ is hydrogen and C １～４ Selected from the group consisting of alkyl groups; R ６ and R ７ is hydrogen and C １～４ Independently selected from the group consisting of alkyls; R ８ C １～４ It is alkyl. The compound according to claim 1, or a pharmaceutically acceptable salt thereof.

6. Each R １ａ However, X-R ４ Independently selected from the group consisting of chlorine, deuterium, methyl and -CN; Each R １ｂ is independently selected from the group consisting of X-R ４ , halogen, deuterium, -C １～４ alkyl, -OCH ３ , heteroaryl, -CN, -NO ２ , -C(O)OH, -C(O)NH ２ , -S(O) ２ CH ３ , and each -C １～４ alkyl and heteroaryl are optionally substituted by one or more substituents selected from the group consisting of fluorine and -C １～４ alkyl; R １ｃ However, deuterium, methyl, -CN, -C(O)OH and -S(O) ２ CH ３ Selected from the group consisting of; X is O, CH ２ , and S(O) ２ Selected from the group consisting of; R ４ However, phenyl and 【Transformation 3】 Selected from the group consisting of; each phenyl is -S(O) ２ CH ３ The compound according to claim 5, or a pharmaceutically acceptable salt thereof, which is optionally substituted by.

7. Each R １ｂ However, X-R ４ F, Cl, Br, deuterium, -methyl, -CF ３ , isopropyl, -OCH ３ , thiazolyl, pyrazolyl, -CN, -NO ２ , -C(O)OH, -C(O)NH ２ , -S(O) ２ CH ３ A compound according to claim 5 or 6, or a pharmaceutically acceptable salt thereof, independently selected from the group consisting of the above, wherein each thiazolyl or pyrazolyl is optionally substituted with methyl. 【Request Item 8】 【Table 1-1】 Table 1-2 Table 1-3 Table 1-4 A compound according to claim 1, or a pharmaceutically acceptable salt thereof, selected from the group consisting of the following.

9. formula: 【Chemistry 4】 The compound according to claim 1, or a pharmaceutically acceptable salt thereof, optionally a hydrochloride salt thereof.

10. A pharmaceutical composition comprising the compound described in claim 1 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient, carrier, or diluent.

11. For inhibiting the activity of any one of the amine oxidases LOX, LOXL1, LOXL2, LOXL3, or LOXL4, or To treat the condition by inhibiting the activity of one of the LOX, LOXL1, LOXL2, LOXL3, and LOXL4 proteins, A pharmaceutical composition according to claim 10, A pharmaceutical composition wherein the aforementioned condition is selected from the group consisting of fibrosis, cancer, myeloid malignancy, and scarring.

12. If the condition is fibrosis, the fibrosis is selected from the group consisting of mediastinal fibrosis, myelofibrosis, retroperitoneal fibrosis, progressive giant fibrosis, nephrogenic systemic fibrosis, Crohn's disease, keloids, scleroderma / systemic sclerosis, articular fibrosis, Dupuytren's contracture, adhesive capsulitis, pancreatic fibrosis, intestinal fibrosis, hepatic fibrosis, pulmonary fibrosis, renal fibrosis, cardiac fibrosis, fibrous stenosis, cystic fibrosis, idiopathic pulmonary fibrosis, radiation-induced fibrosis, Peyronie's disease, and scleroderma, or is related to respiratory diseases, abnormal wound healing and repair, scars, hypertrophic scars, postoperative scars, cardiac arrest, and all conditions in which excess or abnormal deposition of fibrous material is associated with disease, injury, transplantation, or surgery; preferably, the fibrosis is selected from the group consisting of keloids, scars, hypertrophic scars, scleroderma, and Dupuytren's contracture; If the condition is cancer, the cancer is selected from the group consisting of mesenchymal tumors including lung cancer; breast cancer; colorectal cancer; anal cancer; pancreatic cancer; prostate cancer; ovarian cancer; liver cancer and bile duct cancer; esophageal cancer; mesothelioma, non-Hodgkin lymphoma; bladder cancer; uterine cancer; glioma, glioblastoma, medulloblastoma, and other brain tumors; myelofibrosis, kidney cancer; head and neck cancer; gastric cancer; multiple myeloma; testicular cancer; germ cell tumor; neuroendocrine tumor; cervical cancer; oral cancer, carcinoid tumors of the gastrointestinal tract, breast, and other organs; signet ring cell carcinoma; sarcoma, fibrosarcoma, hemangioma, hemangiomatosis, hemangiopericytoma, pseudohemangioma-like hyperplasia of the mammary gland, myofibroblastoma, fibromatosis, inflammatory myofibroblastic tumor, lipoma, angiolipoma, granuloma, neurofibroma, schwannoma, angiosarcoma, liposarcoma, rhabdomyosarcoma, osteosarcoma, leiomyoma, or leiomyosarcoma; The pharmaceutical composition according to claim 11, wherein, if the condition is a myeloid malignant tumor, the myeloid malignant tumor is selected from the group consisting of myelodysplastic syndrome (MDS), myelodysplastic tumor (MPN), MDS / MPS overlap syndrome, and acute myeloid leukemia (AML), including acute promyelocytic leukemia (APL).

13. The pharmaceutical composition according to claim 10 for treating or preventing keloid disease or scarring.

14. The pharmaceutical composition according to claim 13, wherein the keloid disease or scar is a keloid, hypertrophic scar, postoperative scar, burn scar, post-traumatic scar, Dupuytren's contracture, or is caused by trauma or surgical procedure, or is the result of skin injury caused by acne, burns, varicella, infection, perforation, abrasion, surgical incision or vaccination site, and optionally the treatment assists wound healing and improves the skin compliance or appearance of the subject.

15. The pharmaceutical composition according to claim 11, which is administered topically or orally.

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