RU2801092C2 - Haloallylamine sulfone derivative as lysiloxidase inhibitors and their use - Google Patents

Abstract

FIELD: pharmaceuticals.

SUBSTANCE: invention relates to novel compounds capable of inhibiting certain amine oxidase enzymes. The invention relates to a compound — the Z-isomer of formula I, its pharmaceutically acceptable salt or hydrate, which have inhibitory activity against lysyl oxidases LOX, LOXL1, LOXL2, LOXL3 and LOXL4. In this formula, A is C_6-10aryl or 5–10-membered heteroaryl, in which 1–2 ring atoms are heteroatoms independently selected from N and S; each R¹ is independently selected from the group consisting of X-R², halogen, deuterium, C_1-6alkyl, O-C_1-6alkyl, C_6-10aryl, -C(O)NR⁴R⁵ and -S(O)₂R⁶ where each C_1-6alkyl and C_6-10aryl is optionally substituted with one or more halogens; X is selected from the group consisting of O, OCH₂ and CONH; R² is selected from the group consisting of C_3-10cycloalkyl and C_6-10aryl, where each R² is optionally substituted with one or more R⁷; R⁴ and R⁵ are independently selected from the group consisting of hydrogen and C_1-6alkyl; R⁶ is C_1-6alkyl; R⁷ is -S(O)₂NR⁴R⁵; and n is either 0, 1, 2, 3, 4, 5, or 6. The invention also relates to a pharmaceutical composition for inhibiting the amine oxidase activity of any of LOX, LOXL1, LOXL2, LOXL3 and LOXL4, containing a compound of general formula I, its use for inhibiting the above amine oxidase activity, treating a condition associated with any of the LOX, LOXL1, LOXL2 proteins, LOXL3 and LOXL4, and for the manufacture of a medicinal product for the treatment of this condition.

EFFECT: using haloallylamine sulfone derivative as lysiloxidase inhibitors.

20 cl, 11 dwg, 8 tbl, 38 ex

Description

Technical field

[0001] The present invention relates to novel compounds capable of inhibiting certain amine oxidase enzymes. These compounds are useful in the treatment of a variety of indications such as fibrosis, cancer and/or angiogenesis in humans, as well as in domestic animals and livestock. In addition, the present invention relates to pharmaceutical compositions containing these compounds, as well as their various uses.

State of the art

[0002] The enzymes are lysyl oxidase (LOX), the first family member described, and LOX-like1 (LOXL1), LOXL2, LOXL3, and LOXL4 ( J Cell Biochem 2003; 88: 660-672). Lysyl oxidase isoenzymes are copper-dependent amine oxidases that initiate covalent cross-linking of collagen and elastin. The main function of lysyl oxidase isoenzymes is to facilitate the cross-linking of collagen and elastin by oxidative deamination of the amino acid side chains of lysine and hydroxylysine to aldehydes that spontaneously react with neighboring residues. The resulting cross-linked strands contribute to the stability of the extracellular matrix (ECM) and make it less susceptible to proteolytic degradation by enzymes such as matrix metalloproteinases (MMPs). The activity of lysyl oxidase enzymes is critical for maintaining the normal stretch and elasticity properties of the connective tissue of many body organ systems.

[0003] Lysyl oxidase isoenzymes belong to the broader group of amine oxidases, which includes flavin-dependent and copper-dependent oxidases, which are described by the nature of the catalytic cofactor. Flavin-dependent enzymes include monoamine oxidase-A (MAO-A), monoamine oxidase-B (MAO-B), polyamine oxidase and lysine demethylase (LSD1) and copper-dependent enzymes include semicarbazide-sensitive amine oxidase (vascular adhesive protein-1, SSAO/VAP- 1), retinal amine oxidase, diamine oxidase, and lysyl oxidase isoenzymes. Copper-dependent amine oxidases have a second cofactor that varies slightly from enzyme to enzyme. In SSAO/VAP-1, this is an oxidized tyrosine residue (TPQ oxidized to quinone), while in lysyl oxidase isozymes, TPQ is further processed by the addition of an adjacent lysine residue (to form LTQ) ( J Cell Biochem 2003; 88: 660-672).

[0004] Lysyl oxidase isoenzymes exhibit distinct expression patterns in vivo , suggesting that certain isoenzymes will fulfill certain biological roles. Catalytically active forms of LOX have been identified in the cytosolic and nuclear compartments, and research is underway to determine their role in these compartments. LOX itself, for example, plays an important role in the epithelial-mesenchymal transition (EMT), cell migration, adhesion, transformation, and gene regulation. Different patterns of LOX expression/activity have been associated with various pathological processes, including fibrotic diseases, Alzheimer's disease and other neurodegenerative processes, as well as tumor progression and metastasis ( Am J Surg 2005; 189: 297-301).

[0005] The direct replacement of dead or damaged cells with connective tissue after injury is a survival mechanism that persists throughout evolution and appears to be most pronounced in humans, playing a valuable role after traumatic injury, infection, or disease. Progressive scarring may occur after more chronic and/or repetitive trauma that causes dysfunction of parts or the entire affected organ. Various causes such as chronic infections, chronic exposure to alcohol and other toxins, autoimmune and allergic reactions, or surgery, radiotherapy, and chemotherapy can lead to fibrosis. Thus, this pathological process can occur in almost any organ or tissue of the body and is usually the result of situations lasting for several weeks or months in which inflammation, destruction and tissue repair occur simultaneously. In this case, fibrosis most often affects the lungs, liver, skin, kidneys and cardiovascular system.

[0006] Liver fibrosis, for example, 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 can be qualitatively distinguished from that in the normal liver. This fibrosis can progress to cirrhosis, liver failure, cancer, and eventually death ( Pathology - Research and Practice 1994; 190: 910 - 919).

[0007] Fibrous tissues can accumulate in the heart and blood vessels as a result of hypertension, hypertensive heart disease, atherosclerosis, and myocardial infarction, when extracellular matrix accumulation or fibrous deposits lead to stiffness of the vasculature and stiffness of the heart tissue itself ( Am J Physiol Heart Circ Physiol 2010 ; 299: H1 - H9).

[0008] Pulmonary arterial hypertension (PAH) is a rare and rapidly fatal disease characterized by elevated pulmonary arterial pressure and is caused by increased pulmonary vascular resistance. Although it is a heterogeneous condition with a wide range of causes, it is increasingly recognized that PAH is associated with other diseases such as connective tissue disease and scleroderma. Pathological features of PAH include vascular wall remodeling with excessive extracellular matrix (ECM) deposition and cross-linking. Lysyl oxidases deregulate the pulmonary vasculature in patients with idiopathic pulmonary arterial hypertension (IPAH) and promote retention of ECM components and abnormal remodeling of collagen and elastin via cross-linking ( Arterioscler. Thromb Vasc. Biol . 2014; 34: 1446-1458). The prognosis for patients with PAH is poor. Pharmacological action on lysyl oxidases may provide a therapeutic intervention where there is currently little or no such intervention.

[0009] A strong association has been demonstrated between fibrosis and increased lysyl oxidase activity. For example, in experimental liver fibrosis in rats (Proc. Natl. Acad. sci. USA 1978; 75: 2945 - 2949), in models of pulmonary fibrosis (J Pharmacol Exp Ther 1981; 219: 675 - 678), in arterial fibrosis. (Arteriosclerosis 1981; 1: 287 - 291), with skin fibrosis (Br J Dermatol 1995; 133: 710 - 715) and adriamycin-induced kidney fibrosis in rats (Nephron 1997; 76:192-200). Of these experimental human disease models, the most striking increase in enzyme activity was seen in the rat model of CCl-induced liver fibrosis.₄. 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 association between lysyl oxidase activity measured in plasma and the progression of liver fibrosis. The level of lysyl oxidase activity is usually low in the serum of healthy people, but is significantly increased in chronic active hepatitis and even more so in cirrhosis. Therefore, lysyl oxidase can serve as a marker of intrinsic fibrosis.

[0011] Lysyl oxidase isoenzymes are highly regulated by hypoxia-inducible factor 1α (HIF-1α) and TGF-β, the two best known fibrosis-inducing growth factors ( Cell Biol 2009; 29: 4467-4483). Collagen cross-linking occurs in any type of fibrosis, so a lysyl oxidase isoenzyme inhibitor can be used in idiopathic pulmonary fibrosis, scleroderma, renal or hepatic fibrosis.

[0012] In normal wound healing, the formation of granulation tissue is a transient process that provides the basis for re-epithelialization and repair. Subsequently, the tissue is remodeled and a normotrophic scar is formed. However, after an injury, a person cannot restore normal skin. Instead, the repair (or healing) process results in scarring (scarring). Scars are aesthetically and functionally inferior to the skin. Scars are a chronic problem, and excessive or hypertrophic scarring and its accompanying aesthetic, functional, and psychological consequences remain key concerns in the treatment of deep skin lesions and burns. A key factor in the poor appearance and flexibility of scars, especially hypertrophic scars, is changes in collagen in the dermal layer. In scar tissue, collagen (predominantly Collagen I) is more densely packed and aligned in parallel bundles. In normal skin, collagen is not tightly packed and has a basket-weave structure. These changes, both in the structure and in the amount of collagen, largely underlie the poor appearance of the scar and lead to loss of flexibility, discomfort and functional problems.

[0013] Dermal fibrosis or excessive scarring of the skin is a consequence of an excessive healing response and is characterized by disproportionate fibroblast proliferation and extracellular matrix (ECM) formation in the dermis. Clinically, skin fibrosis manifests as thickened, tightened, and hardened areas of the skin. The spectrum of fibrotic skin diseases is broad, including but not limited to: hypertrophic scarring, keloids, scleroderma (diffuse and limited subtypes), scleredema (Buschke's disease), systemic amyloidosis, lipodermatosclerosis, progeroid disorders, stiff skin syndrome, Dupuytren's contracture, nephrogenic fibrosing dermopathy (NFD). ), mixed connective tissue disease, scleromyxedema, graft-versus-host disease (GVHD), and eosinophilic fasciitis. Although each of these disorders has its own etiology and clinical characteristics, they are all associated with excessive collagen production and altered collagen remodeling. One possible mechanism for altered ECM remodeling is covalent cross-linking. The pathogenesis of cutaneous fibrosis is directly affected by the LOX family of enzymes ( Laboratory investigation 2019; 99: 514 - 527). Expression of LOX and LOXL1-4 is upregulated in scar fibroblasts compared to normal skin fibroblasts, with LOX and LOXL1 being the dominant isoforms found in skin tissue.

[0014] Studies included two complementary in vitro skin-like models, human skin equivalent (HSE) and self-organizing stromal tissues, identified LOXL4 as a key isoform mediating TGF-beta-induced fibrotic phenotypes ( Lab. Invest . 2019; 99: 514 - 527) .

[0015] Scarring processes are a serious problem for the eye and surrounding structures. Ocular scarring plays an important role in either primary disease (eg, corneal and conjunctival scarring) or treatment failure (eg, postoperative trabeculectomy) ( Ocular Surgery News US Edition, October 1, 2002).

[0016] Glaucoma is a disease in which the optic nerve is damaged, leading to progressive and irreversible loss of vision. Elevated intraocular pressure (IOP) is one of the main risk factors for the development and progression of glaucoma. Most treatments for glaucoma aim to reduce intraocular pressure, either by reducing the formation of aqueous humor in the eye or, as in the case of glaucoma filtering surgery, by increasing the outflow of fluid from the eye. Trabeculectomy, the current gold standard treatment for IOP, is filtering surgery that creates an opening in the anterior chamber from under a partial scleral flap to allow water to drain from the eye. Postoperative scarring is a major cause of treatment failure. The antimetabolites mitomycin-C (MMC) and 5-fluorouracil (5-FU) are used in modern clinical practice to limit the formation of postoperative scarring on the eyes. Although these agents have been shown to improve IOP outcome in filtering surgery, they do so in a non-selective manner and are associated with significant side effects ( Arch. Ophthalmol . 2002; 120: 297-300). Safer, more targeted anti-fibrotic agents are needed.

[0017] Gingival fibromatosis is a rare and heterogeneous group of diseases that develop as slowly progressive, localized or diffuse, fibrous enlargement of keratinized gingiva (growth or enlargement of the gingiva). In severe cases, excess tissue can cover the crowns of teeth, causing masticatory, aesthetic, phonetic, functional, and periodontal problems. Gingival overgrowth can be inherited, idiopathic in origin, associated with inflammatory diseases of the oral cavity, or associated with other systemic diseases. However, in most cases, this is due to side effects of systemic drugs such as the anticonvulsant drug phenytoin, the immunosuppressant cyclosporine A, and some antihypertensive dihydropyridine calcium channel blockers, in particular nifedipine ( crit rev oral biol 2004; 15: 165-175). The pathological manifestation of gingival overgrowth involves excessive accumulation of extracellular matrix proteins, of which Collagen I is the most predominant. One of the accepted concepts behind the mechanism of drug-induced gingival overgrowth is EMT, a process in which the interaction between gingival cells and extracellular matrix is attenuated as epithelial cells transdifferentiate into fibrogenic fibroblast-like cells ( AJP 2010; 177:208-218). Damaged epithelium, basement membrane, and underlying stroma result in TGF-β stimulation of lysyl oxidase enzyme activity and promote connective tissue fibrosis ( Lab Invest 1999; 79: 1655-1667).

[0018] The rationale behind the consistent and strong inhibition of fibrosis by lysyl oxidase isoenzyme blockers is that the lack of cross-linking activity renders collagen susceptible to degradation by proteolytic enzymes such as MMPs. Therefore, any type of fibrosis should be reversed by treatment with inhibitors of the lysyl oxidase isoenzyme. Given the different involvement of all lysyl oxidase isoenzymes in fibrosis, an inhibitor that exhibits sustained strong inhibition of all lysyl oxidase isoenzymes, i.e. a pan-LOX inhibitor should be the most effective.

[0019] Rheumatoid arthritis (RA) is a systemic autoimmune disease characterized by chronic painful inflammation of the lining of the joints. However, in some people, the condition can progress to include painful swelling and inflammation of surrounding tissue and other body systems, including the skin, eyes, lungs, heart, and blood vessels. Thus, rheumatoid arthritis is a painful and debilitating disease that can lead to significant loss of function and mobility in the hands, wrists, and feet. Active rheumatoid arthritis occurs in a few joints, but can subsequently progress and affect multiple joints. Synovial hyperplasia involving infiltrated immune cells and resident synovial fibroblasts (SF) is a typical feature of RA. Synovial fibroblasts in rheumatoid arthritis (RASF) are the most common cell type at sites of invasion and are a major culprit in joint destruction. Activated RASFs are able to migrate and as such are involved in the spread of arthritis between joints. Cytokines from infiltrated immune cells induce the activation and proliferation of synovial fibroblasts. These activated SFs, in turn, generate pathogenic stroma to perpetuate chronic inflammation, which ultimately leads to cartilage and bone destruction. By implanting RASF along with human cartilage in severe combined immunodeficient mice, it has been demonstrated that activated RASF migrate in vivo , spreading the disease to areas of implanted human cartilage. Moreover, while RAS actively destroy cartilage, controls implanted with synovial fibroblasts from patients with osteoarthritis (OA) and dermal fibroblasts from healthy donors did not ( Nat. Med. 2009; 15: 1414-1420). RASF differ from non-activated healthy fibroblasts in their morphology and gene expression. RASF are characterized by the expression of anti-apoptotic, proto-oncogenes and a lack of expression of tumor suppressor genes. The production of pro-inflammatory cytokines and chemokines by RASF further promotes the recruitment of immune cells to the synovial membrane. In addition, the production of matrix metalloproteinase (MMP) enzymes promotes invasion and destruction of cartilage.

[0020] The type II collagen-induced arthritis (CIA) model is a widely used animal model for RA because it reproduces well the characteristic immunological, pathological, and arthritic manifestations seen in human RA. KIA rats showed high levels of LOX expression in synovial membranes, synovial fluid and serum. Inhibition of LOX by beta-aminopropionitrile (BAPN; a pan inhibitor of LOX) was found to attenuate inflammation, synovial hyperplasia, angiogenesis, and expression of MMP-2 and MMP-9, indicating that LOX promotes synovial hyperplasia and angiogenesis in CIA rats. In addition, knockdown of LOXL2 and anti-LOXL2 antibodies attenuates collagen deposition, proliferation, and RAS invasion ( Mol. Med. Rep. 2017: 6736 - 6742).

[0021] Although there is no cure for RA, there are a number of treatments that alleviate symptoms and alter the progression of the disease. However, such treatment is accompanied by significant side effects, partly associated with suppression of the immune system. Selective drugs targeting RASF represent a more useful therapy for RA.

[0022] Osteoarthritis (OA) is a disease characterized by degeneration of articular cartilage and underlying bone. Mostly as a result of "wear and tear", OA causes pain and stiffness in the joint. The most commonly affected joints are the fingers, knees, back, and hips. Unlike other forms of arthritis (such as RA), osteoarthritis only affects the joints. Often the joints on one side of the body are more affected than the other. OA is a progressive and debilitating disease that can significantly affect work and daily activities.

[0023] Synovial fibrosis is a key factor in OA and is a manifestation of fibroblast proliferation and an imbalance in collagen synthesis and collagen degradation. This imbalance leads to excessive collagen deposition in the extracellular matrix (ECM) and results in synovial membrane thickening and stiffness.

[0024] Genes encoding a number of 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 ( Arthritis and Rheumatology 2014; 66: 647-656) .

[0025] Given the varying contributions of many members of the lysyl oxidase enzyme family to both rheumatoid arthritis and osteoarthritis, a pan LOX inhibitor may provide a potentially more effective therapy.

[0026] BAPN is a widely used irreversible lysyl oxidase inhibitor based on a non-selective mechanism. Since the 1960s, BAPN has been used in animal studies (mainly rats, mice and hamsters) and has been effective in reducing collagen in various models (eg CCl ₄ , bleomycin, quartz, cancer) and tissues (eg liver, lungs and dermis). ( J Cell Biochem 2003; 88: 660-672). However, human studies with scleroderma have shown that BAPN is poorly tolerated and highlights the need for safer alternatives ( Clin. Pharmacol. Ther. 1967: 593-602).

[0027] Collagen cross-linking catalyzed by lysyl oxidase can proceed in two ways: the alisine pathway and the hydroxyalisine pathway. In the hydroxyalisine pathway, immature bivalent crosslinks are formed first, including dehydrodihydroxylysinonorleucine (deH-DHLNL) and dehydrohydroxylysinonorleucine (deH-HLNL), and then further progress ( via lysyl oxidase-independent reactions) to mature trivalent crosslinks between three collagen molecules to form deoxypyridinoline (DPD) and pyridinoline (PYD). These mature and immature cross-links can be measured using LC-MS/MS (PLoS One 2014; 9 (11), e112391).

[0028] Lysyl oxidase isoenzymes are not only involved in the cross-linking of elastin and collagen during wound healing and fibrosis, but also regulate cell movement and signaling. Their intracellular and intranuclear function is related to gene regulation and can lead to oncogenesis and tumor progression (Inflammapharmacol 2011; 19: 117 - 129). Both down- and up-regulation of lysyl oxidase isoenzymes in tumor tissues and cancer cell lines have been described, suggesting a dual role for lysyl oxidase isoenzymes and the LOX propeptide as a metastasis promoter gene and tumor suppressor gene.

[0029] In addition to their role in tissue remodeling, LOX isoenzymes also play an important role in primary cancer and metastasis. Tumor growth is associated with a reactive stroma, which is predominantly composed of fibroblasts; called cancer-associated fibroblasts (RAFs). Mice inoculated subcutaneously with an equal mixture of tumor and RAF cells are known to have a higher growth rate and higher metastasis rate ( Trends Mol Med . 2013; 19(8): 447 - 453). RAF knockout models have been shown to be pro-oncogenic, however this is a rather abstract scenario compared to the patient's tumor microenvironment. RAFs have been shown to have increased LOX expression compared to normal fibroblasts ( Dis Model Mech . 2018; 11(4)). The use of a LOX inhibitor in a cancer setting will potentially affect both the tumor and the stromal compartment, helping to reduce tumor growth and metastasis.

[0030] New evidence suggests an association between idiopathic pulmonary fibrosis and lung cancer, but more research is needed. Chemical or radiation-induced fibrosis in both lung and liver mouse models causes an increase in alpha-smooth muscle actin (a marker of fibroblasts), LOX expression, and metastatic tumor growth that is neutralized by LOX antibody ( Cancer Res. 2013; 73(6) : 1721 - 1732).

[0031] To date, an increase in mRNA and/or protein in lysyl oxidase isoenzymes has been observed in breast cancer cell lines, CNS, head and neck squamous cell carcinoma, esophagus, kidney, lung, prostate, clear cell and lung carcinomas, ovaries, uterus, samples patients with melanoma and osteosarcoma from The Cancer Genome Atlas (TCGA). Table 1 presents the expression data of the patient's TCGA gene for the LOX family. A plus sign indicates that the gene expression is above average in this dataset.

Table 1

Patient TCGA gene expression data for the LOX family

Cancer
LOX
LOXL1
LOXL2
LOXL3
LOXL4 Number of patient samples Invasive breast carcinoma + + + + 1212 Esophageal carcinoma + + + 196 Glioblastoma multiforme + + 171 Squamous cell carcinoma of the head and neck + + + 566 Renal clear carcinoma + + + 606 Papillary cell renal carcinoma + + 323 Squamous cell carcinoma of the lung + + + + 552 Mesothelioma + + + + + 87 Serous cystadenocarcinoma of the ovary + + + + 307 Adenocarcinoma of the pancreas + + + + + 183 Pheochromocytoma and paraganglioma + + 187 Sarcoma + + + + 265 Cutaneous melanoma + + + 473 Uterine carcinoma + + + + + 57 Carcinoma of the endometrium of the body of the uterus + 201

[0032] Statistically significant clinical correlations between lysyl oxidase isoenzyme expression and tumor progression have been observed in breast, head and neck squamous tumors, myelofibrosis, prostatic, pancreatic, ovarian, and clear cell renal cell carcinomas. The role of lysyl oxidase isoenzymes in tumor progression has been most extensively studied in breast cancer using in vitro migration/invasion models and in vivo mouse tumorigenesis and metastasis models (Nature. 2006; 440 (7088): 1222-1226). Increased expression of lysyl oxidase isoenzymes has been found in patients with hypoxia and has been associated with estrogen receptor (ER-) negative status, reduced overall survival in EC patients and patients without lymph node involvement who did not receive adjuvant systemic treatment, as well as a shorter free from bone metastasis to survival in EC patients and patients without lymph node involvement; in vivo models have demonstrated that LOX inhibitors have the potential in breast cancer patients with bone metastases by modulating bone homeostasis independent of RANKL ( Nature. 2015; 522 (7554): 106 - 110). Lysyl oxidase isoenzyme mRNA has been shown to be upregulated in invasive and metastatic cell lines (MDA-MB-231 and Hs578T), as well as in more aggressive breast cancer cell lines and distant metastatic tissues compared to primary cancer tissues ( Cancer Res . 2002; 62 (15): 4478 - 4483).

[0033] Pathogenic processes in primary myelofibrosis include primary megakaryocyte-weighted clonal myeloproliferation and paraneoplastic stromal response, which includes bone marrow fibrosis, osteosclerosis, angiogenesis, and extramedullary hematopoiesis. The bone marrow response 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 lead to reduced hematopoietic capacity. Secondary myelofibrosis may be the result of polycythemia vera or essential thrombocytosis. In myelofibrosis, disease progression correlates with an increase in the number of megakaryocytes that overexpress LOX. In the GATA 1low mouse model of myelofibrosis, disease progression (including increases in megakaryocytes, fibrosis, and spleen size) was significantly attenuated by a pan LOX inhibitor ( J Biol Chem . 2011; 286 (31): 27630-27638).

[0034] For most types of tumors, the first line of treatment is surgical resection. The wound healing response is initiated by surgery and may correlate with increased metastatic spread. Models of breast cancer have shown that abdominal surgery increases lung metastasis. Also, it has been shown to be caused by systemic LOX. Injection of plasma harvested from abdominal surgery mice (which contained LOX) into tumor-bearing mice resulted in an increase in lung metastases. Surgery-induced systemic LOX was blocked by BAPN, reducing metastasis and increasing survival ( Cell Rep . 2017; 19 (4): 774 - 784).

[0035] In models of colon cancer, breast cancer, and melanoma, tumor-associated endothelial cells have been shown to have increased LOX expression, which stimulates angiogenesis and tumor growth ( Cancer Res . 2015; 73 (2): 583 - 594) .

[0036] In patients with pancreatic, breast, lung, ovarian, and colon cancer, high collagen content correlates with high LOX gene expression, resistance to chemotherapy, and a significant reduction in survival ( Oncogene . 2018; 37(36) 4921-4940, EMBO Mol Med . 2015; 7(8) 1063-1076, Oncotarget . 2016; 7(22) 32100-32112). LOX inhibitors (both BAPN and LOX antibody) and standard chemotherapy drugs have been combined in mouse desmoplastic tumor models to reduce tumor interstitial pressure causing vasodilation ( Oncotarget . 2016; 7(22) 32100-32112). Increased vascular blood flow increases the concentration of the chemotherapeutic agent at the site of the primary tumor, resulting in reduced metastatic burden and increased survival ( Oncotarget . 2016 May 31; 7(22) 32100 - 32112).

[0037] Increased expression of the lysyl oxidase isoenzyme associated with CA-IX, a marker of hypoxia, has been found in head and neck squamous cell carcinoma and has been associated with reduced cancer-specific survival, reduced overall survival, and lower metastasis-free survival ( Oncotarget . 2016; 7 (31): 50781 - 50804). In oral squamous cell carcinoma, lysyl oxidase isoenzyme mRNA expression was upregulated compared to normal mucosa.

[0038] Glioma gene expression profiles have revealed overexpression of the lysyl oxidase isoenzyme as part of the molecular signature indicative of invasion and associated with higher grade tumors that are highly correlated with poor patient survival ( PloS ONE . 2015 Mar 19; 10(3) e0119781). Expression of the lysyl oxidase isoenzyme protein was increased in glioblastoma and astrocytoma tissues, as well as in cultured astrocytoma cells invasive U343 and U251.

[0039] In tissues, lysyl oxidase isoenzyme mRNA was upregulated in prostate cancer compared to benign prostatic hypertrophy, correlated with Gleason score, and was associated with both high grade and short time to recurrence ( Oncol Rep 2008; 20:1561-1567).

[0040] In clear cell RCC, smoking has been associated with an allelic imbalance on chromosome 5q23.1, where the LOX gene is located, and may involve gene duplication ( Cancer Genet Cytogenet . 2005; 163(1)7: 7-11).

[0041] SiHa cervical cancer cells have shown increased invasion in vitro under hypoxic/anoxic conditions; this was suppressed by inhibition of extracellular catalytically active lysyl oxidase activity by BAPN treatment as well as LOX antisense oligonucleotides, LOX antibody, LOX shRNA, or extracellular copper chelator ( Oncol Rep . 2013; 29(2), 541-548).

[0042] In a genetically engineered mouse model of ovarian cancer (ApoE knockout), a desmoplastic tumor develops with overexpression of the LOX gene. BAPN treatment significantly increased survival and reduced lung metastasis ( J Exp Clin Cancer Res. 2018; 37:32). Some tumors from patients with ovarian cancer have the G473A single nucleotide polymorphism of the LOX gene. Two independent studies have shown that people with a pronounced G473A polymorphism have an increased chance of developing ovarian cancer (J Int Med Res . 2012; 40(3): 917 - 923; Genet Test Mol Biomarkers . 2012; 16 (8): 915 - 919 ).

[0043] In primary human oral squamous cell carcinoma (SCC), lysyl oxidase enzyme levels (particularly LOX and LOXL2) and lysyl hydroxylase expression are significantly increased and are markedly elevated in advanced tumors, (RLNM)-positive for regional lymph node metastases. Both repairable or immature crosslinks (deH-DHLNL and deH-HLNL) and non-repairable or mature crosslinks (DPD and PYD) are significantly higher in CRCC compared to normal tissues ( J Dent Res 2019; 98(5):517 – 525).

[0044] The results described herein provide a strong rationale for combination therapy involving inhibitors of the LOX isoenzyme and anticancer therapy in patients.

[0045] More recently, CCT365623, a reversible pan LOX inhibitor, has been used in a breast cancer model (MMTV-PyMT) to reduce metastasis and increase survival ( Nat Commun . 2017; 18 (8): 14909).

[0046] The scientific and patent literature describes small molecule inhibitors of lysyl oxidase isoenzymes and antibodies LOX and LOXL2 with therapeutic effects in animal models of fibrosis and cancer metastasis. It is also reported that some known MAO inhibitors inhibit the lysyl oxidase isoenzyme (for example, Mofegiline is an inhibitor of MAO-B, shown below). This inhibitor is a member of the haloallylamine family of MAO inhibitors; the halogen in Mofegiline is fluorine. Fluorallylamine inhibitors are described in US Pat. No. 4,454,158. Patents have been issued for fluoroallylamines and chlorallylamines, such as MDL72274 (illustrated below) as lysyl oxidase inhibitors (US Patents 4,943,593; 4,965,288; 5,021,456; 5,059,714; 5,182,297; 5,252,608). Many of the compounds claimed in these patents are also reported to be potent inhibitors of MAO-B and SSAO/VAP-1.

[0047] Additional fluoroallylamine inhibitors are described in US Pat. No. 4,699,928. Other examples structurally related to Mofegiline can be found in WO 2007/120528.

[0048] WO 2009/066152 describes a family of 3-substituted 3-haloallylamines that are SSAO/VAP-1 inhibitors useful in the treatment of a variety of indications, including inflammatory disease. None of these documents specifically describes the fluoroallylamine compounds of formula (I) according to the present invention.

[0049] Antibodies to LOX and LOXL2 are described in US 2009/0053224 with diagnostic and therapeutic uses. Anti-LOX and anti-LOXL2 antibodies can be used to detect and treat conditions such as fibrosis, angiogenesis, or to prevent the transition from an epithelial cell state to a mesenchymal cell state: US 2011/0044907.

[0050] WO 2017/136871 and WO 2017/136870 describe haloallylamine-indole and azaindole derivatives as lysyl oxidase inhibitors and their use.

[0051] WO 2018/157190 describes haloallylaminpyrazole derivatives as lysyl oxidase inhibitors and their use.

[0052] WO 2017/141049 and WO 2019/073251 describe families of methylamine and bridging homopiperazine derivatives, respectively, as lysyl oxidase inhibitors and their use in the treatment of cancer and diseases associated with fibrosis.

[0053] WO 2003/097612, WO 2006/053555 and US 2008/0293936 describe another class of lysyl oxidase inhibitors.

[0054] WO 2018/048930, WO 2017/015221, WO 2017/003862, WO 2016/144702 and WO 2016/144703 describe other inhibitors of LOXL2.

SUMMARY OF THE INVENTION

[0055] The present invention relates to substituted fluoroallylamine compounds that inhibit lysyl oxidase (LOX), lysyl oxidase-like 2 (LOXL2), and other lysyl oxidase isoenzymes. Surprisingly, the structural modification of 3-substituted-3-fluoroallylamine described earlier led to the discovery of new compounds that are potent inhibitors of the human LOX and LOXL isoenzymes. In addition, some of these new compounds also selectively inhibit certain LOX and LOXL isoenzymes over other enzymes in the amine oxidase family.

[0056] The first aspect of the invention provides a compound of Formula I:

Formula I

or a pharmaceutically acceptable salt, polymorph, solvate, hydrate or tautomeric form thereof; wherein:

A is aryl or heteroaryl;

each R ¹ is independently selected from the group consisting of XR ² , halogen, deuterium, C _1-6 alkyl, OC _1-6 alkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, -CN, -C(O)OR ³ , -C (O)NR ⁴ R ⁵ , -S(O) ₂ NR ⁴ R ⁵ , -S(O) ₂ R ⁶ , -NR ⁸ C(O)R ⁹ and -NR ⁸ S(O) ₂ R ⁹ ; where each C _1-6 alkyl, aryl, heteroaryl, cycloalkyl and heterocycloalkyl is optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -SO ₂ CH ₃ , -C _1-4 alkyl, -OC _{1- 4} alkyl, -CF ₃ , -CH ₂ CF ₃ and -O-CF ₃ ;

X is selected from the group consisting of O, CH ₂ , OCH ₂ , CH ₂ O, CH ₂ S(O) ₂ , CONH and NHCO;

R ² is selected from the group consisting of cycloalkyl, heterocycloalkyl, aryl and heteroaryl; where each R ² is optionally substituted with one or more R ⁷ ;

R ³ is selected from the group consisting of hydrogen, C _1-6 alkyl and C _3-7 cycloalkyl; where each C _1-6 alkyl and C _3-7 cycloalkyl is optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -SO ₂ CH ₃ , -C _1-4 alkyl, -OC _1-4 alkyl , -CF ₃ , -CH ₂ CF ₃ and -O-CF ₃ ;

R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen, C _1-6 alkyl and C _3-7 cycloalkyl; where each C _1-6 alkyl and C _3-7 cycloalkyl is optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -SO ₂ CH ₃ , -C _1-4 alkyl, -OC _1-4 alkyl , -CF ₃ , -CH ₂ CF ₃ and -O-CF ₃ ; or

R ⁴ and R ⁵ , when attached to the same nitrogen atom, are combined to form a 4-7 membered ring having 0 to 1 additional heteroatoms as ring members;

R ⁶ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl; where each C _1-6 alkyl and C _3-7 cycloalkyl are optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -C _1-4 alkyl, -OC _1-4 alkyl, -CF ₃ , - CH ₂ CF ₃ and -O-CF ₃ ;

R ⁷ is selected from the group consisting of halogen, OH, C _1-6 alkyl, OC _1-6 alkyl, C _3-7 cycloalkyl, -C(O)OR ³ , -C(O)NR ⁴ R ⁵ , -NR ⁴ C(O)R ⁶ , -S(O) ₂ NR ⁴ R ⁵ , -NR ⁴ S(O) ₂ R ⁶ and -S(O) ₂ R ⁶ ; where each C _1-6 alkyl is optionally substituted with one or more substituents selected from the group consisting of halogen and -OH;

R ⁸ is hydrogen or C _1-6 alkyl;

R ⁹ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl; where each C _1-6 alkyl and C _3-7 cycloalkyl are optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -C _1-4 alkyl, -OC _1-4 alkyl, -CF ₃ , - CH ₂ CF ₃ and -O-CF ₃ ; or

R⁸ and R⁹ combined to form a 5-7-membered a ring having 0 to 1 additional heteroatoms as ring members;

And

n is 0, 1, 2, 3, 4, 5, or 6.

[0057] A second aspect of the 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.

[0058] A third aspect of the invention provides a method for inhibiting the activity of an amine oxidase of any 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 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.

[0059] A fourth aspect of the invention provides a method for treating a condition associated with any one of the LOX, LOXL1, LOXL2, LOXL3, or LOXL4 proteins, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of the first aspect of the invention, or a pharmaceutically acceptable salt thereof, or solvate, or pharmaceutical composition according to the second aspect of the invention.

[0060] A fifth aspect of the invention provides the use of a compound of the first aspect of the invention, or a pharmaceutically acceptable salt or solvate thereof, for the manufacture of a medicament for the treatment of a condition associated with any of the LOX, LOXL1, LOXL2, LOXL3, or LOXL4 proteins.

[0061] A sixth aspect of the invention provides a compound of the first aspect of the invention, or a pharmaceutically acceptable salt or solvate thereof, for use in the treatment of a condition associated with any of the LOX, LOXL1, LOXL2, LOXL3, or LOXL4 proteins.

[0062] In one embodiment of the methods and uses of the present invention, the condition is selected from fibrosis, cancer, and angiogenesis.

[0063] This document discusses combination therapy, in which the methods further include the co-administration of additional therapeutic agents that are used to treat cancer, fibrosis, angiogenesis, inflammation, hypertension, immunosuppression, and metabolic conditions.

Definitions

[0064] The following are some definitions that may be helpful in understanding the description of the present invention. They are intended as general definitions and should in no way limit the scope of the present invention to these terms only, but are provided for a better understanding of the following description.

[0065] Unless the context otherwise requires or is specifically indicated otherwise, integers, steps, or elements of the invention set forth herein as single integers, steps, or elements clearly encompass the singular and plural forms of the listed integers, steps, or elements.

[0066] Throughout this description, unless the context otherwise requires, the word "comprise" or variations such as "comprises" or "comprising" will be understood to mean the inclusion of a specified step or element, or an integer, or a group of steps or elements , or integers, but not to the exclusion of any other stage, element, integer, or group of elements or integers. Thus, in the context of this description, the term "comprising" means "including mainly, but not necessarily exclusively."

[0067] Those skilled in the art will appreciate that the invention described herein is susceptible to changes and modifications other than those specifically described. It is to be understood that the invention includes all such variations and modifications. The invention also includes all of the steps, features, compositions and compounds mentioned or referred to in this description, individually or together, and any and all combinations or any two or more of these steps or features.

[0068] As used herein, the term "alkyl" includes in its meaning monovalent ("alkyl") and divalent ("alkylene") saturated hydrocarbon radicals with a straight or branched chain containing from 1 to 6 carbon atoms, for example, 1, 2 , 3, 4, 5 or 6 carbon atoms. A straight chain or branched alkyl group attaches at any available point to form a stable compound. For example, the term "alkyl" includes, but is not limited to, methyl, ethyl, 1-propyl, isopropyl, 1-butyl, 2-butyl, isobutyl, t-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.

[0069] The term "alkoxy" or "alkyloxy" as used herein refers to straight or branched chain alkyloxy groups (ie, O-alkyl), where alkyl is defined as defined above. Examples of alkoxy groups include methoxy, ethoxy, n-propoxy and isopropoxy.

[0070] As used herein, the term "cycloalkyl" includes within its meaning monovalent ("cycloalkyl") and divalent ("cycloalkylene") saturated, monocyclic, bicyclic, polycyclic, or fused analogs. In the context of the present description, the cycloalkyl group may have from 3 to 10 carbon atoms. A fused cycloalkyl analog means a monocyclic ring fused to an aryl or heteroaryl group in which the point of attachment is on the non-aromatic moiety. Examples of cycloalkyl and its fused analogs include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, tetrahydronaphthyl, decahydronaphthyl, indanyl, adamantyl, and the like.

[0071] As used herein, the term "aryl" or variants such as "arylene" refers to monovalent ("aryl") and divalent ("arylene") single, polynuclear, conjugated and fused analogues of aromatic hydrocarbons containing from 6 to 10 carbon atoms. A fused aryl analog means an aryl group fused to a monocyclic cycloalkyl or monocyclic heterocyclyl group in which 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. "Substituted aryl" is aryl which is independently substituted with one or more, preferably 1, 2 or 3 substituents attached to any available atom to form a stable compound.

[0072] The term "alkylaryl" as used herein includes in its meaning monovalent ("aryl") and divalent ("arylene"), single, polynuclear, conjugated and fused aromatic hydrocarbon radicals attached to divalent, saturated, linear or branched alkylene radicals. Examples of alkylaryl groups include benzyl.

[0073] The term "heteroaryl" and variants such as "heteroaromatic group" or "heteroarylene" as used herein include monovalent ("heteroaryl") and divalent ("heteroarylene"), single, polynuclear, conjugated and fused heteroaromatic radicals having 5 to 10 atoms, wherein 1 to 4 ring atoms or 1 to 2 ring atoms are heteroatoms independently selected from O, N, NH, and S. Heteroaryl also includes oxidized S or N, such as sulfinyl, sulfonyl and tertiary ring N-nitric oxide. The carbon or nitrogen atom is the site of attachment of the heteroaryl ring structure so that a stable compound is formed. The heteroaromatic group may be C _1-9 heteroaromatic. A fused heteroaryl analog means 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, isoquinolinyl, 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 heteroatoms are N. "Substituted heteroaryl" is a heteroaryl that is independently substituted with one or more, preferably 1, 2, or 3 substituents attached to any available atom to form a stable compound.

[0074] The term "heterocyclyl" and variants such as "heterocycloalkyl" as used herein include monovalent ("heterocyclyl") and divalent ("heterocyclylene"), saturated or partially saturated (non-aromatic), monocyclic, bicyclic, polycyclic or fused hydrocarbon radicals containing from 3 to 10 ring atoms, where from 1 to 4 or from 1 to 2 ring atoms are heteroatoms independently selected from O, N, NH or S, SO or SO ₂ in which the point of attachment may be carbon or nitrogen. A fused heterocyclyl analog means a monocyclic heterocycle fused to an aryl or heteroaryl group in which the point of attachment is on the non-aromatic moiety. The heterocyclyl group may be C _3-8 heterocyclyl. The heterocycloalkyl group may be C _3-6 heterocyclyl. The heterocyclyl group may be C _3-5 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, tetrahydrofur anil, tetrahydropyranyl and the like. The term also includes partially unsaturated monocyclic rings which are not aromatic, such as 2- or 4-pyridones attached via nitrogen or N-substituted uracils.

[0075] The term "halogen" or variants thereof such as "halide" or "halogen" as used herein refers to fluorine, chlorine, bromine, and iodine.

[0076] The term "heteroatom" or variants thereof such as "hetero" or "hetero group" as used herein refers to O, N, NH, and S.

[0077] In general, "substituted" refers to an organic group as defined herein (eg, an alkyl group) in which one or more bonds to a contained hydrogen atom are replaced by a bond to non-hydrogen or non-carbon atoms. Substituted groups also include groups in which one or more bonds to carbon(s) or hydrogen(s) are replaced by one or more bonds, including double or triple bonds, to a heteroatom. Thus, a substituted group will be substituted with one or more substituents unless otherwise indicated. In some embodiments, the substituted group is substituted with 1, 2, 3, 4, 5, or 6 substituents.

[0078] The term "optionally substituted" as used herein means the group to which the term refers, which may be unsubstituted or may be substituted by one or more groups independently selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, heterocycloalkyl , halogen, haloalkyl, hydroxyl, hydroxyalkyl, alkoxy, thioalkoxy, alkenyloxy, haloalkoxy, NO ₂ , NH (alkyl), N (alkyl) ₂ , alkylamino, dialkylamino, acyl, alkenoyl, alkinoyl, acylamino, diacylamino, acyloxy, alkylsulfonyl, alkylsulfonyloxy , sulfonamido, heterocycloxy, heterocycloamino, heterocycloalkyl, alkylsulphenyl, alkylcarbonyloxy, phosphorus-containing groups such as phosphono and phosphinyl, aryl, heteroaryl, alkylaryl, aralkyl, alkylheteroaryl, cyano, CO ₂ H, CO ₂ alkyl, C(O)NH ₂ , - C(O)NH(alkyl) and -C(O)N(alkyl) ₂ . Preferred substituents include halogen, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, hydroxy(C _1-6 )alkyl, C ₃ -C ₆ cycloalkyl, C(O)OH, NHC( O) C ₁ -C ₄ alkyl, C (O) C ₁ -C ₄ alkyl, NH ₂ , NHC ₁ -C ₄ alkyl, N (C ₁ -C ₄ alkyl) ₂ , SO ₂ (C ₁ -C ₄ alkyl ), OH and CN. Particularly preferred substituents include C _1-4 alkyl, C _1-4 alkoxy, SO ₂ (C ₁ -C ₄ alkyl), halogen, OH, hydroxy(C _1-3 )alkyl (eg C(CH ₃ ) ₂ OH) and C _1-3 haloalkyl (eg CF ₃ , CH ₂ CF ₃ ).

[0079] This invention includes compounds 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 described herein.

[0080] The present invention includes isotopes of various atoms. Any atom not specifically designated as a particular isotope means any stable isotope of that atom. Thus, it should be understood that the present description includes the isotopes of hydrogen - deuterium and tritium.

[0081] All references cited in this application are specifically included as cross-references in their entirety. Reference to any such documents should not be construed as an admission that the document is part of the general knowledge or prior art.

[0082] As used herein, the term "administration" and variations of this term, including "administer" and "administration", include bringing into contact, applying, delivering or providing a compound or composition of the invention in the body or on a surface by any suitable means. As used herein, the term "treatment" refers to any and all uses that treat a condition or symptoms of a disease, prevent the onset of a disease, or otherwise prevent, hinder, slow, or reverse the progress of a disease or other undesirable symptoms in any way.

[0083] In the context of this description, the term "topical administration" or variations of this term, including "topical application", includes in its meaning the application, bringing into contact, delivery or provision of a compound or composition of the invention on the skin or localized areas of the body.

[0084] As used herein, the term "topical administration" or variations of this term, including "topical application", includes within its meaning the application, contact, delivery or provision of a compound or composition of the present invention to the skin or localized areas of the body.

[0085] In the context of this description, the term "effective amount" includes in its meaning a sufficient, but non-toxic amount of a compound or composition of the present invention to provide the desired effect. Thus, the term "therapeutically effective amount" includes in its meaning a sufficient but non-toxic amount of a compound or composition of the invention to provide the desired therapeutic effect. The exact amount needed will vary from subject to subject depending on such factors as the species being treated, the sex, age and general condition of the subject, the severity of the condition being treated, the particular agent being administered, the route of administration, and so forth. Thus, it is not possible to specify an exact "effective amount". However, for any particular case, an appropriate "effective amount" can be determined by one of ordinary skill in the art using only routine experimentation.

Brief description of graphic materials

[0086] In FIG. 1(a) and 1(b) show survival curves comparing high and low gene expression in patients with pancreatic adenocarcinoma based on the TCGA dataset. A. Expression of the LOX gene. B. LOXL2 gene expression.

[0087] FIG. 2(a-c) depict dose-dependent blocking of lysyl oxidase enzyme activity by Compounds 1 and 33. Measurement of lysyl oxidase activity compared to untreated control in rat tissues (a) ear (24 hours after a single oral dose of 10 and 30 mg/kg, Compound 1) ; (b) ear (4, 24, 48 and 120 hours after a single 30 mg/kg oral dose of Compound 33) and; (c) aorta (single oral dose of 5, 10 or 30 mg/kg, Compound 33)

[0088] FIG. 3(a) shows the decrease in the amount of collagen in the scar tissue of mice after injury when treated with Compound 1 by topical application.

[0089] FIG. 3(b) histology shows thick parallel collagen bundles in control scar tissue.

[0090] FIG. 3(c) tissue treated with Compound 1 shows reduced tuft density and more "normal" collagen structure.

[0091] FIG. 4(a-b) shows the decrease in LOX activity and total collagen after daily topical treatment for 4 weeks with a 3% solution of Compound 1 compared to control.

[0092] FIG. 4(c-f) general morphology shows similar wounds at the time of injury (c, control; d, treated), and at the time of euthanasia, the wounds appear more healed (e, control, f treated).

[0093] FIG. 4 (g-h) histology shows thick streaks of collagen in untreated scar tissue (arrowed, g). It appears to decrease in treated tissue (h).

[0094] FIG. 5 (ae) Tumor Growth Data in a Xenograft Human Orthotopic Pancreatic Cancer Model: Efficacy Data. A. Growth and treatment strategy diagram. B. Monitoring of tumor growth in vivo using bioluminescent imaging. C. Ex vivo bioluminescent signal of total tumor burden. D. Ex vivo bioluminescent signal of the primary tumor. E. Ex vivo bioluminescent signal of metastatic load.

[0095] FIG. 6(a-c) Histological analysis of a sclerotic mouse skin model treated with Compound 1 topically. A. Evaluation of the composite skin. B. Collagen Average Score. C. Average LOX score.

[0096] FIG. 7(a-e) analysis of the spleen in Compound 19 treated primary myelofibrosis model (GATA-1low). A. Gomory silver staining of the spleen. B. Mass of the spleen. C. Quantification of reticulin fibrosis of the spleen. D. Image of the spleen stained with H&E. E. Quantification of megakaryocytes in the spleen.

[0097] FIG. 8(a-d) analysis of bone marrow from Compound 19 treated primary myelofibrosis model (GATA-1low). A. Bone marrow staining with Gomory silver. B. Quantification of reticulin fibrosis in the bone marrow. C. Bone marrow image stained with H&E. D. Quantification of megakaryocytes in the bone marrow.

[0098] FIG. 9 depicts changes in the area of fibrosis in a mouse PBM model.

[0099] FIG. 10 depicts the ability of Compound 33 to reduce bleomycin-induced pulmonary fibrosis (Ashcroft score) and weight gain.

[0100] FIG. 11(ad) depicts the ability of Compound 33 to reduce fibrosis-associated metastases in a CCl ₄ -induced mouse liver fibrosis model with an orthotopically injected breast cancer cell line (4t). (a) Outline of study design; (b) clinical evaluation of liver fibrosis; (c) the concentration of cross-links in the liver; (d) measurement of liver metastases.

Detailed description

[0101] The present invention relates to substituted fluoroallylamine compounds that inhibit lysyl oxidase (LOX), lysyl oxidase-like 2 (LOXL2), and other lysyl oxidase isoenzymes. In particular, the present invention relates to substituted fluoroallylamine derivatives with a sulfone linker.

[0102] In particular, the present invention relates to compounds of Formula I:

Formula I

or its pharmaceutically acceptable salt polymorphic form, solvate, hydrate or tautomeric form; wherein:

A is aryl or heteroaryl;

each R ¹ is independently selected from the group consisting of XR ² , halogen, deuterium, C _1-6 alkyl, OC _1-6 alkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, -CN, -C(O)OR ³ , -C (O)NR ⁴ R ⁵ , -S(O) ₂ NR ⁴ R ⁵ , -S(O) ₂ R ⁶ , -NR ⁸ C(O)R ⁹ and -NR ⁸ S(O) ₂ R ⁹ ; where each C _1-6 alkyl, aryl, heteroaryl, cycloalkyl and heterocycloalkyl is optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -SO ₂ CH ₃ , -C _1-4 alkyl, -OC _{1- 4} alkyl, -CF ₃ , -CH ₂ CF ₃ and -O-CF ₃ ;

X is selected from the group consisting of O, CH ₂ , OCH ₂ , CH ₂ O, CH ₂ S(O) ₂ , CONH and NHCO;

R ² is selected from the group consisting of cycloalkyl, heterocycloalkyl, aryl and heteroaryl; where each R ² is optionally substituted with one or more R ⁷ ;

R ³ is selected from the group consisting of hydrogen, C _1-6 alkyl and C _3-7 cycloalkyl; where each C _1-6 alkyl and C _3-7 cycloalkyl is optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -SO ₂ CH ₃ , -C _1-4 alkyl, -OC _1-4 alkyl , -CF ₃ , -CH ₂ CF ₃ and -O-CF ₃ ;

R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen, C _1-6 alkyl and C _3-7 cycloalkyl; where each C _1-6 alkyl and C _3-7 cycloalkyl is optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -SO ₂ CH ₃ , -C _1-4 alkyl, -OC _1-4 alkyl , -CF ₃ , -CH ₂ CF ₃ and -O-CF ₃ ; or

R ⁴ and R ⁵ , when attached to the same nitrogen atom, are combined to form a 4-7 membered ring having 0 to 1 additional heteroatoms as ring members;

R ⁶ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl; where each C _1-6 alkyl and C _3-7 cycloalkyl are optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -C _1-4 alkyl, -OC _1-4 alkyl, -CF ₃ , - CH ₂ CF ₃ and -O-CF ₃ ;

R ⁷ is selected from the group consisting of halogen, OH, C _1-6 alkyl, OC _1-6 alkyl, C _3-7 cycloalkyl, -C(O)OR ³ , -C(O)NR ⁴ R ⁵ , -NR ⁴ C(O)R ⁶ , -S(O) ₂ NR ⁴ R ⁵ , -NR ⁴ S(O) ₂ R ⁶ and -S(O) ₂ R ⁶ ; where each C _1-6 alkyl is optionally substituted with one or more substituents selected from the group consisting of halogen and -OH;

R ⁸ is hydrogen or C _1-6 alkyl;

R ⁹ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl; where each C _1-6 alkyl and C _3-7 cycloalkyl are optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -C _1-4 alkyl, -OC _1-4 alkyl, -CF ₃ , - CH ₂ CF ₃ and -O-CF ₃ ; or

R⁸ and R⁹ combined to form a 5-7-membered a ring having 0 to 1 additional heteroatoms as ring members;

And

n is 0, 1, 2, 3, 4, 5, or 6.

[0103] In one embodiment of the compounds of the present invention, A is selected from aryl and heteroaryl. In another embodiment of the compounds of the present invention, A is selected from the group consisting of phenyl, naphthyl, pyridinyl, quinolinyl, benzothiazolyl and indolyl. In a further embodiment of the compounds of the present invention, A is selected from the group consisting of

.

In another further embodiment of the compounds of the present invention, A is selected from the group consisting of In a further embodiment, A is selected from the group consisting of In another further embodiment, A is In another embodiment, A is phenyl. In a further embodiment, A is heteroaryl.

[0104] In one embodiment of the compounds of the present invention, R ¹ is independently selected from the group consisting of XR ² , halogen, deuterium, C _1-6 alkyl, OC _1-6 alkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, -CN, - C(O)OR ³ , -C(O)NR ⁴ R ⁵ , -S(O) ₂ NR ⁴ R ⁵ , -S(O) ₂ R ⁶ , -NR ⁸ C(O)R ⁹ and -NR ⁸ S(O) ₂ R ⁹ ; where each C _1-6 alkyl, aryl, heteroaryl, cycloalkyl and heterocycloalkyl is optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -SO ₂ CH ₃ , -C _1-4 alkyl, -OC _{1- 4} alkyl, -CF ₃ , -CH ₂ CF ₃ and -O-CF ₃ . In another embodiment of the compounds of the present invention, each R ¹ is independently selected from the group consisting of -XR ² , C _1-6 alkyl, C _1-6 haloalkyl, -C(O)OR ³ , -C(O)NR ⁴ R ⁵ , -S(O) ₂ NR ⁴ R ⁵ , -S(O) ₂ R ⁶ . In a further embodiment of the compounds of the present invention, each R ¹ is independently selected from the group consisting of halo, C _1-6 alkyl, C _1-6 haloalkyl, and —S(O) ₂ R ⁶ . In one embodiment of the compounds of the present invention, at least one of R ¹ is XR ² . In another embodiment of the compounds of the present invention, one of R ¹ is XR ² .

[0105] In one embodiment of the compounds of the present invention, X is selected from the group consisting of O, CH ₂ , OCH ₂ , CH ₂ O, CH ₂ S(O) ₂ , CONH and NHCO. In another embodiment of the compounds of the present invention, X is selected from the group consisting of O, CH ₂ , OCH ₂ , CONH and NHCO. In another embodiment of the compounds of the present invention, X is selected from the group consisting of O, OCH ₂ , and CONH. In an additional embodiment of the compounds of the present invention, X is selected from the group consisting of O, CH ₂ and OCH ₂ . In another embodiment of the compounds of the present invention, X is selected from the group consisting of CONH and NHCO. In an additional embodiment of the compounds of the present invention, X is O. In another embodiment of the compounds of the present invention, X is OCH ₂ . In an additional embodiment of the compounds of the present invention, X is CONH.

[0106] In one embodiment of the compounds of the present invention, R ² is selected from the group consisting of cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, where each R ² is optionally substituted with one or more R ⁷ . In another embodiment of the compounds of the present invention, R ² is selected from the group consisting of aryl and cycloalkyl, where each R ² is optionally substituted with one or more R ⁷ . In an additional embodiment of the compounds of the present invention, R ² is cycloalkyl, where each R ² is optionally substituted with one or more R ⁷ . In another embodiment of the compounds of the present invention, R ² is aryl optionally substituted with one or more R ⁷ . In another embodiment of the compounds of the present invention, R ² is phenyl substituted with one R ⁷ . In another embodiment of the compounds of the present invention, R ² is adamantyl or phenyl, where each R ² is optionally substituted with one or more R ⁷ . In another embodiment, R ² is adamantyl or phenyl optionally substituted with -S(O) ₂ NR ⁴ R ⁵ . In a further embodiment, R ² is adamantyl. In another embodiment, R ² is phenyl optionally substituted with -S(O) ₂ NR ⁴ R ⁵ .

[0107] In one embodiment of the compounds of the present invention, R ² is replaced by one R ⁷ . In another embodiment of the compounds of the present invention, R ² is substituted with two R ⁷ . In another embodiment of the compounds of the present invention, R ² is substituted with three R ⁷ . In another embodiment of the compounds of the present invention, R ² is substituted with four or five R ⁷ .

[0108] In one embodiment of the compounds of the present invention, R ³ is selected from the group consisting of hydrogen, C _1-6 alkyl and C _3-7 cycloalkyl; where each C _1-6 alkyl and C _3-7 cycloalkyl is optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -SO ₂ CH ₃ , -C _1-4 alkyl, -OC _1-4 alkyl , -CF ₃ , -CH ₂ CF ₃ and -O-CF _3. In another embodiment of the compounds of the present invention, R ³ is hydrogen. In an additional embodiment of the compounds of the present invention, R ³ is C _1-6 alkyl or C _3-7 cycloalkyl. In another additional embodiment of the compounds of the present invention, R ³ is hydrogen or C _1-6 alkyl. In another embodiment of the compounds of the present invention, R ³ is C _1-6 alkyl. In an additional embodiment of the compounds of the present invention, R ³ is methyl or ethyl. In another embodiment of the compounds of the present invention, R ³ is selected from the group consisting of hydrogen, methyl and ethyl.

[0109] In one embodiment of the compounds of the present invention, R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen, C _1-6 alkyl, and C _3-7 cycloalkyl; where each C _1-6 alkyl and C _3-7 cycloalkyl is optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -SO ₂ CH ₃ , -C _1-4 alkyl, -OC _1-4 alkyl , -CF ₃ , -CH ₂ CF ₃ and -O-CF _3. In another embodiment of the compounds of the present invention, R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen and C _1-6 alkyl. In another embodiment of the compounds of the present invention, R ⁴ and R ⁵ are hydrogen. In an additional embodiment of the compounds of the present invention, R ⁴ and R ⁵ are C _1-6 alkyl. In another embodiment of the compounds of the present invention, R ⁴ and R ⁵ are both methyl. In a further embodiment of the compounds of the present invention, R ⁴ and R ⁵ are both isopropyl. In one embodiment of the compounds of the present invention, R ⁴ is hydrogen and R ⁵ is isopropyl. In an additional embodiment of the compounds of the present invention, R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen and C _3-7 cycloalkyl. In another embodiment of the compounds of the present invention, R ⁴ is hydrogen and R ⁵ is C _1-6 alkyl. In one embodiment of the compounds of the present invention, R ⁴ is hydrogen and R ⁵ is methyl. In an additional embodiment of the compounds of the present invention, R ⁴ is hydrogen and R ⁵ is C _3-7 cycloalkyl.

[0110] In one embodiment of the compounds of the present invention, R ⁴ and R ⁵ , when attached to the same nitrogen atom, are combined to form a 4-7 membered ring having 0 to 1 additional heteroatoms as ring members. In a further embodiment of the compounds of the present invention, R ⁴ and R ⁵ , when attached to the same nitrogen atom, are combined to form a 4-7 membered ring having 1 additional heteroatom as ring members. In another embodiment, R ⁴ and R ⁵ , when attached to the same nitrogen atom, are combined to form a 4-7 membered ring having 0 additional heteroatoms as ring members.

[0111] In one embodiment of the compounds of the present invention, R ⁶ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl; where each C _1-6 alkyl and C _3-7 cycloalkyl is optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -C _1-4 alkyl, -OC _1-4 alkyl, -CF ₃ , - CH ₂ CF ₃ and -O-CF _3. In another embodiment, R ⁶ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl. In another embodiment, R ⁶ is C _1-6 alkyl. In an additional embodiment, R ⁶ is C _3-7 cycloalkyl.

[0112] In one embodiment of the compounds of the present invention, R ⁷ is selected from the group consisting of halo, OH, C _1-6 alkyl, OC _1-6 alkyl, C _3-7 cycloalkyl, -C(O)OR ³ , -C (O)NR ⁴ R ⁵ , -NR ⁴ C(O)R ⁶ , -S(O) ₂ NR ⁴ R ⁵ , -NR ⁴ S(O) ₂ R ⁶ and -S(O) ₂ R ⁶ ; where each C _1-6 alkyl is optionally substituted with one or more substituents selected from the group consisting of halo and -OH. In another embodiment of the compounds of the present invention, R ⁷ is selected from the group consisting of halogen, C _1-6 alkyl, -C(O)NR ⁴ R ⁵ , -S(O) ₂ NR ⁴ R ⁵ and -S(O) ₂ R ⁶ . In a further embodiment of the compounds of the present invention, R ⁷ is selected from the group consisting of -C(O)NR ⁴ R ⁵ , -S(O) ₂ NR ⁴ R ⁵ , and -S(O) ₂ R ⁶ . In another embodiment of the compounds of the present invention, R ⁷ is —S(O) ₂ NR ⁴ R ⁵ . In an additional embodiment of the compounds of the present invention, R ⁷ is —S(O) ₂ N(CH ₃ ) ₂ .

[0113] In one embodiment of the compounds of the present invention, R ⁸ is hydrogen or C _1-6 alkyl. In another embodiment of the compounds of the present invention, R ⁸ is hydrogen. In an additional embodiment of the compounds of the present invention, R ⁸ is selected from the group consisting of hydrogen, methyl and ethyl. In another embodiment of the compounds of the present invention, R ⁸ is hydrogen or methyl.

[0114] In one embodiment of the compounds of the present invention, R ⁹ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl; where each C _1-6 alkyl and C _3-7 cycloalkyl is optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -C _1-4 alkyl, -OC _1-4 alkyl, -CF ₃ , - CH ₂ CF ₃ and -O-CF _3. In another embodiment, R ⁹ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl. In another embodiment, R ⁹ is C _1-6 alkyl. In an additional embodiment, R ⁹ is C _3-7 cycloalkyl.

[0115] In one embodiment of the compounds of the present invention, R⁸ and R⁹ combined to form a 5-7-membered a ring having 0 to 1 additional heteroatoms as ring members. In another implementation of R⁸ and R⁹ combined to form a 5-7 membered ring having 1 additional heteroatom as a ring member. In another implementation of R⁸ and R⁹ combined to form a 5-7 membered ring having 0 additional heteroatoms as ring members.

[0116] In one embodiment of the compounds of the present invention, n is 0, 1, 2, 3, 4, or 5. In another embodiment of the compounds of the present invention, n is 0. In a further embodiment of the compounds of the present invention, n is 0, 1, or 2 In another embodiment of the compounds of the present invention, n is 1, 2, or 3. In another embodiment of the compounds of the present invention, n is 1 or 2. In another embodiment of the compounds of the present invention, n is 1. In another embodiment of the compounds of the present invention, n is 2 In a further embodiment of the compounds of the present invention, n is 3. In another embodiment of the compounds of the present invention, n is 4. In a further embodiment of the compounds of the present invention, n is 5. In another embodiment of the compounds of the present invention, n is 6.

[0117] In one embodiment, the present invention also relates to compounds of Formula Ia:

Formula Ia

or their pharmaceutically acceptable salt, polymorphic form, solvate, hydrate or tautomeric form; wherein:

each R ¹ is independently selected from the group consisting of XR ² , halogen, C _1-6 alkyl, OC _1-6 alkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, -CN, -C(O)OR ³ , -C(O )NR ⁴ R ⁵ , -S(O) ₂ NR ⁴ R ⁵ , -S(O) ₂ R ⁶ , -NR ⁸ C(O)R ⁹ and -NR ⁸ S(O) ₂ R ⁹ ; where each C _1-6 alkyl, aryl, heteroaryl, cycloalkyl and heterocycloalkyl is optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -SO ₂ CH ₃ , -C _1-4 alkyl, -OC _{1- 4} alkyl, -CF ₃ , -CH ₂ CF ₃ and -O-CF ₃ ;

X is selected from the group consisting of O, CH ₂ , OCH ₂ , CH ₂ O, CH ₂ S(O) ₂ , CONH and NHCO;

R ² is selected from the group consisting of cycloalkyl, heterocycloalkyl, aryl and heteroaryl; where each R ² is optionally substituted with one or more R ⁷ ;

R ³ is selected from the group consisting of hydrogen, C _1-6 alkyl and C _3-7 cycloalkyl;

R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen, C _1-6 alkyl and C _3-7 cycloalkyl; or

R ⁴ and R ⁵ , when attached to the same nitrogen atom, are combined to form a 4-7 membered ring having 0 to 1 additional heteroatoms as ring members;

R ⁶ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl;

R ⁷ is selected from the group consisting of halogen, OH, C _1-6 alkyl, OC _1-6 alkyl, C _3-7 cycloalkyl, -C(O)OR ³ , -C(O)NR ⁴ R ⁵ , -NR ⁴ C(O)R ⁶ , -S(O) ₂ NR ⁴ R ⁵ , -NR ⁴ S(O) ₂ R ⁶ and -S(O) ₂ R ⁶ ; where each C _1-6 alkyl is optionally substituted with one or more substituents selected from the group consisting of halogen and -OH;

R ⁸ is hydrogen or C _1-6 alkyl;

R ⁹ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl; where each C _1-6 alkyl and C _3-7 cycloalkyl are optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -C _1-4 alkyl, -OC _1-4 alkyl, -CF ₃ , - CH ₂ CF ₃ and -O-CF ₃ ; or

R⁸ and R⁹ combined to form a 5-7-membered a ring having 0 to 1 additional heteroatoms as ring members;

And

n is 0, 1, 2, or 3.

[0118] In one embodiment, the present invention also relates to compounds of Formula Ib:

Formula Ib

or their pharmaceutically acceptable salts or solvates; wherein:

each R ¹ is independently selected from the group consisting of halogen, C _1-6 alkyl, OC _1-6 alkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, -CN, -C(O)OR ³ , -C(O)NR ⁴ R ⁵ , -S(O) ₂ NR ⁴ R ⁵ , -S(O) ₂ R ⁶ , -NR ⁸ C(O)R ⁹ and -NR ⁸ S(O) ₂ R ⁹ ; where each C _1-6 alkyl, aryl, heteroaryl, cycloalkyl and heterocycloalkyl is optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -SO ₂ CH ₃ , -C _1-4 alkyl, -OC _{1- 4} alkyl, -CF ₃ , -CH ₂ CF ₃ and -O-CF ₃ ;

X is selected from the group consisting of O, CH ₂ , OCH ₂ , CH ₂ O, CH ₂ S(O) ₂ , CONH and NHCO;

R ² is selected from the group consisting of cycloalkyl, heterocycloalkyl, aryl and heteroaryl; where each R ² is optionally substituted with one or more R ⁷ ;

R ³ is selected from the group consisting of hydrogen, C _1-6 alkyl and C _3-7 cycloalkyl;

R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen, C _1-6 alkyl and C _3-7 cycloalkyl; or

R ⁴ and R ⁵ , when attached to the same nitrogen atom, are combined to form a 4-7 membered ring having 0 to 1 additional heteroatoms as ring members;

R ⁶ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl;

R ⁷ is selected from the group consisting of halogen, OH, C _1-6 alkyl, OC _1-6 alkyl, C _3-7 cycloalkyl, -C(O)OR ³ , -C(O)NR ⁴ R ⁵ , -NR ⁴ C(O)R ⁶ , -S(O) ₂ NR ⁴ R ⁵ , -NR ⁴ S(O) ₂ R ⁶ and -S(O) ₂ R ⁶ ; where each C _1-6 alkyl is optionally substituted with one or more substituents selected from the group consisting of halogen and -OH;

R ⁸ is hydrogen or C _1-6 alkyl;

R ⁹ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl; where each C _1-6 alkyl and C _3-7 cycloalkyl are optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -C _1-4 alkyl, -OC _1-4 alkyl, -CF ₃ , - CH ₂ CF ₃ and -O-CF ₃ ; or

R⁸ and R⁹ combined to form a 5-7-membered a ring having 0 to 1 additional heteroatoms as ring members;

And

n is 0, 1, or 2.

[0119] In one embodiment of the compounds of Formula Ib of the invention, X is selected from the group consisting of O, OCH ₂ and CONH; R ² is selected from the group consisting of adamantyl and phenyl; where each R ² is optionally substituted with one or more R ⁷ ; R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen and C _1-6 alkyl; R ⁷ is -S(O) ₂ NR ⁴ R ⁵ ; and n is 0.

[0120] In another embodiment, the present invention also relates to compounds of Formula Ic:

Formula Ic

or their pharmaceutically acceptable salts or solvates; wherein:

each R ¹ is independently selected from the group consisting of halogen, C _1-6 alkyl, OC _1-6 alkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, -CN, -C(O)OR ³ , -C(O)NR ⁴ R ⁵ , -S(O) ₂ NR ⁴ R ⁵ , -S(O) ₂ R ⁶ , -NR ⁸ C(O)R ⁹ and -NR ⁸ S(O) ₂ R ⁹ ; where each C _1-6 alkyl, aryl, heteroaryl, cycloalkyl and heterocycloalkyl is optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -SO ₂ CH ₃ , -C _1-4 alkyl, -OC _{1- 4} alkyl, -CF ₃ , -CH ₂ CF ₃ and -O-CF ₃ ;

X is selected from the group consisting of O, CH ₂ , OCH ₂ , CH ₂ O, CH ₂ S(O) ₂ , CONH and NHCO;

R ² is selected from the group consisting of cycloalkyl, heterocycloalkyl, aryl and heteroaryl; where each R ² is optionally substituted with one or more R ⁷ ;

R ³ is selected from the group consisting of hydrogen, C _1-6 alkyl and C _3-7 cycloalkyl;

R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen, C _1-6 alkyl and C _3-7 cycloalkyl; or

R ⁴ and R ⁵ , when attached to the same nitrogen atom, are combined to form a 4-7 membered ring having 0 to 1 additional heteroatoms as ring members;

R ⁶ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl;

R ⁷ is selected from the group consisting of halogen, OH, C _1-6 alkyl, OC _1-6 alkyl, C _3-7 cycloalkyl, -C(O)OR ³ , -C(O)NR ⁴ R ⁵ , -NR ⁴ C(O)R ⁶ , -S(O) ₂ NR ⁴ R ⁵ , -NR ⁴ S(O) ₂ R ⁶ and -S(O) ₂ R ⁶ ; where each C _1-6 alkyl is optionally substituted with one or more substituents selected from the group consisting of halogen and -OH;

R ⁸ is hydrogen or C _1-6 alkyl;

R ⁹ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl; where each C _1-6 alkyl and C _3-7 cycloalkyl are optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -C _1-4 alkyl, -OC _1-4 alkyl, -CF ₃ , - CH ₂ CF ₃ and -O-CF ₃ ; or

R⁸ and R⁹ combined to form a 5-7-membered a ring having 0 to 1 additional heteroatoms as ring members;

And

n is 0, 1, or 2.

[0121] In one embodiment of the compounds of Formula Ic of the invention, X is selected from the group consisting of OCH ₂ and CONH; R ² is selected from the group consisting of adamantyl and phenyl; where each R ² is optionally substituted with one or more R ⁷ ; R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen and C _1-6 alkyl; R ⁷ is -S(O) ₂ NR ⁴ R ⁵ ; and n is 0.

[0122] In another embodiment, the present invention also relates to compounds of Formula Id:

Formula ID

or their pharmaceutically acceptable salts or solvates; wherein:

each R ¹ is independently selected from the group consisting of halogen, C _1-6 alkyl, OC _1-6 alkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, -CN, -C(O)OR ³ , -C(O)NR ⁴ R ⁵ , -S(O) ₂ NR ⁴ R ⁵ , -S(O) ₂ R ⁶ , -NR ⁸ C(O)R ⁹ and -NR ⁸ S(O) ₂ R ⁹ ; where each C _1-6 alkyl, aryl, heteroaryl, cycloalkyl and heterocycloalkyl is optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -SO ₂ CH ₃ , -C _1-4 alkyl, -OC _{1- 4} alkyl, -CF ₃ , -CH ₂ CF ₃ and -O-CF ₃ ;

X is selected from the group consisting of O, CH ₂ , OCH ₂ , CONH and NHCO;

R ² is selected from the group consisting of cycloalkyl, heterocycloalkyl, aryl and heteroaryl; where each R ² is optionally substituted with one or more R ⁷ ;

R ³ is selected from the group consisting of hydrogen, C _1-6 alkyl and C _3-7 cycloalkyl;

R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen, C _1-6 alkyl and C _3-7 cycloalkyl; or

R ⁴ and R ⁵ , when attached to the same nitrogen atom, are combined to form a 4-7 membered ring having 0 to 1 additional heteroatoms as ring members;

R ⁶ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl;

R ⁷ is selected from the group consisting of halogen, OH, C _1-6 alkyl, OC _1-6 alkyl, C _3-7 cycloalkyl, -C(O)OR ³ , -C(O)NR ⁴ R ⁵ , -NR ⁴ C(O)R ⁶ , -S(O) ₂ NR ⁴ R ⁵ , -NR ⁴ S(O) ₂ R ⁶ and -S(O) ₂ R ⁶ ; where each C _1-6 alkyl is optionally substituted with one or more substituents selected from the group consisting of halogen and -OH;

R ⁸ is hydrogen or C _1-6 alkyl;

R ⁹ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl; where each C _1-6 alkyl and C _3-7 cycloalkyl are optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -C _1-4 alkyl, -OC _1-4 alkyl, -CF ₃ , - CH ₂ CF ₃ and -O-CF ₃ ; or

R⁸ and R⁹ combined to form a 5-7-membered a ring having 0 to 1 additional heteroatoms as ring members;

And

n is 0, 1, or 2.

[0123] In one embodiment of the compounds of Formula Id of the invention, X is selected from the group consisting of OCH ₂ and CONH; R ² is selected from the group consisting of adamantyl and phenyl; where each R ² is optionally substituted with one or more R ⁷ ; R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen and C _1-6 alkyl; R ⁷ is -S(O) ₂ NR ⁴ R ⁵ ; and n is 0.

[0124] In another embodiment, the present invention also relates to compounds of Formula Ie:

Formula Ie

or their pharmaceutically acceptable salts or solvates; wherein:

each R ¹ is independently selected from the group consisting of halogen, C _1-6 alkyl, OC _1-6 alkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, -CN, -C(O)OR ³ , -C(O)NR ⁴ R ⁵ , -S(O) ₂ NR ⁴ R ⁵ , -S(O) ₂ R ⁶ , -NR ⁸ C(O)R ⁹ and -NR ⁸ S(O) ₂ R ⁹ ; where each C _1-6 alkyl, aryl, heteroaryl, cycloalkyl and heterocycloalkyl is optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -SO ₂ CH ₃ , -C _1-4 alkyl, -OC _{1- 4} alkyl, -CF ₃ , -CH ₂ CF ₃ and -O-CF ₃ ;

R ³ is selected from the group consisting of hydrogen, C _1-6 alkyl and C _3-7 cycloalkyl;

R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen, C _1-6 alkyl and C _3-7 cycloalkyl; or

R ⁴ and R ⁵ , when attached to the same nitrogen atom, are combined to form a 4-7 membered ring having 0 to 1 additional heteroatoms as ring members;

R ⁶ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl;

R ⁸ is hydrogen or C _1-6 alkyl;

R ⁹ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl; where each C _1-6 alkyl and C _3-7 cycloalkyl are optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -C _1-4 alkyl, -OC _1-4 alkyl, -CF ₃ , - CH ₂ CF ₃ and -O-CF ₃ ; or

R⁸ and R⁹ combined to form a 5-7-membered a ring having 0 to 1 additional heteroatoms as ring members;

And

n is 0, 1, or 2.

[0125] In another embodiment, the present invention also relates to compounds of Formula If:

If Formula

or their pharmaceutically acceptable salts or solvates; wherein:

each R ¹ is independently selected from the group consisting of halogen, C _1-6 alkyl, OC _1-6 alkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, -CN, -C(O)OR ³ , -C(O)NR ⁴ R ⁵ , -S(O) ₂ NR ⁴ R ⁵ , -S(O) ₂ R ⁶ , -NR ⁸ C(O)R ⁹ and -NR ⁸ S(O) ₂ R ⁹ ; where each C _1-6 alkyl, aryl, heteroaryl, cycloalkyl and heterocycloalkyl is optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -SO ₂ CH ₃ , -C _1-4 alkyl, -OC _{1- 4} alkyl, -CF ₃ , -CH ₂ CF ₃ and -O-CF ₃ ;

R ³ is selected from the group consisting of hydrogen, C _1-6 alkyl and C _3-7 cycloalkyl;

R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen, C _1-6 alkyl and C _3-7 cycloalkyl; or

R ⁴ and R ⁵ , when attached to the same nitrogen atom, are combined to form a 4-7 membered ring having 0 to 1 additional heteroatoms as ring members;

R ⁶ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl;

R ⁸ is hydrogen or C _1-6 alkyl;

R ⁹ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl; where each C _1-6 alkyl and C _3-7 cycloalkyl are optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -C _1-4 alkyl, -OC _1-4 alkyl, -CF ₃ , - CH ₂ CF ₃ and -O-CF ₃ ; or

R⁸ and R⁹ combined to form a 5-7-membered a ring having 0 to 1 additional heteroatoms as ring members;

And

n is 0, 1, or 2.

[0126] In another embodiment, the present invention also relates to compounds of Formula Ig:

Ig formula

or their pharmaceutically acceptable salts or solvates; wherein:

each R ¹ is independently selected from the group consisting of halogen, C _1-6 alkyl, OC _1-6 alkyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, -CN, -C(O)OR ³ , -C(O)NR ⁴ R ⁵ , -S(O) ₂ NR ⁴ R ⁵ , -S(O) ₂ R ⁶ , -NR ⁸ C(O)R ⁹ and -NR ⁸ S(O) ₂ R ⁹ ; where each C _1-6 alkyl, aryl, heteroaryl, cycloalkyl and heterocycloalkyl is optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -SO ₂ CH ₃ , -C _1-4 alkyl, -OC _{1- 4} alkyl, -CF ₃ , -CH ₂ CF ₃ and -O-CF ₃ ;

R ³ is selected from the group consisting of hydrogen, C _1-6 alkyl and C _3-7 cycloalkyl;

R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen, C _1-6 alkyl and C _3-7 cycloalkyl; or

R ⁴ and R ⁵ , when attached to the same nitrogen atom, are combined to form a 4-7 membered ring having 0 to 1 additional heteroatoms as ring members;

R ⁶ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl;

R ⁸ is hydrogen or C _1-6 alkyl;

R ⁹ is selected from the group consisting of C _1-6 alkyl and C _3-7 cycloalkyl; where each C _1-6 alkyl and C _3-7 cycloalkyl are optionally substituted with one or more substituents selected from the group consisting of halogen, -OH, -C _1-4 alkyl, -OC _1-4 alkyl, -CF ₃ , - CH ₂ CF ₃ and -O-CF ₃ ; or

R⁸ and R⁹ combined to form a 5-7-membered a ring having 0 to 1 additional heteroatoms as ring members;

And

n is 0, 1, or 2.

[0127] In another embodiment of the compounds of Formulas Ie, If and Ig of the invention, each R ¹ is C _1-6 alkyl and n is 0 or 1.

[0128] In another embodiment of the compounds of Formula I of the invention, each R¹ independently selected from the group consisting of halogen, C_1-6alkyl, C_1-6haloalkyl, C_1-6alkyloxy, C_1-6haloalkoxy, -CN, -C(O)OR³, -C(O)NR⁴R⁵, -NR⁴C(O)R⁶, -S(O)₂NR⁴R⁵, -NR⁴S(O)₂R⁶ and -S(O)₂R⁶; R³ selected from the group consisting of hydrogen optionally substituted with C_1-6alkyl and optionally substituted C_3-7cycloalkyl; R⁴ and R⁵ independently selected from the group consisting of hydrogen optionally substituted with C_1-6alkyl and optionally substituted C_3-7cycloalkyl; or R⁴ and R⁵, when they are attached to the same nitrogen atom, are combined to form a 4-7 membered a ring having 0 to 1 additional heteroatoms as ring members; R⁶ selected from the group consisting of optionally substituted C_1-6alkyl optionally substituted with C_3-7cycloalkyl and optionally substituted C_1-6haloalkyl; and n is 0, 1, 2, or 3.

[0129] In another embodiment of the compounds of Formula I of the invention, each R ¹ is independently selected from the group consisting of halo, C _1-6 alkyl, C _1-6 haloalkyl, and —S(O) ₂ R ⁶ ; R ⁶ is C _1-6 alkyl; and n is 0, 1, or 2.

[0130] In another embodiment of the compounds of Formula I of the invention, each R ¹ is independently selected from the group consisting of chlorine, fluorine, methyl, isopropyl, OCH ₃ , phenyl, and SO ₂ CH ₃ ; and n is 1 or 2.

[0131] In an additional embodiment of the compounds of Formula I of the invention, A is and n is 0.

[0132] In the context of the present description, any one or more aspects or implementation options can be combined with any other aspects or implementation options.

[0133] Examples of compounds according to the present invention include the compounds listed in Table 2:

table 2

1 ( Z )-3-fluoro-4-(phenylsulfonyl)but-2-en-1-amine 2 ( Z )-4-((3,5-bis(trifluoromethyl)phenyl)sulfonyl)-3-fluorobut-2-en-1-amine 3 ( Z )-3-fluoro-4-((4-(methylsulfonyl)phenyl)sulfonyl)but-2-en-1-amine 4 N -((1 R ,3 R ,5 S )-adamantan-1-yl)-4-((( Z )-4-amino-2-fluorobut-2-en-1-yl)sulfonyl)benzamide 5 ( Z )-4-((2-((4-amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)methyl)- N , N- dimethylbenzenesulfonamide 6 ( Z )-4-(2-((4-Amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)- N , N- dimethylbenzenesulfonamide 7 ( Z )-4-((2-(benzyloxy)phenyl)sulfonyl)-3-fluorobut-2-en-1-amine 8 ( Z )-4-((2-((4-amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)methyl)benzenesulfonamide 9 ( Z )-4-((3-((4-amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)methyl)- N , N- dimethylbenzenesulfonamide 10 ( Z )-4-((4-((4-Amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)methyl)- N , N- dimethylbenzenesulfonamide eleven ( Z )-4-((2-((4-Amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)methyl)- N , N- diisopropylbenzenesulfonamide 12 N -((1 S ,3 R ,5 S )-adamantan-1-yl)-2-((( Z )-4-amino-2-fluorobut-2-en-1-yl)sulfonyl)benzamide 13 N -((1 S ,3 R ,5 S )-adamantan-1-yl)-3-((( Z )-4-amino-2-fluorobut-2-en-1-yl)sulfonyl)benzamide 14 ( Z )-4-((2-((4-amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)methyl)- N- isopropylbenzenesulfonamide 15 ( Z )-2-((4-Amino-2-fluorobut-2-en-1-yl)sulfonyl)- N- (4-( N , N- diisopropylsulfamoyl)phenyl)benzamide 16 ( Z )-4-(3-(4-Amino-2-fluorobut-2-enylsulfonyl)phenoxy)-N,N-dimethylbenzenesulfonamide 17 ( Z )-3-(3-(4-Amino-2-fluorobut-2-enylsulfonyl)phenoxy)-N,N-dimethylbenzenesulfonamide 18 ( Z )-3-(2-(4-Amino-2-fluorobut-2-enylsulfonyl)phenoxy)-N,N-dimethylbenzenesulfonamide 19 ( Z )-4-(2-chlorophenylsulfonyl)-3-fluorobut-2-en-1-amine 20 ( Z )-4-(biphenyl-2-ylsulfonyl)-3-fluorobut-2-en-1-amine 21 ( Z )-3-fluoro-4-(2-isopropylphenylsulfonyl)but-2-en-1-amine 22 ( Z )-3-fluoro-4-(2-methoxyphenylsulfonyl)but-2-en-1-amine 23 ( Z )-3-fluoro-4-(naphthalene-1-ylsulfonyl)but-2-en-1-amine 24 ( Z )-3-fluoro-4-(naphthalene-2-ylsulfonyl)but-2-en-1-amine 25 ( Z )-4-(2,4-dichlorophenylsulfonyl)-3-fluorobut-2-en-1-amine 26 ( Z )-4-(3-chlorophenylsulfonyl)-3-fluorobut-2-en-1-amine 27 ( Z )-4-(4-chlorophenylsulfonyl)-3-fluorobut-2-en-1-amine 28 ( Z )-4-(3,5-dichlorophenylsulfonyl)-3-fluorobut-2-en-1-amine 29 ( Z )-3-fluoro-4-(pyridin-4-ylsulfonyl)but-2-en-1-amine thirty ( Z )-3-fluoro-4-(pyridin-2-ylsulfonyl)but-2-en-1-amine 31 ( Z )-3-fluoro-4-(pyridin-3-ylsulfonyl)but-2-en-1-amine 32 ( Z )-3-fluoro-4-(quinolin-2-ylsulfonyl)but-2-en-1-amine 33 ( Z )-3-fluoro-4-(quinolin-8-ylsulfonyl)but-2-en-1-amine 34 ( Z )-3-fluoro-4-(5-isopropylpyridin-2-ylsulfonyl)but-2-en-1-amine 35 ( Z )-3-fluoro-4-(5-methylpyridin-2-ylsulfonyl)but-2-en-1-amine 36 ( Z )-3-fluoro-4-(6-methylpyridin-2-ylsulfonyl)but-2-en-1-amine 37 ( Z )-3-fluoro-4-(2-fluorophenylsulfonyl)but-2-en-1-amine 38 ( Z )-3-fluoro-4-(3-fluorophenylsulfonyl)but-2-en-1-amine 39 ( Z )-3-fluoro-4-(4-fluorophenylsulfonyl)but-2-en-1-amine 40 ( Z )-3-fluoro-4-(o-tolylsulfonyl)but-2-en-1-amine 41 ( Z )-3-fluoro-4-(m-tolylsulfonyl)but-2-en-1-amine 42 ( Z )-3-fluoro-4-tosylbut-2-en-1-amine 43 ( Z )-3-fluoro-4-(3-methylpyridin-2-ylsulfonyl)but-2-en-1-amine 44 ( Z )-3-fluoro-4-(2-methylpyridin-4-ylsulfonyl)but-2-en-1-amine 45 ( Z )-3-fluoro-4-(2-isopropylpyridin-3-ylsulfonyl)but-2-ene-1-amine 46 ( Z )-3-fluoro-4-(6-methylpyridin-3-ylsulfonyl)but-2-en-1-amine 47 ( Z )-3-fluoro-4-(2-methylpyridin-3-ylsulfonyl)but-2-en-1-amine 48 ( Z )-3-fluoro-4-(4-methylpyridin-3-ylsulfonyl)but-2-en-1-amine 49 ( Z )-3-fluoro-4-(6-isopropylpyridin-3-ylsulfonyl)but-2-en-1-amine 50 ( Z )-3-fluoro-4-((2-methylbenzo[ d ]thiazol-4-yl)sulfonyl)but-2-en-1-amine 51 ( Z )-3-fluoro-4-((3-fluoroquinolin-8-yl)sulfonyl)but-2-en-1-amine 52 ( Z )-4-((2,3-dimethyl-1H-indol-7-yl)sulfonyl)-3-fluorobut-2-en-1-amine 53 ( Z )-3-fluoro-4-((quinolin-8-yl -d ₆ )sulfonyl)but-2-en-1-amine

[0134] In one embodiment, the compound of the present invention is selected from the group consisting of And , or their pharmaceutically acceptable salts or solvates. In another embodiment, the compound of the present invention is or a pharmaceutically acceptable salt or solvate thereof. In an additional embodiment, the compound of the present invention is or a pharmaceutically acceptable salt or solvate thereof.

[0135] In one embodiment, the compound of the present invention is selected from the group consisting of And , or their pharmaceutically acceptable salts or solvates. In another embodiment, the compound of the present invention is or a pharmaceutically acceptable salt or solvate thereof. In an additional embodiment, the compound of the present invention is or a pharmaceutically acceptable salt or solvate thereof.

Preparation of Compounds of Formula I

[0136] Compounds of Formula I can be readily prepared by those skilled in the art using methods and materials known in the art and with reference to standard textbooks such as "Advanced Organic Chemistry" by Jerry March (third edition, 1985, John Wiley and Sons) or “Comprehensive Organic Transformations” by Richard C. Larock (1989, VCH Publishers).

[0137] Compounds of Formula I can be synthesized as described below. The following diagrams provide an overview of typical non-limiting embodiments of the invention. Those skilled in the art will appreciate that analogues of Formula I, including various isomeric forms, can also be prepared from similar starting materials.

Scheme 1:

[0138] The preparation of compounds described by Formula Ib, where X is -OCH ₂ -, is described in Scheme 1 below. One skilled in the art will appreciate that the compounds described by Formulas Ic, Id, Ie, If, and Ig can be prepared using similar synthetic techniques with suitable starting materials.

Scheme 1

[0139] P ¹ is a functional group used to protect the nitrogen group. Examples of P ¹ are carbamate forming groups such as tert- butyloxycarbonyl (BOC), 9-fluorenylmethyloxycarbonyl (FMOC) and benzyloxycarbonyl (CBZ) groups.

[0140] In General Scheme 1, the starting material, R ¹ -substituted hydroxythiophenol, described by Formula II, can be obtained from commercial sources or can be obtained by many methods well known in the art. Although there are many ways to achieve the reaction described by Method A, one convenient method involves reacting the compounds described by Formulas II and III with a base such as potassium carbonate in a solvent such as N,N- dimethylformamide at ambient temperature for several hours. By following standard extraction and purification methods, the product of Formula IV can be obtained in good yield and purity.

[0141] Although there are many ways to achieve the reaction described by Method B, one convenient method involves reacting the compounds described by Formulas IV and V (in which Y is a suitable leaving group such as Br, I, OT and OMs) with a base such as as potassium carbonate in a solvent such as N,N- dimethylformamide at ambient temperature for several hours. The product described by Formula VI can be recovered by standard processing methods.

[0142] One convenient method for converting Formula VI compounds to Formula VII compounds is Method C, which involves treating a solution of a Formula VI compound and a base such as sodium hydrogen carbonate in a solvent such as dichloromethane, an oxidizing agent such as m CPBA (3-chloroperoxybenzoic acid) at 0 °C to ambient temperature for several hours. The product described by Formula VII can be recovered by standard processing methods.

[0143] There are many well-established chemical methods for deprotecting compounds described by Formula VII to compounds described by Formula Ib (Method D). For example, if P ¹ is a BOC protecting group, compounds described by Formula VII can be treated with an acidic agent such as dry hydrogen chloride in a solvent such as diethyl ether to give compounds described by Formula Ib as hydrochloride salts. Typically, free amino compounds are converted to acid addition salts for ease of handling and to improve chemical stability. Examples of acid addition salts include, but are not limited to, hydrochloride, hydrobromide, 2,2,2-trifluoroacetate and methanesulfonate salts.

Scheme 2:

[0144] The preparation of compounds described by Formula Ib, where X is -CONH-, is described in Scheme 2 below. One skilled in the art will appreciate that the compounds described by Formulas Ic, Id, Ie, If, and Ig can be prepared using similar synthetic techniques with suitable starting materials.

Scheme 2

[0145] In General Scheme 2, the starting material, R ¹ -substituted mercaptobenzoic acid, can be obtained from commercial sources or can be obtained by many methods well known in the art.

[0146] Compounds described by Formula XI can be prepared by reacting an appropriately substituted benzoic acid moiety (described by Formula IX) with an amine moiety (Formula X) in the presence of a suitable coupling reagent (such as HATU) and a base (such as triethylamine) in a solvent, such as N,N- dimethylformamide at ambient temperature for several hours (Method E). The product described by Formula XI can be recovered using standard processing methods.

Scheme 3:

[0147] The preparation of compounds described by Formula Ib, where X is -O-, is described in Scheme 3 below. One skilled in the art will appreciate that the compounds described by Formulas Ic, Id, Ie, If, and Ig can be prepared using similar synthetic techniques with suitable starting materials.

Scheme 3

[0148] In General Scheme 3, the starting material, R ¹ -substituted hydroxythiophenol, can be obtained from commercial sources or can be obtained by many methods well known in the art.

[0149] A modification of the copper-catalyzed Ullman reaction can be used to couple the compounds described by Formulas IV and XIII (Method F). Numerous variants of this type of reaction are described in the literature, one of which is the Chan-Evans-Lam modification. The compounds described by Formulas IV and XIII can be dissolved in a solvent such as dichloromethane in the presence of pyridine and then treated with copper (II) acetate at ambient temperature for several hours. By following standard extraction and purification procedures, the bound product described by Formula XIV can be obtained in good yield and purity.

Scheme 4:

[0150] The preparation of compounds described by Formula Ia is described in Scheme 4 below.

Scheme 4

[0151] In General Scheme 4, the starting material, R ¹ -substituted thiol, can be obtained from commercial sources or can be obtained by many methods well known to those skilled in the art.

Scheme 5:

[0152] The preparation of compounds described by Formula Ia is described in Scheme 5 below.

Scheme 5

[0153] In general Scheme 5, the starting material, R ¹ -substituted arylsulfinate, described by Formula XVIII, can be obtained from commercial sources or can be obtained by many methods well known to specialists in this field of technology. One convenient method for achieving the conversion described by Method E involves reacting the compounds described by Formulas XVIII and III with a base such as potassium carbonate in a solvent such as N,N- dimethylformamide at ambient temperature for several hours. By following standard extraction and purification procedures, the product described by Formula XVII can be obtained in good yield and purity.

[0154] The person skilled in the art will understand that the compounds of Formula I, where A is heteroaryl, can be obtained by methods similar to those described above.

[0155] The cis / trans ( E/Z ) isomers can be separated by conventional methods well known to those skilled in the art, such as chromatography and fractional crystallization.

Therapeutic Uses and Formulations

[0156] Another aspect of the present invention relates to a pharmaceutical composition containing a compound of Formula I, or a pharmaceutically acceptable salt or stereoisomer thereof, together with a pharmaceutically acceptable diluent, excipient, or adjuvant.

[0157] The present invention also relates to the use of compounds of Formula I in therapy, in particular for the inhibition of members of the lysyl oxidase family, LOX, LOXL1, LOXL2, LOXL3 and LOXL4. In one embodiment, the invention provides selective inhibition of certain lysyl oxidase isoenzymes. In another embodiment, the invention provides simultaneous inhibition of 2, 3 or 4 LOX isoenzymes. The relative inhibitory abilities of compounds can be determined by the amount needed to inhibit the amine oxidase activity of LOX, LOXL1, LOXL2, LOXL3 and LOXL4 in various ways, for example, in an in vitro assay with a recombinant or purified human protein or with a recombinant or purified non-human enzyme, in cellular assays, expressing the normal rodent enzyme, in cell assays that have been transfected with human protein, in in vivo tests on rodents and other mammalian species, and the like.

[0158] In one embodiment, the compounds of the present invention are inhibitors of long-acting lysyl oxidase family members LOX, LOXL1, LOXL2, LOXL3, and LOXL4. In one embodiment, the compounds of the present invention are long acting inhibitors of the LOX or LOXL1-4 enzymes if the inhibition continues to exceed 50% of the activity of the LOX or LOXL1-4 enzymes after the concentration of the compound has been reduced below the IC50. In one embodiment, the compounds of the present invention exhibit sustained inhibition of LOX or LOXL1-4 enzymes for 24 hours. In one embodiment, the compounds of the present invention are irreversible inhibitors of the lysyl oxidase family members LOX, LOXL1, LOXL2, LOXL3 and LOXL4.

[0159] Accordingly, a further aspect of the invention is directed to a method of inhibiting the activity of an amine oxidase of any 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 Formula I, or a pharmaceutically acceptable salt or solvate thereof, or its pharmaceutical composition.

[0160] In one embodiment, the present invention is directed to a method for inhibiting LOXL2 amine oxidase activity. In another embodiment, the present invention is directed to the inhibition of LOX and LOXL2 amine oxidase activity. In a further embodiment, the present invention is directed to a method for inhibiting LOX amine oxidase activity.

[0161] As previously discussed, the LOX and LOXL1-4 enzymes are members of a large family of flavin-dependent and copper-dependent amine oxidases, which includes SSAO/VAP-1, monoamine oxidase-B (MAO-B), and diamine oxidase (DAO). In one embodiment, the compounds of the present invention selectively inhibit members of the lysyl oxidase isoenzyme family against SSAO/VAP-1, MAO-B, DAO, and other members of the amine oxidase family.

[0162] The present invention also describes methods of using the compounds described by Formula I to inhibit one or more lysyl oxidase isoenzymes (LOX, LOXL1, LOXL2, LOXL3 and LOXL4) in patients suffering from fibrotic disease and methods of treating fibrotic diseases. In addition, the present invention describes methods of using the compounds described by Formula I to inhibit one or more lysyl oxidase isoenzymes (LOX, LOXL1, LOXL2, LOXL3 and LOXL4) in patients suffering from cancer, including metastatic cancer, and methods of treating cancer and metastatic cancer.

[0163] In a further aspect of the invention, there is provided a method for treating a condition associated with any of the LOX, LOXL1, LOXL2, LOXL3, or LOXL4 proteins, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I, or a pharmaceutically acceptable salt or solvate thereof, or its pharmaceutical composition.

[0164] In another aspect, there is provided a method for treating a condition modulated by any of LOX, LOXL1, LOXL2, LOXL3, or LOXL4, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical formulation thereof. compositions.

[0165] In one embodiment of the methods of the present invention, the condition is selected from the group consisting of fibrosis, cancer, and angiogenesis.

[0166] In another aspect, the present invention provides a method of reducing extracellular matrix formation by treating human, domestic, and bovine subjects with fluoroallylamine inhibitors of the Formula I lysyl oxidase family of isoenzymes as described herein.

[0167] The above methods are applicable to fibrosis. As used herein, the term "fibrosis" includes diseases such as cystic fibrosis, idiopathic pulmonary fibrosis, liver fibrosis, renal fibrosis, scleroderma, radiation-induced fibrosis, Peyronie's disease, scarring, and other diseases in which excessive fibrosis contributes to the pathology of the disease.

[0168] In one embodiment, fibrosis is selected from the group consisting of mediastinal fibrosis, myelofibrosis, retroperitoneal fibrosis, progressive massive fibrosis, nephrogenic systemic fibrosis, Crohn's disease, keloid, systemic sclerosis, arthrofibrosis, Dupuytren's contracture, adhesive capsulitis, pancreatic fibrosis, intestinal fibrosis, liver fibrosis, pulmonary fibrosis, kidney 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/keloid , surgical scarring, cardiac arrest, and all conditions in which excessive or aberrant fibrous material deposition is associated with disease, injury, implants, or surgery. In another embodiment, fibrosis is selected from the group consisting of liver fibrosis, pulmonary fibrosis, renal fibrosis, cardiac fibrosis, scarring, and scleroderma. In a further embodiment, fibrosis is selected from the group consisting of myelofibrosis, systemic sclerosis, liver fibrosis, pulmonary fibrosis, renal fibrosis, cardiac fibrosis, and radiation-induced fibrosis.

[0169] 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 mesangiocapillary glomerulonephritis t. In one embodiment, liver fibrosis leads to cirrhosis and includes associated conditions such as chronic viral hepatitis, non-alcoholic fatty liver disease (NAFLD), alcoholic steatohepatitis (ASH), non-alcoholic steatohepatitis (NASH), primary biliary cirrhosis (PBC), biliary cirrhosis and autoimmune hepatitis.

[0170] In one embodiment, fibrosis is selected from keloids, scars, ocular scars, hypertrophic scars, scleroderma, Dupuytren's contracture, and Peyronie's disease. In one embodiment, hypertrophic scarring results from a burn. In one embodiment, hypertrophic scarring is caused by external lesions. In another embodiment, the hypertrophic scarring is caused by surgical procedures. In one embodiment, the implementation of the keloid is caused by external damage. In another embodiment, the keloid is caused by surgical procedures. In a further embodiment, the keloid results from skin damage caused by acne, burns, chickenpox, ear piercing, scrapes, surgical cuts, or vaccination sites.

[0171] 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; colorectal cancer; anal cancer; pancreatic cancer; prostate cancer; ovarian cancer; carcinomas of the liver and bile ducts; esophageal carcinomas; mesothelioma, non-Hodgkin's lymphoma; bladder carcinomas; uterine carcinomas; gliomas, glioblastomas, medullablastomas and other brain tumors; myelofibrosis, kidney cancer; head and neck cancer; stomach cancer; multiple myeloma; testicular cancer; germ cell tumors; neuroendocrine tumor; cervical cancer; cancer of the oral cavity, carcinoids of the gastrointestinal tract, breast and other organs; cricoid cell carcinomas; mesenchymal tumors, including sarcomas, fibrosarcomas, hemangioma, angiomatosis, hemangiopericytoma, pseudoangiomatous stromal hyperplasia, myofibroblastoma, fibromatosis, inflammatory myofibroblastic tumor, lipoma, angiolipoma, granular cell tumor, neurofibroma, schwannomas, angiosarcoma, liposarcoma, rhabdomyo sarcoma, osteosarcoma, leiomyoma, or leiomyosarcoma .

[0172] In one embodiment, the cancer is selected from the group consisting of breast cancer, squamous head and neck cancer, brain cancer, prostate cancer, renal cell carcinoma, liver cancer, lung cancer, oral cancer, cervical cancer, and tumor metastases. .

[0173] In one embodiment, lung cancer includes lung adenocarcinoma, squamous cell carcinoma, large cell carcinoma, bronchoalveolar 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, islet cell carcinoma, and neuroendocrine tumors.

[0174] In one embodiment, the ovarian carcinoma includes an epithelial ovarian carcinoma or a superficial epithelial stromal tumor, including a serous tumor, an endometrioid tumor, and a mucinous cystadenocarcinoma, and an ovarian sex cord stromal tumor. In one embodiment, liver and bile duct carcinoma includes hepatocellular carcinoma, cholangiocarcinoma, and hemangioma. In one embodiment, esophageal carcinoma includes esophageal adenocarcinoma and squamous cell carcinoma. In one embodiment, uterine carcinoma includes endometrial adenocarcinoma, papillary serous carcinoma, uterine clear cell carcinoma, uterine sarcomas and leiomyosarcomas, and mixed Mueller tumors. In one embodiment, kidney cancer includes renal cell carcinoma, clear cell carcinoma, and Wilm's tumor. In one embodiment, the head and neck cancer includes squamous cell carcinomas. In one embodiment, gastric cancer includes gastric adenocarcinoma and stromal tumor of the gastrointestinal tract.

[0175] In one embodiment, the cancer is selected from the group consisting of colon cancer, ovarian cancer, lung cancer, esophageal carcinoma, breast cancer, and prostate cancer. In one embodiment, the cancer is selected from the group consisting of pancreatic cancer, liver cancer, breast cancer, myelofibrosis, and mesothelioma.

[0176] In one embodiment, the compounds of the present invention can be used to treat non-metastatic cancer. In another implementation, the compounds of the present invention can be used to treat metastatic cancer. In an additional embodiment, the compounds of the present invention can be used to prevent or treat tumor metastases.

[0177] The above methods are applicable when the condition is angiogenesis.

[0178] In one embodiment of the methods of the present invention, the subject is selected from the group consisting of humans, domestic animals, and livestock. In another embodiment of the methods of the present invention, the subject is a human.

[0179] A further aspect of the invention provides for the use of a compound of Formula I, or a pharmaceutically acceptable salt or solvate thereof, for the manufacture of a medicament for the treatment of a condition associated with any of the LOX, LOXL1, LOXL2, LOXL3, or LOXL4 proteins.

[0180] Another aspect of the invention provides for the use of a compound of Formula I, or a pharmaceutically acceptable salt or solvate thereof, for the manufacture of a medicament for the treatment of a condition modulated by any of LOX, LOXL1, LOXL2, LOXL3, or LOXL4.

Pharmaceutical and/or therapeutic formulations

[0181] In another embodiment, the present invention provides compositions comprising a compound of Formula I and at least one pharmaceutically acceptable excipient, carrier, or diluent thereof. The compound(s) of Formula I may also be present as suitable salts, including pharmaceutically acceptable salts.

[0182] The phrase "pharmaceutically acceptable carrier" refers to any carrier known to those skilled in the art as suitable for a particular route of administration. In addition, the compounds may be formulated as the sole pharmaceutically active ingredient in a composition, or may be combined with other active ingredients.

[0183] The phrase "pharmaceutically acceptable salt" refers to any salt preparation that is suitable for use in pharmaceuticals. By pharmaceutically acceptable salt is meant those salts which, within the framework of sound medical judgment, are suitable for use in contact with tissues of humans and lower animals without undue toxicity, irritation, allergic reaction, etc., and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art and include acid and base addition salts. Hemi-salts of acids and bases can also be formed. Pharmaceutically acceptable salts include mineral acid amine salts (eg, hydrochlorides, hydrobromides, sulfates, and the like); and amine salts of organic acids (eg, formates, acetates, lactates, malates, tartrates, citrates, ascorbates, succinates, maleates, butyrates, valerates, fumarates, and the like).

[0184] For compounds of Formula (I) having a basic site, suitable pharmaceutically acceptable salts may be acid addition salts. For example, suitable pharmaceutically acceptable salts of such compounds 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, oxalic acid, carbonic acid, tartaric acid or citric acid with the compounds of the invention.

[0185] S.M. Berge et al. describe in detail pharmaceutically acceptable salts in J. Pharmaceutical Sciences, 1977, 66 :1-19. Salts can be generated in situ during the final isolation and purification of the compounds of the invention, or separately by reacting the free base functional group with a suitable organic acid. Exemplary 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-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, 3-phenylpropionate, phosphate, picrate , pivalate, propionate, stearate, succinate, sulfate, tartrate, toluenesulfonate, undecanoate, valerate salts, and the like. Suitable base salts are formed from bases which form non-toxic salts. Examples include salts of arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine and zinc. 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, ethylamine, triethanolamine and the like.

[0186] Pharmaceutically acceptable salts of compounds of Formula I can be prepared by methods known to those skilled in the art, including, for example:

(i) by reacting a compound of formula I with the desired acid or base;

(ii) by removing the acid- or base-labile protecting group from a suitable precursor of the compound of formula I, or by ring-opening a suitable cyclic precursor, eg lactone or lactam, with the desired acid or base; or

(iii) by converting one salt of a compound of formula I into another by reaction with a suitable acid or base, or by using a suitable ion exchange column.

[0187] The above reactions (i)-(iii) are usually carried out in solution. The resulting salt may precipitate and be collected by filtration, or may be isolated by evaporating the solvent. The degree of ionization of the resulting salt can vary from fully ionized to almost non-ionized.

[0188] Thus, for example, suitable pharmaceutically acceptable salts of the compounds of the present 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 the compounds of the invention. Suitable pharmaceutically acceptable salts of the compounds of the present invention therefore include acid addition salts.

[0189] The compounds of the invention may exist in both unsolvated and solvated forms. The term "solvate" is used herein to describe a molecular complex containing a compound of the invention and a stoichiometric amount of one or more pharmaceutically acceptable solvent molecules, such as ethanol. The term "hydrate" is used when the solvent is water.

[0190] In one embodiment, the compounds of Formula I can be administered in the form of a "prodrug". The phrase "prodrug" refers to a compound that, when administered in vivo, is metabolized in one or more steps or processes or otherwise converted to a biologically, pharmaceutically or therapeutically active form of the compound. Prodrugs can be prepared by modifying the functional groups present on a compound such that the modifications are cleaved either by conventional means or in vivo to the compound described herein. For example, prodrugs include compounds of the present invention in which a hydroxy, amino, or carbohydrate group is linked to any group that, when administered to a mammalian subject, can be cleaved to form a free hydroxyl, free amino, or free carboxylic acid group, respectively. Representative prodrugs include, for example, amides, esters, enol ethers, enol esters, acetates, formates, benzoate derivatives, and the like. alcohol and amine functional groups in the compounds of the present invention. The prodrug form can be selected from functional groups such as -C(O)alkyl, -C(O)cycloalkyl, -C(O)aryl, -C(O)-arylalkyl, C(O)heteroaryl, -C(O) -heteroarylalkyl or the like. Based on knowledge of pharmacodynamic processes and in vivo drug metabolism, once the pharmaceutically active compound is known, prodrugs of the compound can be developed by those skilled in the art (see, for example , Nogrady (1985) Medicinal Chemistry A Biochemical Approach , Oxford University Press , New York, pages 388–392).

[0191] the Compositions in this document contain one or more of the compounds presented in this document. In one embodiment, the compounds are formulated into suitable pharmaceutical preparations, such as solutions, suspensions, tablets, creams, gels, dispersible tablets, pills, capsules, powders, sustained release formulations, or elixirs, for oral administration or in sterile solutions or suspensions for parenteral administration, as well as transdermal patch formulations and dry powder inhalers. In one embodiment, the compounds described above are formulated into pharmaceutical compositions using methods and procedures well known in the art (see, for example , Ansel Introduction to Pharmaceutical Dosage Forms, Fourth Edition 1985, 126).

[0192] In the compositions, effective concentrations of one or more compounds or pharmaceutically acceptable derivatives thereof are mixed with a suitable pharmaceutical carrier. The compounds may be derivatized as the appropriate salts, esters, enol ethers or esters, acetals, ketals, orthoesters, hemiacetals, hemiketals, acids, bases, solvates, hydrates or prodrugs prior to formulation as described above. The concentrations of the compounds in the compositions are effective to deliver an amount, when administered, that treats, prevents, or alleviates one or more symptoms of the diseases or disorders being treated.

[0193] In one embodiment, the compositions are formulated to administer a single dose. To formulate the composition, the mass fraction of the compound is dissolved, suspended, dispersed or otherwise mixed in the selected carrier at an effective concentration so that the cured condition is alleviated, prevented or alleviated one or more symptoms.

[0194] The active compound is incorporated in a pharmaceutically acceptable carrier in an amount sufficient to provide a therapeutically beneficial effect in the absence of undesirable side effects in the patient being treated. A therapeutically effective concentration can be determined empirically by testing the compounds in the in vitro and in vivo systems described herein and then extrapolated from there to human doses.

[0195] The concentration of the active compound in the pharmaceutical composition will depend on the rate of absorption, distribution, inactivation and elimination of the active compound, the physicochemical characteristics of the compound, the dosage regimen and the amount administered, as well as other factors known to those skilled in the art.

[0196] In one embodiment, the implementation of a therapeutically effective dose should provide a serum concentration of the active ingredient from about 0.1 ng/ml to about 50-100 μg/ml. In another embodiment, the pharmaceutical compositions should provide a dose of about 0.001 mg to about 2000 mg of the compound per kilogram of body weight per day. Pharmaceutical unit dosage forms are prepared to provide from about 0.01 mg, 0.1 mg, or 1 mg to about 500 mg, 1000 mg, or 2000 mg, and in one embodiment, from about 10 mg to about 500 mg of the active ingredient or combination of essential ingredients per unit dosage form.

[0197] Dosing may occur at intervals of minutes, hours, days, weeks, months, or years, or continuously during any of these periods. Suitable dosages range from about 0.1 ng per kg of body weight to 0.1 g per kg of body weight per dose. The dosage is preferably in the range of 10 μg to 0.1 g per kg of body weight per dose, for example in the range of 0.1 mg to 0.01 g per kg of body weight per dose. Accordingly, the dose is in the range of 10 μg to 50 mg per kg of body weight per dose, for example, 10 μg to 20 mg per kg of body weight per dose, or 10 μg to 10 mg per kg of body weight per dose. Other suitable dosages may be in the range of 0.1 mg to 25 mg per kg of body weight, including 0.1 mg to 10, 20, 50 or 100 mg per kg of body weight per dose.

[0198] Alternatively, an effective dose may be up to about 10 mg/cm ² , or may be up to about 1 mg/cm ² , about 0.5 mg/cm ² , about 0.2 mg/cm ² , about 0.1 mg /cm ² , about 0.05 mg/cm ² , about 0.02 mg/cm ² or about 0.01 mg/cm ² . An effective dose may be, for example, in the range of about 10 µg/cm ² to about 1 mg/cm ² , about 10 µg/cm ² to about 0.1 mg/cm ² , about 10 µg/cm ² to about 0.01 mg/cm ² , about 10 µg/cm ² to about 500 µg/cm ² , about 10 µg/cm ² to about 200 µg/cm ² , about 10 µg/cm ² to about 100 µg/ cm ² , from about 10 µg/cm ² to about 50 µg/cm ² , from about 20 µg/cm ² to about 1 mg/cm ² , from about 50 µg/cm ² to about 1 mg/cm ² , from about 100 µg/cm ² to about 1 mg/cm ² , about 200 µg/cm ² to about 1 mg/cm ² , about 500 µg/cm ² to about 1 mg/cm ² , from about 50 µg/cm ² up to about 500 µg/cm ² , from about 50 µg/cm ² to about 200 µg/cm ² , from about 100 µg/cm ² to about 500 µg/cm ² , or from about 200 µg/cm ² to about 500 µg/ cm ² .

[0199] Suitable dose numbers and dosage regimens may be determined by the attending physician and may depend on the particular condition being treated, the severity of the condition, as well as the general health, age and weight of the subject.

[0200] In cases where compounds exhibit insufficient solubility, methods for solubilizing compounds can be used. Such methods are known to those skilled in the art and include, but are not limited to, the use of co-solvents such as dimethyl sulfoxide (DMSO), the use of surfactants such as ^TWEEN® , dissolution in aqueous sodium bicarbonate, compounding, of interest in the form of nanoparticles and the like. Derivatives of the compounds, such as prodrugs of the compounds, can also be used in the formulation of effective pharmaceutical compositions.

[0201] By mixing or adding the compound(s), the resulting mixture may be a solution, suspension, emulsion, and the like. The form of the resulting mixture depends on a number of factors, including the intended route of administration and the solubility of the compound in the chosen carrier or solvent. An effective concentration is sufficient to alleviate the symptoms of the disease, disorder or condition being treated and can be determined empirically.

[0202] Pharmaceutical compositions are intended for administration to humans and animals 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 suitable amounts of the compound or their pharmaceutically acceptable derivatives. The pharmaceutically therapeutically active compounds and their derivatives are in one embodiment formulated and administered in single dosage forms or multiple dosage forms. The active ingredient may be administered all at once or may be divided into several smaller doses for administration at regular intervals. As used herein, unit dosage forms refer to physically discrete units suitable for humans and animals and individually packaged as is known in the art. Each single dose contains a predetermined amount of a therapeutically active compound sufficient to obtain the desired therapeutic effect, in combination with the necessary pharmaceutical carrier, diluent or diluent. Examples of unit dosage forms include ampoules and syringes and individually packaged tablets or capsules. Single dosage forms may be administered in fractional or multiple doses. A multiple dosage form is a plurality of identical single dose forms packaged in a single container for administration in a separate single dose form. Examples of multiple dose forms include vials, tablets or capsules, or pints or gallons. Therefore, the multiple dose form is a plurality of single doses that are not separated in the package.

[0203] The actual methods for preparing such dosage forms are known or will be apparent to those skilled in the art; for example, see Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 15th Edition, 1975.

[0204] Dosage forms or compositions can be prepared containing the active ingredient in the range from 0.005% to 100% (wt.%), with the rest being a non-toxic carrier. Methods for preparing these compositions are known to those skilled in the art. Intended compositions may contain 0.001% -100% (wt.%) of the active ingredient, in one embodiment, the implementation of 0.1-95% (wt.%), in another embodiment, 75-85% (wt.%).

Modes of administration

[0205] Convenient routes of administration include injection (subcutaneous, intravenous, etc.), oral administration, inhalation, transdermal application, topical creams or gels or powders, 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 environmental conditions that may inactivate the therapeutic activity of the compound. The compound can also be administered parenterally or intraperitoneally.

Compositions for oral administration

[0206] Oral pharmaceutical dosage forms may be solid, gel, or liquid. Solid dosage forms are tablets, capsules, granules and free flowing powders. Types of oral tablets include compressed chewable lozenges and tablets which may be enteric coated, sugar coated or film coated. Capsules may be hard or soft gelatin capsules, while granules and powders may be provided in non-effervescent or effervescent form with a combination of other ingredients known to those skilled in the art.

Solid compositions for oral administration

[0207] In some embodiments, the formulations are solid dosage forms, in one embodiment, capsules or tablets. Tablets, pills, capsules, troche, etc. may contain one or more of the following ingredients or compounds of a similar nature: a binder; lubricating; diluent; sliding; baking powder; dye; sweetener; flavor; wetting agent; vomit cover; and film coating. Examples of binders include microcrystalline cellulose, gum tragacanth, glucose solution, acacia mucus, gelatin solution, molasses, polyvinylpyrrolidine, povidone, crospovidones, 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 sodium croscarmellose, sodium starch glycolate, alginic acid, corn starch, potato starch, bentonite, methylcellulose, agar, and carboxymethylcellulose. Colors include, for example, any approved certified water-soluble colors FD and C, mixtures thereof; and water insoluble FD and C dyes suspended on alumina hydrate. Sweeteners include sucrose, lactose, mannitol and artificial sweeteners such as saccharin and any number of spray dried flavors. Flavors include natural flavors extracted from plants, such as fruits, and synthetic mixtures of compounds that provide a pleasant sensation, such as, but not limited to, 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 phthalates. Film coatings include hydroxyethylcellulose, sodium carboxymethylcellulose, polyethylene glycol 4000, and cellulose acetate phthalate.

[0208] The compound or a pharmaceutically acceptable derivative thereof may be presented in a composition that protects it from the acidic environment of the stomach. For example, the composition may be formulated as an enteric coating that retains its integrity in the stomach and releases the active compound in the intestine. The composition may also be prepared in combination with an antacid or other similar ingredient.

[0209] When the unit dosage form is a capsule, it may contain, in addition to the material of the above type, a liquid carrier such as fatty oil. In addition, unit dosage forms may contain various other materials that modify the physical form of the dosage unit, such as coatings of sugar and other enteric agents. The compounds may also be administered as a component of an elixir, suspension, syrup, implant capsule, powder, chewing gum, and the like. The syrup may contain, in addition to active compounds, sucrose as a sweetening agent, and certain preservatives, dyes, colors and flavors.

[0210] Active materials can also be mixed with other active materials that do not interfere with the desired effect, or with materials that complement the desired effect, such as antacids, H2 blockers, and diuretics. The active ingredient is a compound or a pharmaceutically acceptable derivative thereof, as described herein. Higher concentrations up to about 98% by weight of the active ingredient may be included.

[0211] In all embodiments, tablet and capsule formulations may be coated, known to those skilled in the art, to modify or maintain dissolution of the active ingredient. For example, they can be coated with conventional enteric coatings such as phenyl salicylate, waxes and cellulose acetate phthalate.

Liquid compositions for oral administration

[0212] 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.

[0213] Liquid pharmaceutically administered compositions can, for example, be prepared by dissolving, dispersing or otherwise mixing the active compound as defined above and optional pharmaceutical adjuvants in a carrier such as, for example, water, saline, aqueous dextrose, glycerol, glycols, ethanol, etc. to form a solution or suspension. If desired, the pharmaceutical composition for administration may also contain minor amounts of non-toxic excipients such as wetting agents, emulsifying agents, solubilizing agents, pH buffering agents and the like, for example, acetate, sodium citrate, cyclodextrin derivatives, sorbitan monolaurate, sodium acetate triethanolamine. , triethanolamine oleate and other similar agents.

[0214] Elixirs are clear sweetened hydroalcoholic preparations. Pharmaceutically acceptable carriers for use in elixirs include solvents. Syrups are concentrated aqueous solutions of sugar, such as sucrose, and may contain a preservative. An emulsion is a two-phase system in which one liquid is dispersed as small beads in another liquid. Pharmaceutically acceptable carriers used in emulsions are non-aqueous liquids, emulsifying agents and preservatives. Suspensions use pharmaceutically acceptable suspending agents and preservatives. Pharmaceutically acceptable substances used in non-effervescent granules to be reconstituted into a liquid oral dosage form include diluents, sweeteners and wetting agents. Pharmaceutically acceptable substances used in effervescent granules to be reconstituted into a liquid oral dosage form include organic acids and a source of carbon dioxide. Dyes and flavors are used in all of the above dosage forms.

[0215] Solvents include glycerin, sorbitol, ethyl alcohol, and syrup. Examples of preservatives include glycerin, methyl and propyl paraben, benzoic acid, sodium benzoate and ethanol. Examples of non-aqueous liquids used in emulsions include mineral oil and cottonseed oil. Examples of emulsifiers include gelatin, gum arabic, tragacanth, bentonite, and surfactants such as polyoxyethylene sorbitan monooleate. Suspending agents include sodium carboxymethylcellulose, pectin, tragacanth, Veegum, and gum arabic. Sweeteners include sucrose, syrups, 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 and tartaric acids. Sources of carbon dioxide include sodium bicarbonate and sodium carbonate. Colors include any approved certified FD and C water soluble colors and mixtures thereof. Flavors include natural flavors extracted from plants, such as fruits, and synthetic blends of compounds that provide a pleasant taste sensation.

[0216] For a solid dosage form, a 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, a solution, eg in polyethylene glycol, may be diluted with a sufficient amount of a pharmaceutically acceptable liquid carrier, eg water, to be readily metered for administration.

[0217] Alternatively, liquid or semi-solid 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 vehicles and encapsulating these solutions or suspensions in solid or soft gelatin capsule shells. Other useful formulations include those disclosed in US Pat. Nos. RE28,819 and 4,358,603. 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, where 350, 550 and 750 refer to the approximate average molecular weight of polyethylene glycol, and one or more antioxidants such as butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA ), propyl gallate, vitamin E, hydroquinone, hydroxycoumarins, ethanolamine, lecithin, cephalin, ascorbic acid, malic acid, sorbitol, phosphoric acid, thiodipropionic acid and its esters, and dithiocarbamates.

[0218] Other formulations include, but are not limited to, aqueous-alcoholic solutions, including a pharmaceutically acceptable acetal. The alcohols used in these formulations are any 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, lower alkyl aldehyde di(lower alkyl) acetals such as acetaldehyde diethyl acetal.

Injections, solutions and emulsions

[0219] This document also discusses parenteral administration in one implementation, characterized by injection subcutaneously, intramuscularly or intravenously. Injectable preparations may be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for dissolution or suspension in a liquid prior to injection, or as emulsions. Injections, 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 pharmaceutical compositions administered may also contain minor amounts of non-toxic excipients such as wetting or emulsifying agents, pH buffering agents, stabilizers, solubility enhancers and other such agents, such as, for example, sodium acetate, sorbitan monolaurate, triethanolamine oleate. and cyclodextrins.

[0220] This document also discusses the implantation of a system with a slow or sustained release, so that a constant dosage level is maintained. Briefly, the compound provided herein is dispersed in a solid internal matrix, e.g. , polydimethylsiloxanes, silicone carbonate copolymers, hydrophilic polymers such as hydrogels of acrylic and methacrylic acid esters, collagen, cross-linked polyvinyl alcohol and cross-linked partially hydrolyzed polyvinyl acetate which is surrounded by an outer polymeric membrane such as polyethylene, polypropylene, ethylene/propylene copolymers, copolymers ethylene/ethyl acrylate, ethylene/vinyl acetate copolymers, silicone rubbers, polydimethylsiloxanes, neoprene rubber, chlorinated polyethylene, polyvinyl chloride, vinyl chloride-vinyl acetate copolymers, vinylidene chloride, ethylene and propylene, ionomer polyethylene terephthalate, butyl rubber epichlorohydrin rubbers, ethylene/vinyl alcohol copolymer, ter ethylene/vinyl acetate polymer /vinyl alcohol and an ethylene/vinyloxyethanol copolymer which is insoluble in body fluids. The compound diffuses through the outer polymeric membrane during the release rate control step. The percentage of active compound contained in such parenteral compositions is highly dependent on its particular nature, as well as the activity of the compound and the needs of the patient.

[0221] Parenteral administration of compositions includes intravenous, subcutaneous, and intramuscular administration. Preparations for parenteral administration include sterile solutions ready for injection, sterile dry soluble products such as lyophilized powders ready to be mixed with a solvent immediately before use, including tablets for subcutaneous injection, sterile suspensions ready for injection, sterile dry insoluble products prepared for use in combination with a solvent immediately before use and sterile emulsions. Solutions may be aqueous or non-aqueous.

[0222] When administered intravenously, suitable carriers include saline or phosphate buffered saline (PBS), and solutions containing thickening and solubilizing agents such as glucose, polyethylene glycol and polypropylene glycol, and mixtures thereof.

[0223] Pharmaceutically acceptable carriers used in parenteral formulations include aqueous solvents, non-aqueous solvents, antimicrobial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, emulsifying agents, sequestering or chelating agents, and other pharmaceutically acceptable substances.

[0224] Examples of aqueous vehicles include sodium chloride injection, Ringer's injection, isotonic dextrose injection, sterile water injection, dextrose injection, and Ringer's lactate injection. Non-aqueous solvents for parenteral administration include vegetable fatty oils, olive oil, cottonseed oil, corn oil, sesame oil and peanut oil. Antimicrobial agents at bacteriostatic or fungistatic concentrations should be added to parenteral preparations packaged in multi-dose containers, which include phenols or cresols, mercury, benzyl alcohol, chlorobutanol, methyl and propyl p-hydroxybenzoic acid esters, thimerosal, benzalkonium chloride, and benzethonium chloride. Isotonic agents include sodium chloride and dextrose. Buffers include phosphate and citrate. Antioxidants include sodium bisulfate. Local anesthetics include procaine hydrochloride. Suspension and dispersing agents include sodium carboxymethylcellulose, hydroxypropylmethylcellulose and polyvinylpyrrolidone. Emulsifying agents include Polysorbate 80 (TWEEN® 80). Metal ion sequestering or chelating agents include EDTA. Pharmaceutical carriers also include ethyl alcohol, polyethylene glycol, and propylene glycol for water-miscible solvents; and sodium hydroxide, hydrochloric acid, citric acid or lactic acid for pH adjustment.

[0225] The concentration of the pharmaceutically active compound is adjusted so that the injection provides an effective amount to obtain the desired pharmacological effect. The exact dose depends on the age, weight and condition of the patient or animal, as is known in the art.

[0226] Single doses of parenteral preparations are packaged in ampoules, vials or syringes with a needle. All preparations for parenteral administration must be sterile as known and practiced in the art.

[0227] For example, intravenous or intra-arterial infusion of a sterile aqueous solution containing the active compound is an effective route of administration. Another implementation option is a sterile aqueous or oily solution or suspension containing the active substance, administered as needed to obtain the desired pharmacological effect.

[0228] Injectable preparations are intended for topical and systemic use. In one embodiment, a therapeutically effective dosage is formulated to contain a concentration of at least about 0.1% w/w. up to about 90% wt./wt. or more, in some embodiments, more than 1% wt./wt. active compound to the treated tissue(s).

[0229] The compound may be suspended in a micronized or other suitable form, or may be derivatized to provide a more soluble active product or to obtain a prodrug. The form of the resulting mixture depends on a number of factors, including the intended route of administration and the solubility of the compound in the chosen carrier or solvent. The effective concentration is sufficient to relieve the symptoms of the condition and can be determined empirically.

Freeze-dried powders

[0230] Also of interest herein are lyophilized powders that can be reconstituted for administration as solutions, emulsions, and other mixtures. They may also be reconstituted and obtained as solids or gels.

[0231] A sterile lyophilized powder is prepared by dissolving a compound provided herein, or a pharmaceutically acceptable derivative thereof, in a suitable solvent. The solvent may contain a stability enhancing excipient or other pharmacological component of the powder or reconstituted solution obtained from the powder. Excipients that may be used include, but are not limited to, dextrose, sorbitol, fructose, corn syrup, xylitol, glycerin, glucose, sucrose, or other suitable agent. The solvent may also contain a buffer, such as citrate, sodium or potassium phosphate, or other such buffer known to those skilled in the art, in one embodiment at about neutral pH. Subsequent sterile filtration of the solution, followed by lyophilization under standard conditions known to those skilled in the art, provides the desired composition. In one embodiment, the resulting solution will be dispensed into lyophilization vials. Each vial will contain one or more doses of the compound. The lyophilized powder can be stored under appropriate conditions, for example at about 4°C to room temperature.

[0232] Reconstitution of this lyophilized powder with water for injection provides a formulation for parenteral administration. For reconstitution, the lyophilized powder is added to sterile water or other suitable vehicle. The exact amount depends on the selected compound. This number can be determined empirically.

Local application

[0233] Topical mixtures are prepared as described for topical and systemic use. The resulting mixture can be a solution, suspension, emulsions, etc. and are formulated as creams, gels, ointments, emulsions, solutions, elixirs, lotions, suspensions, tinctures, pastes, foams, aerosols, irrigations, sprays, suppositories, dressings, skin patches or any other formulation suitable for topical application.

[0234] the Compounds or their pharmaceutically acceptable derivatives can be obtained in the form of aerosols for topical use, for example, for inhalation. These respiratory formulations may be in the form of an aerosol or solution for nebulization, or as a fine powder for insufflation, alone or in combination with an inert carrier such as lactose. In this case, the particles of the composition in one implementation will have a diameter of less than 50 microns, in one embodiment, less than 10 microns.

[0235] The compounds can be formulated for topical or topical application, for example, for topical application to the skin and mucous membranes, for example, in the eyes, in the form of gels, creams and lotions, as well as for application to the eyes, or for intracisternal or intraspinal application. Topical application is contemplated for transdermal delivery, and for application to the eye or mucosa, or for inhalation therapy. It is also possible to administer nasal solutions of the active compound alone or in combination with other pharmaceutically acceptable excipients.

[0236] These solutions, especially those intended for ophthalmic use, can be formulated as 0.01-10% (vol.%) isotonic solutions, pH about 5-7, with appropriate salts.

Compositions for other routes of administration

[0237] Other routes of administration are also contemplated herein, such as transdermal patches, including iontophoretic and electrophoretic devices, vaginal and rectal administration.

[0238] Transdermal patches, including iontophoretic and electrophoretic devices, are well known to those skilled in the art. For example, pharmaceutical dosage forms for rectal administration are rectal suppositories, capsules and tablets for systemic action. As used herein, rectal suppositories refer to solid bodies for insertion into the rectum that melt or soften at body temperature to release one or more pharmacologically or therapeutically active ingredients. The pharmaceutically acceptable substances used in rectal suppositories are bases or diluents and melting point raising agents. Examples of bases include cocoa butter (theobroma butter), glycerol-gelatin, carbovax (polyoxyethylene glycol), and appropriate mixtures of mono-, di-, and triglycerides of fatty acids. Combinations of various bases may be used. Agents that increase the melting point of suppositories include spermaceti and wax. Rectal suppositories can be obtained by pressing or molding. In one embodiment, the weight of the rectal suppository is about 2 to 3 grams.

[0239] Tablets and capsules for rectal administration are manufactured using the same pharmaceutically acceptable substance and by the same methods as formulations for oral administration.

Target formulations

[0240] The compounds provided herein, or pharmaceutically acceptable derivatives thereof, can also be formulated to target a specific tissue, receptor, or other area of the subject's body to be treated. Many such targeting methods are well known to those skilled in the art. All such targeting methods are contemplated herein for use in the present compositions.

[0241] In one embodiment, liposomal suspensions, including tissue-targeting liposomes, such as tumor-targeting liposomes, may also be suitable as pharmaceutically acceptable carriers. They can be obtained by methods known to experts in this field. For example, liposome compositions can be prepared as described in US Pat. No. 4,522,811. Briefly, liposomes such as multilamellar vesicles (MLVs) can be formed by drying egg phosphatidylcholine and brain phosphatidylserine (7:3 molar ratio) inside a flask. Add a solution of the compound provided herein in divalent cation-free phosphate buffered saline (PBS) and shake the flask until the lipid film is dispersed. The resulting vesicles are washed to remove unencapsulated compound, pelleted by centrifugation and then resuspended in PBS.

Combined use with other drugs

[0242] In accordance with another aspect of the present invention, it is contemplated that compounds of Formula I, as described herein, can be administered to a subject in need thereof, in combination with a drug considered by those skilled in the art to be the current standard of care for the condition of interest. Such combinations provide one or more benefits to the subject, such as requiring reduced doses to achieve a similar effect, obtaining the desired palliative effect in less time, and the like.

[0243] The compounds of the present invention may be administered as part of a therapeutic regimen with other drugs. It may be desirable to administer a combination of active compounds, for example, for the purpose of treating a particular disease or condition. Accordingly, within the scope of the present invention, two or more pharmaceutical compositions, at least one of which contains a compound of Formula (I) of the present invention, can be combined in the form of a kit suitable for joint use of the compositions.

[0244] In one embodiment of the methods of the present invention, a compound of Formula I may be administered with a second therapeutic agent. In one embodiment, the second therapeutic agent may be selected from one or more of the following categories:

(i) Anti-cancer agents such as cisplatin, oxaliplatin, carboplatin, cyclophosphamide, nitrogen mustard, uracil mustard, bendamustine, melphalan, chlorambucil, chlormethine, busulfan, temozolomide, nitrosoureas, ifosamide, pipobromine, triethylenemelamine, triethylenethiophosphoramine, carmustine, lomustine, strepto zocin and dacarbazine, gemcitabine, phosgemcitabine, palabenamide, 5-fluorouracil, tegafur, raltitrexed, methotrexate, pemetrexed, leucovorin, cytosine arabinoside, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, pentostatin, hydroxyurea, trifluridine, trifur acyl, adriamycin, bleomycin , doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin, mithramycin, vincristine, vinblastine, vindesin and vinorelbine, taxol, taxotere, eribulin, carfilzomib, bortezomib, etoposide, teniposide, amsacrine, topotecan, irinotecan, mitoxantrone, cam ptothecin, dactinomycin , daunorubicin, aldoxorubicin, epirubicin, idarubicin, ara-C, paclitaxel (Taxol™), nabpaclitaxel, docetaxel, deoxycoformycin, L-asparaginase, IFN-alpha, azacitidine, decitabine, vorinostat, MS-275, panobinostat, romidepsin, valproic acid, mocetinostat, pracinostat, belinostat, irabectin, tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene, iodoxifene, bicalutamide, flutamide, nilutamide, cyproterone acetate, goserelin, leuprorelin, buserelin, progestogen, megestrol acetate, anastrozole, letrozole, vorazole, exemestane, finasteride, navelben, capecitabine and droloxafine; and abiraterone, enzalutamide, lanreotide, dasatinib, bosutinib, trastuzumab, pertuzumab, panitumumab, cetuximab, gefitinib, erlotinib, afatinib, vandetanib, osimertinib, rockiletinib, lapatinib, imatinib, nilotinib, sorafenib, tipifarnib, and lonafar nib, vemirafenib, dubrafenib, trametinib, cobimetinib, ponatinib, paldociclib, everolimus, ruxolitinib, pacritinib, jactinib, imetelstat, plitidepsin, pevonedist, ibrutinib, ceritinib, crizotinib, ectinib, cabozantirsib, vismodegiban, sonidegib, regofarenib, vandetanib, vatalanib, sunitinib, axi tinib, pazopanib, lenvatinib, talimogen, laherparepvec, denosumab, obinuluzumab, blinatomumab, dinutuximab, idarucizumab, daratumumab, necitumumab, elotuzumab, olaratumab, alemtuzanab, rituximab, ibritumomab tiuxetan, ofatumumab, peginterferon alfa-2b, Gardasil, Cervarix, Oncophage, Sipuleu cel-T, nivolumab, pembrolizumab, atezolizumab, indoxymod, nivolumab, ipilumumab, brentuximab vedotin, irastuzumab, emtansine, fludaribine, cladribine, pentostatin, idelicib, perifosin, birinapant, bortezomib, ixazomib, carfilzomib, marizomib, olaparib, rucaparib, venetoclax, navitoclax, obatoclax, glasdegib, pacrinostat, buparalisib, memoletinib , itakitinub, umbralisib, gusacitinib, tagraxofusp, ribociclib, abemaciclib, niraparib, trabadectin, pofimer, vinflunin, napabucasin, lurbinectin, tazemetostat, acalabrutinib, levatinib, neratinib, pamiparib, epacadostat, enzastaurin, selinexor, masitin ib, evafosfamide, glufosfamide, roxadustat, streptozocin , devimistat, galunisertib, binimetinib, veliparib, entinostat, pexidartinib, talazoparib, entrectinib.

(II) Anti -inflammatory agents, such as meloxicams, Theoproprofen, oxaprozin, salsalat, ethricoxib, tenoxics, aspirin, nabometon, flurbprofen, mephenamic acid, phenylbutazone, loroxyams, indomethacin, etomacin, diplunizal, ketoprofen, valdexib, hollow, cherpower acid, pyroxics, sulindak, tolmetin, ketorolac, loxoprofen, acetaminophen, bromfenac, diclofenac, ibuprofen, meclofenamate, nabumeton, naproxen, nepafenac, celecoxib, triamcinolone acetonide, hydrocortisone, hydrocortisone acetate, methylprednisolone, aclomethasone dipropionate, emricazan, BI 146 7335, namodenosone, GLPG-1690, terguride.

(iii) An antihypertensive agent 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, nadolol, timolol, pindolol, labetolol, metoprolol, atenolol, esmolol, betaxolol, carvedilol, prazosin, terazosin, doxazosin, phenoxybenzamine, phentolamine, verapamil, diltiazem, nifedipine, felodipine, amlodipine, nimodipine, diaz oxide, minoxidil, pinacidil, nicorandil, hydralazine, sodium nitroprusside, bosentan, epoprostenol, iloprost, beraprost, esuberaprost, ralinepag, macitentan, sitaxentan, ambrisentan, riociguat, treprostinil, ubenimex, selexipag, levosimendan, udenafil, tadalafil, and sildenafil.

(iv) An anti-fibrotic agent such as pirfenidone, nintedanib, cenicriviroc, seloncertib, lanifibranor, nimacimab, nitrazoxanide, NGM282, apararenone, typelukast, Actimmune, ponatinib, lenvatinib, dovitinib, lucitanib, danusertinib, brivatinib, erdafitinib, bel apectin, PD173074, PD166866, AZD4547 BGJ398 LY2874455 TAS-120 ARQ087 BLU9931 FGF401 BAY-1163877 ENMD-2076 IMCA1 FGF401 DEBIO1347 FIIN-2 GP-369 PRO-001 H3B-6527 BAY11 87982, MFGR1877S, FP-1039, BLU554, PRN1371, S49076, SU6668, SU5416, PBI-4050, KD-025.

(v) An anti-angiogenesis agent such as axitinib, bevacizumab, cabozantinib, lenalidomide, lenvatinib, pazopanib, ramucirumab, vandetanib, vatalanib, sunitinib, ziv-aflibercept, thalidomide, pomalidomide, lenalidomide.

(vi) An immunosuppressant 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, daclizumab, dimethyl fumarate, mycophenolate mofetillate.

(VII) Metabolic agent, such as enjoyed acid, elafibrainor, aramkhol, celebradar, MGL-3196, tropicor, MSDC-0602K, BMS-986036, Semaglutide, EDP-305, Gemkaben, PF-068665571, PF-068, PF-068, PF-068, PF-068, PF 35919 , LIK066, LMB763, vitamin E, acarbose, miglitol, pramlintide, alogliptan, linagliptan, saxagliptin, sitagliptin, albiglutide, dulaglutide, exenatide, liraglutide, lixisenatide, insulin, nateglinide, repaglinide, metformin, canagliflozin, dapagliflozin, empa gliflozin, chlorpropamide, glimepiride, glipizide, glyburide, tolazamide, tolbutamide, rosiglitazone, pioglitazone, atorvastatin, amlodipine, simvastatin, ezetimibe, lovastatin, sitagliptin, cholestyramine, colesevelam, colestipol, fenofibrate, gemfibrozil, 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, thaliglucerase alfa, agalsidase beta, sebelipase alfa, westronidase alfa, galsulfase, elosulfase alfa, eliglustat, burosumab, migalastat, sapropterin, metreleptin, nitisinone, pegvaliase, asfotase alfa, inotersen, miglustat, orlistat, sodium phenylbutyrate, glycerol phenylbutyrate.

[0245] 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 may be administered in combination with one or more additional antitumor agents and/or radiotherapy for the treatment of cancer.

[0246] When two or more active ingredients are administered together, the active ingredients may be administered simultaneously, sequentially, or separately. In one embodiment, a compound of Formula I is administered concurrently with a second therapeutic agent. In another embodiment, the compound of Formula I and the second therapeutic agent are administered sequentially. In a further embodiment, the compound of Formula I and the second therapeutic agent are administered separately.

[0247] The invention will now be described in more detail, by way of illustration only, with reference to the following non-limiting examples. The examples are intended to illustrate the invention and should not be construed as limiting the generality of the disclosure in this specification.

Experiment: General Methods

[0248] Commercially available solvents and reagents were used as received. If necessary, the reactions are carried out in an argon atmosphere. Reactions were followed by either analytical thin layer chromatography (TLC) or analytical liquid chromatography-mass spectrometry (LCMS) recorded on either a Shimadzu LCMS 2020 instrument or an Agilent LC/MSD 1200 instrument using phase reversal conditions. If necessary, the purification of intermediates and final compounds was carried out using column chromatography or preparative HPLC. Normal phase column chromatography was performed at medium pressure either on silica gel or on pre-packed silica gel cartridges using a flash chromatography system (CombiFlash Rf200, Teledyne Isco systems, USA). Reverse phase column chromatography was performed at low pressure on pre-packed C18 cartridges using a flash chromatography system (Reveleris® X2). The eluents were controlled by UV light (λ = 254/280 nm). ¹ H-NMR and ¹⁹ F-NMR spectra were recorded using a Bruker 300 MHz NMR spectrometer, a Bruker Avance III plus 400 MHz NMR spectrometer, or a Varian III plus 300 MHz spectrometer. Chemical shifts (δ) are expressed in parts per million (ppm) relative to tetramethylsilane (TMS; internal standard). The following abbreviations are used for multiplicities: s = singlet; ws = wide singlet; d = doublet; u = triplet; k = quartet; m = multiplet; and wm = wide multiplet. Low resolution mass spectra (MS) were obtained as atmospheric pressure electrospray ionization (ES-ADI) mass spectra, which were recorded either on a Shimadzu LCMS 2020 instrument or on an Agilent LC/MSD 1200 instrument using reverse phase processing conditions. All animal experiments were performed in accordance with established guidelines and with the approval of local ethics committees.

EXAMPLE 1

[0249] Preparation of ( Z ) -tert- butyl (4-bromo-3-fluorobut-2-en-1-yl)carbamate

Method A: Getting tert- butyl-2-oxoethylcarbamate

[0250] Di- tert- butyl dicarbonate (55.5 ml, 0.24 mol). After bringing the alkalinity of the solution to pH~9 by adding aq. NaOH (6N), the mixture was allowed to stir at room temperature (RT) for 18 hours. The reaction mixture was cooled to 0-5°C and then acidified to pH~6 before sodium metaperiodate (56.3 g, 0.26 mol) was added. The resulting suspension was stirred at room temperature for 2 hours. The mixture was filtered to remove all solids and the filtrate was transferred to a separating funnel and extracted with ethyl acetate (200 ml). Sodium chloride was added to the aqueous layer until a saturated solution was obtained. Then the aqueous layer was further extracted with ethyl acetate (100 ml). The combined organic layers were dried over Na ₂ SO ₄ and then concentrated in vacuo to give crude tert- butyl 2-oxoethyl carbamate (45.7 g) as a yellow gum. The crude material was used in the next step without purification.

Method B: Getting ( E )-ethyl 4-( tert- butoxycarbonylamino)-2-fluorobut-2-enoate and ( Z )-ethyl 4-( tert- butoxycarbonylamino)-2-fluorobut-2-enoate

Ethyl _{_} 2-fluorophosphonoacetate (55.7 ml, 0.27 mol) and 1,8-diazabicyclo[5.4.0]undec-7-ene (32.8 ml, 0.22 mol). The reaction mixture was allowed to warm to room temperature and stirring was continued for 3 hours. After removal of the solvent under reduced pressure, the residue was taken up in ethyl acetate (200 ml) and then transferred to a separating funnel. The organic layers were washed successively with aq. HCl (2 M; 100 ml x 2), aq. NaOH (2 M; 100 ml x 2) and brine (100 ml). After drying over MgSO ₄ the organic layers were concentrated in vacuo to give the crude desired product as a mixture of E / Z isomers (2:3; 57.0 g). This crude material was used in the next step without purification.

Method C: Getting (E) - tert -butyl-3-fluoro-4-hydroxybut-2-enylcarbamate and (Z) - tert -butyl-3-fluoro-4-hydroxybut-2-enylcarbamate

[0252] To a stirred solution of crude E / Z- ethyl 4-( tert- butoxycarbonylamino)-2-fluorobut-2-enoate (18.0 g, 72.8 mmol) in THF (150 ml) at 0°C under atmosphere N ₂ diiso -butylaluminum hydride (1 M in toluene, 182 ml, 182 mmol) was added dropwise over 45 minutes. After completion of the addition, the mixture was allowed to stir at 0° C. for 3 h. The reaction mixture was transferred to a separating funnel and added dropwise to a stirred mixture of ice (100 g) and aq. NaOH (2 M; 200 ml). After the addition, the mixture was stirred for 2 hours. The quenched reaction mixture was extracted with diethyl ether (100 ml x 2) and the combined organic layers washed with brine (100 ml). After drying over MgSO ₄ , the organic layers were concentrated in vacuo to give the crude alcohol as a mixture of E / Z isomers. This mixture was purified on silica gel (135 g), eluted with 25% ethyl acetate in n- hexane to give ( Z ) -tert- butyl 3-fluoro-4-hydroxybut-2-enylcarbamate (6.20 g, 30% after three steps) and ( E ) -tert -butyl 3-fluoro-4-hydroxybut-2-enylcarbamate (1.85 g, 8.9% after three stages). ( E ) -tert- butyl-3-fluoro-4-hydroxybut-2-enylcarbamate: ¹ H-NMR (200 MHz; CDCl ₃ ) δ ppm: 1.43 (9H, s), 3.72 ( 2H, dd, J 7.5, 5.4 Hz), 4.25 (2H, d, J 21.5 Hz), 4.85 (1H, brs), 5.18 (1H, pcs, J 19, 2, 8.5 Hz). ( Z ) -tert- butyl-3-fluoro-4-hydroxybut-2-enylcarbamate: ¹ H-NMR (300 MHz; CDCl ₃ ) δ ppm: 1.46 (9H, s), 3.84 ( 2H, dd, J 6.2, 6.2 Hz), 4.13 (2H, d, J 13.9 Hz), 4.68 (1H, brs), 5.03 (1H, dt, J 36, 0.7.1 Hz).

Method D: Getting ( Z )- tert- butyl-4-bromo-3-fluorobut-2-enylcarbamate

[0253] To a stirred solution of ( Z ) -tert- butyl-3-fluoro-4-hydroxybut-2-enylcarbamate (6.20 g, 30.2 mmol) and triethylamine (6.32 ml, 45.3 mmol) in acetone (100 ml) at 0 °C was added dropwise methanesulfonyl chloride (2.81 ml, 36.3 mmol). After completion of the addition, the mixture was allowed to stir at 0°C for 30 min. After this time, lithium bromide (13.1 g, 0.15 mol) was added in portions and the resulting suspension was stirred for another 2 hours. The reaction mixture was filtered to remove all solids and the filtrate was concentrated under reduced pressure. The residue was partitioned between water (50 ml) and CH ₂ Cl ₂ (50 ml) and the aqueous layer was further extracted with CH ₂ Cl ₂ (50 ml x 2). The combined organic layers were dried over Na ₂ SO ₄ and concentrated in vacuo . The crude residue was purified on silica gel (100 g) eluting with n- hexane followed by 25% ethyl acetate in n- hexane to give ( Z ) -tert- butyl 4-bromo-3-fluorobut-2-enylcarbamate (7.00 g , 86%) as a colorless solid. ¹ H-NMR (300 MHz; CDCl ₃ ) δ ppm: 1.46 (9H, s), 3.85 (2H, dd, J 6.2, 6.2 Hz), 3.93 (2H , d, J 19.5 Hz), 4.66 (1H, brs), 5.16 (1H, dt, J 34.0, 6.5 Hz).

EXAMPLE 2

[0254] The following compound was prepared according to methods E, F, G, H, and I.

[0255] Preparation of (Z) -4-((2-((4-amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)methyl)-N ,N - diisopropylbenzenesulfonamide hydrochloride (Compound 11 )

Method E: Preparation of 4-(bromomethyl)- N,N -di iso propylbenzenesulfonamide

[0256] Diiso - _{propylamine (} 0.65 ml, 4.63 mmol ) drop by drop. After the addition, the resulting mixture was allowed to stir at this temperature for 30 minutes, after which it was allowed to warm to room temperature and stirred for another 48 hours. The reaction mixture was divided between aq. HCl (1 M, 20 ml) and CH ₂ Cl ₂ (20 ml). The organic layer was washed with aq. HCl (1 M; 20 ml) and water (20 ml), dried over Na ₂ SO ₄ and concentrated in vacuo to give the title compound (yellow oil, 190 mg) as a mixture with 4-(chloromethyl) -N , N - diisopropylbenzenesulfonamide , which was used as such in the next step.

Method F: Getting ( Z )- tert- butyl-(3-fluoro-4-((2-hydroxyphenyl)thio)but-2-en-1-yl)carbamate

[0257] To a solution of 2-mercaptophenol (235 mg, 1.86 mmol) and (Z) -tert -butyl (4-bromo-3-fluorobut-2-en-1-yl) carbamate (500 mg, 1.86 mmol) in acetone (3 ml) at room temperature was added potassium carbonate (387 mg, 2.70 mmol) and the resulting solution was stirred at room temperature for 16 hours. The reaction mixture was then partitioned between EtOAc (20 ml) and water (20 ml) and the phases were separated. The aqueous phase was extracted with EtOAc (20 mL x 2) and then the organic phases were combined and washed (brine; 20 mL), dried (Na ₂ SO ₄ ) and concentrated in vacuo to give ( Z ) -tert- butyl-(3- fluoro-4-((2-hydroxyphenyl)thio)but-2-en-1-yl)carbamate (580 mg, 99%) as a light yellow solid. ¹ H-NMR (300 MHz; CDCl ₃ ) δ ppm: 1.45 (9H, s), 3.31 (2H, d, J = 19.7 Hz), 3.69 (2H, ca. t , J = 6.7 Hz), 4.47 (1H, dt, J = 34.6, 7.2 Hz), 4.49 (1H, brs), 6.67 (1H, s), 6.90 (1H, dd, J = 7.6, 7.6 1.3 Hz), 7.02 (1H, dd, J = 8.2, 1.2 Hz), 7.31 (1H, dd, J = 8.2, 7.3, 1.6 Hz), 7.45 (1H, dd, J = 7.7, 1.7 Hz).

Method G: Getting (Z) - tert -butyl-(4-((2-((4- (N,N -di iso propylsulfamoyl)benzyl)oxy)phenyl)thio)-3-fluorobut-2-en-1-yl)carbamate

[0258] To a stirred solution of ( Z ) -tert- butyl(3-fluoro-4-((2-hydroxyphenyl)thio)but-2-en-1-yl)carbamate (100 mg, 0.32 mmol) and 4 -(bromomethyl) -N,N - diisopropylbenzenesulfonamide (107 mg, 0.32 mmol) in DMF (1 ml) was added potassium carbonate (66 mg, 0.48 mmol) at room temperature. The resulting suspension was stirred at this temperature for 16 hours. The reaction mixture was then partitioned between EtOAc (10 ml) and water (10 ml) and the phases were separated. The aqueous phase was extracted with EtOAc (10 mL×2) and then the organic phases were combined and washed (sat. aq. NH ₄ Cl, then brine), dried (Na ₂ SO ₄ ) and concentrated in vacuo to give ( Z ) -tert - butyl (4-((2-((4- (N,N - di isopropylsulfamoyl ) benzyl) oxy) phenyl) thio) -3-fluorobut-2-en-1-yl) carbamate (180 mg, 99% ) as a yellow gum, which was used in the next step without purification.

Method H: Getting (Z) - tert -butyl-(4-((2-((4- (N,N- di iso propylsulfamoyl)benzyl)oxy)phenyl)sulfonyl)-3-fluorobut-2-en-1-yl)carbamate

[0259] To the stirred slurry (4-((2-((4-(N,N-diisopropylsulfamoyl)benzyl)oxy)phenyl)thio)-3-fluorobut-2-en-1-yl)carbamate (180 mg, 0.32 mmol) and sodium bicarbonate (133 mg, 1.59 mmol) in CH₂Cl₂ (2 ml) and water (2 ml) at 0 °C was added 3-chloroperoxybenzoic acid (178 mg, 0.79 mmol) in three portions over 5 minutes. The resulting suspension was stirred at 0°C for 2 hours before being diluted with sat. aq. NaHCO₃ (15 ml) and extracted with CH₂Cl₂ (10 ml). The aqueous phase was further extracted with CH₂Cl₂ (10 ml x 2) and the organic phases were combined, dried (Na₂SO₄) and concentratedin a vacuum. The crude material was purified by flash column eluting with 40% EtOAc/hexane followed by 2% MeOH in 50% EtOAc/hexane to give (Z)-tert-butyl-(4-((2-((4-(N,N-diisopropylsulfamoyl)benzyl)oxy)phenyl)sulfonyl)-3-fluorobut-2-en-1-yl)carbamate (160 mg, 84%) as a white solid.¹H-NMR (300 MHz; CDCl₃) δ ppm: 1.29 (12 H, d,J = 6.8 Hz), 1.43(9H, s), 3.67-3.80(3H, m), 4.15(2H, d,J = 18.9 Hz), 4.52 (1H, brs), 4.93 (1H, dt,J = 34.4, 6.9 Hz), 5.33 (2H, s), 7.09 (1H, d,J = 8.0 Hz), 7.18 (1H, ddd,J = 8.3, 7.8, 0.8 Hz), 7.63 (1H, ddd,J = 8.4, 7.6, 1.7 Hz), 7.66 (2H, d,J = 8.4), 7.93 (2H, d,J = 8.5 Hz), 7.99 (1H, dd,J = 7.9, 1.7 Hz). As a modification of this procedure to obtain additional compounds, the oxidation was carried out by slowly adding an aqueous solution of OXONE® (4 eq. in 1.2 ml of H₂O per mmol OXONE®) to a solution of the starting sulfanyl ether in MeOH:THF (1 : 1, approx. 3 ml each per mmol sulfanyl ether) at room temperature and allowed to react until LC-MS monitoring showed high conversion to the desired sulfone product. The mixture was then partitioned between excess saturated aqueous sodium metabisulphite and EtOAc, washed with brine, dried (Na₂SO₄), concentratedin a vacuum and purified by column chromatography.

Method I: Preparation of hydrochloride (Z) -4-((2-((4-amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)methyl)- N,N -di iso propylbenzenesulfonamide (Compound 11)

[0260] To a stirred solution of (Z) -tert -butyl-(4-((2-((4- (N,N - diisopropylsulfamoyl )benzyl)oxy)phenyl)sulfonyl)-3-fluorobut-2-ene -1-yl)carbamate (160 mg, 0.27 mmol) in MeOH (1 ml) was added ethereal HCl (2 M; 4.00 ml, 8.00 mmol) at room temperature and the resulting mixture was allowed to stir for 1 hours. After this time, a white solid precipitated, which was collected by filtration and dried under high vacuum to give ( Z )-4-((2-((4-Amino-2-fluorobut-2-en-1-yl)sulfonyl) hydrochloride) phenoxy)methyl)-N ,N -diisopropylbenzene-sulfonamide (79 mg, 55%). White solid; etc. 222-224°C; ¹ H-NMR (300 MHz; CD ₃ OD) δ ppm: 1.27 (12H, d, J = 6.8 Hz), 3.59 (2H, dd, J = 7.4, 1, 8 Hz), 3.79 (2H, hept, J = 6.8 Hz), 4.45 (2H, d, J = 19.2 Hz), 5.16 (1H, dt, J = 32.8, 7.4 Hz), 5.46 (2H, s), 7.23 (1H, ddd, J = 7.4, 7.4, 0.9 Hz), 7.38 (1H, d J = 7, 9 Hz), 7.74 (1H, ddd, J = 8.5, 7.5, 1.7 Hz), 7.79 (2H, d, J = 8.6 Hz), 7.91-7, 95 (3H, m).

EXAMPLE 3

[0261] The following compounds were prepared according to Methods E-I using an appropriately functionalized thiol starting material.

( Z )-4-((2-((4-Amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)methyl) -N , N- dimethylbenzenesulfonamide hydrochloride (Compound 5)

[0262] White solid; etc. 235-236°C; ¹ H-NMR (300 MHz; CD ₃ OD) δ ppm: 2.72 (6H, s), 3.60 (2H, dd, J = 7.4, 1.7 Hz), 4.47 (2H, d, J = 19.2 Hz), 5.18 ( 1H, dt, J = 32.9, 7.4 Hz), 5.49 (2H, s), 7.24 (1H, ddd, J = 7.9, 7.9, 0.9 Hz), 7.39 (1H, dd, J = 8.5, 0.7 Hz), 7.76 (1H, dd, J = 8.4, 7.4, 1.7 Hz), 7.86 (4H, brs), 7.95 (1H, dd, J = 7.9, 1.8 Hz).

( Z )-4-((2-((4-Amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)methyl)benzenesulfonamide hydrochloride (Compound 8)

[0263] Off-white solid; etc. 233-235°C; ¹ H-NMR (300 MHz; CD ₃ OD) δ ppm: 3.59 (2H, dd, J = 7.4, 1.6 Hz), 4.44 (2H, d, J = 19, 2 Hz), 5.14 (1H, dt, J = 32.8, 7.4 Hz), 5.45 (2H, s), 7.23 (1H, dd, J = 7.3, 7.3 Hz), 7.38 (1H, d, J = 8.3 Hz), 7.74 (1H, ddd, J = 8.6, 8.6, 1.7 Hz), 7.78 (2H, d , J = 8.2 Hz), 7.94 (1H, dd, J = 8.1, 1.6 Hz), 7.97 (2H, d, J = 8.5 Hz).

( Z )-4-((3-((4-Amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)methyl) -N , N- dimethylbenzenesulfonamide hydrochloride (Compound 9)

[0264] White solid; etc. 211-213°C; ¹ H-NMR (300 MHz; d ₆ -DMSO) δ ppm: 2.63 (6H, s), 3.48 (2H, brs), 4.65 (2H, d, J = 19.6 Hz), 5.17 (1H, dt, J = 34.6, 7.2 Hz), 5.36 (2H, s), 7.45 (1H, ddd, J = 8.1, 2.5, 1.0 Hz), 7.52-7.57 (2H, m), 7.63 (1H, dd, J = 8.1, 8.1 Hz), 7.75 (2H, d, J = 8 .5 Hz), 7.81 (2H, d, J = 8.6 Hz), 8.11 (3H, brs).

( Z )-4-((4-((4-Amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)methyl) -N , N- dimethylbenzenesulfonamide hydrochloride (Compound 10)

[0265] White solid; etc. 216-218°C; ¹ H-NMR (300 MHz; d ₆ -DMSO) δ ppm: 2.63 (6H, s), 3.48 (2H, brs), 4.55 (2H, d, J = 19.7 Hz), 5.12 (1H, dt, J = 34.8, 7.1 Hz), 5.38 (2H, s), 7.29 (2H, dd, J = 9.0, 1.9 Hz ), 7.74 (2H, dd, J = 8.5, 1.8 Hz), 7.81 (2H, dd, J = 8.5, 1.9 Hz), 7.88 (2H, dd, J = 8.9, 2.0 Hz), 8.03 (3H, br).

( Z )-4-((2-((4-Amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)methyl) -N -isopropylbenzenesulfonamide hydrochloride (Compound 14)

[0266] White solid; etc. 248-250°C; ¹ H-NMR (300 MHz; CD ₃ OD) δ ppm: 1.05 (6H, d, J = 6.6 Hz), 3.40 (1H, hept, J = 6.6 Hz), 3.59 (2H, approx. d, J = 7.3 Hz), 4.45 (2H, d, J = 19.1 Hz), 5.16 (1H, dt, J = 33.0, 7, 4 Hz), 5.46 (2H, s), 7.23 (1H, ddd, J = 8.0, 8.0, 1.0 Hz. ), 7.38 (1H, d, J = 8, 1 Hz), 7.74 (1H, ddd, J = 8.4, 7.4, 1.8 Hz), 7.80 (2H, d, J = 8.7 Hz), 7.91-7, 95 (3H, m).

EXAMPLE 4

[0267] The following compound was prepared according to Methods F-I using the appropriate thiol starting material.

( Z )-4-((2-(Benzyloxy)phenyl)sulfonyl)-3-fluorobut-2-en-1-amine hydrochloride (Compound 7)

[0268] White solid; etc. 205-207°C; ¹ H-NMR (300 MHz; CD ₃ OD) δ ppm: 3.57 (2H, approx. d, J = 7.0 Hz), 4.44 (2H, d, J = 19.1 Hz ), 5.14 (1H, dt, J = 32.8, 7.3 Hz), 5.36 (2H, s), 7.20 (1H, dd, J = 7.4, 0.9 Hz) , 7.35-7.46 (4H, m), 7.55-7.60 (2H, m), 7.73 (1H, ddd, J = 8.5, 7.4, 1.7 Hz) , 7.92 (1H, dd, J = 7.9, 1.7 Hz).

EXAMPLE 5

[0269] The following compounds were prepared according to methods F, H, and I using the appropriately functionalized thiol starting material.

( Z )-4-((3,5-bis(trifluoromethyl)phenyl)sulfonyl)-3-fluorobut-2-en-1-amine hydrochloride (Compound 2)

[0270] White solid; etc. 217-220°C; ¹ H-NMR (300 MHz; d ₆ -DMSO) δ ppm: 3.48 (2H, approx. d, J = 7.1 Hz), 4.96 (2H, d, J = 19.6 Hz), 5.19 (1H, dt, J = 34.8, 7.2 Hz), 8.10 (3H, brs), 8.55 (2H, s), 8.67 (1H, s).

( Z )-4-(Biphenyl-2-ylsulfonyl)-3-fluorobut-2-en-1-amine hydrochloride (Compound 20)

[0271] White solid; etc. 170°C; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 8.18 (dd, J = 8.0, 1.4 Hz, 1H), 7.81 (td, J = 7.5, 1 .4 Hz, 1H), 7.68 (td, J = 7.6, 1.6 Hz, 1H), 7.52 - 7.44 (m, 6H), 5.03 (dt, J = 32, 9, 7.4 Hz, 1H), 3.83 (d, J = 18.9 Hz, 2H), 3.56 (dd, J = 7.4, 1.3 Hz, 2H).

( Z )-3-Fluoro-4-(2-isopropylphenylsulfonyl)but-2-en-1-amine hydrochloride (Compound 21)

[0272] White solid; etc. 205-215°C; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 8.00-7.95 (m, 1H), 7.77-7.67 (m, 2H), 7.48-7.41 (m, 1H), 5.25 (dt, J = 32.8, 7.4 Hz, 1H), 4.34 (d, J = 19.2 Hz, 2H), 3.88 (hept, J = 6.9 Hz, 1H), 3.63 (dd, J = 7.5, 1.9 Hz, 2H), 1.36 (d, J = 6.8 Hz, 6H).

( Z )-3-Fluoro-4-(2-methoxyphenylsulfonyl)but-2-en-1-amine hydrochloride (Compound 22)

[0273] White solid; etc. 228-221°C; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 7.89 (dd, J = 7.9, 1.7 Hz, 1H), 7.75 (dd, J = 8.4, 7 .4, 1.8 Hz, 1H), 7.32 (dd, J = 8.5, 0.9 Hz, 1H), 7.19 (td, J = 7.6, 1.0 Hz, 1H) , 5.23 (dt, J = 32.8, 7.4 Hz, 1H), 5.17 (t, J = 7.4 Hz, 0H), 4.48 (d, J = 19.3 Hz, 2H), 4.05 (s, 3H), 3.61 (dd, J = 7.4, 1.9 Hz, 2H).

( Z )-3-Fluoro-4-(naphthalene-1-ylsulfonyl)but-2-en-1-amine hydrochloride (Compound 23)

[0274] White solid; etc. 230–240°C; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 8.76 (ddd, J = 8.7, 1.2, 0.6 Hz, 1H), 8.33 (dd, J = 7 .8 Hz, 2H), 8.13 (ddd, J = 8.2, 1.5, 0.8 Hz, 1H), 7.81 (ddd, J = 8.6, 6.9, 1.5 Hz, 1H), 7.72 (td, J = 8.0, 1.9 Hz, 2H), 5.13 (dt, J = 32.7, 7.4 Hz, 1H), 4.49 (d , J = 19.2 Hz, 2H), 3.57 (dd, J = 7.4, 1.9 Hz, 2H).

( Z )-3-Fluoro-4-(naphthalene-2-ylsulfonyl)but-2-en-1-amine hydrochloride (Compound 24)

[0275] White solid; etc. 215-220°C; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 8.60 (d, J = 1.9 Hz, 1H), 8.18 - 8.11 (m, 2H), 8.06 ( dd, J = 8.2, 1.4 Hz, 1H), 7.95 (dd, J = 8.7, 1.9 Hz, 1H), 7.81 - 7.68 (m, 2H), 5 .19 (dt, J = 32.8, 7.4 Hz, 1H), 4.46 (d, J = 19.2 Hz, 2H), 3.63 (dt, J = 7.4, 1.3 Hz, 2H).

( Z )-4-(2,4-Dichlorophenylsulfonyl)-3-fluorobut-2-en-1-amine hydrochloride (Compound 25)

[0276] White solid; etc. 220°C; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 8.10 (d, J = 8.6 Hz, 1H), 7.85 (d, J = 2.0 Hz, 1H), 7 .66 (dd, J = 8.6, 2.0 Hz, 1H), 5.28 (dt, J = 32.9, 7.4 Hz, 1H), 4.60 (d, J = 19.1 Hz, 2H), 3.63 (ddd, J = 7.4, 2.0, 0.6 Hz, 2H).

( Z )-4-(3-Chlorophenylsulfonyl)-3-fluorobut-2-en-1-amine hydrochloride (Compound 26)

[0277] White solid; etc. 225-235°C; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 8.00 (t, J = 1.9 Hz, 1H), 7.92 (ddd, J = 7.8, 1.8, 1 .1 Hz, 1H), 7.82 (ddd, J = 8.1, 2.1, 1.1 Hz, 1H), 7.68 (d, J = 8.3 Hz, 1H), 5.22 (dt, J = 32.9, 7.4 Hz, 1H), 4.44 (d, J = 19.1 Hz, 2H), 3.65 (dd, J = 7.4, 1.9 Hz, 2H)

( Z )-4-(4-Chlorophenylsulfonyl)-3-fluorobut-2-en-1-amine hydrochloride (Compound 27)

[0278] White solid; etc. 240°C; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 7.96 (dt, J = 8.8, 2.3 Hz, 2H), 7.71 (dt, J = 8.3, 1 .9 Hz, 2H), 5.20 (dt, J = 32.9, 7.4 Hz, 1H), 4.40 (d, J = 19.1 Hz, 2H), 3.65 (dd, J = 7.4, 1.9 Hz, 2H).

( Z )-4-(3,5-Dichlorophenylsulfonyl)-3-fluorobut-2-en-1-amine hydrochloride (Compound 28)

[0279] White solid; etc. 250°C; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 7.97 (d, J = 1.8 Hz, 2H), 7.93 (t, J = 1.9 Hz, 1H), 5 .27 (dt, J = 33.0, 7.4 Hz, 1H), 4.50 (d, J = 19.0 Hz, 2H), 3.67 (dd, J = 7.4, 2.0 Hz, 2H).

( Z )-3-Fluoro-4-(pyridin-4-ylsulfonyl)but-2-en-1-amine dihydrochloride (Compound 29)

[0280] White solid; etc. 162-164°C; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 9.07 (dd, J = 4.9, 1.7 Hz, 2H), 8.22 (dd, J = 4.6, 1 .6 Hz, 2H), 5.31 (dt, J = 33.1, 7.4 Hz, 1H), 4.63 (d, J = 19.0 Hz, 2H), 3.67 (d, J = 7.4Hz, 2H)

( Z )-3-Fluoro-4-(quinolin-2-ylsulfonyl)but-2-en-1-amine dihydrochloride (Compound 32)

[0281] White solid; etc. 203-205°C; ¹ H NMR (300 MHz, d ₆ -DMSO) δ ppm: 8.82 (d, J = 8.6 Hz, 1H), 8.29 - 8.20 (m, 2H), 8.16 (d, J = 8.5 Hz, 1H), 8.12 - 7.93 (m, 3H), 7.92 - 7.83 (m, 1H), 5.26 (dt, J = 34.8 , 7.2 Hz, 1H), 4.92 (d, J = 19.6 Hz, 2H), 3.48 (s, 2H).

( Z )-3-Fluoro-4-(quinolin-8-ylsulfonyl)but-2-en-1-amine dihydrochloride (Compound 33)

[0282] White solid; etc. 150-153°C; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 9.18 (d, J = 4.7 Hz, 1H), 8.70 (dd, J = 8.4, 2.6 Hz, 1H), 8.57 (d, J = 7.4 Hz, 1H), 8.45 (d, J = 8.5 Hz, 1H), 7.99 - 7.68 (m, 2H), 5, 22 (dt, J = 32.9, 7.4 Hz, 1H), 5.00 (d, J = 19.4 Hz, 2H), 3.60 (d, J = 7.7 Hz, 2H); LCMS: for C ₁₃ H ₁₃ FN ₂ O ₂ S calculated 280.1, found 281.1 [M+1] ⁺ .

( Z )-3-Fluoro-4-(2-fluorophenylsulfonyl)but-2-en-1-amine hydrochloride (Compound 37)

[0283] Off-white solid; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 8.01-7.90 (m, 1H), 7.89-7.77 (m, 1H), 7.53-7.39 (m, 2H), 5.29 (dt, J = 32.8, 7.4 Hz, 1H), 4.49 (d, J = 19.1 Hz, 2H), 3.63 (dd, J = 7.4, 1.9 Hz, 2H).

( Z )-3-Fluoro-4-(3-fluorophenylsulfonyl)but-2-en-1-amine hydrochloride (Compound 38)

[0284] Off-white solid; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 7.83 (ddd, J = 7.8, 1.7, 1.1 Hz, 1H), 7.81-7.66 (m , 2H), 7.56 (tdd, J = 8.4, 2.6, 1.1 Hz, 1H), 5.23 (dt, J = 32.9, 7.4 Hz, 1H), 4, 44 (d, J = 19.1 Hz, 2H), 3.65 (dd, J = 7.4, 1.9 Hz, 2H).

( Z )-3-Fluoro-4-(4-fluorophenylsulfonyl)but-2-en-1-amine hydrochloride (Compound 39)

[0285] Off-white solid; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 8.13-7.99 (m, 2H), 7.51-7.34 (m, 2H), 5.19 (dt, J = 32.8, 7.5 Hz, 1H), 4.39 (d, J = 19.1 Hz, 2H), 3.62 (ddt, J = 7.4, 1.9, 0.6 Hz, 2H).

( Z )-3-Fluoro-4-( o- tolylsulfonyl)but-2-en-1-amine hydrochloride (Compound 40)

[0286] Off-white solid; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 7.99 (dd, J = 7.6, 1.1 Hz, 1H), 7.71-7.59 (m, 1H), 7.53-7.39 (m, 2H), 5.22 (dt, J = 32.8, 7.4 Hz, 1H), 4.35 (dd, J = 19.3, 0.5 Hz, 2H), 3.63 (ddt, J = 7.4, 2.0, 0.6 Hz, 2H), 2.73 (s, 3H).

( Z )-3-Fluoro-4-( m- tolylsulfonyl)but-2-en-1-amine hydrochloride (Compound 41)

[0287] Off-white solid; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 7.84-7.72 (m, 2H), 7.67-7.49 (m, 2H), 5.44-5.08 (m, 1H), 4.35 (dc, J = 19.1, 0.5 Hz, 2H), 3.64 (ddt, J = 7.4, 2.0, 0.6 Hz, 2H), 2.56 - 2.38 (m, 3H).

( Z )-3-Fluoro-4-( p- tolylsulfonyl)but-2-en-1-amine hydrochloride (Compound 42)

[0288] Off-white solid; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 7.88-7.82 (m, 2H), 7.54-7.45 (m, 2H), 5.17 (dt, J = 32.8, 7.4 Hz, 1H), 4.32 (dd, J = 19.2, 0.5 Hz, 2H), 3.64 (ddt, J = 7.4, 2.0, 0 .6 Hz, 2H), 2.49 (s, 3H)

( Z )-3-Fluoro-4-((3-fluoroquinolin-8-yl)sulfonyl)but-2-en-1-amine dihydrochloride (Compound 51)

[0289] ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 9.09 (dd, J = 2.9, 0.6 Hz, 1H), 8.49 (dd, J = 7, 4, 1.4, 0.4 Hz, 1H), 8.38 (ddd, J = 8.3, 1.4, 0.4 Hz, 1H), 8.31 (dd, J = 8.8, 2.9 Hz, 1H), 7.87 (ddd, J = 8.3, 7.3, 0.8 Hz, 1H), 5.20 (dd, J = 32.9, 7.4 Hz, 1H ), 4.99 (d, J = 19.3 Hz, 2H), 3.59 (d, J = 7.4 Hz, 2H).

EXAMPLE 6

[0290] The following compounds were prepared according to methods F, J, H, and I using the appropriately functionalized thiol starting material.

Method J: Getting ( Z )- tert- butyl-(3-fluoro-4-((4-(methylsulfonyl)phenyl)sulfonyl)but-2-en-1-yl)carbamate

[0291] To a stirred suspension of ( Z ) -tert -butyl(3-fluoro-4-((4-(methylthio)phenyl)thio)but-2-en-1-yl)carbamate (120 mg, 0.35 mmol ) and sodium bicarbonate (150 mg, 1.79 mmol) in CH ₂ Cl ₂ (4 ml) and water (2 ml) at 0 ° C was added 3-chloroperoxybenzoic acid (378 mg, 2.19 mmol) in three portions during 5 minutes. The resulting suspension was stirred at 0°C for 1.5 hours before diluting with aq. NaOH (2 M; 1 ml), water (10 ml) and CH ₂ Cl ₂ . Then the phases were separated and the aqueous phase was extracted with CH ₂ Cl ₂ (10 ml × 2). The organic phases were combined, dried (Na ₂ SO ₄ ) and concentrated in vacuo . The crude material was purified by flash column eluting with 30% EtOAc/hexane to give ( Z ) -tert- butyl-(3-fluoro-4-((4-(methylsulfonyl)phenyl)sulfonyl)but-2-ene-1- yl)carbamate (17 mg, 12%) as a white solid. ¹ H-NMR (300 MHz; d ₆ -DMSO) δ ppm: 1.37 (9H, s), 3.33 (3H, s), 3.54 (2H, ca. t, J = 5 .6 Hz), 4.62 (2H, d, J = 19.4 Hz), 4.93 (1H, dt, J = 36.4, 6.8 Hz), 7.05 (1H, t, J = 5.8 Hz), 8.15 (2H, dd, J = 8.7, 2.1 Hz), 8.21 (2H, dd, J = 8.7, 2.1 Hz).

( Z )-4-((3,5-bis(trifluoromethyl)phenyl)sulfonyl)-3-fluorobut-2-en-1-aminotrifluoroacetate (Compound 3)

[0292] White solid; etc. 155-157°C; ¹ H-NMR (300 MHz; d ₆ -DMSO) δ ppm: 3.35 (3H, s), 3.50 (2H, approx. d, J = 6.7 Hz), 4.80 ( 2H, d, J = 19.7 Hz), 5.12 (1H, dt, J = 34.7, 7.3 Hz), 7.88 (3H, brs), 8.19 (2H, dd, J = 8.8, 2.5 Hz), 8.24 (2H, dd, J = 8.9, 2.4 Hz).

EXAMPLE 7

[0293] The following compound was prepared according to methods K, L, and M, then F, N, H, and I.

Preparation of ( Z )-4-(2-((4-Amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy) -N , N- dimethylbenzenesulfonamide hydrochloride (Compound 6)

Procedure K: Preparation of 4-bromo- N,N ,-dimethylbenzenesulfonamide

[0294] To a stirred solution of dimethylamine (12 ml, 40% w/w aqueous solution) in THF (20 ml) at 5°C was added a solution of 4-bromobenzenesulfonyl chloride (5.00 g, 19.6 mmol) in THF ( 10 ml) for 5 minutes. After the addition, the mixture was allowed to stir at room temperature for 1 hour. Then the reaction mixture was concentrated in vacuo and the resulting residue was divided between water (25 ml) and CH ₂ Cl ₂ (20 ml), and the aqueous layer was extracted with additional CH ₂ Cl ₂ (20 ml x 2). The combined organic layers were dried over Na ₂ SO ₄ and concentrated in vacuo to give 4-bromo- N,N -dimethylbenzenesulfonamide (4.83 g, 93%) as a white solid. ¹ H-NMR (300 MHz; CDCl ₃ ) δ ppm: 2.74 (6H, s), 7.64-7.73 (4H, m).

Method L: Getting N,N- dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide

[0295] A stirred suspension of 4-bromo- N,N -dimethylbenzenesulfonamide (1.00 g, 3.79 mmol), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (1.15 g, 4.54 mmol) and potassium acetate (1.11 g, 11.4 mmol) in 1,4-dioxane (25 ml) was purged with nitrogen for 15 minutes before adding the complex of 1,1'-bis(diphenylphosphino)ferrocene-palladium (II) dichloride and dichloromethane (155 mg, 0.19 mmol). The resulting suspension was heated at 80°C under nitrogen for 16 hours. The mixture was cooled to room temperature, partitioned between EtOAc (40 ml) and water (30 ml) and filtered through celite. The organic layer was separated and the aqueous layer was further extracted with EtOAc (20 ml x 2). The combined organic layers were then washed with brine, dried (Na ₂ SO ₄ ) and concentrated in vacuo to give N,N- dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl )benzenesulfonamide (1.60 g, 68%) as a gray solid. ¹ H-NMR (300 MHz; CDCl ₃ ) δ ppm: 1.38 (12H, s), 2.71 (6H, s), 7.77 (2H, dd, J = 8.4, 1 .0 Hz), 7.98 (2H, dd, J = 8.4, 0.9 Hz).

Method M: Getting (4-( N , N- dimethylsulfamoyl)phenyl)boronic acid

[0296] To a stirred solution of N , N- dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide (1.00 g, 2.25 mmol) in THF (20 ml) and water (5 ml) were added sodium periodate (2.06 g, 9.64 mmol) at 0°C. The mixture was stirred for 5 minutes at this temperature and then allowed to warm to room temperature and stirred for another 30 minutes. Aqueous HCl (1 M; 1.57 ml, 1.57 mmol) was added and the resulting mixture was stirred at room temperature for an additional 1 hour. The reaction mixture was diluted with water (30 ml) and extracted with EtOAc (20 ml x 3). The organic layers were then combined and washed (brine), dried (Na ₂ SO ₄ ) and concentrated in vacuo . The crude material was purified by flash column eluting with 50% EtOAc/hexane followed by 10% MeOH in 50% EtOAc/hexane to give (4-( N , N- dimethylsulfamoyl)phenyl)boronic acid (470 mg, 91%) in as a brown solid. ¹ H-NMR (300 MHz; CD ₃ OD) δ ppm: 2.69 (6H, s), 7.75 (2H, d, J = 8.2 Hz), 7.88-7.98 (2H, m).

Method N: Getting ( Z )- tert- butyl(4-((2-(4-( N , N- dimethylsulfamoyl)phenoxy)phenyl)thio)-3-fluorobut-2-en-1-yl)carbamate

[0297] To a stirred solution of ( Z ) -tert -butyl (3-fluoro-4- ((2-hydroxyphenyl) thio) but-2-en-1-yl) carbamate (150 mg, 0.48 mmol), ( 4-( N , N- dimethylsulfamoyl)phenyl)boronic acid (219 mg, 0.96 mmol) and pyridine (0.19 ml, 2.39 mmol) in CH ₂ Cl ₂ (6 ml) copper acetate was added at room temperature (II) (87 mg, 0.48 mmol) in one serving. The resulting mixture was stirred at this temperature for 16 hours. After this time, the reaction mixture was diluted with CH ₂ Cl ₂ (30 ml), filtered through celite and washed with aq. HCl (1 M; 20 ml) followed by sat. aq. NaHCO ₃ (20 ml) and brine (20 ml). The organic phase was then dried (Na ₂ SO ₄ ) and concentrated in vacuo . The crude material was purified by flash column eluting with 25% EtOAc/hexane to give ( Z )-t tert- butyl-(4-((2-(4-( N , N- dimethylsulfamoyl)phenoxy)phenyl)thio)-3 -fluorobut-2-en-1-yl)carbamate (80 mg, 34%) as a yellow oil. ¹ H-NMR (300 MHz; CDCl ₃ ) δ ppm: 1.45 (9H, s), 2.73 (6H, s), 3.55 (2H, d, J = 17.1 Hz) , 3.73 (2H, about t, J = 5.6 Hz), 4.46 (1H, brs), 4.80 (1H, dt, J = 34.8, 6.8 Hz), 7.02 (2H, d, J = 8.7 Hz), 7.06 (1H, dd, J = 8.2, 1.0 Hz), 7.24 (1H, ddd, J = 7.5, 7.5 , 1.1 Hz), 7.35 (1H, dd, J = 7.6, 7.6, 1.6 Hz), 7.52 (1H, dd, J = 7.7, 1.5 Hz) , 7.75 (2H, d, J = 8.6 Hz).

( Z )-4-(2-((4-Amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy) -N , N- dimethylbenzenesulfonamide hydrochloride (Compound 6)

[0298] White solid; etc. 153-156°C; ¹ H-NMR (300 MHz; CD ₃ OD) δ ppm: 2.73 (6H, s), 3.64 (2H, approx. d, J = 6.9 Hz), 4.57 (2H , d, J = 19.1 Hz), 5.30 ( 1H, dt, J = 32.8, 7.3 Hz), 7.25 (1H, dd, J = 8.3, 0.7 Hz) , 7.30 (2H, dd, J = 8.9, 2.0 Hz), 7.49 (1H, dd, J = 7.9, 7.9, 1.0 Hz), 7.81 (1H , dd, J = 7.8, 8.3, 1.7 Hz), 7.86 (2H, dd, J = 8.6, 2.0 Hz), 8.07 (1H, dd, J = 7 .9, 1.6 Hz).

EXAMPLE 8

[0299] The following compound was prepared according to methods K, L, M, N, H, and I.

( Z )-4-(3-(4-Amino-2-fluorobut-2-enylsulfonyl)phenoxy) -N , N- dimethylbenzenesulfonamide hydrochloride (Compound 16)

[0300] White solid; 1H NMR (300 MHz, CD ₃ OD) δ ppm: 7.89 - 7.82 (m, 3H), 7.76 (td, J = 8.0, 0.5 Hz, 1H), 7 .68 (t, J = 2.4, 1.7 Hz, 1H), 7.53 (ddd, J = 8.1, 2.5, 1.1 Hz, 1H), 7.29 - 7.22 (m, 2H), 5.28 (dt, J = 33.2, 7.4 Hz, 1H), 4.90 (s, 6H), 4.43 (d, J = 19.1 Hz, 2H) , 3.65 (dd, J = 7.4, 1.9 Hz, 2H).

( Z )-3-(3-(4-Amino-2-fluorobut-2-enylsulfonyl)phenoxy) -N , N- dimethylbenzenesulfonamide hydrochloride (Compound 17)

[0301] White solid; etc. 220°C; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 7.81 (ddd, J = 7.8, 1.7, 1.2 Hz, 1H), 7.77-7.69 (m , 2H), 7.66-7.59 (m, 2H), 7.50 (ddd, J = 8.0, 2.5, 1.2 Hz, 1H), 7.46-7.38 (m , 2H), 5.24 (dt, J = 32.9, 7.4 Hz, 1H), 4.90 (s, 6H), 4.41 (d, J = 19.1 Hz, 2H), 3 .65 (dd, J = 7.4, 1.9 Hz, 2H).

( Z )-3-(2-(4-Amino-2-fluorobut-2-enylsulfonyl)phenoxy) -N , N- dimethylbenzenesulfonamide hydrochloride (Compound 18)

[0302] White solid; etc. 205°C; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 8.06 (ddd, J = 7.9, 1.7, 0.3 Hz, 1H), 7.83 - 7.73 (m , 1H), 7.70 (dd, J = 7.7, 0.7 Hz, 1H), 7.65 (dt, J = 7.8, 1.4 Hz, 1H), 7.51 - 7, 42 (m, 3H), 7.20 (dd, J = 8.3, 1.0 Hz, 1H), 5.31 (dt, J = 33.0, 7.4 Hz, 1H), 4.90 (s, 6H), 4.60 (d, J = 19.2 Hz, 2H), 3.64 (dd, J = 7.4, 1.9 Hz, 2H).

EXAMPLE 9

[0303] The following compound was prepared according to methods O and P, then F, Q, H, and I.

Preparation of (Z) -2-((4-Amino-2-fluorobut-2-en-1-yl)sulfonyl) -N- (4- (N,N - diisopropylsulfamoyl )phenyl)benzamide (Compound 15)

Method O: Receiving N,N - diiso -propyl-4 nitrobenzenesulfonamide

[0304] To a stirred solution of diisopropylamine (3.16 ml, 22.6 mmol) in THF (10 ml) at 0-5 °C was added a solution of 4-nitrobenzenesulfonyl chloride (2.00 g, 9.02 mmol) in THF (5 ml) for 5 minutes. After the addition, the mixture was allowed to stir at room temperature for 16 hours. Then the reaction mixture was concentrated in vacuo and the resulting residue was divided between water (25 ml) and CH ₂ Cl ₂ (20 ml). The phases were separated and the aqueous layer was further extracted with CH ₂ Cl ₂ (20 ml x 2). The combined organic layers were then dried over Na ₂ SO ₄ and concentrated in vacuo. The crude material was purified by flash column eluting with 20% EtOAc/hexane to give N,N - diisopropyl -4-nitrobenzenesulfonamide (285 mg, 11% yield) as a yellow solid. ¹ H-NMR (300 MHz; CDCl ₃ ) δ ppm: 1.30 (12H, d, J = 6.7 Hz), 3.79 (1H, hept, J = 6.9 Hz), 8 .07 (2H, dd, J = 9.2, 2.2 Hz), 8.35 (2H, dd, J = 8.9, 1.9 Hz).

Method P: Preparation of 4-amino- N,N- di iso propylbenzenesulfonamide

[0305] To a stirred solution of N,N -di isopropyl -4-nitrobenzenesulfonamide (260 mg, 0.91 mmol) in methanol (10 ml) at room temperature was added a suspension of palladium on carbon (10% w/w; 50 mg) into water (50 μl) under a nitrogen atmosphere. The resulting mixture was stirred under a hydrogen atmosphere for 3 hours. The mixture was filtered through celite and concentrated in vacuo to give 4-amino- N,N- di- isopropylbenzenesulfonamide (200 mg, 86%) as a brown solid. ¹ H-NMR (300 MHz; CDCl ₃ ) δ ppm: 1.27 (12H, d, J = 6.8 Hz), 3.66 (2H, hept, J = 6.8 Hz), 4 .04 (2H, brs), 6.67 (2H, dd, J = 8.7, 2.1 Hz), 7.65 (2H, dd, J = 8.6, 1.9 Hz).

Procedure Q: Preparation of (Z)-tert-butyl-(4-((2-((4-(N,N-diisopropylsulfamoyl)phenyl)carbamoyl)phenyl)thio)-3-fluorobut-2-en-1-yl ) carbamate

[0306] To a stirred solution of ( Z )-2-((4-(( tert- butoxycarbonyl)amino)-2-fluorobut-2-en-1-yl)thio)benzoic acid (180 mg, 0.53 mmol) , 4-amino- N,N- di -isopropylbenzenesulfonamide (203 mg, 0.79 mmol) and triethylamine (0.26 ml, 1.85 mmol) in DMF (0.8 ml) HATU (301 mg) was added at room temperature , 0.79 mmol) and the resulting solution was stirred at this temperature for 16 hours Then the reaction mixture was divided between water (10 ml) and EtOAc (10 ml) and the phases were separated. The aqueous phase was back extracted with EtOAc and the organic phases were combined and washed with HCl (1 M; 20 ml), then water (20 ml x 3) and brine (20 ml). The organic phase was dried (Na ₂ SO ₄ ) and concentrated in vacuo . The crude product was purified by flash chromatography eluting with 50-100% EtOAc/hexanes to give (Z) -tert -butyl-(4-((2-((4- (N,N - diisopropylsulfamoyl)phenyl)carbamoyl) phenyl)thio)-3-fluorobut-2-en-1-yl)carbamate (93 mg, 30%) as a pale lilac oil. ¹ H-NMR (300 MHz; CDCl ₃ ) δ ppm: 1.31 (12H, d, J = 6.7 Hz), 1.44 (9H, s), 3.57 (2H, d, J = 18.5 Hz), 3.63 (2H, approx. t, J = 5.6 Hz), 3.74 (2H, hept, J = 6.7 Hz), 4.40 (1H, dt, J = 34.8, 6.9 Hz), 4.75 (1H, brs), 7.41 (1H, dd, J = 7.1, 7.1 Hz), 7.47 (1H, dd, J = 7.8, 7.8, 1.8 Hz), 7.62 (1H, dd, J = 7.8, 1.2 Hz), 7.75 (1H, dd, J = 7.3 Hz), 7.88 (4H, br).

(Z) -2-((4-Amino-2-fluorobut-2-en-1-yl)sulfonyl) -N- (4- (N,N - diisopropylsulfamoyl )phenyl)benzamide hydrochloride (Compound 15)

[0307] White solid; etc. 248-250°C; ¹ H-NMR (300 MHz; CD ₃ OD) δ ppm: 1.29 (12H, d, J = 6.9 Hz), 3.64 (2H, dd, J = 7.4, 1, 5 Hz), 3.78 (2H, hept, J = 6.7 Hz), 4.69 (2H, d, J = 19.2 Hz), 5.23 (1H, dt, J = 32.4, 7.4 Hz), 7.76-7.82(2H, m), 7.85-7.93(5H, m), 8.10-8.13(1H, m).

EXAMPLE 10

[0308] The following compounds were prepared according to methods Q, H, and I using the appropriate starting thiol and amino materials.

N- (adamantan-1-yl)-4-((( Z )-4-amino-2-fluorobut-2-en-1-yl)sulfonyl)benzamide hydrochloride (Compound 4)

[0309] Off-white solid; etc. 231-233°C; ¹ H-NMR (300 MHz; d ₆ -DMSO) δ ppm: 1.67 (6H, brs), 2.08 (9H, brs), 3.46 (2H, brs), 4.67 ( 2H, d, J = 19.7 Hz), 5.10 (1H, dt, J = 34.6, 7.1 Hz), 7.97 (2H, dd, J = 8.6, 1.5 Hz ), 8.00 (2H, dd, J = 8.8, 1.4 Hz), 8.06 (3H, brs).

N- (adamantan-1-yl)-2-(((Z)-4-amino-2-fluorobut-2-en-1-yl)sulfonyl)benzamide hydrochloride (Compound 12)

[0310] White solid; ¹ H-NMR (300 MHz; CD ₃ OD) δ ppm: 1.77-1.80 (6H, m), 2.09-2.15 (3H, m), 2.16-2, 21(6H, m), 3.61(2H, ca. q, J =7.2 Hz), 4.62(2H, d, J =19.5 Hz), 5.16(1H, dt, J = 32.6, 7.4 Hz), 7.56 (1H, dd, J = 7.5, 1.0 Hz), 7.67 (1H, dd, J = 7.7, 7.7, 1, 2 Hz), 7.78 (1H, dd, J = 7.5, 7.5, 1.1 Hz), 8.01 (1H, dd, J = 7.8, 1.0 Hz).

N- (adamantan-1-yl)-3-((( Z )-4-amino-2-fluorobut-2-en-1-yl)sulfonyl)benzamide hydrochloride (Compound 13)

[0311] White solid; etc. 230-232°C; ¹ H-NMR (300 MHz; d ₆ -DMSO) δ ppm: 1.68 (6H, brs), 2.05-2.12 (9H, m), 3.49 (2H, brs), 4.68 (2H, d, J 19.8 Hz), 5.12 (1H, dt, J 34.7, 7.0 Hz), 7.73 (1H, dd, J 7.7, 7.7 Hz), 7.98 (3H, brs), 8.03 (1H, ddd, J 7.8, 1.7, 0.9 Hz), 8.16 (1H, ddd, J 7.8, 1, 4, 1.0 Hz), 8.28 (1H, dd, J 1.6, 1.1 Hz).

EXAMPLE 11

[0312] The following compounds were prepared according to methods R and I.

( Z )-3-Fluoro-4-(phenylsulfonyl)but-2-en-1-amine hydrochloride (Compound 1)

Method R: Getting tert- butyl ( Z )-(3-fluoro-4-(phenylsulfonyl)but-2-en-1-yl)carbamate

[ 0313 ] Sodium benzenesulfinate ( 44.1 g, 269 mmol) in portions at room temperature for 15 minutes. The resulting reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with water (2.7 L) and continued stirring at room temperature for another 15 minutes. The resulting precipitated solid was filtered, washed with water (50 ml x 2) and then dried in an oven at 60° C. to give t- butyl (Z)-(3-fluoro-4-(phenylsulfonyl)but-2-ene-1- yl)carbamate (74.0 g, 100%) as a white solid, which was used as such in the next step. ¹ H NMR (300 MHz, CDCl ₃ ) δ ppm: 7.98-7.91 (m, 2H), 7.76-7.67 (m, 1H), 7.61 (ddt, J = 8.3, 6.6, 1.3 Hz, 2H), 4.94 (dt, J = 34.3, 7.0 Hz, 1H), 4.59 (s, 1H), 3.94 (d , J = 18.4 Hz, 2H), 3.79 (t, J = 6.5 Hz, 2H), 1.46 (s, 9H).

( Z )-3-Fluoro-4-(phenylsulfonyl)but-2-en-1-amine hydrochloride (Compound 1)

[0314] Off-white solid; etc. 209-211°C; ¹ H-NMR (300 MHz; CD ₃ OD) δ ppm: 3.64 (2H, dd, J = 7.3, 1.2 Hz), 4.36 (2H, d, J = 19, 1 Hz), 5.18 (1H, dt, J = 32.7, 7.4 Hz), 7.65-7.71 (2H, m), 7.79 (1H, tt, J = 7.4 , 1.2 Hz), 7.96-8.00 (2H, m); LCMS: for C ₁₀ H ₁₂ FNO ₂ S calculated 229.1, found 230.1 [M+1] ⁺ .

EXAMPLE 12

[0315] The following compounds were prepared according to methods R and I.

( Z )-4-(2-Chlorophenylsulfonyl)-3-fluorobut-2-en-1-amine hydrochloride (Compound 19)

[0316] White solid; etc. 205-207°C; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 8.13 (ddd, J = 7.9, 1.5, 0.6 Hz, 1H), 7.81-7.69 (m , 2H), 7.62 (ddd, J = 7.9, 6.4, 2.2 Hz, 1H), 5.27 (dt, J = 32.8, 7.4 Hz, 1H), 4, 59 (d, J = 19.1 Hz, 2H), 3.62 (dd, J = 7.4, 1.9 Hz, 2H); LCMS: for C ₁₀ H ₁₁ ClFNO ₂ S calculated 263.0, found 264.0 [M+1] ⁺ .

( Z )-3-Fluoro-4-(pyridin-2-ylsulfonyl)but-2-en-1-amine dihydrochloride (Compound 30)

[0317] White solid; etc. 153-155°C; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 8.88-8.73 (m, 1H), 8.25-8.10 (m, 2H), 7.77 (ddd, J = 6.8, 4.7, 1.9 Hz, 1H), 5.26 (dt, J = 32.9, 7.4 Hz, 1H), 4.59 (d, J = 19.1 Hz, 2H), 3.63 (dd, J = 7.3, 1.8 Hz, 2H).

( Z )-3-Fluoro-4-(pyridin-3-ylsulfonyl)but-2-en-1-amine dihydrochloride (Compound 31)

[0318] White solid; etc. 168-170°C; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 9.23 (dd, J = 2.3, 0.8 Hz, 1H), 9.04 (dd, J = 5.2, 1 .5 Hz, 1H), 8.64 (dt, J = 8.2, 1.9 Hz, 1H), 7.94 (ddd, J = 8.2, 5.2, 0.8 Hz, 1H) , 5.30 (dt, J = 33.1, 7.4 Hz, 1H), 4.59 (d, J = 19.1 Hz, 2H), 3.68 (d, J = 7.4 Hz, 2H).

EXAMPLE 13

[0319] The following compound was prepared according to methods S, F, H, and I.

( Z )-3-Fluoro-4-(3-methylpyridin-2-ylsulfonyl)but-2-en-1-amine dihydrochloride (Compound 43)

Method S: Preparation of 3-methylpyridine-2-thiol

[0320] 2-Chloro-3-methylpyridine (500 mg, 3.93 mmol) and sodium hydrosulfide (2.21 g, 39.36 mmol) were taken up in DMF (2.0 ml). The resulting mixture was heated at 120°C for 12 h. After completion of the reaction (TLC), the reaction mixture was cooled to room temperature, diluted with water (20 ml), acidified (pH = 5) with acetic acid and extracted with EtOAc (20 ml x 3) . The combined organic extract was dried over Na ₂ SO ₄ and concentrated under reduced pressure to give 3-methylpyridine-2-thiol (1.50 g, 50.5%). ¹ H NMR (600 MHz, d ₆ -DMSO) δ ppm: 13.5-13.3 (m, 1H), 7.6-7.58 (m, 1H), 7.51 (d, J = 6 Hz, 1H), 2.21 (s, 3H).

( Z )-3-Fluoro-4-(3-methylpyridin-2-ylsulfonyl)but-2-en-1-amine dihydrochloride (Compound 43)

[0321] Off-white solid; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 8.53 (dd, J = 4.7, 1.3 Hz, 1H), 7.92 (d, J = 7.6 Hz, 1H), 7.60 (dd, J = 7.8, 4.5 Hz, 1H), 5.37 (dt, J = 32.9, 7.2 Hz, 1H), 4.75 (d, J = 19.0 Hz, 2H), 3.67 (d, J = 7.1 Hz, 2H), 2.68 (s, 3H).

EXAMPLE 14

[0322] The following compounds were prepared according to methods S, F, H, and I.

( Z )-3-Fluoro-4-(2-methylpyridin-4-ylsulfonyl)but-2-en-1-amine dihydrochloride (Compound 44)

[0323] Pale yellow solid; ¹ H NMR (300 MHz, CD ₃ OD) δ 9.00 (d, J = 5.6 Hz, 1H), 8.35 (s, 1H), 8.23 (d, J = 5.4 Hz, 1H), 5.36 (dt, J = 33.0, 7.1 Hz, 1H), 4.68 (d, J = 18.9 Hz, 2H), 3.69 (d, J = 6.7 Hz, 2H), 2.90 (s, 3H).

( Z )-3-Fluoro-4-(5-isopropylpyridin-2-ylsulfonyl)but-2-en-1-amine dihydrochloride (Compound 34)

[0324] White glassy solid; ¹ H NMR (300 MHz, CD ₃ OD) δ 8.70 (t, J = 1.3 Hz, 1H), 8.12-7.94 (m, 2H), 5.28 (dt, J = 32 .9, 7.4 Hz, 1H), 4.55 (d, J = 19.1 Hz, 2H), 3.64 (dd, J = 7.6, 1.7 Hz, 2H), 3.15 (hept, J = 6.9 Hz, 1H), 1.36 (d, J = 6.9 Hz, 6H).

( Z )-3-Fluoro-4-(5-methylpyridin-2-ylsulfonyl)but-2-en-1-amine dihydrochloride (Compound 35)

[0325] White solid; etc. 155-157°C; ¹ H NMR (300 MHz, d ₆ -DMSO) δ 8.67 (d, J = 0.9 Hz, 1H), 8.09 (s, 3H), 7.98 (t, J = 1.4 Hz , 2H), 5.20 (dt, J = 34.8, 7.2 Hz, 1H), 4.70 (d, J = 19.6 Hz, 2H), 3.46 (t, J = 5, 9 Hz, 2H), 2.45 (s, 3H).

( Z )-3-Fluoro-4-(6-methylpyridin-2-ylsulfonyl)but-2-en-1-amine dihydrochloride (Compound 36)

[0326] Pale yellow solid; etc. 174-176°C; ¹ H NMR (300 MHz, d ₆ -DMSO) δ 8.18 (s, 3H), 8.06 (t, J = 7.8 Hz, 1H), 7.88 (d, J = 7.7 Hz , 1H), 7.66 (d, J = 7.5 Hz, 1H), 5.23 (dt, J = 34.8, 7.1 Hz, 1H), 4.70 (d, J = 19, 6 Hz, 2H), 3.46 (t, J = 5.9 Hz, 2H), 2.61 (s, 3H).

EXAMPLE 15

[0327] The following compound was prepared according to methods T, U and V, then F, H and I.

( Z )-3-Fluoro-4-(6-isopropylpyridin-3-ylsulfonyl)but-2-en-1-amine dihydrochloride (Compound 49)

Method T: Preparation of methyl 3-((6-chloropyridin-3-yl)thio)propanoate

[0328] In a 50 ml resealable reaction tube, 2-chloro-5-iodopyridine (2.50 g, 10.5 mmol) was dissolved in degassed 1,4-dioxane (25 ml) at room temperature under nitrogen. Pd ₂ (dba) ₃ (100 mg, 0.11 mmol), xanthos (125 mg, 0.22 mmol), methyl 3-mercaptopropanoate (1.25 g, 10.5 mmol) and DIPEA (2.50 ml, 14.4 mmol) under nitrogen atmosphere. The solution was degassed by purging with nitrogen gas for 15 min and then gradually heated to 70°C. The resulting reaction mixture was stirred at this temperature for 6 hours. Upon completion of the reaction (TLC), the reaction mixture was cooled to room temperature, diluted with cold water and extracted with ethyl acetate (3×30 ml). The combined organic layers were washed with brine and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (100-200 mesh) eluting with 10% EtOAc-hexanes to give methyl 3-((6-chloropyridin-3-yl)thio)propanoate (2.40 g, 99%) in form of an off-white solid. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm: 8.36 (d, J = 2.4 Hz, 1H), 7.67 (dd, J = 8.4, 2.4 Hz, 1H ), 7.28 (d, J = 9.2 Hz, 1H), 3.69 (s, 3H), 3.17 (t, J = 7.2 Hz, 2H), 2.64 (t, J = 7.2 Hz, 2H).

Method U: Preparation of methyl 3-((6-isopropylpyridin-3-yl)thio)propanoate

[0329] In a 1000 ml round bottom flask was dissolved methyl 3-((6-chloropyridin-3-yl)thio)propanoate (5.00 g, 21.6 mmol) in anhydrous THF (200 ml) and 1-methyl- 2-pyrrolidinone (25 ml) under nitrogen atmosphere. The solution was cooled to -55°C, and a solution of iron (III) acetylacetonate (1.70 g, 4.80 mmol) in THF (50 ml) was added, maintaining a nitrogen atmosphere. The resulting mixture was stirred at -55°C for 15 min, during which time a solution of isopropylmagnesium chloride in THF (2 M, 50 ml) was added dropwise at -55°C under nitrogen atmosphere. The resulting reaction mixture was stirred at -40°C for another 1 hour. After completion of the reaction (TLC), the reaction mixture was cooled to 0°C, quenched with saturated NH ₄ Cl solution (50 ml) and the product was extracted with ethyl acetate (100 ml x 3). The combined organic layers were dried over Na ₂ SO ₄ and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (100-200 mesh) eluting with 10% EtOAc-hexanes to give methyl 3-((6-isopropylpyridin-3-yl)thio)propanoate (2.60 g, 50%) in form of a pale yellow liquid.

Procedure V: Preparation of 6-isopropylpyridine-3-thiol

[0330] In a 100 ml round bottom flask, dissolve methyl 3-((6-isopropylpyridin-3-yl)thio)propanoate (860 mg, 3.59 mmol) in anhydrous THF (20 ml) and cool the solution to -78° C in a nitrogen atmosphere. To the above mixture was added a solution of potassium tert- butoxide in THF (1.0 M, 3.5 ml, 3.59 mmol) under a nitrogen atmosphere. The resulting mixture was stirred at -78°C for 1 h. After completion of the reaction (TLC), the reaction mixture was warmed to room temperature and concentrated under reduced pressure. The resulting residue was washed with n- hexane to give 6-isopropylpyridine-3-thiol (655 mg) as a light brown solid. The compound was used in the next step without further purification.

( Z )-3-Fluoro-4-(6-isopropylpyridin-3-ylsulfonyl)but-2-en-1-amine dihydrochloride (Compound 49)

[0331] ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 9.29 (dd, J = 2.1, 0.7 Hz, 1H), 9.02 (ddd, J = 8, 6, 3.5, 2.1 Hz, 1H), 8.32 (dd, J = 8.6, 3.2 Hz, 1H), 5.48 (t, J = 7.3 Hz, 1H), 4.74 (d, J = 19.0 Hz, 2H), 3.81 - 3.65 (m, 2H), 3.55 (dc, J = 7.0, 2.2 Hz, 1H), 1 .53 (dd, J = 7.0, 0.8 Hz, 6H).

EXAMPLE 16

[0332] The following compound was prepared according to methods T, U, and V, then F, H, and I.

( Z )-3-Fluoro-4-(2-isopropylpyridin-3-ylsulfonyl)but-2-en-1-amine dihydrochloride (Compound 45)

[0333] ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 8.95 (dd, J = 5.2, 1.7 Hz, 1H), 8.66 (dd, J = 8, 2, 1.8 Hz, 1H), 7.78 (dd, J = 8.0, 5.0 Hz, 1H), 5.39 (dt, J = 33.2, 7.3 Hz, 1H), 4.58 (d, J = 19.0 Hz, 2H), 4.08 (p, J = 6.8 Hz, 1H), 3.72 - 3.62 (m, 2H), 1.45 (d , J = 6.7 Hz, 6H).

EXAMPLE 17

[0334] The following compound was prepared according to methods T and V, then F, H and I.

( Z )-3-Fluoro-4-(6-methylpyridin-3-ylsulfonyl)but-2-en-1-amine dihydrochloride (Compound 46)

[0335] ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 9.17 (d, J = 2.2 Hz, 1H), 8.67 (d, J = 8.5 Hz, 1H ), 7.96 (d, J = 8.4 Hz, 1H), 5.33 (dt, J = 33.0, 7.3 Hz, 1H), 4.62 (d, J = 19.1 Hz , 2H), 3.75 - 3.63 (m, 2H), 2.85 (s, 3H).

( Z )-3-Fluoro-4-(2-methylpyridin-3-ylsulfonyl)but-2-en-1-amine dihydrochloride (Compound 47)

[0336] ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 9.00 (dd, J =5.6, 1.5 Hz, 1H), 8.91 (dd, J =8, 1, 1.5 Hz, 1H), 8.05 (dd, J = 8.1, 5.6 Hz, 1H), 5.42 (dt, J = 33.2, 7.3 Hz, 1H), 4.67 (d, J = 19.0 Hz, 2H), 3.79 - 3.64 (m, 2H), 3.12 (s, 3H).

( Z )-3-Fluoro-4-(4-methylpyridin-3-ylsulfonyl)but-2-en-1-amine dihydrochloride (Compound 48)

[0337] Off-white solid; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 9.25 (s, 1H), 8.97 (d, J = 5.8 Hz, 1H), 8.07 (d, J = 5.9 Hz, 1H), 5.42 (dt, J = 33.2, 7.3 Hz, 1H), 4.68 (d, J = 19.1 Hz, 2H), 3.70 (dd, J = 7.4, 1.9 Hz, 2H), 2.99 (d, J = 0.6 Hz, 3H).

( Z )-3-Fluoro-4-((2-methylbenzo[ d ]thiazol-4-yl)sulfonyl)but-2-en-1-amine hydrochloride (Compound 50)

[0338] Pale yellow solid; etc. 243-245°C; ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 8.37 (dd, J = 8.1, 1.2 Hz, 1H), 8.10 (dd, J = 7.7, 1 .2 Hz, 1H), 7.63 (t, J = 7.9 Hz, 1H), 5.20 (dt, J = 32.8, 7.4 Hz, 1H), 4.82 (d, J = 19.0 Hz, 2H), 3.60 (d, J = 7.4 Hz, 2H), 2.97 (s, 3H).

EXAMPLE 18

[0339] The following compound was prepared according to methods W, X, Y, Z, then H and I.

( Z )-4-((2,3-dimethyl- 1H- indol-7-yl)sulfonyl)-3-fluorobut-2-en-1-amine hydrochloride (Compound 52)

Method W: Preparation of (2-iodophenyl)hydrazine

[0340] In a 1 L round bottom flask, a solution of 2-iodaniline (40.0 g, 0.182 mmol) in conc. HCl (250 ml) was treated with a solution of NaNO ₂ (15.1 g, 0.22 mmol) in water (40 ml) at 0°C. The reaction mixture was stirred at 0°C for 2 h and slowly treated with SnCl ₂ (86.6 g, 0.46 mmol) at 0°C. The reaction temperature was gradually raised to room temperature and stirred for another 6 hours. After completion of the reaction (TLC), the reaction mixture was filtered and washed with n-pentane (50 ml) and diethyl ether (50 ml) to give the title compound (42 g, 98.26%). ¹ H NMR (300 MHz, d ₆ -DMSO) δ ppm: 10.2 (br, 2H), 7.79-7.76 (m, 1H), 7.7-7.45 (br, 1H), 7.36 (t, J = 8.1 Hz, 1H), 7.02 (d, J = 7.8 Hz, 1H), 6.75 (t, J = 7.8 Hz, 1H ).

Method X: Preparation of 7-iodo-2,3-dimethyl-1 H -indole

[0341] In a 250 ml round bottom flask, a solution of (2-iodophenyl)hydrazine (10 g, 42.73 mmol) in acetic acid (80 ml) was gradually heated to 60° C. and stirred for 1 hour. 2-Butanone (6.18 g, 85.36 mmol) was added slowly at 60°C. The resulting reaction mixture was then heated at 80° C. for 5 hours. After completion of the reaction (TLC), the reaction mixture was cooled to room temperature and then concentrated under reduced pressure. The resulting residue was diluted with cold water and extracted with ethyl acetate (100 ml x 2). The combined organic extract was washed with brine and concentrated under reduced pressure. The resulting residue was purified on silica gel eluting with 10% EtOAc-hexanes to give the title compound as an off-white solid (2.00 g, 18%). ¹ H NMR (400 MHz, d ₆ -DMSO) δ ppm: 10.54 (s, 1H), 7.37-7.24 (m, 2H), 6.73 (t, J = 8 Hz , 1H), 2.32 (s, 3H), 2.12 (s, 3H).

Method Y: Preparation of methyl-3-((2,3-dimethyl-1 H- indol-7-yl)thio)propanoate

[0342] In a 50 ml resealable reaction tube, a solution of 7-iodo-2,3-dimethyl- 1H -indole (1.60 g, 10.5 mmol) in 1,4-dioxane (10 ml) was degassed under atmosphere nitrogen. Pd ₂ (dba) ₃ (50.0 mg, 0.06 mmol), xanthos (100 mg, 0.18 mmol), methyl 3-mercaptopropanoate (0.70 g, 5.90 mmol) and DIPEA (2 .00 ml, 11.80 mmol) under nitrogen atmosphere. The solution was degassed by purging with argon gas for 15 min, gradually heated to 110°C and stirred at this temperature for 12 hours. After completion of the reaction (TLC), the reaction mixture was cooled to room temperature, diluted with cold water and extracted with EtOAc (2×20 ml). The combined organic extract was washed with brine, dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure. The resulting residue was purified on silica gel eluting with 10% EtOAc-hexanes to give the title compound as an off-white solid (1.50 g, 96%). ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm: 8.6 (br, 1H), 7.44 (d, J = 8 Hz, 1H), 7.22 (d, J = 7.6 Hz, 1H), 7.03 (t, J = 7.6 Hz, 1H), 3.7 (s, 3H), 3.08 (t, J = 6.8 Hz, 2H), 2.57 ( t, J = 7.2 Hz, 2H), 2.41 (s, 3H), 2.22 (s, 3H).

Method Z: Receiving tert- butyl ( Z )-(4-((2,3-dimethyl-1 H- indol-7-yl)thio)-3-fluorobut-2-en-1-yl)carbamate

[0343] In a 100 ml round bottom flask, cesium carbonate (1.49 g, 4.56 mmol) was added to a stirred solution of methyl 3-((2,3-dimethyl-1H-indol-7-yl)thio)propanoate (0 .40 g 1.52 mmol) and tert- butyl ( Z )-(4-bromo-3-fluorobut-2-en-1-yl)carbamate (0.41 g, 1.53 mmol) in DMF (15 ml ) at room temperature. The reaction mixture was stirred at room temperature for 5 hours. Upon completion of the reaction (TLC), the reaction mixture was quenched by adding ice-cold water (10 ml) and extracted with ethyl acetate (10 ml x 3). The combined organic extract was dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure. The crude material was purified on silica gel eluting with 20% ethyl acetate in hexanes to give the title compound as a pale yellow liquid (350 mg, 63%). ¹ H NMR (300 MHz, d ₆ -DMSO) δ ppm: 10.65 (s, 1H), 7.31 (d, J = 7.5 Hz, 1H), 7.07 (d, J = 7.8 Hz, 1H), 6.93-6.83 (m, 2H), 4.66 (dt, J = 36.3, 7.2 Hz, 1H), 3.67 (d, J = 19.5 Hz, 2H), 3.47 (t, J = 6 Hz, 2H), 2.32 (s, 3H), 2.13 (s, 3H), 1.34 (s, 9H).

( Z )-4-((2,3-dimethyl- 1H- indol-7-yl)sulfonyl)-3-fluorobut-2-en-1-amine hydrochloride (Compound 52)

[0344] ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 10.25 (s, 1H), 7.80 (dd, J = 7.8, 1.0 Hz, 1H), 7 .55 (dd, J = 7.7, 1.1 Hz, 1H), 7.20 (t, J = 7.7 Hz, 1H), 5.05 (dt, J = 32.7, 7.4 Hz, 1H), 4.34 (d, J = 19.2 Hz, 2H), 3.55 (dd, J = 7.4, 1.9 Hz, 2H), 2.49 - 2.39 (m , 3H), 2.26 (d, J = 0.8 Hz, 3H).

EXAMPLE 19

Preparation of ( Z ) -tert -butyl (4-bromo-3-fluorobut-2-en-1-yl) carbamate

AA Method: Getting tert- butyl-2-oxoethylcarbamate

[0345] The vessel loaded with 3-amino-1,2-propanediol (50.0 kg, 549 mol) and ethyl acetate (150 L) was cooled to 0-5°C. Di- tert- butyl dicarbonate (120 kg, 550 mol) was added portionwise to the vessel. The resulting mixture was stirred at 20-25°C for 12 hours. After cooling the reaction mixture to 0-5 °C, sodium periodate (120 kg, 561 mol) was added in portions. The suspension was stirred at this temperature for 1 hour. The reaction mixture was then filtered and the filter cake washed with ethyl acetate (180 L). The combined filtrate was washed with aqueous NaCl (10% w/w, 300 L), dried over Na ₂ SO ₄ and then concentrated in vacuo to give crude tert- butyl 2-oxoethyl carbamate (70.0 kg, 80%). The crude material was used in the next step without purification.

Method AB: Getting ( E )-ethyl-4-( tert- butoxycarbonylamino)-2-fluorobut-2-enoate

[0346] Vessel loaded with ethyl 2-fluorophosphonoacetate (46.0 kg, 190 mol), acetonitrile (250 L) and 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) (38.0 kg, 250 mol), cooled to 0 - 10 °C. tert- Butyl 2-oxoethylcarbamate (68.0 kg, 427 mol) was added dropwise to the vessel, and the resulting mixture was stirred at 0-10°C for 4 hours. The reaction mixture was diluted with tert- butyl methyl ether (500 L) and water (500 L) and continued to stir for 30 minutes. After another 30 min of settling, the aqueous layer was removed. The organic layer was washed with water (250 L) and then concentrated in vacuo . The residue was dissolved in ethyl acetate (56 L) and petroleum ether (240 L) and purified on silica gel (40 kg, mesh size: 100-200) eluting with ethyl acetate in petroleum ether (1:10) to give the crude product ( E )- ethyl 4-( tert- butoxycarbonylamino)-2-fluorobut-2-enoate (90 kg). The crude material was used in the next step without further purification.

AC Method: Getting ( Z )-T ret- butyl 3-fluoro-4-hydroxybut-2-enylcarbamate

[0347] The reaction vessel was charged with crude ( E )-ethyl 4-( tert- butoxycarbonylamino)-2-fluorobut-2-enoate (90.0 kg, 364 mol), THF (314 L) and methanol (36 L) at 0 - 10 °C, sodium borohydride (16.6 kg, 439 mol) was added in portions over 4 hours. After completion of the addition, the resulting mixture was stirred at 0-10°C for 3 hours. The reaction was quenched by adding aqueous HCl (0.5 N, 900 L). The product was then extracted with ethyl acetate (720 L x 2). The combined organic layers were washed with water (450 L) and concentrated in vacuo . The remaining water was removed by coevaporation with THF (200 L×3) to give crude ( Z ) -tert- butyl-3-fluoro-4-hydroxybut-2-enylcarbamate (85 kg). This material was used in the next step without further purification.

AD technique: Getting ( Z )-4-((tert-butoxycarbonyl)amino)-2-fluorobut-2-en-1-yl 3,5-dinitrobenzoate

[0348] To a reaction vessel containing crude ( Z ) -tert- butyl-3-fluoro-4-hydroxybut-2-enylcarbamate (170 kg, 828 mol), THF (765 L) and triethylamine (174 L, 1245 mol) was added portions of 3,5-dinitrobenzoyl chloride (190 kg, 824 mol) at 0 - 20 °C. The resulting mixture was stirred at 0-20°C for 2 hours and then diluted with water (850 L) and ethyl acetate (1700 L). The aqueous layer was removed and the organic layers were washed with aqueous Na ₂ CO ₃ (10% wt., 850 L x 2) and then water (850 L). After concentration in vacuo (to about 190 L), ethyl acetate (245 L) was added. The mixture was stirred at 55°C to obtain a clear solution. The mixture was cooled to 10-20°C, n- heptane (730 L) was added and the mixture was stirred for 6 hours. The resulting solid was isolated by filtration and washed with n- heptane/ethyl acetate (4:1, 170 L). HPLC analysis showed an E- isomer content of 30%. The content of the E- isomer is reduced to 4.3% by subsequent precipitation. The solid was dissolved in ethyl acetate (570 L). n- heptane was added to the solution, and the resulting suspension was stirred at 20-30°C for 4-6 hours. The solid was separated by filtration and the filter cake was washed with n- heptane/ethyl acetate (4:1). This process is repeated one more time to give ( Z )-4-(( tert- butoxycarbonyl)amino)-2-fluorobut-2-en-1-yl-3,5-dinitrobenzoate (84 kg as a wet "cake") . This material was used in the next step.

Method AE: Getting ( Z )-T ret- butyl 3-fluoro-4-hydroxybut-2-enylcarbamate

[0349] Reaction vessel loaded with ( Z )-4-(( tert- butoxycarbonyl)amino)-2-fluorobut-2-en-1-yl-3,5-dinitrobenzoate (84 kg, 210 mol), THF (187 L ) and aqueous NaOH (1.00 M, 420 L) were stirred at 15-25°C for 2-5 hours. The progress of the reaction was monitored by TLC. The reaction mixture was diluted with isopropyl acetate (966 L) and stirring continued for 10-30 minutes. After settling for 10-30 minutes and subsequent separation of the layers, the aqueous layer was further extracted with isopropyl acetate (483 L). The combined organic layers were washed with aqueous Na ₂ CO ₃ (10% w/w, 420 L x 2), aqueous NaCl (10% w/w, 420 L) and then concentrated to about 84 L. The remaining water was removed by co-evaporation with THF (468 L) to give ( Z ) -tert- butyl-3-fluoro-4-hydroxybut-2-enylcarbamate (29.0 kg, 67%). HPLC analysis showed that the content of the E- isomer was 3.8%.

AF Methodology: Getting ( Z )- tert- butyl-4-bromo-3-fluorobut-2-enylcarbamate

[0350] Reaction vessel loaded with (Z) -tert -butyl 3-fluoro-4-hydroxybut-2-enylcarbamate (29.0 kg, 141 mol) in THF (290 L) and diisopropylethylamine (DIPEA) (74.0 l, 425 mol) was cooled to 0 - 10 °C. A solution of methanesulfonic anhydride (50.0 kg, 287 mol) in THF (145 L) was added dropwise to the solution. After completion of the addition, lithium bromide (74.0 kg, 852 mol) was added in portions while maintaining the temperature between 0-10°C. The resulting mixture was stirred at 0 - 10°C for 4 hours. TLC after this time indicates complete consumption of the starting material. The reaction mixture was diluted with water (290 L) and the product was extracted with ethyl acetate (290 L + 145 L). The combined organic layers were washed with water (150 L) and then concentrated to dryness. The crude residue was dissolved in ethyl acetate (67 L) and n- heptane (440 L) and purified on silica gel (40 kg; mesh size 100-200) eluting with ethyl acetate/ n- heptane (1:5). All fractions containing the desired product were concentrated to dryness. Ethyl acetate (56 L) was added and stirring continued at 40-50°C until a clear solution was obtained. n- heptane (280 L) was added dropwise to the solution. The mixture was cooled to 10-15°C and stirred for 8 hours. The resulting solid was collected by filtration. HPLC analysis showed that the content of the E- isomer is 0.9%. To further reduce the content of the E- isomer, the precipitation process is repeated as follows. Ethyl acetate (33 L) was added and stirring continued at 40-50°C until a clear solution was obtained. n- heptane (164 L) was added dropwise to the solution. The mixture was cooled to 10-15°C and stirred for 8 hours. The resulting solid was collected by filtration and dried to give ( Z ) -tert- butyl 4-bromo-3-fluorobut-2-enylcarbamate (29.5 kg, 68%). HPLC analysis showed that the content of the E- isomer is 0.1%. ¹ H-NMR (300 MHz; CDCl ₃ ) δ ppm: 1.46 (9H, s), 3.85 (2H, dd, J 6.2, 6.2 Hz), 3.93 (2H , d, J 19.5 Hz), 4.66 (1H, brs), 5.16 (1H, dt, J = 34.0, 6.5 Hz).

EXAMPLE 20

[0351] The following compound was prepared according to Methods AG and AH.

Preparation of ( Z )-3-fluoro-4-(phenylsulfonyl)but-2-en-1-amine hydrochloride (Compound 1)

AG Method: Receiving tert- butyl ( Z )-(3-fluoro-4-(phenylsulfonyl)but-2-en-1-yl)carbamate

[0352] Vessel loaded with tert- butyl ( Z )-(4-bromo-3-fluorobut-2-en-1-yl)carbamate (2.40 kg, 8.95 mol) and DMF (12.0 L), cooled to 15 - 20 °C. Sodium benzenesulfinate (2.20 kg, 13.4 mol) was added to the solution, and the resulting mixture was stirred at 15-20°C for 4 hours. The reaction mixture was diluted with water (12.0 L) and continued stirring at room temperature for another 1 hour. The resulting solid was filtered off and the filter cake was further washed with water (6.0 L). The solid was then dried in vacuo at 50-55°C for 20 hours to give tert- butyl( Z )-(3-fluoro-4-(phenylsulfonyl)but-2-en-1-yl)carbamate (2, 70 kg, 92%). retention time (RT) = 13.75 min; Method - Agilent LC/MSD 1200 Series; column: ZORBAX SB-C18, ODS 2000 (50 × 4.6 mm, 5 µm) operating in ES (+) or (-) ionization mode; flow rate 1.5 ml/min; Temperature (T) = 30 °C; detection wavelength: 214 nm; mobile phase: 5% acetonitrile (containing 0.05% trifluoroacetic acid (TFA)) and 95% water (containing 0.05% TFA) to 90% acetonitrile and 10% water over 24 min.

Method AH: preparation of hydrochloride ( Z )-3-fluoro-4-(phenylsulfonyl)but-2-en-1-amine (Compound 1)

[0353] To a vessel loaded with HCl (4.00 M in ethyl acetate; 13.5 L, 54.0 mol) and cooled to 10-20°C was added a filtered solution of (hyflo) tert- butyl ( Z )-(3 -fluoro-4-(phenylsulfonyl)but-2-en-1-yl)carbamate (2.70 kg, 8.20 mol) in ethyl acetate (27.0 L). The reaction mixture was stirred at 10–20°C for 6 hours. The resulting solid was filtered off and the filter cake was washed with ethyl acetate (8.0 L). The solid was then dried in vacuo at 50-55°C for 40 hours to give ( Z )-3-fluoro-4-(phenylsulfonyl)but-2-en-1-amine hydrochloride (Compound 1) (2.10 kg ; 96%). ¹ H-NMR (300 MHz; CD ₃ OD) δ ppm: 3.64 (2H, dd, J = 7.3, 1.2 Hz), 4.36 (2H, d, J = 19, 1 Hz), 5.18 (1H, dt, J = 32.7, 7.4 Hz), 7.65-7.71 (2H, m), 7.79 (1H, tt, J = 7.4 , 1.2 Hz), 7.96-8.00 (2H, m); LCMS: for C ₁₀ H ₁₂ FNO ₂ S calculated 229.1, found 230.1 [M+1] ⁺ .

EXAMPLE 21

[0354] The following compound was prepared according to methods AI, AJ and AK.

Preparation of ( Z )-3-fluoro-4-(quinolin-8-ylsulfonyl)but-2-en-1-amine dihydrochloride monohydrate (Compound 33)

Method AI: Preparation of sodium quinoline-8-sulfinate

[0355] A vessel loaded with Na ₂ SO ₃ (6.70 kg, 53.2 mol) and water (21.0 L) was stirred at room temperature for 20 minutes. Na ₂ CO ₃ (5.50 kg, 51.9 mol) was added to the vessel and stirring continued at room temperature for 20 minutes. Quinoline-8-sulfonyl chloride (6.00 kg, 26.4 mol) was then added in portions while maintaining the temperature below 40°C. The resulting mixture was stirred at room temperature for 3 hours. The reaction mixture was filtered and the filter cake washed with methanol (7.0 L). The filtrate was concentrated to dryness in vacuo and methanol (7.0 L) was added to the resulting residue. After stirring at room temperature for 1 hour, the mixture was filtered and the filtrate was concentrated to dryness. In the second and last wash cycle, the residue was dissolved in methanol (10.0 L) and continued stirring at room temperature for 1 hour. The mixture was filtered and the filtrate concentrated in vacuo to give sodium quinoline-8-sulfinate (4.10 kg, 72%).

AJ Methodology: Receiving tert- butyl ( Z )-(3-fluoro-4-(quinolin-8-ylsulfonyl)but-2-en-1-yl)carbamate

[0356] Vessel loaded with tert- butyl ( Z )-(4-bromo-3-fluorobut-2-en-1-yl)carbamate (3.50 kg, 13.1 mol), sodium quinoline-8-sulfinate (4 .20 kg, 19.5 mol) and DMF (17.5 L) were cooled to 15 - 20 °C. The resulting mixture was stirred at this temperature for 20 hours. The mixture was then diluted with ethyl acetate (35.0 L) and water (35.0 L) and stirring was continued for another 10 minutes. The organic layer was then separated and washed with water (20.0 L×2). After the organic layer was concentrated to about 20 L, n- heptane (42.0 L) was added dropwise. The resulting suspension was stirred at 20 - 30 °C for 20 hours. The solid was isolated by filtration, washed with n- heptane, and then dried in vacuo at 50-55°C for 20 hours to give tert- butyl ( Z )-(3-fluoro-4-(quinolin-8-ylsulfonyl)but- 2-en-1-yl) carbamate (3.80 kg, 77%). RT = 12.97 min; Method - Agilent LC/MSD 1200 Series; column: ZORBAX SB-C18, ODS 2000 (50 × 4.6 mm, 5 µm) operating in ES (+) or (-) ionization mode; flow rate 1.5 ml/min; Temperature (T) = 30 °C; detection wavelength: 214 nm; mobile phase: 5% acetonitrile (containing 0.05% trifluoroacetic acid (TFA)) and 95% water (containing 0.05% TFA) to 90% acetonitrile and 10% water over 24 min.

Method AK: Preparation of dihydrochloride monohydrate ( Z )-3-fluoro-4-(quinolin-8-ylsulfonyl)but-2-en-1-amine (Compound 33)

[0357] To a vessel loaded with HCl (1.5 M in ethyl acetate; 53 L) at 10-20°C was added tert- butyl ( Z )-(3-fluoro-4-(quinolin-8-ylsulfonyl)but- 2-en-1-yl) carbamate (5.3 kg, 14 mol). The mixture was stirred at 15°C for 4 hours. The resulting solid was isolated by filtration and washed with ethyl acetate (20 L). Ethyl acetate (53 L) was added to the flask containing the solid. Then the suspension was stirred at 10–20°C for 2 hours. The solid was isolated by filtration and washed with ethyl acetate (20 L). The solid was dissolved in methanol (53 L) and the solution was filtered. Water (500 ml) and tert- butyl methyl ether (53 L) were then added dropwise to the material, and stirring was continued at 15°C for another 20 hours. The solid was collected by filtration and dried in vacuo at 55-60°C to give ( Z )-3-fluoro-4-(quinolin-8-ylsulfonyl)but-2-en-1-amine dihydrochloride monohydrate (Compound 33) (4 .4 kg, 90%). ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 9.18 (d, J = 4.7 Hz, 1H), 8.70 (dd, J = 8.4, 2.6 Hz, 1H), 8.57 (d, J = 7.4 Hz, 1H), 8.45 (d, J = 8.5 Hz, 1H), 7.99 - 7.68 (m, 2H), 5, 22 (dt, J = 32.9, 7.4 Hz, 1H), 5.00 (d, J = 19.4 Hz, 2H), 3.60 (d, J = 7.7 Hz, 2H); LCMS: for C ₁₃ H ₁₃ FN ₂ O ₂ S calculated 280.1, found 281.1 [M+1] ⁺ .

EXAMPLE 22

[0358] The following compound was prepared according to methods AL and AM.

Preparation of ( Z )-3-fluoro-4-((quinolin-8-yl -d ₆ )sulfonyl)but-2-en-1-amine dihydrochloride (Compound 53)

AL Method: Getting tert- butyl ( Z )-(3-fluoro-4-((quinolin-8-yl- d ₆ )sulfonyl)but-2-en-1-yl)carbamate

[ 0359 ] 2 ,3,4,5,6,7-hexadeuterioquinoline-8-sulfinate sodium (500 mg, 2.26 mmol) per serving. The reaction mixture was stirred at room temperature for 2 hours. Then the reaction mixture was diluted with water (40 ml) and the product was extracted with ethyl acetate (20 ml x 3). The combined organic layer was washed with sat. aq. NH ₄ Cl (20 ml × 3) and brine (20 ml), dried over Na ₂ SO ₄ and then concentrated in vacuo. The crude product was purified by normal phase chromatography (Reveleris) eluting with 20-50% ethyl acetate in hexane to give t- butyl ( Z )-(3-fluoro-4-((quinolin-8-yl _- d6 )sulfonyl)but -2-en-1-yl)carbamate (470 mg, 53%) as an off-white solid.

Method AM: Preparation of dihydrochloride ( Z )-3-fluoro-4-((quinolin-8-yl- d ₆ )sulfonyl)but-2-en-1-amine (Compound 53)

[0360] To a stirred solution of tert- butyl ( Z )-(3-fluoro-4-((quinolin-8-yl -d ₆ )sulfonyl)but-2-en-1-yl)carbamate (450 mg, 1, 22 mmol) in methanol (1.0 ml) at room temperature was added HCl (2.0 M in diethyl ether; 4.0 ml, 8.0 mmol). The reaction mixture was stirred at room temperature for 1 hour, during which time a precipitate formed. The reaction mixture was transferred to a vessel and centrifuged (4000 rpm, 4 min). The supernatant was carefully decanted and the remaining solid cake was dried under high _vacuum to give ( Z )-3-fluoro-4-((quinolin-8-yl- d6 )sulfonyl)but-2-en-1-amine dihydrochloride ( 355 mg, 81%) as a white solid. ¹ H NMR (300 MHz, CD ₃ OD) δ ppm: 5.20 (dt, J = 32.8, 7.4 Hz, 1H), 5.08 - 4.96 (m, 2H), 3.59 (d, J = 7.4 Hz, 2H).

EXAMPLE 23

Method for determining the ability of the compounds of the invention to inhibit LOX and LOXL1-4 from different sources

[0361] Lysyl oxidase (LOX) is an extracellular copper-dependent enzyme that oxidizes peptidyl lysine and hydroxylysine residues in collagen and lysine residues in elastin to form peptidyl-alpha-aminoadipine-delta-semi-aldehydes. This catalytic reaction can be irreversibly inhibited by β-aminopropionitrile (BAPN), which binds to the LOX active site (Tang SS, Trackman PC and Kagan HM, Reaction of aortic lysyl oxidase with beta-aminoproprionitrile. J Biol Chem 1983; 258: 4331-4338 ). There are five members of the LOX family; they are LOX, LOXL1, LOXL2, LOXL3 and LOXL4. The LOX and LOXL family members can be purchased as recombinant active proteins from commercial sources or derived from animal tissues such as aorta, tendons, porcine skin; or obtained from cell cultures. The inhibitory effects of the compounds of the present invention were tested against this LOX-LOXL preparation using a method based on the detection of hydrogen peroxide by the Amplex Red oxidation assay (Zhou et al . 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 1997;253, 162-168). The assay was designed using a 384 or 96 well format. Briefly, in a standard assay, in a 384-well black bottom plate with 25 µl of a dilution of any of the isoenzymes and orthologues 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 and MAO-B and MAO-A, respectively; not required if the enzyme is from a recombinant or purified form). Test compounds were dissolved in DMSO and tested on a concentration versus concentration curve (CPC) with 11 data points, typically in the micromolar or nanomolar range, after incubation with the enzyme for 30 minutes at 37°C. Twenty-five µl of a reaction mixture containing twice the concentration of K _M putrescine (Sigma Aldrich, e.g., 20 mM for LOX, or 10 mM for LOXL2 and LOXL3), 120 µM Amplex Red (Sigma Aldrich), and 1.5 U/ml horseradish peroxidase ( Sigma Aldrich) prepared in 1.2 M urea, 50 mM sodium borate buffer (pH 8.2) was then added to the appropriate wells. The above volumes were doubled for the 96-well plate. Fluorescence (RFU) was read every 2.5 min for 30 min over a temperature range of 37 °C, excitation 565 nm and emission 590 (Optima; BMG labtech). The slope of the kinetics for each well was calculated using MARS data analysis software (BMG labtech) and this value was used to determine the IC ₅₀ value (Dotmatics). The ability of the compounds of the invention to inhibit the activity of LOX amine oxidase and other family members is shown in Table 3.

Table 3

[0362] LOX and LOXL2 inhibitory activities by exemplary compounds of the invention

Compound Bovine LOX activity IC ₅₀ (micromolar) Human LOXL2 activity IC ₅₀ (micromolar) BAPN <10 <1 1 <10 <10 2 <10 <10 3 <10 <10 4 <10 <10 5 <1 <1 6 <10 <1 7 <10 <1 8 <1 <1 9 <1 <1 10 <10 <10 eleven <1 <1 12 <10 <1 13 <10 <10 14 <1 <1 15 <10 <1 16 <1 <1 17 <10 <1 18 <1 <1 19 <10 <1 20 <10 <1 21 <10 <1 22 <10 <1 23 <10 <1 24 <10 <1 25 <10 <1 26 <10 <1 27 <10 <1 28 <10 <1 29 <10 <1 thirty <10 <1 31 <10 <1 32 <10 <1 33 <10 <1 34 <10 <1 35 <10 <1 36 <10 <1 37 <10 <1 38 <10 <1 39 <10 <1 40 <10 <1 41 <10 <1 42 <10 <1 43 <10 <1 44 <10 <1 45 <10 <1 46 <10 <1 47 <10 <1 48 <10 <1 49 <10 <1 50 <10 <1 51 <10 <1 52 <10 <1

EXAMPLE 24

Compounds of the present invention exhibit sustained inhibition of LOXL1 and LOXL2

[0363] For a significant pharmacological effect in the presence of a high substrate concentration, compounds that exhibit sustained, sustained inhibition of LOX and LOXL1-4 have a strong advantage over competitive inhibitors because the pharmacological effect lasts longer than the presence of an unbound inhibitor. In a preferred embodiment, the compounds of the present invention exhibit sustained inhibition of LOX and LOXL1-4.

Method for determining sustained inhibition of LOX and LOXL1-4 by the compounds of the invention

[0364] Step Dilution Experiment: The assay was designed using a 96-well format and the initial enzyme concentration was set to 100 times higher than for inhibition assays. The enzyme is incubated for 40 minutes at 37°C in the presence of 10 times or (if necessary to ensure that the inhibitor concentration exceeds the enzyme concentration) 30 times the IC ₅₀ concentration of the inhibitor being tested. After the incubation, the mixture is diluted 50-fold in assay buffer followed by an additional 2-fold dilution in the Amplex Red-horseradish peroxidase-putrescine reaction mixture (as in inhibition assays) prior to fluorescence measurement. The results were expressed as % signal recovery after a given time compared to uninhibited controls. The results are presented in Table 4.

Table 4

[0365] Measurement of long-term inhibition of LOXL1 and LOXL2 by the compounds of the invention

Compound LOXL1 - Restoring activity. (%) LOXL2 - Activity recovery. (%) Reverse standard (control) 80.9 96.8 1 2.4 9.5 4 16.7 21.2 5 3.6 10.0 6 2.5 10.4 7 3.0 8 35.1 9 4.4 15.1 10 15.2 eleven 10.2 12 2.6 9.3 13 3.5 8.9 14 1.9 15 26.1 16 14.4 17 5.3 18 6.9 19 7.8 7.9 29 8.6 thirty 1.9 31 8.2 33 4.5 6.9 38 1.6 45 6.6 46 1.0

EXAMPLE 25

Pharmacokinetics in vivo

[0366] Test compounds (10 mg/kg and 30 mg/kg in PBS, po n=3) were administered to male Wistar rats at 7-9 weeks of age. Test compound blood samples were collected at time points from 0.25 h to 8.00 h (Compound 1); 0.5 h - 8.00 h after injection (Compound 33) from the tail vein.

[0367] Sample preparation: 20 µl of calibration samples (in one copy), quality control samples (in duplicate) and rat plasma samples were mixed with 60 µl of acetonitrile containing an internal standard (IS; 200 ng/ml tolbutamide and 50 ng/ ml of propanolol) in Eppendorf tubes. The mixture was shaken for 1 min, then centrifuged for 10 min, 50 µl of the supernatant was transferred into a 96-well plate with 100 µl of water. After shaking for 10 minutes, 10 μl was injected into a liquid chromatography and mass spectrometry (LC-MS/MS) system for analysis.

LC method (Compound 1)

HPLC: Agilent 1100; Mass spectrometer: API 4000

Column: Phenomenex Gemini C18 5 µm 50 x 4.6 mm

Mobile phase: 0.1% formic acid in water, 0.1% formic acid in acetonitrile

LC Method (Compound 33)

HPLC: Shimadzu LC30AD; Mass spectrometer: API 4000

Column: Agilent SB C18 1.8 µm 50 x 2.1 mm

Mobile phase: 0.1% formic acid in water, 0.1% formic acid in acetonitrile

[0368] For dilution samples: 10 µl of sample was added to 90 µl of blank rat plasma. The mixture was precipitated with 300 μl of acetonitrile containing IS in Eppendorf tubes. The mixture was shaken for 1 min, then centrifuged for 10 min, 50 µl of the supernatant was transferred into a 96-well plate with 100 µl of water. After shaking for 10 minutes, 10 μl was injected into the LC-MS/MS system for analysis. The determined mean plasma concentrations of Compounds 1 and 33 are shown in Table 5 and Table 6, respectively.

Table 5

Mean plasma concentration of Compound 1 after oral administration of 10 and 30 mg/kg

Compound 1 Oral administration (mg/kg) 10 thirty Time
(watch) Average plasma concentration (ng/ml) Std. Off Average plasma concentration (ng/ml) Std. Off 0.25 379.0 201.0 1057.3 9.4 0.50 736.2 127.5 2088.0 180.9 1.00 449.6 0.7 1202.9 186.7 2.00 133.6 38.5 657.4 60.8 3.00 66.4 18.0 238.5 14.2 5.00 27.2 11.8 41.3 20.6 8.00 6.4 2.7 13.5 n.a.

Table 6

Mean plasma concentration of Compound 33 after oral administration of 10 and 30 mg/kg

Compound 33 Oral administration (mg/kg) 10 thirty Time
(watch) Average plasma concentration (ng/ml) Std. Off Average plasma concentration (ng/ml) Std. Off 0.50 771.3 465.4 3714.2 352.1 1.00 548.3 243.4 3013.6 309.2 2.00 246.0 100.2 1474.8 187.1 4.00 56.4 49.4 184.7 23.0 8.00 8.4 5.2 13.7 0.7

EXAMPLE 26

Target interaction

[0369] A single dose of a mechanism-based inhibitor may be sufficient to block enzymatic activity in vivo for an extended period.

Lysyl oxidase activity measurement - ear slice analysis

[0370] Ears collected immediately after sacrificing rats (treated or untreated with the LOX inhibitor) are quickly frozen in dry ice for storage at -80°C. For analysis, 5x5 mm samples (still half thawed) are cut out, embedded in agarose gel and cut into 200 µm thick cross sections (vibrate). Thin sections were collected in ice-cold PBS containing protease inhibitors and left in the bath for 2-3 hours to allow soluble contaminants to diffuse prior to analysis. For the 96-well plate format: three thin sections were collected, gently blotted dry on Kimwipe, and transferred to each well containing 100 µl of assay buffer (1.2 M urea, 50 mM sodium borate buffer, pH 8.2) at 30 -minute pre-incubation at 37°C in the presence of 1 µM mofegiline and 500 µM pargyline. BAPN (500 μM) was added to all "low control" wells. A minimum of three repetitions were performed for each condition.

[0371] After pre-incubation, 100 µl of a mixture of Amplex Red and horseradish peroxidase (HRP) (120 µM Amplex Red, 1.5 U/ml HRP, 20 mM putrescine) was added to all wells, and data from the plate were read at 37 ° C every 2.5 minutes, 13 times (544 nm excitation and 590 nm emission, on BMG Clariostar in top-reading orbital mode). The slope of the kinetic curves, subtracted from the values obtained in the low control (in the presence of BAPN), was considered a measure of the specific activity of lysyl oxidase in the sample.

EXAMPLE 27

Measurement of lysyl oxidase activity in the aorta

[0372] Sample preparation: all preparation steps were performed on ice with buffers in the presence of protease inhibitors (PMSF: Sigma P7626, 0.25 mM, aprotinin: Sigma A6279, 1 μl per ml) ( Methods in Cell Biology , 2018; 143: 147 - 156). Surrounding adventitia and muscle/fat from a frozen aortic sample was removed in ice-cold wash buffer (0.15 M NaCl, 50 mM sodium borate, pH = 8.0, with protease inhibitors) using fine forceps under a dissecting microscope. The aorta was blotted on Kimwipe, weighed in a 1.5 ml Eppendorf tube and left on ice. After flash freezing the aorta in liquid nitrogen, a mortar and pestle (stored at -80°C) was used to grind the tissue. The minced tissue was transferred into a special tube containing metal balls, 10 x v/w. wash buffer + protease inhibitors. The tissues were homogenized for 5 seconds using a bead mill. The homogenate was centrifuged at 10,000 g for 10 minutes at 4°C, and the supernatant was discarded. The homogenization and washing steps were repeated twice more. The resulting pellet was resuspended in 3 x vol./wt. 6 M urea buffer (50 mM sodium borate, 6 M urea, pH = 8.2) + protease inhibitors, and then shake. The sample was then left at 4°C on a roller for 3 hours of extraction. After extraction, the sample was centrifuged (10,000 g at 4°C for 20 minutes) and the supernatant was saved. An equal volume of all samples was transferred to a fresh dilution tube (minimum 33 µl). Samples were diluted to 2.4 M urea (sample at this stage is ~4.5 M urea) sodium borate buffer (50 mM sodium borate, pH 8.2) + protease inhibitors and used in subsequent analysis.

[0373] Analysis: Pargyline (final concentration 0.5 mM) and mofegiline (final concentration 1 μM) were added to the samples. Duplicate wells were placed in a 384-well black plate, 25 µl per well. To one duplicate was added 0.5 μl of 30 mM BAPN (pan-lox inhibitor); providing a background value (low control). The samples are then incubated for 30 minutes at 37°C. The reaction mixture (25 μl) was added to each well (concentrations were shown as twice the final in the analysis: 120 μm Amplex red, 1.5 u/ml HRP, 20 mm putrescine). The plate fluorescence was then measured every 2.5 min (Ex: 544 nm/ Em: 590 nm/ Gain: 1260; 37°C) on Fluostar™.

[0374] Young male Wistar rats were given a single oral dose (10 or 30 mg/kg) of Compound 1 or 33 (Tables 5 and 6, respectively) and enzyme activity was measured in tissues with high basal activity at that age, in the ear (Fig. 2a and b; Compound 1 and 33, respectively) and aorta (Fig. 3c; Compound 33 only). Responses were normalized to activity measured in animals treated with saline (control).

[0375] Described herein, Compound 1 exerts long-term inhibition of LOX. While Compound 1 plasma concentrations are well below the IC ₅₀ at 8 hours (Table 5) in rat plasma, a dose-dependent sustained decrease in LOX activity can be measured 24 hours after a single oral dose (FIG. 2a); > 20 hours after Compound 1 plasma concentration has fallen below the IC _50.

[0376] A single high (30 mg/kg) dose of Compound 33 was found to completely abolish lysyl oxidase activity. Although Compound 33 plasma concentrations are well below the IC ₅₀ at 8 hours (Table 6), the half-life ranges from 2-3 days (ear) to 24 hours (aorta) (FIGS. 2b and 2c). Thus, Compound 33 causes long-term inhibition that persists beyond the presence of the active compound in plasma.

EXAMPLE 28

Method for determining the ability of compounds of Formula I to inhibit recombinant human SSAO/VAP-1

[0377] Recombinant human amine oxidase SSAO/VAP-1 activity is determined using a combined colorimetric method as described for monoamine oxidase, copper-containing amine oxidases, and related enzymes (Holt A. and Palcic M., A peroxidase-coupled continuous absorbance plate-reader assay for flavin monoamine oxidases, copper-containing amine oxidases and related enzymes Nat Protoc 2006;1: 2498-2505). Briefly, a cloned cDNA template was assembled corresponding to residues 34-763 of human SSAO/VAP-1 and comprising the mouse Ig kappa (κ) signal sequence, the N-terminal epitope tag flag, and the tobacco etch virus (TGV) cleavage site in a mammalian expression vector (pLO -CMV) from Geneart AG. This vector containing 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 isolated by immunoaffinity chromatography. This was used as a source for SSAO/VAP-1 activity. A high throughput colorimetric assay was developed using a 96 or 384 well format. Briefly, in a standard 96-well assay plate, 50 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 tested on a concentration-dependence curve (CPC) with 4-11 data points, typically in the micromolar or nanomolar range after incubation with human SSAO/VAP-1 for 30 minutes at 37°C. After 30 min incubation, a 50 µl reaction mixture containing 600 µM benzylamine (Sigma Aldrich), 120 µM Amplex Red (Sigma Aldrich) and 1.5 U/ml horseradish peroxidase (Sigma Aldrich) prepared in 0.1 M sodium phosphate buffer (pH 7.4) was added to the appropriate well. The fluorescence unit (RFU) was read every 2.5 min for 30 min at 37°C, excitation 565 nm and emission 590 (Optima; BMG labtech). The slope of the kinetics for each well was calculated using MARS data analysis software (BMG labtech) and this value was used to determine the IC ₅₀ value (Dotmatics). The ability of compounds of Formula I to inhibit SSAO/VAP-1 is shown in Table 7.

EXAMPLE 29

Method for determining the ability of compounds of Formula I to inhibit recombinant human MAO-B

[0378] The specificity of the compounds of the present invention is tested by determining their ability to inhibit MAO-B activity in vitro . Recombinant human MAO-B (0.06 mg/ml; Sigma Aldrich) was used as a source of MAO-B enzyme activity. The assay was carried out in the same manner as for human SSAO/VAP-1 (Example 28), except that the substrate benzylamine was used at a concentration of 100 μM. The ability of compounds of Formula I to inhibit MAO-B is shown in Table 7.

Table 7

[0379] Selectivity of Compounds of Formula I for LOX and LOXL2 versus SSAO/VAP-1 and MAO-B

Compound Activity IC ₅₀ SSAO/VAP-1 (micromoles) Activity IC ₅₀ MAO-B (micromoles) BAPN >30 >30 1 >30 >30 2 >30 >30 3 >30 >30 4 >30 <10 5 >30 >30 6 >30 >30 7 >30 >30 8 >30 >30 9 >30 <10 10 >30 >30 eleven >30 >30 12 >30 >30 13 >30 <10 14 >30 >30 15 >30 >30 19 >30 >30 23 >30 - 28 >30 - 31 >30 - 32 >30 33 >10 >30 45 >30 50 >30 51 >10 >30 53 >30

[0380] LOX and LOXL1-4 enzymes are members of a large family of flavin-dependent and copper-dependent amine oxidases, which includes SSAO/VAP-1 and monoamine oxidase-B (MAO-B). The compounds of the present invention selectively inhibit members of the LOX enzyme family against SSAO/VAP-1, MAO-B, and other members of the amine oxidase family. Examples of selectivity values can be seen in Table 7.

EXAMPLE 30

Rodent injury model

[0381] Mice were injured by cutting the skin tissue. In the treatment group, a 1% solution of Compound 1 was applied topically from 24 hours post-injury to 1 week post-injury. The wounds were then left to heal for another week. Mice are sacrificed 14 days post-injury and tissue is analyzed for collagen I content and changes in overall morphology and histology.

[0382] Collagen I (as measured by LCMS and normalized for protein content) was reduced in treated tissue compared to control (FIG. 3a). Histology showed thick parallel bundles of collagen in the control scar tissue. Tissue treated with Compound 1 showed reduced tuft density and more "normal" collagen structure (FIGS. 3b and 3c).

EXAMPLE 31

Pig burn model

[0383] Pigs received 4 deep skin burns of 25 cm ² , 2 on each side. From the time of re-epithelialization, in each pig, 2 wounds were treated with 3% Compound 1 cream daily for 4 weeks and 2 wounds received a control cream. The pigs were sacrificed and the tissues were processed for analysis.

[0384] LOX activity was significantly reduced in the treated tissue (FIG. 4a), as was total collagen I protein (FIG. 4b). General morphology (Fig. 4c-f) and histology (Fig. 4g-h) confirm the decrease in the density of thick collagen fibers in the treated wounds.

EXAMPLE 32

Mouse model of pancreatic cancer

[0385] Female athymic mice (4-5 weeks old) were inoculated with MiaPaca-2luc (human pancreatic cell line) directly into the pancreas. The measured bioluminescent signal intensity and body weight were used to stratify tumors into treatment groups at day 14 after initial inoculation. Treatment groups included 1. solvent, ip. 3 times a week. 2. Abraxane (10 mg/kg ip twice a week) and gemcitabine (initial dose 100 mg/kg ip, for the second dose it was reduced to 60 mg/kg ip due to acute toxicity and was taken twice a week). 3. Abraxane (dosed as described above) and gemcitabine (dosed as described above) and Compound 33 (10 mg/kg ip 3 times a week) (see Fig. 5a). Mice were sacrificed 43 days after tumor inoculation. Mice were monitored twice a week for condition and weight changes. Tumor growth was monitored in vivo by bioluminescent imaging throughout the studies (FIG. 5b) and organs ex vivo during tissue harvest (FIGS. 5c-e).

EXAMPLE 33

Mouse model of sclerosis

[0386] Subcutaneous bleomycin was administered every other day (for a total of 20 days) to female C57BL/6 to induce skin fibrosis as a model for sclerosis. Lesions were treated with either vehicle or 0.5%, 1%, or 3% Compound 1. Histology was completed after 21 days. Histological analysis is shown in Fig. 6 (a-c).

EXAMPLE 34

Mouse model of primary myelofibrosis

[0387] Fifteen to sixteen week old male and female GATA1low mice were intraperitoneally injected with either vehicle (olive oil) or Compound 19 at 15 mg/kg four times a week for 10 weeks. Mice were then sacrificed and spleens and femurs were harvested for histology and analysis (Fig. 7(a-e) and Fig. 8(a-d)).

[0388] Mice treated with Compound 19 had a significantly lower body weight-adjusted spleen weight compared to the solvent group (242.25 ± 18 mg vs. 305.11 ± 22.4 mg, p < 0.05) . There were no differences in pre-treatment hematological parameters, however, mice in the treatment group had significantly lower platelet counts compared to vehicle-treated mice after treatment (77.5 ± 4.4 C/μl vs 106 ± 12, p < 0, 05). Bone marrow (BM) and spleen fibrosis were significantly reduced in mice in the treatment group compared to vehicle. Morphological megakaryocytes (MK) of BM on H&E staining were counted and were reduced in mice in the treatment group compared to vehicle (24.65 ± 0.6 at 20x vs 32.91 ± 0.71, p < 0.001 ).

EXAMPLE 35

Model of unilateral ureteral obstruction (UOM)

[0389] A 14-day unilateral ureteral obstruction (OOM) of acute renal fibrosis in mice was performed to mimic various stages of obstructive nephropathy leading to tubulointerstitial fibrosis in an accelerated fashion. OOM operation was performed by ligation of the left ureter and induced reduction in left kidney thickness and atrophy (indicated by a significant decrease in kidney weight and kidney/body weight ratio compared to the contralateral side), as well as a significant increase in renal fibrosis. The treatment groups received Compound 33 (10 mg/kg per day) orally throughout the study. Compound 33 increased kidney weight and thickness and reduced fibrosis area as measured by Picrosirius Red (FIG. 9). Captopril (~32 mg/kg/day in drinking water) was used as a positive control.

EXAMPLE 36

Model of pulmonary fibrosis induced by bleomycin

[0390] C57Bl/6 mice are injected with 1.5 units/kg of total clinical bleomycin (Blenoxan) via the oropharynx. Compound 33 was administered daily by gavage for 21 days, after which tissues were collected and analyzed. As shown in FIG. 10, Compound 33 significantly reduces Ashcroft's score and lung weight. As expected for the lysyl oxidase inhibitor, the number of immature (DHLNL) and mature (PYD) cross-links per lung also decreased by 37 and 45 percent, respectively.

EXAMPLE 37

Compound 33 Reduces Metastases Associated with Fibrosis

[0391] Increasing evidence suggests that for the occurrence of cancer metastases, it is necessary to create a pre-metastatic niche to promote extravasation and metastatic colonization. The fibrous microenvironment is thought to enhance tumor invasion and metastasis.

[0392] Liver fibrosis was induced by treating BALB/c mice twice a week with carbon tetrachloride (CCl ₄ ) at 1 mg/kg for 8 weeks. In parallel, treatment with Compound 33 (20 mg/kg daily ip) was performed, which significantly reduced liver fibrosis (Fig. 11a and b). At the end of week 4, the mouse breast cancer cell line (4t1) was orthotopically introduced (as shown in Fig. 11a). Treatment with Compound 33 significantly reduced hepatic fibrosis (FIG. 11b), collagen crosslinking, and hepatic metastatic burden (FIGS. 11c and 11d). No differences in primary tumor growth or collagen crosslinking were observed.

EXAMPLE 38

Compounds of the present invention have reduced substrate activity in SSAO compared to E- alkene, isomeric analogs

[0393] Minimizing off-target activity is key in planning and developing compounds for therapeutic use. Compounds including E- 1 have been reported as semicarbazide-sensitive amine oxidase (SSAO) inhibitors [WO2009/066152]. It has been reported that molecules of this type can also become substrates for SSAO, generating potentially toxic metabolites (Foot et. Al., 2012). In a preferred embodiment, the compounds of the present invention are neither inhibitors nor substrates of SSAO.

Substrate turnover measurement by SSAO

[0394] Briefly, the assay used determines the propensity of a compound to a substrate relative to background (dimethyl sulfoxide only). The oxidation of rhSSAO compounds is measured by fluorometric analysis (Holt and Palcic, 2006). Briefly, rhSSAO is incubated for 2 hours at 37°C in HEPES buffer before adding an equal volume of Amplex Red (20 mM), horseradish peroxidase (4 U/ml) and Compound (2.5 mM) in the same buffer. The kinetics of resorufin formation is immediately measured using an Optima reader (BMG Labtech GmbH, Ortenburg, Germany) at 37°C. Representative results are shown in Table 8.

Table 8

Structure Compound Inhibition of recombinant human SSAO (rhSSAO) (IC ₅₀ ) Substrate activity for rhSSAO 1 >100 µM no significant turnover E -1 - turned around like a substrate 31 27 µM no significant turnover E -31 - turned around like a substrate

Claims (51)

1. Compound-Z-isomer of Formula I Formula I or a pharmaceutically acceptable salt or hydrate thereof, wherein: A is C _6-10 aryl or 5-10 membered heteroaryl in which 1-2 ring atoms are heteroatoms independently selected from N and S; each R ¹ is independently selected from the group consisting of XR ² , halogen, deuterium, C _1-6 alkyl, OC _1-6 alkyl, C _6-10 aryl, -C(O)NR ⁴ R ⁵ and -S(O) ₂ R ⁶ where each C _1-6 alkyl and C _6-10 aryl is optionally substituted with one or more halogens; X is selected from the group consisting of O, OCH ₂ and CONH; R ² is selected from the group consisting of C _3-10 cycloalkyl and C _6-10 aryl, where each R ² is optionally substituted with one or more R ⁷ ; R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen and C _1-6 alkyl; R ⁶ is C _1-6 alkyl; R ⁷ is -S(O) ₂ NR ⁴ R ⁵ ; And n is 0, 1, 2, 3, 4, 5, or 6. 2. A compound according to claim 1 wherein A is selected from the group consisting of phenyl, naphthyl, pyridinyl, quinolinyl, benzothiazolyl and indolyl. 3. The compound according to claim 1 or 2, characterized in that A is selected from the group consisting of: 4. A compound according to claim 1, wherein A is a 5-10 membered heteroaryl in which 1-2 ring atoms are heteroatoms independently selected from N and S. 5. Connection according to claim 1, characterized in that A is selected from the group consisting of And ; R ¹ is methyl or isopropyl; And n is 0 or 1. 6. The compound according to claim. 1, characterized in that A is and n is 0. 7. Compound according to claim 1 of Formula Ia Formula Ia or a pharmaceutically acceptable salt thereof, wherein: each R ¹ is independently selected from the group consisting of XR ² , halogen, C _1-6 alkyl, OC _1-6 alkyl, C _6-10 aryl, -C(O)NR ⁴ R ⁵ and -S(O) ₂ R ⁶ where each C _1-6 alkyl and C _6-10 aryl is optionally substituted with one or more halogens; X is selected from the group consisting of O, OCH ₂ and CONH; R ² is selected from the group consisting of C _3-10 cycloalkyl and C _6-10 aryl, where each R ² is optionally substituted with one or more R ⁷ ; R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen and C _1-6 alkyl; R ⁶ is C _1-6 alkyl; R ⁷ is -S(O) ₂ NR ⁴ R ⁵ ; And n is 0, 1, 2, or 3. 8. Connection according to claim 7, characterized in that n is equal to 0. 9. Connection according to claim 7, characterized in that each R ¹ is independently selected from the group consisting of halo, C _1-6 alkyl, OC _1-6 alkyl, C _6-10 aryl, and —S(O) ₂ R ⁶ , wherein each C _1-6 alkyl is optionally substituted with one or several halogens; R ⁶ is C _1-6 alkyl; And n is 1 or 2. 10. The compound according to claim 1, selected from the group consisting of 1 ( Z )-3-fluoro-4-(phenylsulfonyl)but-2-en-1-amine 2 ( Z )-4-((3,5-bis(trifluoromethyl)phenyl)sulfonyl)-3-fluorobut-2-en-1-amine 3 ( Z )-3-fluoro-4-((4-(methylsulfonyl)phenyl)sulfonyl)but-2-en-1-amine 4 N -((1 R ,3 R ,5 S )-adamantan-1-yl)-4-((( Z )-4-amino-2-fluorobut-2-en-1-yl)sulfonyl)benzamide 5 ( Z )-4-((2-((4-amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)methyl)- N , N- dimethylbenzenesulfonamide 6 ( Z )-4-(2-((4-Amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)- N , N- dimethylbenzenesulfonamide 7 ( Z )-4-((2-(benzyloxy)phenyl)sulfonyl)-3-fluorobut-2-en-1-amine 8 ( Z )-4-((2-((4-amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)methyl)benzenesulfonamide 9 ( Z )-4-((3-((4-amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)methyl)- N , N- dimethylbenzenesulfonamide 10 ( Z )-4-((4-((4-Amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)methyl)- N , N- dimethylbenzenesulfonamide eleven ( Z )-4-((2-((4-Amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)methyl)- N , N- diisopropylbenzenesulfonamide 12 N -((1 S ,3 R ,5 S )-adamantan-1-yl)-2-((( Z )-4-amino-2-fluorobut-2-en-1-yl)sulfonyl)benzamide 13 N -((1 S ,3 R ,5 S )-adamantan-1-yl)-3-((( Z )-4-amino-2-fluorobut-2-en-1-yl)sulfonyl)benzamide 14 ( Z )-4-((2-((4-amino-2-fluorobut-2-en-1-yl)sulfonyl)phenoxy)methyl)- N- isopropylbenzenesulfonamide 15 ( Z )-2-((4-Amino-2-fluorobut-2-en-1-yl)sulfonyl)- N- (4-( N , N- diisopropylsulfamoyl)phenyl)benzamide 16 ( Z )-4-(3-(4-Amino-2-fluorobut-2-enylsulfonyl)phenoxy)-N,N-dimethylbenzenesulfonamide 17 ( Z )-3-(3-(4-Amino-2-fluorobut-2-enylsulfonyl)phenoxy)-N,N-dimethylbenzenesulfonamide 18 ( Z )-3-(2-(4-Amino-2-fluorobut-2-enylsulfonyl)phenoxy)-N,N-dimethylbenzenesulfonamide 19 ( Z )-4-(2-chlorophenylsulfonyl)-3-fluorobut-2-en-1-amine 20 ( Z )-4-(biphenyl-2-ylsulfonyl)-3-fluorobut-2-en-1-amine 21 ( Z )-3-fluoro-4-(2-isopropylphenylsulfonyl)but-2-en-1-amine 22 ( Z )-3-fluoro-4-(2-methoxyphenylsulfonyl)but-2-en-1-amine 23 ( Z )-3-fluoro-4-(naphthalene-1-ylsulfonyl)but-2-en-1-amine 24 ( Z )-3-fluoro-4-(naphthalene-2-ylsulfonyl)but-2-en-1-amine 25 ( Z )-4-(2,4-dichlorophenylsulfonyl)-3-fluorobut-2-en-1-amine 26 ( Z )-4-(3-chlorophenylsulfonyl)-3-fluorobut-2-en-1-amine 27 ( Z )-4-(4-chlorophenylsulfonyl)-3-fluorobut-2-en-1-amine 28 ( Z )-4-(3,5-dichlorophenylsulfonyl)-3-fluorobut-2-en-1-amine 29 ( Z )-3-fluoro-4-(pyridin-4-ylsulfonyl)but-2-en-1-amine thirty ( Z )-3-fluoro-4-(pyridin-2-ylsulfonyl)but-2-en-1-amine 31 ( Z )-3-fluoro-4-(pyridin-3-ylsulfonyl)but-2-en-1-amine 32 ( Z )-3-fluoro-4-(quinolin-2-ylsulfonyl)but-2-en-1-amine 33 ( Z )-3-fluoro-4-(quinolin-8-ylsulfonyl)but-2-en-1-amine 34 ( Z )-3-fluoro-4-(5-isopropylpyridin-2-ylsulfonyl)but-2-en-1-amine 35 ( Z )-3-fluoro-4-(5-methylpyridin-2-ylsulfonyl)but-2-en-1-amine 36 ( Z )-3-fluoro-4-(6-methylpyridin-2-ylsulfonyl)but-2-en-1-amine 37 ( Z )-3-fluoro-4-(2-fluorophenylsulfonyl)but-2-en-1-amine 38 ( Z )-3-fluoro-4-(3-fluorophenylsulfonyl)but-2-en-1-amine 39 ( Z )-3-fluoro-4-(4-fluorophenylsulfonyl)but-2-en-1-amine 40 ( Z )-3-fluoro-4-(o-tolylsulfonyl)but-2-en-1-amine 41 ( Z )-3-fluoro-4-(m-tolylsulfonyl)but-2-en-1-amine 42 ( Z )-3-fluoro-4-tosylbut-2-en-1-amine 43 ( Z )-3-fluoro-4-(3-methylpyridin-2-ylsulfonyl)but-2-en-1-amine 44 ( Z )-3-fluoro-4-(2-methylpyridin-4-ylsulfonyl)but-2-en-1-amine 45 ( Z )-3-fluoro-4-(2-isopropylpyridin-3-ylsulfonyl)but-2-en-1-amine 46 ( Z )-3-fluoro-4-(6-methylpyridin-3-ylsulfonyl)but-2-en-1-amine 47 ( Z )-3-fluoro-4-(2-methylpyridin-3-ylsulfonyl)but-2-en-1-amine 48 ( Z )-3-fluoro-4-(4-methylpyridin-3-ylsulfonyl)but-2-en-1-amine 49 ( Z )-3-fluoro-4-(6-isopropylpyridin-3-ylsulfonyl)but-2-en-1-amine 50 ( Z )-3-fluoro-4-((2-methylbenzo[ d ]thiazol-4-yl)sulfonyl)but-2-en-1-amine 51 ( Z )-3-fluoro-4-((3-fluoroquinolin-8-yl)sulfonyl)but-2-en-1-amine 52 ( Z )-4-((2,3-dimethyl-1H-indol-7-yl)sulfonyl)-3-fluorobut-2-en-1-amine 53 ( Z )-3-fluoro-4-((quinolin-8-yl -d ₆ )sulfonyl)but-2-en-1-amine or a pharmaceutically acceptable salt or hydrate thereof. 11. The compound according to claim 1, selected from the group consisting of And , or their pharmaceutically acceptable salts. 12. The compound according to claim 1, selected from the group consisting of And , or their pharmaceutically acceptable salts or hydrates. 13. Pharmaceutical composition for inhibiting amine oxidase activity of any of LOX, LOXL1, LOXL2, LOXL3 or LOXL4, containing an effective amount of the compound according to any one of paragraphs. 1-12 or a pharmaceutically acceptable salt or hydrate thereof and at least one pharmaceutically acceptable excipient, carrier or diluent. 14. The use of an effective amount of a compound according to any one of paragraphs. 1-12 or a pharmaceutically acceptable salt or hydrate thereof or a pharmaceutically acceptable composition according to claim 13 for inhibiting amine oxidase activity of any of LOX, LOXL1, LOXL2, LOXL3 or LOXL4. 15. The use of an effective amount of a compound according to any one of paragraphs. 1-12 or a pharmaceutically acceptable salt or hydrate thereof or a pharmaceutically acceptable composition according to claim 13 for the treatment of a condition associated with any of the LOX, LOXL1, LOXL2, LOXL3 or LOXL4 proteins. 16. Use according to claim 15, characterized in that the condition is selected from the group consisting of fibrosis, cancer and angiogenesis. 17. The use according to claim 16, characterized in that in the case of fibrosis, the fibrosis is selected from the group consisting of mediastinal fibrosis, myelofibrosis, retroperitoneal fibrosis, progressive massive fibrosis, nephrogenic systemic fibrosis, Crohn's disease, keloid, systemic sclerosis, arthrofibrosis, contracture Dupuytren's disease, adhesive capsulitis, pancreatic fibrosis, intestinal fibrosis, liver fibrosis, pulmonary fibrosis, kidney fibrosis, cardiac fibrosis, fibrostenosis, cystic fibrosis, idiopathic pulmonary fibrosis, radiation-induced fibrosis, Peyronie's disease, and scleroderma, or associated with respiratory disease, abnormal healing, and repair of wounds, scarring, hypertrophic scarring/keloid, post-surgical scarring, cardiac arrest, and all conditions in which excessive or aberrant fibrous deposition is associated with disease, trauma, implants, or surgery; preferably the fibrosis is selected from the group consisting of myelofibrosis, systemic sclerosis, hepatic fibrosis, pulmonary fibrosis, renal fibrosis, cardiac fibrosis, and radiation-induced fibrosis; And in case the disease is cancer, the cancer is selected from the group consisting of lung cancer; breast cancer; colorectal cancer; anal cancer; pancreatic cancer; prostate cancer; ovarian cancer; carcinomas of the liver and bile ducts; esophageal carcinomas; mesothelioma; non-Hodgkin's lymphoma; bladder carcinomas; uterine carcinoma; gliomas, glioblastomas, medullablastomas and other brain tumors; myelofibrosis, kidney cancer; head and neck cancer; stomach cancer; multiple myeloma; testicular cancer; germ cell tumors; neuroendocrine tumor; cervical cancer; cancer of the oral cavity, carcinoids of the gastrointestinal tract, breast and other organs; cricoid cell carcinomas; mesenchymal tumors, including sarcomas, fibrosarcomas, hemangiomas, angiomatosis, hemangiopericytoma, pseudoangiomatous stromal hyperplasia, myofibroblastomas, fibromatosis, inflammatory myofibroblastic tumors, lipomas, angiolipomas, granular cell tumors, neurofibromas, schwannomas, angiosarcomas, liposarcomas, rhabdomes iosarcoma, osteosarcoma, leiomyoma, or leiomyosarcoma . 18. Application according to any one of paragraphs. 15-17, further comprising administering a second therapeutic agent. 19. Use according to claim 18, wherein the second therapeutic agent is selected from the group consisting of an anticancer agent, an anti-inflammatory agent, an antihypertensive agent, an antifibrotic agent, an antiangiogenic agent, an immunosuppressant, and a metabolic agent. 20. The use of an effective amount of a compound according to any one of paragraphs. 1-12 or a pharmaceutically acceptable salt or hydrate thereof for the manufacture of a medicament for the treatment of a condition associated with any of the LOX, LOXL1, LOXL2, LOXL3 or LOXL4 proteins.