EA043371B1 - 1-METHYL-4-[(4-PHENYLPHENYL)SULPHONYLMETHYL]CYCLOHEXANOL AND 1-METHYL-4-[[4-(2-PYRIDYL)PHENYL]SULPHONYLMETHYL]CYCLOHEXANOL COMPOUNDS AND THEIR THERAPEUTIC APPLICATION - Google Patents

Description

Related applications

This application relates to United Kingdom (UK) Patent Application No. 1813312.4, filed August 15, 2018, the contents of which are incorporated herein by reference in their entirety.

Field of technology

In general, the present invention relates to the field of therapeutic compounds. More particularly, the present invention relates to certain substituted 1-methyl-4-[(4phenylphenyl)sulfonylmethyl]cyclohexanol and 1-methyl-4-[[4-(2-pyridyl)phenyl]sulfonylmethyl]cyclohexanol compounds (generally designated herein document as CHMSA compounds) that are useful, for example, in the treatment of disorders (e.g., diseases), including, for example, multiple myeloma, diffuse large B-cell lymphoma, acute myeloid leukemia, eosinophilic leukemia, glioblastoma, melanoma, ovarian cancer, resistant to chemotherapy cancer, radiation-resistant cancer, inflammatory arthritis, rheumatoid arthritis, psoriatic arthritis, psoriasis, ulcerative colitis, Crohn's disease, systemic lupus erythematosus (SLE), lupus nephritis, asthma, chronic obstructive pulmonary disease (COPD), non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), autoimmune hepatitis, hidradenitis suppurativa, etc. The present invention also relates to pharmaceutical compositions containing such compounds, and to the use of such compounds and compositions, for example, in therapy.

State of the art

A number of publications are cited herein to more fully describe and disclose the invention and the technical field to which the invention relates. Each of these publications is incorporated herein by reference in its entirety to the same extent as if each individual publication were specifically and separately stated to be incorporated by reference.

Throughout this specification, including the appended claims, unless the context otherwise requires, the word contain and its variants such as contains and containing are to be understood to include the specified integer or step or group of integers or steps, but not to imply the exclusion of any other integers or stages or groups of integers or stages.

It should be noted that in the specification and in the accompanying claims, a singular noun also refers to a plural noun unless the context clearly indicates otherwise. Thus, for example, reference to a pharmaceutical carrier includes mixtures of two or more such carriers, and the like.

As used herein, ranges are often expressed as from about one particular value and/or to about another particular value. When such a range is indicated, another embodiment includes a range from one particular value and/or to another particular value. Likewise, if values are expressed as approximations, using the preceding characteristic of approximately, then it should be understood that the particular value constitutes another embodiment.

The present description includes information that may be useful for understanding the present invention. This is not an admission that any information presented herein is prior art or relevant to the claimed invention, or that any publication specifically or impliedly referred to is prior art.

Cellular metabolism.

Cellular metabolism is the set of complex sequences of biochemical reactions that occur in the cells of living organisms to support life. Each sequence of reactions is known as a metabolic pathway, and these pathways act in concert to provide energy, synthesize new molecules, and break down and remove other molecules within the cell. One key metabolic pathway is known as oxidative phosphorylation, a process that produces energy in the form of adenosine triphosphate (ATP) by transferring electrons through carriers known as electron transport complexes. Other examples of metabolic pathways include glycolysis, the process of breaking down glucose to release ATP, and beta-oxidation, the process of breaking down fatty acids.

Glycolysis occurs in the cytoplasm. Glucose, the substrate for glycolysis, is converted to pyruvate through a series of reactions catalyzed by ten enzymes. Pyruvate in turn is converted to lactic acid, the end product of glycolysis. ATP is directly produced by the transfer of phosphate from a substrate to ATP, or phosphorylation of the substrate. Some of the pyruvate enters the tricarboxylic acid (TCA) cycle, while most of the end product, lactic acid, is flushed out of the cell. Oxidative phosphorylation occurs in cell mitochondria. Glutamine, glucose or fatty acids are suppliers to the electron transport chain, and ATP is formed through a series of redox reactions involving oxygen as the final electron acceptor. A series of redox reactions occur through

- 1 043371 four complexes of the electron transport chain, which then creates an electrochemical gradient in the inner mitochondrial membrane. Protons are returned to the mitochondrial matrix through ATP synthase, and this process is associated with ATP synthesis. In total, 1 mole of glucose produces 36 moles of ATP.

The metabolic properties of some cell types can vary greatly. For example, energy production in cancer cells is abnormally biased toward aerobic glycolysis (a process known as the Warburg effect) and is accompanied by increased fatty acid synthesis and an increased rate of metabolism of the amino acid glutamine. In addition, changes in the metabolism of cancer cells can lead to their resistance to therapy, and a number of studies have shown that chemoresistance is, at least in part, due to mitochondrial metabolism and oxidative phosphorylation, while high ATP levels in cancer cells can lead to to increase the efflux of chemotherapeutic agents and contribute to the development of hypoxia-related drug resistance.

Like cancer cells, immune cells exhibit changes in metabolism depending on their activation status and the stimulatory signals they receive. The study of immunometabolism involves the study of the interaction between immunology and metabolism as it relates to both the control of immune cell functions and their role in chronic inflammatory diseases and cancer, among other things.

Chronic inflammatory disease.

Inflammation is an immune response of tissues to bodily injury. Acute inflammation is a normal protective response that protects and heals the body after physical injury or infection, and is characterized by heat, swelling and redness at the site of injury. However, if the inflammation persists for a long period, it becomes chronic. Chronic inflammation is a feature and contributing factor to a number of disease conditions, including diseases such as rheumatoid arthritis, inflammatory bowel disease, systemic lupus erythematosus, multiple sclerosis and psoriasis.

The inflammatory process is complex and involves a biological cascade of molecular and cellular signals that alter physiological responses. At the site of injury, cells release molecular signals such as cytokines and interleukins, which cause a number of changes in the affected area, including dilation of blood vessels, increased blood flow, increased vascular permeability, invasion of leukocytes (white blood cells), and the release of fluids containing proteins such as immunoglobulins (antibodies). Several different types of leukocytes are involved in the inflammatory cascade, including granulocytes, monocytes, and lymphocytes. However, chronic inflammation is primarily mediated by monocytes and long-lived macrophages; monocytes mature into macrophages when they leave the bloodstream and enter tissues. Macrophages engulf and digest microorganisms, foreign agents, and senescent cells, and macrophages also release several different chemical mediators, including tumor necrosis factor alpha (TNFa), interleukins (eg, IL-1, IL-6, IL-12, and IL-23) and prostaglandins, which promote the inflammatory response. In later stages, other cells, including lymphocytes, invade the affected tissue. Recent evidence has shown that many aberrant immune responses result from metabolic disturbances and that changes in cellular metabolism can either enhance or decrease immune responses. Therefore, changes in the metabolism of monocytes, macrophages and lymphocytes (immunometabolism) are critical for the development of diseases.

Thus, there is a common pathology underlying many chronic inflammatory conditions. In addition, signs of chronic inflammation are also observed in other diseases, including cancer and metabolic diseases such as obesity, atherosclerosis and diabetes.

One of the most common chronic inflammatory conditions is rheumatoid arthritis (RA), a condition that affects up to 2% of the population worldwide. Although it is a complex disease, there are a number of physiological, cellular and biochemical factors associated with the progression of RA that are also common to a number of other diseases, including diseases with an autoimmune component (eg, multiple sclerosis), inflammatory components (eg, atherosclerosis and cancer), bone loss (eg, osteoporosis), and proliferative activity (eg, hematologic malignancies). Understanding RA is therefore important not only for studying a much wider range of diseases, but it also suggests that pharmaceutical agents that act through modification of these general processes may have applications other than RA. The latter is confirmed by clinical practice, in which drugs for the treatment of RA have been shown to have wide applicability in many other conditions.

Rheumatoid arthritis and related autoimmune/inflammatory diseases.

Rheumatoid arthritis (RA) is an autoimmune disorder characterized by chronic inflammation of the synovial lining of multiple joints coupled with progressive joint degradation. RA typically affects the joints of the wrist and hands, and can also affect the elbows, shoulders, hips, neck and knees, leading to severe pain and disability (see, for example, Scott et al.,

2010). The World Health Organization (WHO) Global Burden of Disease 2010 project estimates that 23.7 million people have RA, with incidence rising due to the association between the condition and age.

The exact cause of RA, like all autoimmune diseases, remains unclear, although possible triggers include decreased autotolerance, abnormal responses to environmental factors, infectious agents, and hormonal stimuli (see, for example, Klareskog et al., 2006; Firestem et al., 2005). The main hallmark of this condition is dysregulation of innate and adaptive immunity, accompanied by an imbalance of pro-inflammatory and anti-inflammatory cytokines and an altered balance between osteoclast-mediated degradation and osteoblast-mediated deposition in the bone marrow compartment (see, e.g., Kleyer et al., 2014; Jung et al., 2014). al., 2014).

At the cellular level, the development of RA usually begins with the infiltration of T cells into the synovial membrane lining the affected joint; this then leads to the activation of monocytes, macrophages and synovial fibroblasts through cell-cell contact and subsequent release of various cytokines including tumor necrosis factor alpha (TNFa) and pro-inflammatory interleukins such as IL-1, IL-6, IL-12 and IL-23 (see, for example, Astry et al., 2011). These pro-inflammatory cytokines then play an important role in organizing several complex signaling cascades, including NFkB, interferon regulatory factor (IRF), Toll-like receptor (TLR) and the Jak/STAT pathway (see, for example, Malemud et al., 2010). , which lead to the induction of genes encoding various products that propagate the inflammatory response and also contribute to tissue destruction. These products include tissue-degrading enzymes such as collagenases, matrix metalloproteinases (MMPs), cathepsins and other pro-inflammatory factors such as selectins, integrins, leukotrienes, prostaglandins, chemokines and other cytokines (see, for example, McNenes et al., 2011 ; Chimenti et al., 2015). In addition, these cells also increase the production of MMPs, which leads to degradation of the extracellular matrix and loss of cartilage in the joint (see, for example, Sun, 2010), a process also involving a special class of cells known as osteoclasts and a factor known as receptor activator of nuclear factor kappa B ligand (RANKL) (see, for example, Takayanagi, 2009).

RANKL is an essential factor for osteoclast formation, and increased production of RANKL leads to increased osteoclast differentiation and ultimately bone destruction (see, e.g., Long et al., 2012). The inflammatory response in RA results in the accumulation of lymphocytes, dendritic cells and macrophages, which act locally to produce cytokines and other proinflammatory mediators such as TNFa and IL-6, and these further enhance the effects of RANKL on bone destruction. In addition, the inflammatory cascade causes hyperplasia of synoviocytes (see, e.g., Takayanagi, 2009), which in turn leads to thickening and vascularization of the synovium and its transformation into destructive and aggressive tissue known as pannus. The pannus contains both osteoclasts, which destroy bone, and metalloproteinases, which are involved in the destruction of cartilage. Thus, the RANKL axis plays a critical role in the progression and pathology of RA and is of great importance to the osteoimmune system (the interaction between the immune and skeletal systems), which is critical to the pathology of a number of different disease states.

The role of immune metabolism in RA.

All cells produce adenosine triphosphate (ATP), a high-energy molecule of which serves as fuel, and synthesize macromolecules to support their basic cellular functions, whether they are active, replicating, or quiescent (see, e.g., Spies et al., 2012 ). These bioenergetic needs are met by interconnected metabolic pathways within the cell: glycolysis (the first step in the breakdown of glucose), the tricarboxylic acid cycle (a series of reactions that release stored energy from carbohydrates, fats, and proteins), and oxidative phosphorylation (the process of producing ATP through electron transfer). Changes in these pathways control the effector functions of immune cells from lymphocytes to monocytes, macrophages and dendritic cells, and can also influence cell fate.

In chronic inflammatory diseases, including RA, a very large amount of energy is spent on activation of the immune system (up to 2000 kJ/day) (see, for example, Straub et al., 2010). This energy is used, at least in part, by the immune system to maintain a state of chronic inflammation in response to environmental cues (see, e.g., Procaccini et al., 2012; Nutsch et al., 2011) and hence the interaction between immunology and metabolism plays a central role in the pathophysiology of chronic inflammatory diseases (see, for example, Perl, 2017; Ganeshan et al., 2014).

Several metabolic changes in cells that are involved in inflammation are observed in immune cells in RA (see, for example, Weyand et al., 2017a). Chronic stimulation and the synovial microenvironment alter the metabolism of T cells and macrophages in RA. For example, T cells from RA patients show decreased expression of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3), an enzyme involved in ATP generation and autophagy (see, e.g., Yang et al., 2013), while

- 3 043371 time, macrophages from RA patients produce higher levels of ATP than cells from healthy individuals (see, for example, Weyand et al., 2017b). In addition to direct cellular changes, the hypoxic environment in the RA synovium (see, e.g., Fearon et al., 2016) creates chronic mitochondrial hyperpolarization, which is also observed in systemic lupus erythematosus (SLE) and in fibroblast-like synoviocytes of RA patients; there is a transition to glycolysis compared to cells under non-inflammatory conditions (see, for example, Garcia-Carbonnel et al., 2016). Thus, agents that modulate ATP or alter immune cell metabolism are highly promising, as they may be useful in the treatment of chronic inflammatory diseases such as RA, SLE, inflammatory bowel disease (IBD), psoriasis, and atherosclerosis.

Cellular metabolism and cancer.

Cellular energy in the form of ATP is generated by two main pathways: mitochondrial oxidative phosphorylation and cytoplasmic glycolysis. In normal cells, glycolysis is followed by the oxidation of pyruvate using the mitochondrial oxidative phosphorylation mechanism, and this pathway is the major pathway for ATP production. However, in cancer cells, glycolysis is increased and lactic acid is fermented in the cytosol of the cell in a process known as the Warburg effect. Thus, reprogrammed metabolism is a hallmark of cancer and enhances cell growth and proliferation under stressful conditions.

Mitochondrial metabolism is also important for creating the building blocks necessary for cancer cell proliferation, and mitochondrial oxidative metabolism is also required to maintain redox balance in cancer cells. Most cancer cells have functional mitochondria and are able to generate ATP through mitochondrial metabolism (see, e.g., Koppenol, 2011). Depending on the cellular environment, mitochondria make a significant contribution to the generation of cellular reactive oxygen species (ROS) as a natural byproduct of mitochondrial ATP production. ROS generation occurs due to incomplete reduction of molecular oxygen, and in cancer cells, ROS has been shown to promote tumor development and progression through the induction of oncogenic signaling, genetic instability, and DNA mutations (see, e.g., Weinberg et al., 2010) . However, when ROS production exceeds the capacity of intracellular systems to detoxify ROS, cellular toxicity occurs. Thus, cancer cells have to strictly control their metabolic machinery to maintain a balance between ROS production and removal.

Therefore, changes in cellular and mitochondrial metabolism are critical for the growth and spread of tumors. Indeed, mitochondrial biogenesis and the associated increase in oxidative phosphorylation have been shown to promote tumor metastasis (see, e.g., LeBleu et al., 2014 ); however, reduction of oxidative phosphorylation has been proposed as a means to target cancer stem cells (see, e.g., Fiorillo et al., 2016). Data also suggest that targeting components of the mitochondrial electron transport chain may have anticancer effects. For example, inhibition of complex I by the antidiabetic metformin suppresses tumorigenesis (see, e.g., Evans et al., 2005; Pollak et al., 2014; Wheaton et al., 2014; Bridges et al., 2014), while new small molecule electron transport inhibitors also exhibit antitumor activity in cancer xenograft models (see, e.g., Ellinghaus et al., 2013). Thus, altering cellular metabolism becomes a means to prevent cancer growth and progression, as well as overcome chemotherapy resistance and prevent metastasis.

Osteoimmune system and bone disorders.

The term osteoimmune system refers to the combined and interconnected interaction between the immune and skeletal systems.

Under normal physiological conditions, the skeletal system provides support, mobility, protection of vital organs, and a mineral reservoir for calcium and phosphate. To achieve and adapt these functions, the skeleton is in a dynamic equilibrium characterized by continuous osteoclast-mediated bone resorption and osteoblast-mediated bone deposition (see, e.g., Karsenty et al., 2002). This biological process has been termed bone remodeling and involves osteoblasts producing key osteoclast differentiation factors, including RANKL described above, and osteoclasts promoting bone formation by producing osteoblast mediators as osteoclasts break down bone.

Cells of both the innate and adaptive immune systems influence osteoclasts and osteoblasts through a variety of cell surface and secreted mediators (see, for example, Takayanagi, 2009). Activation of the RANKL receptor (RANK) on osteoclast precursors triggers a cascade of transcriptional changes that leads to the formation of osteoclasts and the expression of what is necessary for bone resorption, including molecules required for bone attachment, acid secretion, and proteolysis. Many transcription factors important for osteoclast differentiation are key regulators of immune responses, such as NFkB and nuclear factor of activated T cells c1 (NFATc1), and this process is also enhanced by factors involved in

- 4 043371 inflammation, such as TNFa and IL-6.

In addition to its critical role in the progression and pathogenesis of RA, the osteoimmune system plays a critical role in a number of other diseases, including osteoporosis and other bone diseases, as well as cancer (see, for example, Dallas et al., 2011).

Osteoporosis is a common disease characterized by decreased bone density, bone breakdown, and an increased risk of fractures. Many factors contribute to the pathogenesis of osteoporosis, including poor diet, lack of exercise, smoking, and excessive alcohol consumption. Osteoporosis also occurs in association with inflammatory diseases such as rheumatoid arthritis, endocrine diseases such as thyrotoxicosis, and in association with treatment with certain medications such as glucocorticoids. Indeed, pathological fractures associated with osteoporosis represent one of the most serious complications that can occur in patients with rheumatic diseases such as RA, systemic lupus erythematosus and ankylosing spondylitis.

Paget's disease is a common condition of unknown cause characterized by increased bone turnover and disorganized bone remodeling, with areas of increased osteoclast and osteoblast activity. Although bone affected by Paget's disease is often denser than healthy bone, the abnormal structure causes the bone to become mechanically weak, resulting in bone deformation and increased susceptibility to pathological fractures.

IL-6, TNFa, and RANKL signaling have been shown to play important roles in increased osteoclast activity and consequent increased bone loss (see, e.g., Tapaka et al., 2003; Roodman, 2006). The effectiveness of drugs targeting these pathways has been supported by completed clinical trials of the anti-RANKL monoclonal antibody, AMG162 (Denosumab®, Amgen) for the treatment of osteoporosis/multiple myeloma, and by growing evidence that anti-RANKL TNFa therapy and anti-III-6 therapy also prevent bone loss in arthritic diseases (see, for example, Ogata et al., 2012; Billiau, 2010).

Osteoimmune system and cancer.

Bone is affected by many types of cancer. Cancer-associated bone disease may manifest as the occurrence of hypercalcemia or the development of osteolytic and/or osteosclerotic metastases. Increased osteoclastic bone resorption plays a key role in the pathogenesis of both conditions. Although almost any cancer can be complicated by bone metastases, the most common sources are multiple myeloma, breast carcinoma, and prostate carcinoma. The most common tumors associated with hypercalcemia are multiple myeloma, breast carcinoma, and lung carcinoma.

As described above, RANK/RANKL signaling is of great importance for osteoclast formation and bone resorption that occurs during bone remodeling. Physiological levels of RANK/RANKL signaling stimulate proliferation and survival of mammary epithelial cells, but aberrant RANK/RANKL signaling in these tissues has recently been shown to influence the initiation and progression of mammary tumorigenesis, and blocking RANKL signaling has also been shown to with denosumab (Xgeva®, Amgen) effectively prevents secondary complications of bone metastases, such as pathological fracture and hypercalcemia, in patients with breast cancer (see, for example, Steger et al., 2011).

Therapy that blocks RANK/RANKL signaling may also reduce the ability of osteotropic cancers to form bone metastases. Signaling through RANK on the surface of human epithelial tumor cells as well as melanoma cells has been shown to induce a chemotactic response in these tumor cells, while in a mouse model of melanoma metastasis, therapeutic treatment of mice with osteoprotegrin, which neutralizes the RANKL receptor, RANK, significantly reduces tumor burden in bones, but not in other organs.

In addition to the role of RANKL in cancer development, there is growing evidence that activation of NFkB through molecules such as TNFa may play an important role in the promotion and progression of both hematological malignancies such as myeloma and lymphomas and solid tumors such as cancer breast, prostate and lung (see, for example, Baud et al., 2009). There is also a growing understanding of the role and importance of inflammation and the osteoimmune system in the development of cancer and in the development of resistance to radiation therapy and chemotherapeutic agents. In addition, it has been suggested that inflammation is in fact one of the main hallmarks of cancer (see, for example, Mantovani, 2009). Thus, increasing the effectiveness of anticancer treatment by preventing NFkB activation is a promising strategy to improve existing therapeutic regimens and is currently being explored, especially for the treatment of multiple myeloma.

Defects in normal apoptotic pathways are also involved in the development and progression of tumor growth.

- 5 043371 chole cells, as well as in inflammation. Apoptosis (programmed cell death) plays a key role in the removal of abnormal cells; defects in the signaling cascades that normally lead to its induction play a key role in tumorigenesis. Radiation therapy and many chemotherapy agents act by causing cell damage, which typically causes apoptosis; therefore, pathway defects will also reduce the effectiveness of such agents. The most important effector molecules in the signaling pathway leading to apoptosis are known as caspases, which can be triggered by a number of stimuli, including the binding of TNFa to its receptor. Mutations in the genes that encode caspases have been found in a number of tumor types, including gastric cancer, breast cancer, renal cell cancer and cervical cancer, and also commonly in lymphoblastic T-cell lymphoma and basal cell ameloblastomas (see, for example, Philchenkov et al., 2004). Compounds that activate caspases and thereby sensitize cells to apoptosis will be highly effective as cancer treatments, either as stand-alone agents or as agents to enhance the effectiveness of existing chemotherapy and radiotherapy for cancer.

Agents that modulate cellular and immune metabolism, prevent inflammation and modify the osteoimmune system.

The inventors of the present invention have identified new compounds that, for example, modulate cellular and immune metabolism, prevent inflammation and modify the osteoimmune system, and are accordingly useful in the treatment of related disorders as described herein.

Without wishing to be bound by any particular theory, the present inventors believe that this action may occur through a mechanism that involves modulation of cellular and immune cell metabolism by reducing cellular ATP with subsequent effects on inflammatory signaling.

Known compounds.

Greig et al., 2010a, describe certain biphenyl-4-sulfonic acid amides for the treatment of inflammation and/or joint destruction and/or bone loss; disorders mediated by excessive and/or inappropriate and/or prolonged activation of the immune system; inflammatory and autoimmune disorders, for example, rheumatoid arthritis, psoriasis, psoriatic arthritis, chronic obstructive pulmonary disease (COPD), atherosclerosis, inflammatory bowel disease and ankylosing spondylitis; disorders associated with bone loss, such as bone loss associated with excessive osteoclast activity in rheumatoid arthritis, osteoporosis, cancer-related bone disease and Paget's disease; and cancers such as hematologic malignancies and solid tumors. Examples of connections described in this document include the following:

F

F

Patel et al., 2014 and Patel et al., 2016 describe compounds of the following formulas for the treatment of inflammation and/or joint destruction and/or bone loss; disorders mediated by excessive and/or inappropriate and/or prolonged activation of the immune system; inflammatory and autoimmune disorders, such as rheumatoid arthritis; psoriasis; psoriatic arthritis; chronic obstructive pulmonary disease (COPD); asthma; atherosclerosis; inflammatory bowel disease; ankylosing spondylitis; multiple sclerosis; systemic lupus erythematosus; Sjögren's syndrome; a bone loss disorder, such as bone loss associated with excessive osteoclast activity in rheumatoid arthritis, osteoporosis, cancer-related bone disease, or Paget's disease; cancer, such as a hematological malignancy such as multiple myeloma, leukemia or lymphoma, or a solid tumor cancer such as bladder cancer, breast cancer (in women and/or men), colon cancer, renal cell carcinoma, kidney cancer, lung cancer, pancreatic cancer, stomach cancer, prostate cancer, brain cancer, skin cancer, thyroid cancer, basal cell ameloblastoma or melanoma; fibrosis-related disorders such as systemic sclerosis or scleroderma; or rare vasculitis such as Behçet's disease.

- 6 043371

- 7 043371

New compounds with improved properties.

In addition to superior biological properties, e.g., as or better than the corresponding sulfonamide compounds (e.g., as described in Greig et al., 2010a, Patel et al., 2014, and Patel et al., 2016), the CHMSA compounds described herein have the additional advantage of producing little or no unwanted sulfonamide metabolite.

For example, as demonstrated by the data presented herein, corresponding sulfonamide compounds (eg, reference compound HMC-C-01-A) produce a biarylsulfonamide metabolite (eg, MET-001) that has a long half-life and therefore persists in the bloodstream. In addition, the biarylsulfonamide metabolite acts as a metabolic inducer in rats, which complicates the assessment of toxicity in rodents and, in turn, may affect the usefulness of these compounds in humans. Therefore, compounds with a lower propensity to form the biarylsulfonamide metabolite have a higher potential suitability for use in humans.

As demonstrated by the data presented herein, CHMSA compounds exhibit a significantly reduced propensity to form a biarylsulfonamide metabolite and are thus significantly more amenable to development for human use compared to known sulfonamide compounds.

In addition, the CHMSA compounds described herein have other beneficial properties similar to, and often better than, the corresponding sulfonamide compounds, including, for example, improved metabolism and cardiovascular safety.

If a drug is to be used clinically, it must have a suitable safety and efficacy profile. The drug must demonstrate adequate short-term safety so that a dose can be administered to a person without the expectation of serious overall side effects. A clinically acceptable drug must also not inhibit hERG, an ion channel that, when inhibited, can cause the fatal cardiac disorder known as long QT syndrome. In addition to these safety characteristics, the drug must have a minimum potential for interaction with drug-metabolizing enzymes in the body in order to: ensure reliable delivery of the drug; minimize the ability of a drug to affect the metabolism of other drugs (the so-called drug-drug interaction); and prevent serious adverse reactions that may be caused by drug-drug interactions.

The CHMSA compounds described herein largely exclude the possibility of inhibiting the human cardiac specific potassium channel gene (hERG), and inhibition of hERG poses a serious threat to the cardiovascular system.

The CHMSA compounds described herein also demonstrate significant benefits in minimizing potential drug-drug interactions due to their in vitro metabolic profile and the reduced propensity of the compounds to form a sulfonamide metabolite that acts as a metabolic inducer, such as MET001.

Reducing the toxicological properties (adverse effects) of a drug is a scientific and production limiting factor, representing the same problem and having the same importance as optimizing the pharmacodynamics (effects of the drug).

- 8 043371 va on the body) and pharmacokinetic (the effect of the body on the drug) properties. The CHMSA compounds described herein provide significant advantages as oral therapeutic agents (compared to known compounds) by improving cardiovascular safety and providing an improved metabolic profile with little or no loss of activity on the biological target.

The CHMSA compounds described herein combine the necessary characteristics of agents for the treatment of, for example, autoimmune/inflammatory conditions and cancer, as described herein.

Disclosure of the invention

One aspect of the present invention relates to compounds of formula (I) n ₃ with

or a pharmaceutically acceptable salt thereof, wherein =X- independently represents -CH= or -N=;

- R ¹ is independently H or -R ^1X ;

- R ^1X is independently -F, -Cl or -CN;

- R ² is independently -H or -R ^2X ;

- R ^2X is independently -F, -Cl or -CN;

- R ³ is independently -H or -R ^3X ;

- R ^3X is independently -F, -Cl, -R ^3C , -R ^3F or -CN;

- R ^3C independently represents a saturated straight or branched C _1-3 alkyl;

- R ^3F independently represents a saturated straight or branched C _1-3 fluoroalkyl;

- R ⁴ is independently -H or -R ^4X ;

- R ^4X is independently -F, -Cl, -R ^4C , -R ^4F or -CN;

- R ^4C independently represents a saturated straight or branched C _1-3 alkyl;

- R ^4F independently represents a saturated straight or branched C _1-3 fluoroalkyl;

- R ⁵ represents -H; And

- R ⁶ represents -H.

Another aspect of the present invention relates to a composition for treating a disorder comprising a compound of formula (I), and a carrier, diluent or excipient; the disorder being inflammatory arthritis.

Another aspect of the present invention relates to a composition for treating a disorder comprising a compound of formula (I), and a carrier, diluent or excipient; wherein said disorder is rheumatoid arthritis; psoriatic arthritis; ankylosing spondylitis; spondyloarthritis; reactive arthritis; infectious arthritis; systemic lupus erythematosus; scleroderma; gout; acquired Still's disease; or juvenile idiopathic arthritis.

Another aspect of the present invention relates to a composition for treating a disorder comprising a compound of formula (I), and a carrier, diluent or excipient; wherein said disorder is rheumatoid arthritis.

Another aspect of the present invention relates to a method for preparing a composition, comprising the step of mixing a compound of formula (I) described herein and a carrier, diluent or excipient.

Another aspect of the present invention relates to the use of a compound of formula (I) for the treatment of a disorder, wherein said disorder is inflammatory arthritis.

Another aspect of the present invention relates to the use of a compound of formula (I) for the treatment of a disorder, wherein said disorder is rheumatoid arthritis; psoriatic arthritis; ankylosing spondylitis; spondyloarthritis; reactive arthritis; infectious arthritis; systemic lupus erythematosus; scleroderma; gout; acquired Still's disease; or juvenile idiopathic arthritis.

Another aspect of the present invention relates to the use of a compound of formula (I) for the treatment of a disorder, wherein said disorder is rheumatoid arthritis.

Another aspect of the present invention relates to a method of treating a disorder, comprising administering to a patient in need of treatment a therapeutically effective amount of a compound

- 9 043371 formula (I), where the specified disorder is inflammatory arthritis.

Another aspect of the present invention relates to a method of treating a disorder, comprising administering to a patient in need of treatment a therapeutically effective amount of a compound of formula (I), wherein the disorder is rheumatoid arthritis; psoriatic arthritis; ankylosing spondylitis; spondyloarthritis; reactive arthritis; infectious arthritis; systemic lupus erythematosus; scleroderma; gout; acquired Still's disease; or juvenile idiopathic arthritis.

Another aspect of the present invention relates to a method of treating a disorder, comprising administering to a patient in need of treatment a therapeutically effective amount of a compound of formula (I), wherein the disorder is rheumatoid arthritis.

As one skilled in the art will appreciate, features and preferred embodiments of one aspect of the present invention will also be relevant to other aspects of the present invention.

Brief description of graphic materials

In fig. 1 shows a graph of plasma concentration (ng/ml) versus time after dosing (h) for the reference compound NMC-S-01-A (dark round marks) and the corresponding biarylsulfonamide metabolite (MET-001) (light round marks), obtained using the methods described herein. The metabolite was formed in large quantities and accumulated over time.

In fig. 2 is a plot of plasma concentration (ng/mL) versus time post-dose (h) for compound CHMSA-10-B (closed circle marks) and the corresponding biarylsulfonic acid metabolite (MET-002) (open circle marks), obtained with using the methods described herein. The metabolite was detected temporarily, 0.5 hours after administration.

In fig. 3 is a graph of the mean arthritis index versus time (day of dosing) for test compound CHMSA-01-A at 10 mg/kg/day by gavage (light circles) and control (dark circles) obtained using the methods described in this document.

In fig. 4 is a graph of the mean arthritis index versus time (day of dosing) for test compound CHMSA-03-A at 10 mg/kg/day by gavage (light circles) and control (dark circles) obtained using the methods described in this document.

In fig. 5 is a graph of the Arthritis Index versus time (dose day) for reference compound ABD899 at 10 mg/kg/day (open circle, open square), control (closed circle), and marketing-registered positive control. etanercept (triangular labels) prepared using the methods described herein.

In fig. 6 shows a graph of the arthritis index versus time (day of dosing) for reference compound NMC-S-01-A at a dose of 10 mg/kg/day (light round marks) and control (dark round marks), obtained using the methods described in this document.

Detailed Description of the Invention

Connections.

One aspect of the present invention relates to certain cyclohexylmethylsulfonylaryl compounds.

More specifically, these compounds are certain 1-methyl-4[arylsulfonylmethyl]cyclohexanol compounds related to the following biphenyl and pyridylphenyl compounds:

1-methyl-4-[[4-(2-pyridyl)phenyl]sulfonylmethyl]cyclohexanol

Thus, one aspect of the present invention relates to a compound of the following formula, or a pharmaceutically acceptable salt or solvate thereof, wherein =X-, -R ¹ , -R ² , -R ³ , -R ⁴ , -R ⁵ and -R ⁶ are significant - 10 043371 definitions defined in this document (for convenience, collectively referred to herein as

1-methyl-4-[arylsulfonylmethyl]cyclohexanol compounds and CHMSA compounds):

Conformation of the cyclohexyl ring.

It is worth noting that the OH and CH3 substituents on the cyclohexyl ring can be trans/cis or cis/trans, respectively, with respect to the rest of the molecule (that is, with the cyclohexyl ring to which they are attached, relative to the rest of the compound that attached in the para position of the cyclohexyl ring).

Unless otherwise indicated, all such conformations are intended to be covered by the compound reference in which no specific conformation is specified.

Configuration of the cyclohexyl ring.

Compounds in the trans-OH conformation can be designated as follows:

Compounds in the cis-OH conformation can be designated as follows:

Note also that the cyclohexane ring can adopt a chair, boat, or twist conformation, and that interconversion between conformations is possible. Again, unless otherwise noted, all such conformations (e.g., chair, boat, twist, OH axial, OH equatorial, etc.) are assumed to be covered by the compound reference, which does not specify a specific conformation.

The configuration of the carbon atom to which -R ⁵ and -R ⁶ are attached.

It should be noted that depending on what the -R ⁵ and -R ⁶ groups are, the carbon atom to which they are attached may be chiral and therefore may be in the (R) or (S) configuration.

Unless otherwise noted, all such configurations are assumed to be covered by the connection reference that does not specify a specific configuration.

Connections in one configuration can be designated as follows:

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Connections in a different configuration may be designated as follows:

Conformation of the biaryl group.

It should be noted that depending on what the -R ¹ , -R ² , -R ³ , -R ⁴ and X groups are, there may be free rotation about the single bond connecting the two aryl groups.

For the avoidance of doubt, all such rotational conformations (ie, various rotations around a single bond connecting two aryl groups) are intended to be included within the scope of the present invention. For example, the following formulas are assumed to be equivalent and represent the same group:

Implementation options

Some embodiments of the present invention include the following: 1. A compound of the following formula:

or a pharmaceutically acceptable salt or solvate thereof;

where =X- independently represents -CH= or -N=;

- R ¹ is independently -H or -R ^1X ;

- R ^1X is independently -F, -Cl, -R ^1C , -R ^1F or -CN;

- R ^1C independently represents a saturated straight or branched C _1-3 alkyl;

- R ^1f independently represents a saturated straight or branched C _1-3 fluoroalkyl;

- R ² is independently -H or -R ^2X ;

- R ^2x is independently -F, -Cl, -R ^2C , -R ^2F or -CN;

- R ^2C independently represents a saturated straight or branched C _1-3 alkyl;

- R ^2f independently represents a saturated straight or branched C _1-3 fluoroalkyl;

- R ³ is independently -H or -R ^3X ;

- R ^3X is independently -F, -Cl, -R ^3C , -R ^3F or -CN;

- R ^3C independently represents a saturated straight or branched C _1-3 alkyl;

- R ^3f independently represents a saturated straight or branched C _1-3 fluoroalkyl;

- R ⁴ is independently -H or -R ^4X ;

- R ^4X is independently -F, -Cl, -R ^4C , -R ^4F or -CN;

- R4C independently represents a saturated straight or branched C _1-3 alkyl;

- R ^4f independently represents a saturated straight or branched C _1-3 fluoroalkyl;

- R ⁵ is independently -H or -R ^5X ;

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-R ^5X is independently -F, -R ^5C or -R ^5F ;

-R ^5C independently represents a saturated straight or branched C _1-3 alkyl;

-R5F independently represents a saturated straight or branched C _1-3 fluoroalkyl;

- R ⁶ is independently -H or -R ^6X ;

- R ^6X is independently -F, -R ^6C or -R ^6F ;

- R ^6C independently represents a saturated straight or branched C1_ ₃ alkyl; And

- R ^6f independently represents a saturated straight or branched C1_ ₃ fluoroalkyl;

or -R ⁵ and -R ⁶ together with the carbon atom to which they are attached form a saturated C _3-6 cycloalkyl.

Unless otherwise indicated, if a compound is shown or described that has one or more chiral centers and two or more stereoisomers are possible, all such stereoisomers are considered to be described herein and within the scope of the invention, individually (e.g., isolated from another stereoisomer(s). stereoisomers)), and in the form of mixtures (for example, in the form of equimolar or nonequimolar mixtures of two or more stereoisomers). For example, unless otherwise indicated, if a compound has a single chiral center, each of the (R)- and (S)-enantiomers is considered to be described herein and covered by the invention, either individually (e.g., isolated from the other enantiomer) or as mixtures (for example, in the form of equimolar or nonequimolar mixtures of two enantiomers).

The term saturated straight or branched C _1-3 alkyl means -CH ₃ (methyl), -CH2CH3 (ethyl), -CH2CH2CH3 (n-propyl) and -CH(CH ₃ ) ₂ (isopropyl).

The term saturated straight or branched C _1-3 fluoroalkyl means a saturated straight or branched C _1-3 alkyl group substituted with one or more fluorines. Accordingly, C _1-3 fluoroalkyl includes, for example, -CF ₃ , -CH2F, -CHF2, -CH ₂ CF ₃ , -CH2CH2F, etc.

The term saturated C _{3_6} cycloalkyl means cyclopropyl, cyclobutyl, cyclopentyl and _cyclohexyl .

Group =X-.

2. The compound according to claim 1, where =X- represents -CH=.

3. The connection according to claim 1, where =X- represents -N=.

Group -R ¹ .

4. A compound according to any one of claims 1 to 3, where -R ¹ is -R ^1X

5. A compound according to any one of claims 1 to 3, where -R ¹ represents -H.

Group -R ^1X

6. A compound according to any one of claims 1 to 5, wherein -R ^1X , if present, is independently -F, -Cl or -CN.

7. A compound according to any one of claims 1 to 5, wherein -R ^1X , if present, represents -F.

8. A compound according to any one of claims 1 to 5, wherein -R ^1X , if present, represents -Cl.

9. A compound according to any one of claims 1 to 5, wherein -R ^1X , if present, represents -CN.

10. A compound according to any one of claims 1 to 5, wherein -R ^1X , if present, is -R ^1C .

11. A compound according to any one of claims 1 to 5, wherein -R ^1X , if present, is -R ^1F .

Group -R ^1C .

12. A compound according to any one of claims 1 to 11, wherein -R ^1C , if present, represents -CH ₃ .

Group -R ^1F .

13. A compound according to any one of claims 1 to 12, wherein -R ^1F , if present, is -CF ₃ .

Group -R ² .

14. A compound according to any one of claims 1 to 13, where -R ² is -R ^2X .

15. A compound according to any one of claims 1 to 13, where -R ² represents -H.

Group -R ^2X .

16. A compound according to any one of claims 1 to 15, wherein -R ^2X , if present, independently represents F, -Cl or -CN.

17. A compound according to any one of claims 1 to 15, wherein -R ^2X , if present, represents -F.

18. A compound according to any one of claims 1 to 15, wherein -R ^2X , if present, represents -Cl.

19. A compound according to any one of claims 1 to 15, wherein -R ^2X , if present, represents -CN.

20. A compound according to any one of claims 1 to 15, wherein -R ^2X , if present, is -R ^2C .

21. A compound according to any one of claims 1 to 15, wherein -R ^2X , if present, is -R ^2F .

Group -R ^2C .

22. A compound according to any one of claims 1 to 21, wherein -R ^2C , if present, is -CH ₃ .

Group -R ^2F .

23. A compound according to any one of claims 1 to 22, wherein -R ^2F , if present, is -CF ₃ .

Group -R ³ .

24. A compound according to any one of claims 1 to 23, where -R ³ is -R ^3X .

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25. A compound according to any one of claims 1 to 23, where -R ³ represents -H.

Group -R ^3X .

26. A compound according to any one of claims 1 to 25, wherein -R ^3X , if present, independently represents F, -Cl or -CN.

27. A compound according to any one of claims 1 to 25, wherein -R ^3X , if present, is -F.

28. A compound according to any one of claims 1 to 25, wherein -R ^3X , if present, represents -Cl.

29. A compound according to any one of claims 1 to 25, wherein -R ^3X , if present, represents -CN.

30. A compound according to any one of claims 1 to 25, wherein -R ^3X , if present, is -R ^3C .

31. A compound according to any one of claims 1 to 25, wherein -R ^3X , if present, is -R ^3F .

Group -R ^3C .

32. A compound according to any one of claims 1 to 31, wherein -R ^3C , if present, represents -CH3.

Group -R ^3F .

33. A compound according to any one of claims 1 to 32, wherein -R ^3F , if present, is -CF3.

Group -R ⁴ .

34. A compound according to any one of claims 1 to 33, where -R ⁴ is -R ^4X

35. A compound according to any one of claims 1 to 33, where -R ⁴ represents -H.

Group -R ^4X .

36. A compound according to any one of claims 1 to 35, wherein -R ^4X , if present, independently represents F, -Cl or -CN.

37. A compound according to any one of claims 1 to 35, wherein -R ^4X , if present, is -F.

38. A compound according to any one of claims 1 to 35, wherein -R ^4X , if present, represents -Cl.

39. A compound according to any one of claims 1 to 35, wherein -R ^4X , if present, represents -CN.

40. A compound according to any one of claims 1 to 35, wherein -R ^4X , if present, is -R ^4C .

41. A compound according to any one of claims 1 to 35, wherein -R ^4X , if present, is -R ^4F .

Group -R ^4C .

42. A compound according to any one of claims 1 to 41, wherein -R ^4C , if present, is -CH3.

Group -R ^4F .

43. A compound according to any one of claims 1 to 42, wherein -R ^4F , if present, is -CF ₃ .

Group -R ⁵ .

44. A compound according to any one of claims 1 to 43, where -R ⁵ is -R ^5X .

45. A compound according to any one of claims 1 to 43, where -R ⁵ represents -H.

Group -R ^5X .

46. A compound according to any one of claims 1 to 45, wherein -R ^5X is independently -F, -R5C or -R ^5F .

47. A compound according to any one of claims 1 to 45, wherein -R ^5X , if present, is -F.

48. A compound according to any one of claims 1 to 45, wherein -R ^5X , if present, is -R ^5C .

49. A compound according to any one of claims 1 to 45, wherein -R ^5X , if present, is -R ^5F .

Group -R ^5C .

50. A compound according to any one of claims 1 to 49, wherein -R ^5C , if present, is -CH3.

Group -R ^5F .

51. A compound according to any one of claims 1 to 50, wherein -R ^5F , if present, is -CF3.

Group -R ⁶ .

52. A compound according to any one of claims 1 to 51, where -R ⁶ is -R ^6X .

53. A compound according to any one of claims 1 to 51, where -R ⁶ represents -H.

Group -R ^6X .

54. A compound according to any one of claims 1 to 53, wherein -R ^6X is independently -F, -R ^6C or -R ^6F .

55. A compound according to any one of claims 1 to 53, wherein -R ^6X , if present, is -F.

56. A compound according to any one of claims 1 to 53, wherein -R ^6X , if present, is -R ^6C

57. A compound according to any one of claims 1 to 53, wherein -R ^6X , if present, is -R ^6F .

Group -R ^6C .

58. A compound according to any one of claims 1 to 57, wherein -R ^6C , if present, represents -CH3.

Group -R ^6F .

59. A compound according to any one of claims 1 to 58, wherein -R ^6F , if present, is -CF3.

Groups -R ⁵ and -R ⁶ together.

60. A compound according to any one of claims 1 to 43, wherein -R ⁵ and -R ⁶ together with the carbon atom to which they are attached form a saturated C ₃ - ₆ cycloalkyl.

61. A compound according to any one of claims 1 to 43, wherein -R ⁵ and -R ⁶ together with the carbon atom to which they are attached form cyclopropyl.

62. A compound according to any one of claims 1 to 43, wherein -R ⁵ and -R ⁶ together with the carbon atom to which they are attached form cyclobutyl.

63. A compound according to any one of claims 1 to 43, wherein -R ⁵ and -R ⁶ together with the carbon atom to which they are attached form a cyclopentyl.

64. A compound according to any one of claims 1 to 43, where -R ⁵ and -R ⁶ together with the carbon atom to which they

- 14 043371 are added to form cyclohexyl.

Conformation of the cyclohexyl ring.

65. A compound according to any one of claims 1 to 64, wherein said compound is a compound of the following formula, or a pharmaceutically acceptable salt or solvate thereof:

66. A compound according to any one of claims 1 to 64, wherein said compound is a compound of the following formula, or a pharmaceutically acceptable salt or solvate thereof:

H ₃ C

The configuration of the carbon atom to which -R ⁵ and -R ⁶ are attached.

67. A compound according to any one of claims 1 to 66, wherein -R ⁵ and -R ⁶ are different, and said compound is a compound of the following formula, or a pharmaceutically acceptable salt or solvate thereof:

68. A compound according to any one of claims 1 to 66, where -R ⁵ is a compound of the following formula,

-R ⁶ or are different, and the said compound is its pharmaceutically acceptable salt or solvate:

Some preferred connections.

69. A compound according to claim 1, which is a compound of one of the following formulas or a pharmaceutically acceptable salt or solvate thereof:

- 15 043371

- 16 043371

CHMSA-07 H ₃ C .A '—'2—' o Cl CHMSA-08 H ₃ C •<k/ CF ₃ CHMSA-09 H ₃ C '—n '—ς o CF ₃ CHMSA-10 h ₃ c У\ ^OHCN /\<H>r CHMSA-11 H ₃ C \—f \—/ _o CHMSA-12 H ₃ C '—')--' 0 F

- 17 043371

Code designation

Structure

- 18 043371

71. A compound according to claim 1, which is a compound of one of the following formulas or a pharmaceutically acceptable salt or solvate thereof:

- 19 043371

- 20 043371

Combinations.

It is understood that certain features of the invention, which for clarity are described in the context of individual embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the present invention, which for brevity are described in the context of a single embodiment, may also be presented separately or in any suitable subcombination. All combinations of embodiments related to the chemical groups represented by the variables (for example, =X-, -R ¹ , -R ^1X , -R ^1C , -R ^1F , -R ² , -R ^2X , -R ^2C , -R ^2F , -R ³ , -R ^3X , -R ^3C , -R ^3F , -R ⁴ , -R ^4x , -R ^4C , -R ^4f , -R ⁵ , -R ^{5X , -R 5C ,} -R ^5F , - R ⁶ , -R ^6x , -R ^6C , -R ^6f , etc.) are specifically covered by the present invention and are described herein as if each combination were described separately and in detail, to the extent that such combinations cover compounds that are stable compounds (ie, compounds that can be isolated, characterized and tested for biological activity). In this context, one skilled in the art will readily appreciate that certain combinations of groups (eg, substituents) can produce compounds that cannot be easily synthesized and/or are chemically unstable. In addition, all subcombinations of chemical groups listed in the embodiments describing such variables are also specifically covered by the present invention and are described herein as if each such subcombination of chemical groups were separately and explicitly described herein.

Essentially purified forms.

One aspect of the present invention relates to CHMSA compounds as described herein in substantially purified form and/or in a form substantially free of impurities.

In one embodiment of the present invention, said substantially purified form is at least 50% by weight, such as at least 60% by weight, such as at least 70% by weight, such as at least 80% by weight, for example at least 90% by weight, for example at least 95% by weight, for example at least 97% by weight, for example at least 98% by weight, for example at least 99% by weight.

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Unless otherwise indicated, substantially purified form refers to the compound in any stereoisomeric or enantiomeric form. For example, in one embodiment of the present invention, said substantially purified form refers to a mixture of stereoisomers, i.e. it is purified in relation to other compounds. In one embodiment of the present invention, the substantially purified form is of a single stereoisomer, for example an optically pure stereoisomer. In one embodiment of the present invention, said substantially purified form is a mixture of enantiomers. In one embodiment of the present invention, the substantially purified form refers to an equimolar mixture of enantiomers (ie, a racemic mixture, racemate). In one embodiment of the present invention, said substantially purified form is of a single enantiomer, for example, an optically pure enantiomer.

In one embodiment of the present invention, the impurities are no more than 50 wt.%, for example no more than 40 wt.%, for example no more than 30 wt.%, for example no more than 20 wt.%, for example no more than 10 wt.%, for example no more 5 wt.%, for example not more than 3 wt.%, for example not more than 2 wt.%, for example not more than 1 wt.%.

Unless otherwise stated, impurities refer to other compounds, e.g. compounds other than stereoisomers or enantiomers. In one embodiment of the present invention, the term impurities refers to other compounds and other stereoisomers. In one embodiment of the present invention, the term impurities refers to other compounds and other enantiomers.

In one embodiment of the present invention, said substantially purified form has an optical purity of at least 60% (i.e., 60% of the compound (per mole) is the desired stereoisomer or enantiomer and 40% is the undesired stereoisomer or enantiomer) , for example, optical purity of at least 70%, for example, optical purity of at least 80%, for example, optical purity of at least 90%, for example, optical purity of at least 95%, for example, optical purity of at least 97% , for example, an optical purity of at least 98%, for example, an optical purity of at least 99%.

Isomers.

Certain compounds may exist in one or more specific geometric, optical, enantiomeric, diastereomeric, epimeric, stereoisomeric, tautomeric, conformational or anomeric forms, including, but not limited to, cis and trans forms; E- and Z-forms; c-, t- and r-forms; endo- and exoforms; R-, S- and meso forms; D- and L-forms; d- and I-forms; (+) and (-) forms; keto, enol and enolate forms; syn- and anti-forms; synclinal and anticlinal forms; α- and βforms; axial and equatorial shapes; boat, chair, twist, envelope and half-chair forms, as well as combinations thereof, hereinafter collectively referred to as isomers (or isomeric forms).

A reference to a class of structures may also include structurally isomeric forms falling within that class (e.g., C _1-3 alkyl includes n-propyl and isopropyl; butyl includes n-butyl, isobutyl, sec-butyl, and tert-butyl; methoxyphenyl includes ortho-, meta - and paramethoxyphenyl). However, reference to a specific group or substitution pattern is not intended to include other structures (or structural isomers) that differ in the bonds between atoms rather than in the position in space. For example, a reference to a methoxy group, -OCH3, should not be construed as a reference to its structural isomer, the hydroxymethyl group, -CH2OH.

The above exception does not apply to tautomeric forms, such as the keto, enol and enolate forms, as in the following tautomeric pairs: keto/enol (shown below), imine/enamine, amide/iminoalcohol, amidine/amidine, nitroso/oxime , thioketone/enethiol, N-nitroso/hydroxyazo and nitro/acy-nitro. In this document, reference to one tautomer is intended to cover both tautomers.

n ,o \ _z oh -n ⁺ _h o ^- -s-s's=s' - s=s' | \ / \ _n ⁺ / \ keto enol enolate

It should be noted that the term isomer specifically includes compounds with one or more isotopic substitutions. For example, H can be in any isotopic form, including 1H, ^2H (D) and ^3H (T); C can be in any isotopic form, including ¹² C, ¹³ C and ¹⁴ C; O can be in any isotopic form, including ¹⁶ O and ¹⁸ O; etc.

Unless otherwise indicated, reference to a specific compound includes all such isomeric forms, including mixtures thereof (eg, racemic mixtures). Methods for preparing (eg, asymmetric synthesis) and separating (eg, fractional crystallization and chromatographic methods) such isomeric forms are either known in the art or can be readily prepared by adapting the methods described herein in a known manner.

Salt.

It may be convenient or desirable to prepare, purify and/or process the corresponding salt of the compound, for example, a pharmaceutically acceptable salt.

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Examples of pharmaceutically acceptable salts are described in Berge et al., 1977, Pharmaceutically Acceptable Salts, J. Pharm. Sci., Vol. 66, p. 1-19.

For example, if the compound is anionic, or contains a functional group that can be anionic (eg, -COOH can be ^-COO- ), then a salt can be formed with a suitable cation. Examples of suitable inorganic cations include, but are not limited to, alkali metal ions, such as Na ⁺ and K ⁺ , alkaline earth cations such as Ca ²⁺ and Mg ²⁺ , and other cations such as Al ³⁺ , as well as ammonium ion (i.e. NH4 ⁺ ). Examples of suitable organic cations include, but are not limited to, substituted ammonium ions (e.g., NH3R ⁺ , NH2R2 ⁺ , NHR3 ⁺ , NR4 ⁺ ), for example, where each R is independently a straight or branched saturated C1_ ₁₈ alkyl, C _3-8 cycloalkyl, C _3-8 cycloalkyl-C ₁ . ₆ alkyl and phenyl-C ₁ . ₆ alkyl, where the phenyl group is optionally substituted. Examples of some suitable substituted ammonium ions are those derived from: ethylamine, diethylamine, dicyclohexylamine, triethylamine, butylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, benzylamine, phenylbenzylamine, choline, meglumine and tromethamine, as well as amino acids such as lysine and arginine An example of a common quaternary ammonium ion is N(CH ₃ ) ₄ ⁺ .

If the compound is cationic or has a functional group that can become cationic when protonated (for example, -NH ₂ can become -NH ₃ ⁺ ), then a salt can be formed with the appropriate cation.

For example, if the starting structure contains a cationic group (eg -NMe ₂ ⁺ ) or contains a functional group that can become cationic when protonated (eg -NH2 can become -NH3 ⁺ ) then a salt with the appropriate anion can be formed. In the case of a quaternary ammonium compound, a counteranion is usually always present to balance the positive charge. If, in addition to a cationic group (eg -NMe2 ⁺ , _-NH3 ⁺ ), the compound also contains a group capable of forming an anion (eg -COOH), then an internal salt (also called a zwitterion) can form.

Examples of suitable inorganic anions include, but are not limited to, those derived from the following inorganic acids: hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfurous acid, nitric acid, nitrous acid, phosphoric acid and phosphorous acid.

Examples of suitable organic anions include, but are not limited to, those derived from the following organic acids: 2-acetyloxybenzoic acid, acetic acid, trifluoroacetic acid, ascorbic acid, aspartic acid, benzoic acid, camphorsulfonic acid, cinnamic acid, citric acid, edetic acid, 1 ,2-ethanedisulfonic acid, ethanesulfonic acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, hydroxymaleic acid, hydroxynaphthalenecarboxylic acid, isethionic acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, methanesulfonic acid , mucinic acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, pantothenic acid, phenylacetic acid, phenylsulfonic acid, propionic acid, pyruvic acid, salicylic acid, stearic acid, succinic acid, sulfanilic acid, tartaric acid, toluenesulfonic acid and valeric acid acids. Examples of suitable polymeric organic anions include, but are not limited to, those derived from the following polymeric acids: tannic acid, carboxymethylcellulose.

Examples of suitable counterions that are particularly suitable for quaternary ammonium compounds (eg those with ^a -NMe ₂₊ group) include 1-adamantane sulfonate, benzene sulfonate, bisulfate, bromide, chloride, iodide, methanesulfonate, methyl sulfate, 1,5-naphthalene bis-sulfonate , 4nitrobenzenesulfonate, formate, tartrate, tosylate, trifluoroacetate, trifluoromethylsulfonate, sulfate. Again, if the compound also contains a group capable of forming an anion (eg -COOH), then an internal salt can be formed.

Unless otherwise indicated, reference to a specific compound also includes its salt forms.

Solvates and hydrates.

It may be convenient or desirable to prepare, purify and/or process the corresponding solvate of the compound. The term solvate is used here in the usual sense to refer to a complex of a solute (eg, a compound, a salt of a compound) and a solvent. If the solvent is water, the solvate is conveniently called a hydrate, for example, monohydrate, dihydrate, trihydrate, etc.

Unless otherwise indicated, reference to a specific compound also includes its solvate and hydrate forms.

Chemically protected forms.

It may be convenient or desirable to obtain, purify and/or process the compound in a chemically protected form. The term chemically protected form is used herein in its generally accepted chemistry sense and refers to a compound in which one or more reactive functional groups are protected from unwanted chemical interactions under certain conditions (e.g., pH, temperature, radiation, solvent, etc.) .P.). In practice, well-known chemical methods are used to reversibly convert a functional group into a non-reactive group that would otherwise be reactive under certain conditions. In a chemically protected form, one or more reactive functional groups are in the form of a protected or protecting group (alternatively, as a masked or masking group, or a blocked or blocking group). By protecting a reactive functional group, reactions involving other unprotected reactive functional groups can be performed without affecting the protected group; the protecting group can be removed or the masking group can be converted, usually in a subsequent step, without significantly affecting the rest of the molecule. See, for example, Protective Groups in Organic Synthesis (T. Green and P. Wuts; ^4th Edition; John Wiley and Sons, 2006).

A wide range of such shielding, blocking or masking techniques are widely used and well known in organic synthesis. For example, a compound that has two non-equivalent reactive functional groups, both of which will be reactive under specified conditions, can be derivatized in such a way as to make one of the functional groups protected and therefore non-reactive under specified conditions; a compound thus protected can be used as a reagent that actually has only one reactive functional group. Once the desired interaction (involving another functional group) is completed, the protected group can be unprotected to return it to its original functionality.

For example, the hydroxyl group may be protected as an ether (-OR) or an ester (-OC(=O)R), for example: tert-butyl ether; benzyl, benzhydryl (diphenylmethyl) or trityl (triphenylmethyl) ether; trimethylsilyl or tert-butyldimethylsilyl ether; or acetyl ester (-OC(=O)CH ₃ , -OAc).

Prodrugs.

It may be convenient or desirable to prepare, purify and/or process the compound as a prodrug. As used herein, the term prodrug refers to a compound that produces the desired active compound in vivo. Typically, a prodrug is inactive, or less active, than the target active compound, but may provide processing or administration advantages or advantageous metabolic properties.

For example, some prodrugs are esters of the active compound (eg, a physiologically acceptable metabolically labile ester). When metabolized, the ester group (-C(=O)OR) is cleaved to form the active drug. Such esters can be formed by esterifying, for example, any of the carboxylic acid groups (-C(=O)OH) in the parent compound, optionally pre-protecting any other reactive groups present in the parent compound, followed by deprotection groups if necessary.

In addition, some prodrugs are activated enzymatically to form the active compound or compound that, through a further chemical reaction, produces the active compound (eg, as in antibody-directed enzyme prodrug therapy (ADEPT), gene-directed enzyme prodrug therapy (GDEPT), lipid-directed enzyme prodrug therapy prodrug therapy (LIDEPT), etc.). For example, the prodrug may be a sugar derivative or other glycoside conjugate, or may be an amino acid ester derivative.

General chemical synthesis.

This section describes methods for the chemical synthesis of CHMSA compounds. These and/or other well known methods (see, for example, Greig et al., 2010a; Bahmanyar et al., 2010; Patel et al., 2014; Patel et al., 2016) can be modified and/or modified in known ways. adapted to provide alternative or improved synthesis methods.

In one approach, the cyclohexanone-4-ester (1) is protected with ethylene glycol (eg, in toluene with p-toluenesulfonic acid (PTSA)) (2) and reduced with lithium aluminum hydride to give the corresponding alcohol derivative (3). The said alcohol (3) is converted into bromide (4) using triphenylphosphine (PPh ₃ ) and carbon tetrabromide (CBr ₄ ). The bromide group of bromide (4) is replaced with a suitable aromatic thiolate anion (e.g. from 4-bromobenzenethiol (5)) using cesium carbonate (Cs ₂ CO ₃ ) as a base to give the corresponding sulfide derivative (6), which is then oxidized to give bromophenylsulfone (7) using mchloroperbenzoic acid (m-CPBA). The said bromophenylsulfone (7) is coupled with a suitable aromatic boronic acid ester (8) using transition metal catalysis, for example tetrakis(triphenylphosphine)palladium(0) (Pd(PPh3)4), to obtain the corresponding biarylsulfone (9). Ketone (10) is regenerated by deprotection using dilute aqueous hydrochloric acid (HCl) and then reacted with methyl magnesium bromide (MeMgBr) to produce a pair of isomeric tertiary alcohols (11A, 11B), which are then separated with

- 24 043371 using preparative HPLC.

This approach is illustrated in the following diagram.

Scheme 1

If the corresponding aromatic thiol (eg (5) above) is not commercially available, it can be prepared by reducing the corresponding sulfonyl chloride (RSO2Cl) with a reducing agent such as triphenylphosphine (PPh ₃ ).

This approach is illustrated in the following diagram.

Scheme 2

In another approach, the corresponding aniline can be diazotized, for example, with sodium nitrite (NaNO2) and hydrochloric acid (HCl). The resulting diazonium salt is then reacted with potassium ethyl xanthate (CH3CH2OCS2K) and then hydrolyzed with potassium hydroxide (KOH) to produce the corresponding aromatic thiol (eg (5) above).

This approach is illustrated in the following diagram.

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Scheme 3

If one of the substituents (R ³ and R ⁴ ) is a nitrile group (-CN), then it is possible that the nitrile group is hydrated to a primary amide group during hydrolysis using potassium hydroxide. In this case, an aromatic thiol containing a primary amide group is combined with a bromide (for example, (4)+(5)), and then treated with a dehydrating agent, for example, trifluoroacetic anhydride (CF ₃ C(=O)OC(=O) CF ₃ ) with regeneration of the nitrile group from the primary amide group (for example, (6)).

In the second approach, bromophenylsulfone (7) is converted to boronic acid ester (13) using 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2 -yl)-1,3,2dioxaborolane (12) and transition metal catalysis, for example, using bis(triphenylphosphine)palladium(P) dichloride. The boronic acid ester (13) is then coupled with a suitable aromatic bromide (14) using transition metal catalysis, for example tetrakis(triphenylphosphine)palladium(0) (Pd(PPh ₃ ) ₄ ), to give the corresponding biarylsulfone (15 ). Ketone (16) is regenerated by deprotection using dilute aqueous hydrochloric acid (HCl) and then reacted with methyl magnesium bromide (MeMgBr) to produce a pair of isomeric tertiary alcohols (17A, 17B), which are then separated by preparative HPLC.

This approach is illustrated in the following diagram.

Scheme 4

Alternatively, the boronic acid ester (13) is coupled with a suitable aromatic triflate (14') using transition metal catalysis, for example tetrakis(triphenylphosphine)palladium(0) (Pd(PPh3)4), to give the corresponding biarylsulfone (15 '). Ketone (16') is regenerated by deprotection using dilute aqueous hydrochloric acid (HCl) and then reacted with methyl magnesium bromide (MeMgBr) to produce a pair of isomeric tertiary alcohols (17A', 17B'), which are then separated by preparative HPLC .

This approach is illustrated in the following diagram.

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Scheme 5

In another approach, the bromine group of said bromide (4) is replaced with a suitable biarylthiolate anion (for example, from 4-(2,4-dichlorophenyl)benzenethiol (18)) using cesium carbonate (CsCO ₃ ) as a base to give the corresponding sulfide derivative ( 19), which is then oxidized to give bromophenylsulfone (20) using m-chloroperbenzoic acid (mCPBA). Ketone (21) is regenerated by deprotection using dilute aqueous hydrochloric acid (HCl) and then reacted with methyl magnesium bromide (MeMgBr) to produce a pair of isomeric tertiary alcohols (22A, 22B), which are then separated by preparative HPLC.

This approach is illustrated in the following diagram.

Scheme 6

If the corresponding biarylthiol (eg (18) above) is not available for purchase, it can be prepared, for example, as follows. The combination of a suitable boronic acid (23) with a suitable bromobenzene (24) is carried out using the Suzuki combination. The resulting biaryl (25) is sulfonylated using chlorosulfonic acid (CISO3H) to give the corresponding sulfonic acid, which is then reacted with thionyl chloride (SOCl ₂ ) to give the corresponding arylsulfonyl chloride (26). Reduction of sulfonyl chloride (26), for example, with triphenylphosphine (PPh ₃ ), gives the biarylthiol derivative (18).

This approach is illustrated in the following diagram.

Scheme 7

In another approach, biarylsulfone (9) is treated with a base, such as lithium diisopropylamide (LDA), and then either a fluorinating agent, such as N-fluorobenzenesulfonimide (NFSI), or an alkylating agent, such as methyl iodide (MeI), to produce biarylsulfone (27), where R ¹ = fluorine or R ¹ = alkyl (eg methyl), respectively. The ketone (28') is regenerated by deprotection using dilute aqueous hydrochloric acid (HCl) and then reacted with methyl magnesium bromide (MeMgBr) to produce isomeric tertiary alcohols (29A, 29B), then some or all of them are separated by preparative HPLC.

This approach is illustrated in the following diagram.

Scheme 8

Additionally, said biarylsulfone (27) can be treated with a base, such as LDA, and then either a fluorinating agent, such as NFSI, or an alkylating agent, such as MeI, to obtain biarylsulfone (30), where R ² = fluorine or R ² = alkyl ( for example, methyl), respectively. In this way, compounds can be obtained in which R ¹ and R ² are different (for example, F and Me; Me and Et, etc.). Ketone (31') is regenerated by deprotection using dilute aqueous hydrochloric acid (HCl) and then reacted with methyl magnesium bromide (MeMgBr) to produce isomeric tertiary alcohols (32A, 32B). In the case where R ¹ and R ² are the same, the isomeric tertiary alcohol pair is separated using preparative HPLC. In the case where R ¹ and R ² are different, some or all of the isomeric tertiary alcohols are separated using preparative HPLC.

This approach is illustrated in the following diagram.

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Scheme 9

Alternatively, biarylsulfone (9) is treated with a base, such as LDA, and then with a terminally dihalogenated alkane, such as 1-bromo-2-chloroethane (32). The resulting biarylsulfone (33) is treated with a second equivalent of a base, eg LDA, to produce biarylsulfone (34), in which R ¹ and R ² together with the carbon atom to which they are attached form a cycloalkyl ring, eg a cyclopropyl ring. Ketone (35) is regenerated by deprotection using dilute aqueous hydrochloric acid (HCl) and then reacted with methyl magnesium bromide (MeMgBr) to produce a pair of isomeric tertiary alcohols (36A, 36B), which are then separated using preparative HPLC.

This approach is illustrated in the following diagram.

Scheme 10

In another approach, bromo-monoarylsulfone (37) (another example is (7)) is deprotected with dilute aqueous hydrochloric acid (HCl) to yield the corresponding ketone (38), which is then reacted with methylmagnesium bromide (MeMgBr) to form a pair isomeric tertiary alcohols (39A, 39B). These alcohols are then coupled with a suitable aromatic boronic ester (8) using transition metal catalysis, such as tetrakis(triphenylphosphine)palladium(0) (Pd(PPh ₃ ) ₄ ), to produce the corresponding pair of isomeric tertiary alcohols ( 40A, 40B). These isomers are then separated using preparative HPLC.

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This approach is illustrated in the following diagram.

Scheme 11

F F

These and/or other well known methods can be modified and/or adapted in known manner to facilitate the synthesis of additional compounds described herein. See for example:

Comprehensive Organic Transformations: A Guide to Functional Group Preparations,

^2nd Edition (Wiley) 2010. Ed. RCLarock. ISBN: 978-1-118-03758-4.

Comprehensive Organic Synthesis, 2nd Edition (Elsevier) 2014. Editor in Chiefs P.

Knochel, G. A. Molander. eBook ISBN: 9780080977430. Hardcover version, ISBN: 9780080977423.

Science of Synthesis: Cross Coupling and Heck-Type Reactions, Workbench Edition (Thieme) 2013. Ed. G. Molander, J.P. Wolfe, Mats Larhed. ISBN 9783131734112.

Greene's Protective Groups in Organic Synthesis, ^4th Edition (Wiley) 2006. PGM

Wuts, T.W. Greene. Printed version, ISBN: 9780471697541. Online ISBN:

9780470053485.

e-EROS Encyclopedia of Reagents for Organic Synthesis, (Wiley). Online ISBN:

9780470842898. DOI: 10.1002/047084289X.

Organic Reactions: Electrophilic Fluorination with N-F Reagents (Wiley) 2008.

J. Baudoux, D. Cahard. DOI: 10.1002/0471264180.or069.02.

Compositions.

One aspect of the present invention relates to a composition (eg, a pharmaceutical composition) containing a CHMSA compound described herein and a carrier, diluent or excipient (eg, a pharmaceutically acceptable carrier, diluent or excipient).

In one embodiment of the present invention, said composition further comprises one or more (eg, 1, 2, 3, 4) additional therapeutic agents described herein.

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Another aspect of the present invention relates to a method for preparing a composition (eg, a pharmaceutical composition) comprising mixing a CHMSA compound described herein and a carrier, diluent or excipient (eg, a pharmaceutically acceptable carrier, diluent or excipient).

Another aspect of the present invention relates to a method for preparing a composition (eg, a pharmaceutical composition), comprising mixing a CHMSA compound described herein; one or more (eg, 1, 2, 3, 4) additional therapeutic agents described herein; and a carrier, diluent or excipient (eg, a pharmaceutically acceptable carrier, diluent or excipient).

Application.

The CHMSA compounds described herein are useful, for example, in the treatment of disorders (eg, diseases), including, for example, the disorders (eg, diseases) described herein.

Application in methods of therapy.

Another aspect of the present invention relates to the compound CHMSA, as described herein, for use in a method of treating a human or animal body through therapy, for example, for use in a method of treating a disorder (eg, a disease) described herein.

Another aspect of the present invention relates to the compound CHMSA described herein in combination with one or more (e.g., 1, 2, 3, 4) additional therapeutic agents described herein for use in a method of treating a human or animal body by therapy, for example, for use in a method of treating a disorder (eg, disease) described herein.

Application for the production of medicines.

Another aspect of the present invention relates to the use of a CHMSA compound as described herein for the manufacture of a medicament for treating, for example, treating a disorder (eg, a disease) described herein.

In one embodiment, said drug comprises a CHMSA compound.

Another aspect of the present invention relates to the use of a CHMSA compound described herein and one or more (e.g., 1, 2, 3, 4) additional therapeutic agents described herein for the manufacture of a medicament for treating, for example, treating a disorder (eg, a disease) described herein.

In one embodiment of the present invention, the drug comprises a CHMSA compound and one or more (eg, 1, 2, 3, 4) additional therapeutic agents.

Methods of treatment.

Another aspect of the present invention relates to a method of treating, for example, a disorder (eg, a disease) described herein, comprising administering to a patient in need of treatment a therapeutically effective amount of a CHMSA compound described herein, preferably in the form of a pharmaceutical composition.

Another aspect of the present invention relates to a method of treating, for example, a disorder (e.g., a disease) as described herein, comprising administering to a patient in need of treatment a therapeutically effective amount of a CHMSA compound as described herein, preferably in the form of a pharmaceutical composition, and one or more (eg, 1, 2, 3, 4) additional therapeutic agents as described herein, preferably in the form of a pharmaceutical composition.

Conditions to be treated: disorders associated with changes in cellular metabolism.

In one embodiment of the present invention, the treatment is a treatment for a disorder associated with changes in cellular metabolism.

In one embodiment of the present invention, the treatment is a treatment for a disorder in which the regulation of cellular metabolism is disturbed.

Examples of such disorders include many of those described below, including, for example, an autoimmune/inflammatory disorder; cancer and osteoclast-mediated disorder.

In one embodiment of the present invention, the treatment is the treatment of multiple myeloma, diffuse large B-cell lymphoma, acute myeloid leukemia, eosinophilic leukemia, glioblastoma, melanoma, ovarian cancer, chemotherapy-resistant cancer, radiation-resistant cancer, inflammatory arthritis, rheumatoid arthritis, psoriatic arthritis, psoriasis, ulcerative colitis, Crohn's disease, systemic lupus erythematosus (SLE), lupus nephritis, asthma, chronic obstructive pulmonary disease (COPD), non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), autoimmune hepatitis or hidradenitis suppurativa.

Conditions Treated: Autoimmune/Inflammatory Disorders.

In one embodiment of the present invention, the treatment is a treatment for an autoimmune/inflammatory disorder.

In one embodiment of the present invention, said treatment is a treatment for an autoimmune disorder.

In one embodiment of the present invention, said treatment is a treatment for an inflammatory disorder.

In one embodiment of the present invention, said treatment is a treatment for: inflammatory arthritis (including, for example, rheumatoid arthritis; psoriatic arthritis; ankylosing spondylitis; spondyloarthritis; reactive arthritis; infectious arthritis; systemic lupus erythematosus; scleroderma; gout; acquired Still's disease; juvenile idiopathic arthritis); psoriasis; systemic lupus erythematosus; lupus nephritis; systemic sclerosis; scleroderma; hepatitis A; endometriosis; Sjögren's syndrome; inflammatory bowel disease; ulcerative colitis; Crohn's disease; multiple sclerosis; asthma; atherosclerosis; chronic obstructive pulmonary disease (COPD); hidradenitis suppurativa; autoimmune hepatitis; uveitis; pulmonary fibrosis; non-alcoholic fatty liver disease (NAFLD); non-alcoholic steatohepatitis (NASH); allergic disease (including, for example, atopy, allergic rhinitis, atopic dermatitis, anaphylaxis, allergic bronchopulmonary aspergillosis, allergic gastroenteritis, hypersensitivity pneumonitis); allergies; type I diabetes; rheumatic fever; celiac disease; encephalitis; oophoritis; primary biliary cirrhosis; insulin-resistant diabetes; autoimmune adrenal insufficiency (Addison's disease); autoimmune oophoritis; autoimmune orchitis; autoimmune hemolytic anemia; paroxysmal cold hemoglobinuria; Behçet's disease; autoimmune thrombocytopenia; autoimmune neutropenia; pernicious anemia; congenital aplastic anemia; autoimmune coagulopathy; myasthenia gravis; autoimmune polyneuritis; pemphigus; rheumatic carditis; Goodpasture's syndrome; postcardiotomy syndrome; polymyositis; dermatomyositis; irritable bowel syndrome; pancreatitis; gastritis, lichen planus; delayed type hypersensitivity; chronic pneumonia; pulmonary alveolitis; pulmonary granuloma; inflammation of the gums; endodontic diseases; periodontal diseases; hypersensitivity pneumonitis; hay fever; anaphylaxis; skin allergies; hives; gout; polycystic kidney disease; cryopyrin-associated periodic syndrome (CAPS); Muckle-Wells syndrome; Guillain-Barré syndrome; chronic inflammatory demyelinating polyneuropathy; organ or transplant rejection; chronic allograft rejection; acute or chronic graft versus host disease; dermatitis; atopic dermatomyositis; Graves' disease; autoimmune thyroiditis (Hashimoto's thyroiditis); pemphigus; vasculitic syndrome; immune complex mediated vasculitis; bronchitis; cystic fibrosis; pneumonia; pulmonary edema; pulmonary embolism; sarcoidosis; hypertension; emphysema; respiratory failure; acute respiratory distress syndrome; BENTA disease or polymyositis.

In one embodiment of the present invention, said treatment is a treatment for: inflammatory arthritis (including, for example, rheumatoid arthritis; psoriatic arthritis; ankylosing spondylitis; spondyloarthritis; reactive arthritis; infectious arthritis; systemic lupus erythematosus; scleroderma; gout; acquired Still's disease; juvenile idiopathic arthritis); psoriasis; systemic lupus erythematosus; lupus nephritis; systemic sclerosis; scleroderma; hepatitis A; endometriosis; Sjögren's syndrome; inflammatory bowel disease; ulcerative colitis; Crohn's disease; multiple sclerosis; asthma; atherosclerosis; chronic obstructive pulmonary disease (COPD); hidradenitis suppurativa; autoimmune hepatitis; uveitis; pulmonary fibrosis; non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH).

In one embodiment of the present invention, said treatment is a treatment for: inflammatory arthritis (including, for example, rheumatoid arthritis; psoriatic arthritis; ankylosing spondylitis; spondyloarthritis; reactive arthritis; infectious arthritis; systemic lupus erythematosus; scleroderma; gout; acquired Still's disease; juvenile idiopathic arthritis).

In one embodiment of the present invention, said treatment is a treatment for psoriasis; psoriatic arthritis; systemic lupus erythematosus; lupus nephritis; systemic sclerosis; scleroderma; hepatitis A; endometriosis; Sjögren's syndrome; inflammatory bowel disease; ulcerative colitis; Crohn's disease; multiple sclerosis; asthma; atherosclerosis; chronic obstructive pulmonary disease (COPD); hidradenitis suppurativa; autoimmune hepatitis; uveitis; pulmonary fibrosis; non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH).

In one embodiment of the present invention, said treatment is a treatment for: inflammatory arthritis (including, for example, rheumatoid arthritis; psoriatic arthritis; systemic lupus erythematosus; juvenile idiopathic arthritis); psoriasis; lupus nephritis; systemic sclerosis; inflammatory bowel disease; ulcerative colitis; Crohn's disease or multiple sclerosis.

In one embodiment of the present invention, said treatment is a treatment for inflammatory arthritis.

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In one embodiment of the present invention, said treatment is a treatment for rheumatoid arthritis.

In one embodiment of the present invention, said treatment is a treatment for psoriatic arthritis.

In one embodiment of the present invention, said treatment is a treatment for systemic lupus erythematosus.

In one embodiment of the present invention, said treatment is a treatment for juvenile idiopathic arthritis.

In one embodiment of the present invention, said treatment is a treatment for psoriasis.

In one embodiment of the present invention, said treatment is a treatment for lupus nephritis.

In one embodiment of the present invention, said treatment is a treatment for systemic sclerosis.

In one embodiment of the present invention, said treatment is a treatment for inflammatory bowel disease.

In one embodiment of the present invention, said treatment is a treatment for ulcerative colitis.

In one embodiment of the present invention, said treatment is a treatment for Crohn's disease.

In one embodiment of the present invention, said treatment is a treatment for multiple sclerosis.

Conditions to be treated: cancer.

In one embodiment of the present invention, said treatment is a treatment for cancer.

In one embodiment of the present invention, said treatment is a treatment for multiple myeloma; lymphomas; leukemia; carcinomas or sarcomas.

Multiple myeloma.

In one embodiment of the present invention, said treatment is a treatment for multiple myeloma.

Lymphoma.

In one embodiment of the present invention, said treatment is a treatment for lymphoma.

In one embodiment of the present invention, said treatment is a treatment for Hodgkin's lymphoma; non-Hodgkin's lymphoma; lymphocytic lymphoma; granulocytic lymphoma; monocytic lymphoma; diffuse large B-cell lymphoma (DLBCL); mantle cell lymphoma (MCL); follicular lymphoma (FL); mucosal lymphoid tissue (MALT) lymphoma; marginal zone lymphoma; T-cell lymphoma; marginal zone lymphoma or Burkitt lymphoma.

In one embodiment of the present invention, said treatment is a treatment for lymphocytic lymphoma; granulocytic lymphoma; monocytic lymphoma or diffuse large B-cell lymphoma (DLBCL).

In one embodiment of the present invention, the treatment is a treatment for diffuse large B cell lymphoma (DLBCL).

Leukemia.

In one embodiment of the present invention, said treatment is a treatment for leukemia.

In one embodiment of the present invention, said treatment is a treatment for: chronic lymphocytic leukemia (CLL); acute myeloid leukemia (AML); acute lymphocytic leukemia (ALL); lymphoblastic T-cell leukemia; chronic myelogenous leukemia (CML); hairy cell leukemia; acute lymphoblastic T-cell leukemia; acute eosinophilic leukemia; immunoblastic large cell leukemia; megakaryoblastic leukemia; acute megakaryocytic leukemia; promyelocytic leukemia; erythroleukemia or plasmacytoma.

In one embodiment of the said invention, said treatment is a treatment for: chronic lymphocytic leukemia (CLL); acute myeloid leukemia (AML); acute lymphocytic leukemia (ALL); lymphoblastic T-cell leukemia; chronic myelogenous leukemia (CML) or acute eosinophilic leukemia.

In one embodiment of the present invention, said treatment is a treatment for chronic lymphocytic leukemia (CLL).

In one embodiment of the present invention, said treatment is a treatment for acute myeloid leukemia (AML).

In one embodiment of the present invention, said treatment is a treatment for acute lymphocytic leukemia (ALL).

In one embodiment of the present invention, the treatment is a treatment for lymphoblastic T-cell leukemia.

In one embodiment of the present invention, said treatment is a treatment for chronic myelogenous leukemia (CML).

Carcinoma.

In one embodiment of the present invention, said treatment is a treatment for carcinoma.

In one embodiment of the present invention, said treatment is a treatment for: colon cancer; breast cancer; ovarian cancer; lung cancer (including, for example, small cell lung carcinoma and non-small cell lung carcinoma); prostate cancer; cancer of the oral cavity or pharynx (including, for example, cancer of the lip, tongue, mouth, larynx, pharynx, salivary gland, oral mucosa); esophageal cancer; stomach cancer; small intestine cancer; colon cancer; rectal cancer; liver duct cancer; biliary tract cancer; pancreatic cancer; bone cancer; connective tissue cancer; skin cancer; cervical cancer; uterine cancer; cancer of the uterus; endometrial cancer; vulvar cancer; vaginal cancer; testicular cancer; bladder cancer; kidney cancer; ureteral cancer; urethral cancer; urachus cancer; eye cancer; gliomas; spinal cord cancer; cancer of the central nervous system; cancer of the peripheral nervous system; meningeal cancer; thyroid cancer; adrenocarcinoma; astrocytomas; acoustic neuroma; anaplastic astrocytoma; basal cell carcinoma; blastogliomas; choriocarcinomas; chordomas; craniopharyngiomas; cutaneous melanoma; cystadenocarcinoma; embryonal carcinoma; ependymomas; epithelial carcinoma; stomach cancer; genitourinary tract cancer; glioblastoma multiforme; head and neck cancer; hemangioblastoma; hepatocellular carcinoma, renal cell carcinoma (RCC); hepatomas; large cell carcinoma; medullary thyroid carcinoma; medulloblastomas; meningiomas, mesothelioma; myelomas; neuroblastoma; oligodendrogliomas; epithelial ovarian cancer; papillary carcinoma; papillary adenocarcinoma; paragangliomas; parathyroid tumors; pheochromocytomas; pinealomas; plasmacytomas; retinoblastoma; carcinoma of the sebaceous glands; seminomas; melanoma; squamous cell carcinoma; sweat gland carcinomas; synoviomas; thyroid cancer; uveal melanoma or Wilms tumor.

In one embodiment of the present invention, said treatment is a treatment for: colon cancer; breast cancer; ovarian cancer; lung cancer (including, for example, small cell lung carcinoma and non-small cell lung carcinoma); prostate cancer; stomach cancer; pancreatic cancer; bone cancer; skin cancer; cervical cancer; uterine cancer; endometrial cancer; testicular cancer; bladder cancer; kidney cancer; eye cancer; liver cancer; gliomas; thyroid cancer; adrenocarcinoma; astrocytomas; acoustic neuroma; anaplastic astrocytoma; cutaneous melanoma; stomach cancer; glioblastoma multiforme; head and neck cancer; hepatocellular carcinoma; renal cell carcinoma (RCC); melanoma or squamous cell carcinoma.

In one embodiment of the present invention, said treatment is a treatment for: colon cancer; breast cancer; ovarian cancer; lung cancer (including, for example, small cell lung carcinoma and non-small cell lung carcinoma); prostate cancer; pancreatic cancer; bone cancer; liver cancer; glioblastoma multiforme; head and neck cancer or melanoma.

In one embodiment of the present invention, said treatment is a treatment for melanoma.

In one embodiment of the present invention, said treatment is a treatment for glioblastoma multiforme.

In one embodiment of the present invention, said treatment is a treatment for breast cancer.

In one embodiment of the present invention, said treatment is a treatment for prostate cancer.

In one embodiment of the present invention, said treatment is a treatment for bone cancer.

In one embodiment of the present invention, said treatment is a treatment for pancreatic cancer.

In one embodiment of the present invention, said treatment is a treatment for head and neck cancer.

In one embodiment of the present invention, said treatment is a treatment for lung cancer (including, for example, small cell lung carcinoma and non-small cell lung carcinoma).

In one embodiment of the present invention, said treatment is a treatment for ovarian cancer.

In one embodiment of the present invention, said treatment is a treatment for liver cancer.

Sarcoma.

In one embodiment of the present invention, said treatment is a treatment for sarcoma.

In one embodiment of the present invention, said treatment is a treatment for Askin's tumor; pampiniform sarcoma; chondrosarcomas; endothelial sarcoma; Ewing's sarcoma; malignant hemagioendothelioma; malignant schwannoma; osteosarcomas; gastrointestinal stromal tumor (GIST); myxosarcoma; alveolar soft tissue sarcoma; angiosarcomas; leaf-shaped cystosarcoma; dermatofibrosarcoma; desmoid tumor; desmoplastic small round cell tumor; extraskeletal chondrosarcoma; osteosarcomas; fibrosarcomas; hemagiopericytomas; hemangiosarcomas; Kaposi's sarcoma; leiomyosarcoma; liposarcoma; lymphangiosarcomas; lymphangioendothelial sarcoma; lymphosarcoma; malignant tumor of the peripheral nerve sheath; neurofibrosarcomas; plexiform fibrohistiocytic tumor; rhabdomyosarcoma or synovial sarcoma.

Treatment-resistant cancer.

In one embodiment of the present invention, said treatment is the treatment of: treatment-resistant cancer (including, for example, chemotherapy-resistant cancer and radiation-resistant cancer); metastatic cancer; metastases or recurrent cancer.

In one embodiment of the present invention, said treatment is the treatment of: chemotherapy-resistant cancer (including, for example, chemotherapy-resistant multiple myeloma, lymphoma, leukemia, carcinoma and sarcoma).

In one embodiment of the present invention, said treatment is the treatment of: radiation-resistant cancer (including, for example, radiation-resistant multiple myeloma, lymphoma, leukemia, carcinoma and sarcoma).

In one embodiment of the present invention, said treatment is a treatment for metastatic cancer.

In one embodiment of the present invention, said treatment is a treatment for metastases.

In one embodiment of the present invention, said treatment is a treatment for recurrent cancer.

In one embodiment of the present invention, the treatment is used to: prevent, reduce or overcome resistance to radiation therapy or chemotherapy (eg, due to changes in cellular metabolism); preventing or reducing tumor invasion; preventing or reducing tumor metastasis; improving the action of antitumor agents and/or enhancing the action of immunomodulators.

In one embodiment of the present invention, the treatment is used to: prevent, reduce or overcome resistance to radiation therapy.

In one embodiment of the present invention, the treatment is used to: prevent, reduce or overcome resistance to chemotherapy.

In one embodiment of the present invention, the treatment is used to: prevent or reduce tumor invasion or tumor metastasis; improving the action of antitumor agents and/or enhancing the action of immunomodulators.

In one embodiment of the present invention, said treatment is used to: improve the effect of antineoplastic agents and/or enhance the effect of immunomodulators.

In one embodiment of the present invention, the treatment is used to: improve the effect of immunomodulators.

Conditions to be treated: Osteoclast-mediated diseases.

In one embodiment of the present invention, said treatment is a treatment for: an osteoclast-mediated disorder.

In one embodiment of the present invention, said treatment is a treatment for: rheumatoid arthritis; osteoporosis; Paget's disease; osteopetrosis; osteoarthritis; ectopic bone formation; bone loss associated with endometriosis; bone neoplasia (including, for example, a primary tumor or metastases, including, for example, bone cancer; osteosarcoma or osteoma); cancer-related bone disease (including, for example, metastatic bone disease associated with, for example, breast cancer, lung cancer, prostate cancer, or multiple myeloma; changes in bone mineralization and density associated with cancer, including, for example, hypercalcemia associated with cancer); bone metastases (including, for example, osteolytic bone metastases); hypercalcemia (including, for example, hypercalcemia associated with cancer; hypercalcemia caused by conditions associated with increased bone resorption (including, for example, hypercalcemia caused by vitamin D intoxication, primary or tertiary hyperparathyroidism, immobilization, or sarcoidosis); or aseptic loosening of prosthetic implants ( e.g. artificial joints such as knees, hips, etc.).

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In one embodiment of the present invention, said treatment is a treatment for: rheumatoid arthritis; osteoporosis; bone neoplasia (including, for example, a primary tumor or metastases, including, for example, bone cancer; osteosarcoma or osteoma); cancer-related bone disease (including, for example, metastatic bone disease associated with, for example, breast cancer, lung cancer, prostate cancer, or multiple myeloma; changes in bone mineralization and density associated with cancer, including, for example, hypercalcemia associated with cancer); or bone metastases (including, for example, osteolytic bone metastases).

In one embodiment of the present invention, said treatment is a treatment for rheumatoid arthritis.

In one embodiment of the present invention, said treatment is a treatment for osteoporosis.

In one embodiment of the present invention, said treatment is a treatment for: bone neoplasia (including, for example, a primary tumor or metastases, and including, for example, bone cancer; osteosarcoma or osteoma).

In one embodiment of the present invention, said treatment is a treatment for: bone cancer; osteosarcoma or osteoma.

In one embodiment of the present invention, said treatment is treatment of: cancer-related bone disease (including, for example, metastatic bone disease associated with, for example, breast cancer, lung cancer, prostate cancer or multiple myeloma; changes in bone mineralization and density cancer-related, including for example cancer-related hypercalcemia).

In one embodiment of the present invention, said treatment is a treatment for bone metastases.

Treatment.

The term treatment, as used herein in the context of treating a condition, generally refers to treatments and therapies in humans or animals (e.g., in veterinary applications) that achieve some desired therapeutic effect, such as inhibiting the progression of a condition, and includes reducing the rate of progression, stopping the rate of progression, relieving the symptoms of the condition, improving the condition and curing the condition. Treatment as a preventive measure (eg, prophylaxis) is also included. For example, the term treatment also covers use in patients who have not yet developed the condition but who are at risk of developing it.

For example, treating inflammation includes preventing inflammation, reducing the frequency of inflammation, reducing the severity of inflammation, alleviating symptoms of inflammation, etc.

The term therapeutically effective amount as used herein refers to that amount of a compound, material, composition, or dosage form containing a compound that is effective to produce some desired therapeutic effect and has a reasonable risk/benefit ratio when administered in accordance with the desired treatment regimen.

Combination therapy.

The term treatment includes combination treatments and therapies in which two or more treatments or therapies are combined, for example, sequentially or simultaneously. For example, the compounds described herein can also be used in combination therapy, for example, in combination with other agents, such as anti-inflammatory agents, etc. Examples of treatment and therapy include chemotherapy (administration of active agents including, for example, drugs, antibodies (eg, as in immunotherapy), prodrugs (eg, as in photodynamic therapy, GDEPT, ADEPT, etc.); surgery; radiation therapy , photodynamic therapy, gene therapy and controlled diets.

One aspect of the present invention relates to a compound described herein in combination with one or more additional therapeutic agents.

The specific combination will be chosen at the discretion of the physician, who will select the dosage using his general knowledge and dosing regimens known to the qualified practitioner.

The agents (ie, a compound described herein plus one or more other agents) may be administered simultaneously or sequentially, and may be administered in individually varying dosage schedules and by different routes. For example, when administered sequentially, the agents may be administered at close intervals (for example, over a period of 5-10 minutes) or at longer intervals (for example, at intervals of 1, 2, 3, 4 or more hours, or at even longer intervals, if necessary), with the precise dosage regimen being commensurate with the properties of the therapeutic agent(s).

The agents (i.e., the compound described herein plus one or more other agents) may be part of a single dosage form together, or, alternatively, the individual agents may be formulated as separate formulations and presented together in kit form, optionally with instructions for use.

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Other uses.

The CHMSA compounds described herein may also be used as part of an in vitro assay, for example, to determine whether treatment with the subject compound is likely to benefit a candidate patient.

The CHMSA compounds described herein can also be used as a standard, for example, in an assay to identify other compounds, other anti-inflammatory agents, etc.

Sets.

One aspect of the present invention relates to a kit containing:

(a) a CHMSA compound as described herein, or a composition containing a CHMSA compound as described herein, for example, preferably provided in a suitable container and/or with suitable packaging; and (b) instructions for use, for example, written instructions on how to administer the specified compound or composition.

In one embodiment of the present invention, the kit further comprises one or more (eg, 1, 2, 3, 4) additional therapeutic agents described herein.

The written instructions may also include a list of indications for which the active ingredient is an appropriate treatment.

Routes of administration.

The CHMSA compound or a pharmaceutical composition containing the CHMSA compound can be administered to a subject by any convenient route of administration, systemically/peripherally or locally (ie, at the site of desired action).

Routes of administration include oral (eg, oral administration); buccal; sublingual; transdermal (including, for example, patch, patch, etc.); transmucosal (including, for example, patch, patch, etc.); intranasal (eg, via nasal spray, drops, nebulizer, or dry powder delivery device); ophthalmic (for example, using eye drops); inhalation (for example, by inhalation or insufflation therapy using, for example, an aerosol, for example, through the mouth or nose); rectal (for example, using a suppository or enema); vaginal (for example, using a pessary); parenteral, for example, by injection, including subcutaneous, intradermal, intramuscular, intravenous, intraarterial, intracardiac, intrathecal, intraspinal, intracapsular, subcapsular, intraocular, intraperitoneal, intratracheal, intradermal, intraarticular, subarachnoid and intrasternal; by implanting a depot or reservoir, for example, subcutaneously or intramuscularly.

In one preferred embodiment of the present invention, the route of administration is an oral route of administration (eg, oral administration). In one preferred embodiment of the present invention, the route of administration is a parenteral route of administration (eg, by injection).

Subject/patient.

The subject/patient may be a chordate, vertebrate, mammal, placental mammal, marsupial (e.g., kangaroo, wombat), rodent (e.g., guinea pig, hamster, rat, mouse), murine (e.g., mouse), lagomorph (e.g., rabbit), birds (e.g. poultry), canids (e.g. dog), felids (e.g. cat), equidae (e.g. horse), porciniformes (e.g. pig), ovine (e.g. sheep), cattle (e.g. , cow), primate, ape (e.g., marmoset or ape), member of the marmoset (e.g., marmoset, baboon), ape (e.g., gorilla, chimpanzee, orangutan, gibbon), or human. Additionally, the subject/patient may be in any of its developmental forms, such as a fetus.

In one preferred embodiment of the present invention, said subject/patient is a human.

Compositions.

Although the CHMSA compound can be administered separately, it is preferably provided in the form of a pharmaceutical composition (e.g., composition, preparation, drug) containing at least one CHMSA compound described herein together with one or more other pharmaceutically acceptable ingredients well known those skilled in the art, including pharmaceutically acceptable carriers, diluents, excipients, adjuvants, excipients, buffers, preservatives, antioxidants, lubricants, stabilizers, solubilizers, surfactants (eg, wetting agents), masking agents, colorants, flavorings, and sweeteners. Said composition may further contain other active agents, for example other therapeutic or prophylactic agents.

Thus, the present invention further relates to pharmaceutical compositions, as defined herein, and methods for preparing a pharmaceutical composition,

- 37 043371 comprising mixing at least one CHMSA compound described herein with one or more other pharmaceutically acceptable ingredients well known to those skilled in the art, for example, carriers, diluents, excipients, etc. If the formulation is presented in the form of discrete units (eg, tablets, etc.), each unit contains a predetermined amount (dosage) of the compound.

As used herein, the term pharmaceutically acceptable refers to those compounds, materials, compositions, dosage forms, etc., which, within the bounds of sound medical judgment, are suitable for use in contact with the tissues of a subject (e.g., human) without undue toxicity, irritation, allergic reaction or other problems and complications, and which have a reasonable risk/benefit ratio. Each carrier, diluent, excipient, etc. must also be acceptable, i.e. compatible with other ingredients of the composition.

Suitable carriers, diluents, auxiliaries, etc. can be found in standard pharmaceutical texts, for example, Remington's Pharmaceutical Sciences, ^18th edition, Mack Publishing Company, Easton, Pa., 1990; and Handbook of Pharmaceutical Excipients, ^5th edition, 2005.

The compositions can be prepared by any methods well known in the field of pharmacy. Such methods include the step of combining the compound with a carrier that consists of one or more accessory ingredients. Typically, formulations are prepared by uniformly and thoroughly combining the compound with carriers (eg, liquid carriers, finely divided solid carriers, etc.), then molding the product as necessary.

The formulation may be formulated to provide fast or slow release; immediate, delayed, timed or sustained release; or combinations thereof.

The compositions may also suitably be presented in the form of liquids, solutions (e.g., aqueous, non-aqueous), suspensions (e.g., aqueous, non-aqueous), emulsions (e.g., oil-in-water, water-in-oil), elixirs, syrups, elixirs, mouthwashes, drops, tablets (including, for example, coated tablets), granules, powders, lozenges, lozenges, capsules (including, for example, hard and soft gelatin capsules), wafers, pills, ampoules, boluses, suppositories, pessaries, tinctures, gels, pastes, ointments, creams, lotions, oils, foams, sprays, fine aerosols or aerosols.

The compositions may suitably be presented in the form of a patch, adhesive, bandage, dressing, or the like, impregnated with one or more compounds and optionally one or more other pharmaceutically acceptable ingredients, including, for example, penetration, permeability and absorption enhancers. The compositions may also suitably be presented in the form of a depot or reservoir.

The compound may be dissolved, suspended, or mixed with one or more other pharmaceutically acceptable ingredients. The compound may be presented in the form of liposomes or other microparticles that are designed to target the compound, for example, to blood components or one or more organs.

Formulations suitable for oral administration (eg, oral administration) include liquids, solutions (eg, aqueous, non-aqueous), suspensions (eg, aqueous, non-aqueous), emulsions (eg, oil in water, water in oil), elixirs, syrups , medicinal porridges, tablets, granules, powders, capsules, wafers, pills, ampoules, boluses.

Formulations suitable for buccal administration include mouthwashes, lozenges, lozenges, as well as patches, adhesives, depots and reservoirs. The candies usually contain the compound in a flavored base, usually sucrose and gum arabic or tragacanth. Lozenges usually contain the compound in an inert matrix such as gelatin and glycerin or sucrose and gum arabic. Mouthwashes typically contain the compound in a suitable liquid carrier.

Formulations suitable for sublingual administration include tablets, lozenges, lozenges, capsules and pills.

Formulations suitable for oral transmucosal administration include liquids, solutions (e.g., aqueous, non-aqueous), suspensions (e.g., aqueous, non-aqueous), emulsions (e.g., oil-in-water, water-in-oil), mouthwashes, lozenges, lozenges, as well as patches, adhesives, depots and reservoirs.

Formulations suitable for non-oral transmucosal administration include liquids, solutions (eg, aqueous, non-aqueous), suspensions (eg, aqueous, non-aqueous), emulsions (eg, oil-in-water, water-in-oil), suppositories, pessaries, gels, pastes , ointments, creams, lotions, oils, as well as patches, adhesives, depots and reservoirs.

Formulations suitable for transdermal administration include gels, pastes, ointments, creams, lotions and oils, as well as patches, adhesives, dressings, dressings, depots and reservoirs.

Tablets can be prepared by conventional methods, for example by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared by compressing in a suitable compounding device into a bulk form such as powder or

- 38 043371 granules, optionally mixed with one or more binders (eg povidone, gelatin, gum arabic, sorbitol, tragacanth, hydroxypropyl methylcellulose); fillers or diluents (eg lactose, microcrystalline cellulose, calcium hydrogen phosphate); lubricants (for example, magnesium stearate, talc, silicon dioxide); disintegrants (eg sodium starch glycolate, cross-linked povidone, cross-linked sodium carboxymethylcellulose); surfactants or dispersing or wetting agents (eg sodium lauryl sulfate); preservatives (for example, para-hydroxybenzoic acid methyl ester, para-hydroxybenzoic acid propyl ester, sorbic acid); flavorings, flavor enhancers and sweeteners. Molded tablets can be prepared by molding in a suitable device a mixture of said powdered compound moistened with an inert liquid diluent. Tablets may optionally be coated or scored and may be formulated to provide a slow or controlled release of the compound contained therein using, for example, hydroxypropyl methylcellulose in varying proportions to provide the desired release profile. Tablets may optionally contain a coating, for example, a release modifying coating, such as an enteric coating to provide release in parts of the intestine other than the stomach.

Ointments are usually prepared from a compound and a paraffin or water-miscible ointment base.

Creams are usually prepared from a compound and an oil-in-water cream base. If desired, the aqueous phase of the cream base may include, for example, at least 30% w/w. polyhydric alcohol, i.e. an alcohol containing two or more hydroxyl groups, such as propylene glycol, butane-1,3diol, mannitol, sorbitol, glycerol and polyethylene glycol and mixtures thereof. Topical formulations may optionally include a compound that enhances penetration or absorption of the compound through the skin or other affected areas. Examples of such skin penetration enhancers include dimethyl sulfoxide and related analogs.

Emulsions are typically prepared from a compound and an oil phase, which may optionally contain only an emulsifier (otherwise known as an emulsifier), or it may contain a mixture of at least one emulsifier with a fat or oil, or both fat and oil. Preferably, the hydrophilic emulsifier is included together with a lipophilic emulsifier which acts as a stabilizer. It is preferable to also include both oil and fat. Together, the emulsifier(s) with or without stabilizer(s) form the so-called emulsifying wax, and the wax together with the oil and/or fat form the so-called emulsifying ointment base, which forms the oil dispersed phase of the composition in the form of a cream.

Suitable emulgents and emulsion stabilizers include Tween 60, Span 80, cetostearyl alcohol, myristyl alcohol, glycerol monostearate and sodium lauryl sulfate. The selection of suitable oils or fats for the composition is based on achieving the desired cosmetic properties, since the solubility of the compound in most oils likely to be used in pharmaceutical emulsion compositions may be very low. The cream should preferably be a non-greasy, non-staining and washable product with a suitable consistency to avoid leakage from tubes or other containers. Straight or branched mono or dibasic alkyl esters may be used, such as diisoadipate, isocetyl stearate, propylene glycol diester of coconut oil fatty acids, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate or a mixture of branched chain fatty acid esters known as Crodamol CAP, with the last three being the preferred esters. They can be used individually or in combination depending on the required properties. Alternatively, high melting point lipids such as white soft paraffin and/or liquid paraffin or other mineral oils can be used.

Formulations suitable for intranasal administration, where the carrier is a liquid, include, for example, nasal spray, nasal drops or formulations for aerosol administration by nebulizer, including aqueous or oily solutions of the compound.

Formulations suitable for nasal administration where the carrier is a solid include those in the form of a coarse powder with a particle size, for example, in the range of from about 20 to about 500 microns, which is administered in the same manner as snuff is consumed, i.e. . The powder in the container is held close to the nose and quickly inhaled through the nasal passage.

Formulations suitable for inhalation administration (eg, via inhalation or insufflation therapy) include those that are delivered as an aerosol spray from a sealed container using a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gases.

Formulations suitable for ophthalmic administration include eye drops in which the compound is dissolved or suspended in a suitable vehicle, especially an aqueous vehicle for the compound.

Formulations suitable for rectal administration may be presented in the form of suppositories with a suitable base containing, for example, natural or hardened oils, waxes, fats, semi-liquid or liquid polyols, for example cocoa butter or salicylate; or in the form of a solution or suspension for enema treatment.

Formulations suitable for vaginal administration may be in the form of vaginal suppositories, tampons, creams, gels, pastes, foams or sprays containing, in addition to said compound, carriers known to be suitable in the art.

Formulations suitable for parenteral administration (eg, by injection) include aqueous or non-aqueous, isotonic, pyrogen-free, sterile liquids (eg, solutions, suspensions) in which the compound is dissolved, suspended, or otherwise present (eg, liposomes or other microparticles). Such liquids may further contain other pharmaceutically acceptable ingredients, such as antioxidants, buffers, preservatives, stabilizers, bacteriostatic agents, suspending agents, thickening agents and solutes that render the formulation isotonic with the blood (or other appropriate body fluid) of the intended recipient. Examples of excipients include, for example, water, alcohols, polyols, glycerin, vegetable oils and the like. Examples of suitable isotonic vehicles for use in such formulations include sodium chloride injection, Ringer's solution, or lactated Ringer's solution. Typically, the concentration of the compound in the liquid is from about 1 ng/ml to about 10 μg/ml, for example, from about 10 ng/ml to about 1 μg/ml. The formulations may be presented in single-dose or multi-dose sealed containers, such as sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) state requiring only the addition of a sterile liquid carrier, such as water for injection, immediately prior to use. Injectable solutions and suspensions prepared for immediate use can be prepared from sterile powders, granules and tablets.

Dosage.

One skilled in the art will appreciate that suitable dosages of CHMSA compounds and compositions containing CHMSA compounds may vary from patient to patient. Determining the optimal dosage usually involves balancing the level of therapeutic benefit with any risks or harmful side effects. The dosage level chosen will depend on a variety of factors, including the potency of the particular CHMSA compound, route of administration, time of administration, rate of elimination of the CHMSA compound, duration of treatment, other drugs, compounds and/or materials used in combination, severity of the condition, and the type gender, age, weight, condition, general health and previous medical history of the patient. The amount of CHMSA compound and route of administration remains ultimately at the discretion of the physician, veterinarian or treating personnel, although generally the dosage will be selected to obtain local concentrations at the site of action that achieve the desired effect without causing significant adverse effects or harmful side effects.

Administration may be administered in a single dose, continuously or intermittently (eg, in divided doses at appropriate intervals) throughout the course of treatment. Methods for determining the most effective agents and dosage to administer are well known to those skilled in the art and will vary depending on the formulation used for therapy, the purpose of therapy, the target cell(s) being treated, and the subject being treated. Single or multiple administrations may be performed at the dosage level and schedule selected by the attending physician, veterinarian, or treating personnel.

In general, a suitable dose of a CHMSA compound is in the range of about 10 μg to about 20 mg (more typically from about 100 μg to about 10 mg) per kilogram of subject body weight per day. If the compound is a salt, ester, amide, prodrug or the like, the amount administered is calculated based on the parent compound and therefore the actual mass to be used is increased proportionally.

Chemical synthesis.

Acronyms and abbreviations.

aq.: aquatic;

B ₂ pin ₂ : bis(pinacolato)diborane;

DCM: dichloromethane;

DEA: diethylamine;

DMF: dimethylformamide;

DMSO: dimethyl sulfoxide;

Eq: equivalent;

ESI: electrospray ionization;

EtOAc: ethyl acetate;

FID: flame ionization detector;

GC: gas chromatography;

HPLC: high performance liquid chromatography;

KOAc: potassium acetate;

- 40 043371

LAH: lithium aluminum hydride;

LCMS: liquid chromatography-mass spectrometry;

LiAlH4: lithium aluminum hydride;

m-CPBA: meta-chloroperoxybenzoic acid;

m/z: mass to charge ratio;

MeOH: methanol;

NaH: sodium hydride;

NMR: nuclear magnetic resonance (spectroscopy);

p-TSA: p-toluenesulfonic acid;

PdCl2(PPh ₃ )2: bis(triphenylphosphine)palladium(P) dichloride;

Pd(dppf)Cl2: (1,1'-bis(diphenylphosphino)ferrocene)dichloropalladium(II);

Pd(Ph ₃ ) ₄ : tetrakis(triphenylphosphine)palladium(0);

PPh3: triphenylphosphine;

RT: room temperature;

SLC: supercritical liquid chromatography;

TFA: trifluoroacetic acid;

TFAA: trifluoroacetic anhydride;

Tf2O: trifluoromethanesulfonic acid anhydride;

THF: tetrahydrofuran;

TLC: thin layer chromatography;

TPP: triphenylphosphine.

Analytical HPLC.

Analytical HPLC characterization of the final compounds was carried out as follows:

Column: X-select CSH C18, 4.6 mm x 150 mm, internal diameter 3.5 µm.

Injection volume: 5 µl.

Flow rate: 1 ml/min.

Solvents:

A: 0.1% formic acid in water: acetonitrile (95:5).

B: acetonitrile.

Gradient (V% increases linearly from 1 to 8 min):

- 41 043371

Intermediate, conn. 3

Intermediate, conn. 4

Intermediate, conn. 5 bis(pinacolato)diboron, Pd(PPh h) ₂ C12, COAc, dioxane, 100°C

Intermediate, conn. 6

Intermediate 1. Ethyl 1,4-dioxaspiro[4.5]decane-8-carboxylate OSTO

CTOet

To a stirred solution of ethyl 4-oxocyclohexane-1-carboxylate (40.00 g, 235.00 mmol) in toluene (400 ml), add ethylene glycol (16.05 g, 258.50 mmol) and p-toluenesulfonic acid (monohydrate ) (0.45 g, 2.35 mmol). The reaction mixture was heated in a Dean-Stark apparatus at a temperature of 110°C for 18 hours while continuously removing water. The reaction progress was monitored by TLC [(silica gel TLC plate), 30% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was cooled to room temperature. The reaction was quenched with saturated NaHCO ₃ solution (100 ml) and extracted with EtOAc (3x200 ml). The combined organic layer was washed with brine (1 x 200 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the title intermediate 1 (48.32 g, crude) as a yellow oil, which was used in the next step without further purification .

Intermediate 2. (1,4-dioxaspiro[4.5]decan-8-yl)methanol

To a stirred solution of intermediate 1 ethyl-1,4-dioxaspiro[4.5]decane-8-carboxylate (48.32 g, 225.52 mmol) in THF (300 mL), a 1 M solution of LAH (225.52 mL, 225.52 mmol) at 0°C. The reaction mixture was allowed to warm to room temperature and stirred for 12 hours. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 30% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was quenched with a saturated aqueous solution of Na ₂ SO ₄ (240 ml). The precipitated solid was filtered and washed with EtOAc (250 ml). The organic layer was separated from the filtrate and the aqueous layer was extracted with EtOAc (2x100 ml). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the title intermediate 2 (33.45 g, crude) as a colorless oil, which was used in the next step without further purification.

Analytical data: ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm: 3.95 (s, 4H), 3.49 (d, J=6.4 Hz, 2H), 1.80 1.75 (m, 4H), 1.59 - 1.49 (m, 3H), 1.32 - 1.23 (m, 2H).

Intermediate 3. 8-(bromomethyl)-1,4-dioxaspiro[4.5]decane

To a stirred solution of intermediate 2 (1,4-dioxaspiro[4.5]decane-8

- 42 043371 yl)methanol (33.45 g, 194.23 mmol) in DCM (400 ml) added triphenylphosphine (50.95 g, 194.23 mmol), carbon tetrabromide (67.63 g, 203.94 mmol) at a temperature of 0°C. The reaction mixture was allowed to warm to room temperature and stirred for 12 hours. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 30% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was quenched with water (200 ml) and extracted with DCM (3x200 ml). The combined organic layer was washed with brine (2x150 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by column chromatography (silica gel 230-400 mesh, gradient 1-10% EtOAc in n-hexane) to give the title intermediate 3 (35.00 g, 77%) as a colorless oil.

Analytical data: LCMS (ESI) m/z=237.10 [M+1]+ ( ⁸¹ Br).

Intermediate 4. 8-(((4-bromophenyl)thio)methyl)-1,4-dioxaspiro[4.5]decane

To a stirred solution of 4-bromobenzenethiol (11.77 g, 62.26 mmol) in THF:MeOH (2:1, 150 ml) was added CsCO ₃ (33.81 g, 103.78 mmol) at 0°C and the reaction the mixture was stirred for 20 min at 0°C. Intermediate 3 8-(bromomethyl)1,4-dioxaspiro[4.5]decane (12.20 g, 51.89 mmol) was added to the mixture and the reaction mixture was stirred at 60°C for 12 hours. The reaction progress was monitored by TLC [(silica gel TLC plate), 20% EtOAc in n-hexane]. After intermediate 3 8-(bromomethyl)-1,4dioxaspiro[4.5]decane was completely consumed, the reaction mixture was concentrated under reduced pressure, the resulting residue was dissolved in water (150 ml) and extracted with EtOAc (3x150 ml). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude residue. The residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 1-10% EtOAc in n-hexane) to give the title intermediate 4 (12.00 g, 67%) as a colorless oil.

Analytical data: LCMS (ESI) m/z=343.10 [M+H]+.

Intermediate 5. 8-(((4-bromophenyl)sulfonyl)methyl)-1,4-dioxaspiro[4.5]decane

3-chloroperoxybenzoic acid ( ~ 60% in water) (30.16 g, 104.87 mmol) at 0°C in portions over 30 min. The reaction mixture was allowed to warm to room temperature and stirred for 16 hours. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 20% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was cooled to 0°C, saturated aqueous NaHCO ₃ solution (100 ml) was slowly added and the layers were separated. The separated organic layer was cooled to 0°C, and a saturated aqueous solution of Na ₂ S ₂ O ₃ (100 ml) was slowly added. The organic layer was separated, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude residue. The residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 1-20% EtOAc in n-hexane) to give the title intermediate 5 (11.00 g, 84%) as a white solid.

Analytical data: LCMS (ESI) m/z=377.10 [M+1]+ ( ⁸¹ Br).

Intermediate 6. 2-(4-(((1,4-dioxaspiro[4.5]decan-8-yl)methyl)sulfonyl)phenyl)4,4,5,5-tetramethyl-1,3,2-dioxaborolane

- 43 043371

To a stirred solution of intermediate 5 8-(((4-bromophenyl)sulfonyl)methyl)1,4-dioxaspiro[4.5]decane (11.00 g, 29.31 mmol) in 1,4-dioxane (120 ml), Potassium acetate (8.63 g, 87.93 mmol) and bis(pinacolato)diboron (9.68 g, 38.10 mmol) were added at room temperature, then the reaction mixture was degassed using argon for 15 min. Bis(triphenylphosphine)palladium(II) dichloride (0.31 g, 0.44 mmol) was added to the mixture, the reaction mixture was degassed for 10 min and stirred at 100°C for 4 h in a sealed tube. The reaction progress was monitored by TLC [(silica gel TLC plate), 30% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, the resulting residue was dissolved in EtOAc (250 ml) and washed with water (100 ml). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude residue. The residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 1-20% EtOAc in n-hexane) to give the title intermediate 6 (10.00 g, 81%) as a white solid.

Analytical data: LCMS (ESI) m/z=423.30 [M+H]+ and 341.10 [M+H] ⁺ (corresponding boronic acid).

Synthesis scheme 2

A. Pd(PPh ₃ )4, Na ₂ CO ₃ , H] dioxane:H20 (3:1), 100 °C T I ψ F Intermediate, conn. 5 Intermediate, conn. 7 о=з=о i) ZM MeMgBr, THF, FROM -20°С TO CT A. ...----------------------- ί J i) separation of diastereomers kJ F Intermediate, conn. 8 rn ,,σ 0=3=0 O

Intermediate dioxaspiro[4.5]decane Mixture of intermediate r r F F Synthesized Synthesized connection 1 connection 2 7. 8-(((2',4'-difluoro-(1,1'-biphenyl)-4-yl)sulfonyl)methyl)-1,4- o=s=o ό V F compound 5 8-(((4-bromophenyl)sulfonyl)methyl)-1,4-

- 44 043371 dioxaspiro[4.5]decane (3.0 g, 8.0 mmol), 2-(2,4-difluorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2, 3 g, 9.6 mmol) and sodium carbonate (2.5 g, 24 mmol) in dioxane-water (3:1, 30 ml) were degassed with argon for 30 min. Tetrakis(triphenylphosphine)palladium(0) (0.9 g, 0.78 mmol) was added to the mixture, the reaction mixture was degassed for 10 min and stirred at 100°C for 12 h in a sealed tube. The reaction progress was monitored by TLC [(silica gel TLC plate), 60% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the resulting residue was diluted with water and extracted with EtOAc. The organic layer was washed with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography (silica gel 100-200 mesh) to give the title intermediate 7 (2.8 g, 86%) as a white solid.

Analytical data: LCMS (ESI) m/z=409.20 [M+H] ⁺ .

Intermediate 8. 4-(((2',4'-difluoro-(1,1'-biphenyl)-4-yl)sulfonyl)methyl)cyclohexan1-one o=s=o ό

V F

Intermediate 7 8-(((2',4'-difluoro-(1,1'-biphenyl)-4-yl)sulfonyl)methyl)-1,4dioxaspiro[4.5]decane (2.8 g, 6.9 mmol) in 0.5 M aqueous HCl solution (20 ml) was stirred at 70°C for 4 hours. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 1% MeOH in DCM]. After completion of the reaction, the reaction mixture was neutralized to pH 7 with 5% aqueous sodium hydroxide, stirred for 30 min and extracted with 10% MeOH in DCM. The organic layer was washed with brine (1x200 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the title intermediate 8 (2.2 g, crude), which was used in the next step without further purification.

Analytical data: ¹ H NMR (400 MHz, DMSOC) δ ppm, 8.04 (d, J=8.4 Hz, 2H), 7.85 - 7.81 (m, 2H), 7.73 - 7, 65 (m, 1H), 7.49 - 7.42 (m, 1H), 7.3 - 7.23 (m, 1H), 3.47 (d, J=6.4 Hz, 2H), 2 .42 - 2.3 (m, 3H), 2.25 - 2.16 (m, 2H), 2.15 - 2.05 (m, 2H), 1.63 - 1.51 (m, 2H) .

Synthesized compound 1.

cis-4-((((2',4'-difluoro-(1,1'-biphenyl)-4-yl)sulfonyl)methyl)-1-methylcyclohexan-1-ol (CHMSA-04-A)

X

0=5=0 O

Tj

F

Synthesized compound 2.

trans-4-((((2',4'-difluoro-(1,1'-biphenyl)-4-yl)sulfonyl)methyl)-1-methylcyclohexan-1-ol (CHMSA-04-B)

Intermediate solution

4-(((2',4'-difluoro-(1,1'-biphenyl)-4-45 043371 yl)sulfonyl)methyl)cyclohexan-1-one (2.70 g, crude) in THF (30 ml ) in an argon atmosphere was cooled to a temperature of -20°C and 3M methylmagnesium bromide (2.96 ml, 8.9 mmol) was added dropwise over 30 minutes. The reaction mixture was stirred at room temperature for 12 hours. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was quenched with saturated ammonium chloride solution and extracted with EtOAc. The organic layer was washed with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 2.5 g of material. 1 g of this crude material was purified by preparative HPLC (see details below) to give the title compounds synthesized 1 (0.10 g) and synthesized 2 (0.10 g) as white solids.

Preparative HPLC method: column - X-Select CSH 250 mmx30 mm, 5 µm; flow rate: 30 ml/min; Detection wavelength: 210-400 nm; mobile phases: A: 0.1% formic acid in water and B: Acetonitrile.

Analytical data (synthesized compound 1):

LCMS (ESI) m/z=363.05 [M-H ₂ O+1]+.

HPLC (see general method): retention time=8.69 min. Purity=99.5%.

1H NMR (400 MHz, DMSO-06) δ ppm: 8.00 (d, J=8.0 Hz, 2H), 7.83 - 7.78 (m, 2H), 7.72 - 7.65 ( m, 1H), 7.48 - 7.41 (m, 1H), 7.29 - 7.23 (m, 1H), 3.94 (s, 1H), 3.26 (d, J=6, 0 Hz, 2H), 1.78 - 1.65 (lat s, 1H), 1.6 - 1.35 (m, 6H), 1.26 - 1.16 (m, 2H), 1.05 (s, 3H).

Analytical data (synthesized compound 2):

LCMS (ESI) m/z=363.0 [M-H3O+1] ⁺ .

HPLC (see general method): retention time=8.43 min. Purity=98.5%.

^1H NMR (400 MHz, DMSO-d6) δ ppm: 8.00 (d, J=8.0 Hz, 2H), 7.83 - 7.79 (m, 2H), 7.72 - 7.65 (m, 1H), 7.48 - 7.41 (m, 1H), 7.29 - 7.22 (m, 1H), 4.17 (s, 1H), 3.34 - 3.30 (m , 2H), 1.89 - 1.7 (m, 3H), 1.5 - 1.44 (m, 2H), 1.33 - 1.15 (m, 4H), 1.06 (s, 3H ).

Synthesis scheme 3

Intermediate, conn. 6 Intermediate, conn. 9 Intermediate, conn. 10

- 46 043371

Intermediate 9. 4-chloro-4'-((1,4-dioxaspiro[4.5]decan-8-yl)methanesulfonyl)-(1,1'biphenyl)-2-carbonitrile

To a stirred solution of intermediate 6 2-(4-(((1,4-dioxaspiro[4.5]decane-8yl)methyl)sulfonyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (3.27 g, 7.7 mmol) to 1,4dioxane:water (3:1) (40 ml) was added sodium carbonate (2.46 g, 23.2 mmol) followed by the addition of 2-bromo-5- chlorobenzonitrile (1.67 g, 7.7 mmol). The reaction mixture was degassed for 20 minutes under argon, tetrakis(triphenylphosphine)palladium(0) (0.9 g, 0.78 mmol) was added and the reaction mixture was degassed again for 10 minutes. Then the specified reaction mixture was stirred at a temperature of 100°C for 12 hours in a sealed test tube. The reaction progress was monitored by TLC [(silica gel TLC plate), 20% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was filtered through Celite® and the filtrate was evaporated under reduced pressure. The resulting residue was diluted with water and extracted with EtOAc. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting residue was purified by column chromatography (silica gel 100-200 mesh) to obtain the title intermediate 9 (3.0 g) as a white solid.

Analytical data: LCMS (ESI) m/z=432.45[M+H] ⁺ .

Intermediate 10. 4-chloro-4'-((4-oxocyclohexyl)methanesulfonyl)-(1,1'-biphenyl)-2carbonitrile

Intermediate 9 4-chloro-4'-((1,4-dioxaspiro[4.5]decan-8-yl)methanesulfonyl)-(1,Gbiphenyl)-2-carbonitrile (3.0 g, 7.0 mmol) was stirred in 0.5 M aqueous HCl (30 ml) at 70°C for 4 hours. The reaction progress was monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was neutralized to pH 7 with 5% aqueous sodium hydroxide solution and stirred for 30 minutes. The product was extracted with 10% MeOH in DCM. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the title intermediate 10 (2.3 g, crude).

Analytical data: LCMS (ESI) m/z=388.15 [M+H]+.

Synthesized compound 3.

4-chloro-4'-((trans-4-hydroxy-4-methylcyclohexyl)methanesulfonyl)-(1,1'-biphenyl)-2-carbonitrile

- 47 043371

Synthesized compound 4.

4-chloro-4'-((cis-4-hydroxy-4-methylcyclohexyl)methanesulfonyl)-(1,1'-biphenyl)-2-carbonitrile (CHMSA-10-A)

A stirred solution of intermediate 10 4-chloro-4'-((4oxocyclohexyl)methanesulfonyl)-(1,1'-biphenyl)-2-carbonitrile (2.3 g, crude) in dry THF under argon was cooled to -20 °C, and 3M methylmagnesium bromide (2.3 mL, 6.9 mmol) was added dropwise over 30 min. After addition was complete, the reaction mixture was allowed to warm to room temperature and stirred at room temperature for 12 hours. The reaction progress was monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was quenched with saturated ammonium chloride solution and extracted with EtOAc. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a residue, which was purified by preparative HPLC (see details below) to obtain the title compounds synthesized compound 3 (0.110 g) and synthesized compound 4 (0.115 g) as white solids.

Preparative HPLC method: column - X-Select CSH 250 mmx30 mm, 5 µm; flow rate: 30 ml/min; Detection wavelength: 210-400 nm; mobile phases: A: 0.1% formic acid in water and B: Acetonitrile.

Analytical data (synthesized compound 3):

LCMS (ESI) m/z =386.20 [M+1-H ₂ O]+.

HPLC (see general method): retention time=8.37 min. Purity=99.0%.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm: 8.22 (d, J=2.4 Hz, 1H), 8.06 (d, J=8.4 Hz, 2H), 7.93 (dd, J=8.8 Hz, J =2.4 Hz, 1H), 7.87 (d, J=8.4 Hz, 2H), 7.72 (d, J=8.8 Hz, 1H ), 4.18 (s, 1H), 3.38 - 3.33 (m, 2H), 1.9 - 1.8 (m, 1H), 1.78 - 1.70 (m, 2H), 1.51 - 1.42 (m, 2H), 1.33 - 1.16 (m, 4H), 1.05 (s, 3H).

Analytical data (synthesized compound 4):

LCMS (ESI) m/z =386.05 [M+1-H ₂ O] ⁺ .

HPLC (see general method): retention time=8.60 min. Purity=99.8%.

1H NMR (400 MHz, DMSO-d6) δ ppm: 8.22 (d, J=2 Hz, 1H), 8.06 (d, J=8.4 Hz, 2H), 7.92 (dd, j =8.4, J=2.4 Hz, 1H), 7.86 (d, J=8.4 Hz, 2H), 7.72 (d, J=8.4 Hz, 1H), 3.94 (s, 1H), 3.32 - 3.27 (m, 2H), 1.79 - 1.67 (m, 1H),

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1.57 - 1.36 (m, 6H), 1.26 - 1.17 (m, 2H), 1.05 (s, 3H).

Synthesis scheme 4

Intermediate 11: 4-bromo-3-fluorobenzenethiol

To a stirred solution of triphenylphosphine (43.16 g, 164.53 mmol) in DCM (200 ml) and DMF (30 ml), 4-bromo-3-fluorobenzenesulfonyl chloride (15.00 g, 54.84 mmol) was added dropwise at room temperature, and the reaction mixture was stirred for 16 hours. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 5% EtOAc in n-hexane]. After completion of the reaction, 1M HCl aqueous solution (150 ml) was added, the layers were separated, and the organic layer was concentrated under reduced pressure to obtain a crude residue. The resulting residue was dissolved in 1M aq. NaOH solution (150 ml), the resulting suspension was filtered through Celite® and the filtrate was washed with Et ₂ O (2x100 ml). The aqueous layer was neutralized with 1M aqueous HCl (150 ml) and extracted with Et ₂ O (3x150 ml). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title intermediate 11 (7.00 g, crude) as a colorless oil, which was used in the next step without further purification.

Analytical data: ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm: 7.39 - 7.35 (m, 1H), 7.03 (dd, J=8.8, 2.4 Hz, 1H), 6 .91 (dd, J=8.0, 2.0 Hz, 1H), 3.52 (s, 1H).

Intermediate 12. 8-(((4-bromo-3-fluorophenyl)thio)methyl)-1,4-dioxaspiro[4.5]decane

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Cs ₂ CO ₃ (16.64 g, 51.07 mmol) at 0°C, and the reaction mixture was stirred for 20 minutes. Intermediate 3 8-(bromomethyl)-1,4dioxaspiro[4.5]decane (6.00 g, 25.54 mmol) dissolved in MeOH:THF (1:1, 15 ml) was added dropwise at the same temperature, The reaction mixture was allowed to warm to room temperature, then heated to 60°C for 12 hours. The reaction was monitored by TLC [(silica gel TLC plate), 10% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain a crude residue. The resulting residue was diluted with water (50 ml) and extracted with EtOAc (3x50 ml). The combined organic layer was washed with brine (2x50 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 5% EtOAc in n-hexane) to give the title intermediate 12 (6.12 g) as a colorless oil.

Analytical data: LCMS (ESI) m/z=361.10 [M+1]+ ( ⁷⁹ Br).

Intermediate 13. 8-(((4-bromo-3-fluorophenyl)sulfonyl)methyl)-1,4dioxaspiro[4.5]decane

G Γό

To a stirred solution of intermediate 12 8-(((4-bromo-3fluorophenyl)thio)methyl)-1,4-dioxaspiro[4.5]decane (6.10 g, 16.88 mmol) in DCM (50 ml), was added 3chloroperoxybenzoic acid (~70% in water) (10.41 g, 42.21 mmol) at 0°C in portions over 30 minutes, the reaction mixture was allowed to warm to room temperature and stirred for 12 hours. The progress of the reaction was monitored with by TLC [(silica gel TLC plate), 20% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was quenched with saturated Na ₂ S ₂ O ₃ solution (70 ml) and extracted with DCM (3x100 ml). The organic layer was separated, washed with brine (2x50 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 10% EtOAc in n-hexane) to give the title intermediate 13 (5.80 g, 87%) as a colorless solid. Analytical data: LCMS (ESI) m/z=395.00 [M+1] ⁺ ( ⁸¹ Br).

Intermediate 14 dioxaborolane

To a stirred solution of intermediate 13 8-(((4-bromo-3fluorophenyl)sulfonyl)methyl)-1,4-dioxaspiro[4.5]decane (5.70 g, 14.49 mmol) in 1,4-dioxane (60 ml), potassium acetate (4.27 g, 43.48 mmol) and bis(pinacolato)diboron (4.78 g, 18.84 mmol) were added at room temperature and the reaction mixture was degassed using argon for 15 min. Bis(triphenylphosphine)palladium(II) dichloride (0.15 g, 0.22 mmol) was added to the mixture, the reaction mixture was degassed for 10 min and heated at 100°C for 4 h in a sealed tube. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 30% EtOAc in n-50 043371 hexane]. After completion of the reaction, the reaction mixture was filtered, the residue was washed with EtOAc (100 ml) and the filtrate was concentrated under reduced pressure to give the title intermediate 14 (7.45 g, crude) as a black solid, which was directly used in the next step without further cleaning.

Analytical data: LCMS (ESI) m/z=359.20 [M+1]+ (corresponding boronic acid).

Intermediate 15: 4-(((1,4-dioxaspiro[4.5]decan-8-yl)methyl)sulfonyl)-2'-fluoro[1,1'-biphenyl]-4-carbonitrile

X o=s=o and

CN

To a stirred solution of intermediate 14 2-(4-(((1,4-dioxaspiro[4.5]decane-8yl)methyl)sulfonyl)-2-fluorophenyl)-4,4,5,5-tetramethyl-1,3, 2-dioxaborolane (3.00 g, 6.81 mmol) in dioxane-water (3:1, 40 ml), sodium carbonate (2.17 g, 20.44 mmol) and 4-bromobenzonitrile (1.24 g, 6.81 mmol) and the reaction mixture was degassed using argon for 20 min. Tetrakis(triphenylphosphine)palladium(0) (0.79 g, 0.68 mmol) was added to the mixture, the reaction mixture was degassed for 10 min and stirred at 100°C for 12 h in a sealed tube. The reaction progress was monitored by TLC [(silica gel TLC plate), 40% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was cooled to room temperature, filtered through Celite® and the filtrate was concentrated under reduced pressure to obtain a crude residue. The resulting residue was dissolved in EtOAc (50 ml) and washed with water (50 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 20% EtOAc in n-hexane) to give the title intermediate 15 (2.20 g) as a colorless solid.

Analytical data: LCMS (ESI) m/z=416.00 [M+1] ⁺ .

Intermediate 16. 2'-fluoro-4'-(((4-oxocyclohexyl)methyl)sulfonyl)-[1,1'-biphenyl]4-carbonitrile o=s=o ό ^F f CN

A solution of intermediate 15 4'-(((1,4-dioxaspiro[4.5]decan-8-yl)methyl)sulfonyl)-2'fluoro-[1,1'-biphenyl]-4-carbonitrile (2.20 g , 5.30 mmol) in 0.5 M aqueous HCl solution (25 ml) was stirred at 70°C for 4 hours. The reaction progress was monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane ]. After completion of the reaction, the reaction mixture was cooled to 0°C, neutralized to pH 7 with 5% aqueous NaOH (~20 ml) and extracted with EtOAc (3x30 ml). The organic layer was separated, washed with brine (50 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by flash column chromatography (silica gel 230-400 mesh, 30% EtOAc in n-hexane) to give the title intermediate 16 (1.70 g, 86%) as an off-white solid.

Analytical data: LCMS (ESI) m/z=372.10 [M+1] ⁺ .

Synthesized compound 5.

2'-fluoro-4'-(((trans-4-hydroxy-4-methylcyclohexyl)methyl)sulfonyl)-[1,1'-biphenyl]-4carbonitrile

- 51 043371 (CHMSA-12-B)

A

O=S=O (jL

CN

Synthesized compound 6.

2'-fluoro-4'-(((cis-4-hydroxy-4-methylcyclohexyl)methyl)sulfonyl)-[1,1'-biphenyl]-4-carbonitrile (CHMSA-12-A)

To a stirred solution of intermediate 16 2'-fluoro-4'-(((4oxocyclohexyl)methyl)sulfonyl)-[1,1'-biphenyl]-4-carbonitrile (1.70 g, 4.58 mmol) in anhydrous THF (20 ml), 3M methylmagnesium bromide (1.83 ml, 5.49 mmol) was added at -20°C and the reaction mixture was stirred at -20°C for 1 hour. The reaction mixture was allowed to warm to room temperature and stirred for 4 hours. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was quenched with saturated aqueous NH4Cl (20 ml) and extracted with EtOAc (3x20 ml). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by SLC chromatography (see details below) to obtain the title compounds, synthesized compound 5 (0.10 g, 6%) and synthesized compound 6 (0.18 g, 10%) as white solids.

Analytical data (synthesized compound 5):

LCMS (ESI) m/z=370.10 [M-H ₂ O+1]+.

HPLC (see general method): retention time=8.07 min. Purity=98.65%.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm: 8.01 (d, J=8.4 Hz, 2H), 7.94 - 7.82 (m, 5H), 4.18 (s, 1H), 3.40 (d, J=6.0 Hz, 2H), 1.85 (lat s, 1H), 1.77 - 1.76 (m, 2H), 1.50 - 1.43 (m, 2H), 1.33 - 1.18 (m, 4H), 1.06 (s, 3H).

Analytical data (synthesized compound 6):

LCMS (ESI) m/z=370.10 [M-H ₂ O+1] ⁺ .

HPLC (see general method): retention time=8.33 min. Purity=99.15%.

1H NMR (400 MHz, DMSO-d ₆ ) δ ppm: 8.01 (d, J=8.4 Hz, 2H), 7.94 - 7.82 (m, 5H), 3.95 (s, 1H ), 1.74 (br. s, 1H), 1.57 - 1.39 (m, 6H), 1.26 - 1.20 (m, 2H), 1.05 (s, 3H). (2H combined at solvent peak).

SKZHH conditions:

Mobile phases: A: CO ₂ ; B: 0.1% NH ₃ in MeOH.

Gradient: Started at 10% B, increased to 40% B over 5 min, held at 40% B for 4 min, decreased to 10% B over 1 min, and held at 10% B for 2 min.

Column: Chiralpak IA (250 mm x 4.6 mm, 5 µm). Wavelength: 260 nm. Flow rate: 3 ml/min.

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Synthesis scheme 5

Intermediate 17: 4-cyano-2-fluorophenyltrifluoromethanesulfonate

To a stirred solution of 3-fluoro-4-hydroxybenzonitrile (5.00 g, 36.47 mmol) in DCM (50 ml) was added triethylamine (10.17 ml, 72.93 mmol) at 0°C, followed by the addition of triflate anhydride (7.35 ml, 43.76 mmol) and the reaction mixture was stirred at 0°C for 30 minutes. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 20% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was diluted with DCM (25 ml) and washed with water (50 ml). The organic layer was separated, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by flash column chromatography (silica gel 60-120 mesh, gradient 1-10% EtOAc in nhexane) to give the title intermediate 17 (7.00 g, 71%) as a colorless oil.

Analytical data: ^1H NMR (400 MHz, DMSO^) δ ppm: 8.35 (dd, J=10.0, 2.0 Hz, 1H), 8.02 - 7.94 (m, 2H).

Intermediate 18. 4'-(((1,4-dioxaspiro[4.5]decan-8-yl)methyl)sulfonyl)-2-fluoro[1,1'-biphenyl]-4-carbonitrile

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To a stirred solution of intermediate 6 2-(4-(((1,4-dioxaspiro[4.5]decane-8yl)methyl)sulfonyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.50 g, 5.92 mmol) in dioxane:water (3:1, 40 ml), add sodium carbonate (1.88 g, 17.76 mmol), intermediate 17 4cyano-2-fluorophenyltrifluoromethanesulfonate (3 .20 g, 11.83 mmol) and the reaction mixture was degassed with argon for 20 min. Tetrakis(triphenylphosphine)palladium(0) (0.68 g, 0.59 mmol) was added to the mixture, the reaction mixture was degassed for 10 min and stirred at 100°C for 12 h in a sealed tube. The reaction progress was monitored by TLC [(silica gel TLC plate), 30% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was filtered through Celite® and the filtrate was concentrated under reduced pressure to obtain a crude residue. The resulting residue was dissolved in EtOAc (50 ml) and washed with water (50 ml). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude residue. The residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 1-30% EtOAc in n-hexane) to give the title intermediate 18 (2.20 g, 89%) as an off-white solid.

Analytical data: LCMS (ESI) m/z=416.00 [M+H]+.

Intermediate 19. 2-fluoro-4'-(((4-oxocyclohexyl)methyl)sulfonyl)-[1,1'-biphenyl]-4carbonitrile

A solution of intermediate 18 4'-(((1,4-dioxaspiro[4.5]decan-8-yl)methyl)sulfonyl)-2fluoro-[1,1'-biphenyl]-4-carbonitrile (2.20 g, 5 .30 mmol) in 0.5 M aqueous HCl solution (40 ml) was stirred at 70°C for 4 hours. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 30% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was cooled to 0°C, neutralized to pH 7 with 5% aqueous NaOH and extracted with EtOAc (3x50 ml). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the title intermediate 19 (1.80 g, crude) as a white solid, which was used in the next step without further purification.

Analytical data: LCMS (ESI) m/z=371.80 [M+H] ⁺ .

Synthesized compound 7.

2-fluoro-4'-(((trans-4-hydroxy-4-methylcyclohexyl)methyl)sulfonyl)-[1,1'-biphenyl]-4-carbonitrile (CHMSA-01-B)

Synthesized compound 8.

2-fluoro-4'-(((cis-4-hydroxy-4-methylcyclohexyl)methyl)sulfonyl)-[1,1'-biphenyl]-4-carbonitrile

- 54 043371

(CHMSA-01-A)

To a stirred solution of intermediate 19 2-fluoro-4'-(((4oxocyclohexyl)methyl)sulfonyl)-[1,1'-biphenyl]-4-carbonitrile (1.80 g, 4.85 mmol) in anhydrous THF ( 25 ml), 3M methylmagnesium bromide (1.94 ml, 5.82 mmol) was added at -20°C, then the reaction mixture was allowed to warm to room temperature and stirred for 12 hours. The progress of the reaction was monitored by TLC [( silica gel TLC plate), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was cooled to 0°C and quenched with saturated aqueous NH ₄ Cl (30 ml) and extracted with EtOAc (3x50 ml). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 1-30% EtOAc in n-hexane) to obtain 1.00 g of a mixture of diastereomers, which was purified by SLC chromatography (see details below) to obtain the indicated in the title compounds, synthesized compound 7 (0.16 g, 9%) and synthesized compound 8 (0.20 g, 11%) as white solids.

Analytical data (synthesized compound 7):

LCMS (ESI) m/z=370.10 [M-H ₂ O+1]+.

HPLC (see general method): retention time=7.94 min. Purity=98.33%.

1H NMR (400 MHz, DMSO-d6) δ ppm: 8.07 - 8.03 (m, 3H), 7.89 - 7.82 (m, 4H), 4.19 (s, 1H), 3, 35 (s, 1H), 1.84 (broad s, 1H), 1.78 - 1.74 (m, 2H), 1.52 - 1.46 (m, 2H), 1.33 - 1, 17 (m, 4H), 1.06 (s, 3H) (1H combined with solvent peak).

Analytical data (synthesized compound 8):

LCMS (ESI) m/z=370.10 [M-H ₂ O+1] ⁺ .

HPLC (see general method): retention time=8.21 min. Purity=98.33%.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm: 8.07 - 8.03 (m, 3H), 7.90 - 7.82 (m, 4H), 3.95 (s, 1H), 3.28 (d, J=6.0 Hz, 2H), 1.74 (broad s, 1H), 1.57 - 1.54 (m, 2H), 1.49 - 1.37 (m, 4H), 1.25 - 1.17 (m, 2H), 1.05 (s, 3H).

SKZHH conditions:

Mobile phases: A: CO ₂ ; B: 0.1% NH ₃ in MeOH.

Gradient: started at 10% B, increased to 40% B over 5 min, held at 40% B for 4 min, decreased to 10% B over 1 min, and held at 10% B for 2 min.

Column: Chiralpak IA (250 mm x 4.6 mm, 5 µm). Wavelength: 260 nm. Flow rate: 3 ml/min.

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Synthesis scheme 6

Synthesized Synthesized compound 9 compound 10

Intermediate 20. 4'-(((1,4-dioxaspiro[4.5]decan-8-yl)methyl)sulfonyl)-2-chloro[1,1'-biphenyl]-4-carbonitrile

0=5=0 f

CN

To a stirred solution of intermediate 6 2-(4-(((1,4-dioxaspiro[4.5]decane-8yl)methyl)sulfonyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (3.30 g, 7.81 mmol) to a dioxane:water (3:1, 40 ml) mixture was added sodium carbonate (2.48 g, 23.44 mmol) and 4-bromo-3-chlorobenzonitrile (1.69 g, 7.81 mmol) and the reaction mixture was degassed with argon for 20 min. Tetrakis(triphenylphosphine)palladium(0) (0.90 g, 0.78 mmol) was added to the mixture, the reaction mixture was degassed for 10 min and stirred at 100°C for 12 h in a sealed tube. The reaction progress was monitored by TLC [(silica gel TLC plate), 30% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was filtered through Celite® and concentrated under reduced pressure. The resulting residue was dissolved in EtOAc (100 ml) and washed with water (50 ml). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude residue. The residue was purified by flash column chromatography (silica gel 230400 mesh, gradient 1-20% EtOAc in n-hexane) to give the title intermediate 20 (3.00 g, 89%) as a white solid.

Analytical data: LCMS (ESI) m/z=432.20 [M+1]+.

Intermediate 21. 2-chloro-4'-(((4-oxocyclohexyl)methyl)sulfonyl)-[1,1'-biphenyl]-4carbonitrile

- 56 043371

A solution of intermediate 20 4'-(((1,4-dioxaspiro[4.5]decan-8-yl)methyl)sulfonyl)-2chloro-[1,1'-biphenyl]-4-carbonitrile (3.00 g, 6 .95 mmol) in 0.5 M aqueous HCl solution (40 ml) was stirred at 70°C for 4 hours. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 30% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was cooled to 0°C, neutralized to pH 7 with 5% aqueous NaOH and extracted with EtOAc (3x50 ml). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude residue. The resulting residue was triturated with n-hexane to give the title intermediate 21 (2.50 g, 93%) as a white solid.

Analytical data: LCMS (ESI) m/z=388.20 [M+1]+.

Synthesized compound 9.

2-chloro-4'-(((trans-4-hydroxy-4-methylcyclohexyl)methyl)sulfonyl)-[1,1'-biphenyl]-4-carbonitrile (CHMSA-05-B)

Synthesized compound 10.

2-chloro-4'-(((cis-4-hydroxy-4-methylcyclohexyl)methyl)sulfonyl)-[1,1'-biphenyl]-4-carbonitrile (CHMSA-05-A)

To a stirred solution of intermediate 21 2-chloro-4'-(((4oxocyclohexyl)methyl)sulfonyl)-[1,1'-biphenyl]-4-carbonitrile (2.50 g, 6.45 mmol) in THF (30 ml) 3M methylmagnesium bromide (2.60 ml, 7.73 mmol) was added dropwise at -20°C over 30 minutes, then the reaction mixture was allowed to warm to room temperature and stirred for 12 hours. The progress of the reaction was monitored with by TLC [(silica gel TLC plate), 40% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was quenched with saturated NH ₄ Cl solution (30 ml) and extracted with EtOAc (3x50 ml). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue (2 g). The resulting residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 1-30% EtOAc in n-hexane) to obtain 1.00 g of a mixture of diastereomers, which was purified by SLC chromatography (see details below) to obtain the title compounds synthesized compound 9 (0, 13 g, 5%) and synthesized compound 10 (0.20 g, 8%) as white solids.

Analytical data (synthesized compound 9):

LCMS (ESI) m/z=386.10 [M-H2O+1] ⁺ .

HPLC (see general method): retention time=8.12 min. Purity=99.66%.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ ppm: 8.25 (d, J=1.2 Hz, 1H), 8.04 - 8.01 (m, 2H), 7.96 (dd, J=7.6, 1.4

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Hz, 1H), 7.76 - 7.74 (m, 2H), 7.68 (d, J=8.0 Hz, 1H), 4.19 (s, 1H), 3.34 (d, J =6.4 Hz, 2H), 1.85 (lat. s, 1H),

1.77 - 1.73 (m, 2H), 1.50 - 1.44 (m, 2H), 1.32 - 1.17 (m, 4H), 1.05 (s, 3H).

Analytical data (synthesized compound 9):

LCMS (ESI) m/z=386.15 [M-H ₂ O+1]+.

HPLC (see general method): retention time=8.38 min. Purity=98.48%.

1H NMR (400 MHz, DMSO-d ₆ ) δ ppm: 8.24 (br s, 1H), 8.03 (d, J=8.0 Hz, 2H), 7.96 (d, J=8 ,0 Hz, 1H), 7.75 (d, J=8.4 Hz, 2H), 7.68(d, J=7.6 Hz, 1H), 3.95 (s, 1H), 3, 28 (d, J=6.4 Hz, 2H), 1.75 (broad s, 1H), 1.56 - 1.39 (m, 6H), 1.25 - 1.19 (m, 2H) , 1.05 (s, 3H).

SKZHH conditions:

Mobile phases: A: CO ₂ ; B: 0.1% NH ₃ in MeOH.

Gradient: Isocratic mode: 30% B.

Column: Chiralpak IA (250 mm x 4.6 mm, 5 µm). Wavelength: 256 nm. Flow rate: 3 ml/min.

Synthesis scheme 7

Intermediate, conn. 3

Intermediate 22. 2-bromo-5-mercaptobenzamide t-CPBA, DCM, OC to RT

ZM MeMgBr, THF, from -20 C to RT

Synthesized compound 12

Synthesized

I) HCl, NaNO ₂ , H ₂ o, -5 °C

m) K-O-ethyl xanthate.

from -5 C to 75 C

Hi) KOH, EtOH, arr. COOL.

I) Pd(PPh h) 4, Na ₂ CO h, dioxane:HgO (3:1), 100 °C

i) separation

CS2CO3, THF:MeOH 2:1, from O C to RT compound 11

To a suspension of 5-amino-2-bromobenzonitrile (4.00 g, 20.30 mmol) in concentrated HCl (5.50 ml) and ice (10 g) was added NaNO ₂ (1.46 g, 21.11 mmol) and H2O (10 ml) at 5°C, and the resulting reaction mixture was added to a solution of potassium O-ethyl xanthate (6.51 g, 40.60 mmol) in H2O (10 ml).

- 58 043371

The resulting mixture was stirred at 75°C for 1.5 hours. The reaction mixture was cooled to room temperature, the pH was adjusted to 8 with 5% NaHCO ₃ solution (50 ml) and extracted with Et ₂ O (3x200 ml). The organic layer was separated, washed with water (200 ml) and concentrated under reduced pressure to obtain a crude residue. The resulting residue was dissolved in EtOH (35 ml), KOH (4.56 g, 81.20 mmol) was added and the resulting reaction mixture was refluxed for 17 hours. The reaction was monitored by TLC [(silica gel TLC plate) , 30% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain a crude residue. Water (70 ml) was added to the crude residue and extracted with Et ₂ O (2x50 ml). The aqueous layer was acidified to pH 1-2 with 3N. H ₂ SO ₄ (60 ml) and extracted with DCM (3x100 ml). The combined organic layer was separated, washed with brine (200 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the title intermediate 22 (1.63 g, crude) as a pale brown oil, which was used in the next step without additional cleaning.

Analytical data: LCMS (ESI) m/z=233.90 [M+1] ⁺ ( ⁸¹ Br).

Intermediate 23. 5-(((1,4-dioxaspiro[4.5]decan-8-yl)methyl)thio)-2-bromobenzamide

To a stirred solution of intermediate 22 2-bromo-5-mercaptobenzamide (1.63 g, 7.03 mmol) in MeOH:H ₂ O (2:1, 30 ml) was added CsCO ₃ (4.58 g, 14.06 mmol) at 0°C, and the reaction mixture was stirred for 15 minutes. Intermediate 3 8-(bromomethyl)-1,4-dioxaspiro[4.5]decane (1.65 g, 7.03 mmol) was added dropwise to this mixture, then the reaction mixture was allowed to warm to room temperature and stirred for 12 hours The reaction progress was monitored by TLC [(silica gel TLC plate), 40% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain a crude residue. Water (10 ml) was added to the residue and extracted with EtOAc (3x20 ml). The combined organic layer was washed with brine (2x20 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by flash column chromatography (silica gel 230-400 mesh, 40% EtOAc in n-hexane) to give the title intermediate 23 (0.80 g) as an off-white solid.

Analytical data: LCMS (ESI) m/z=385.93 [M+1]+.

Intermediate 24. 5-(((1,4-dioxaspiro[4.5]decan-8-yl)methyl)thio)-2-bromobenzonitrile

To a stirred solution of intermediate 23 5-(((1,4-dioxaspiro[4.5]decane-8yl)methyl)thio)-2-bromobenzamide (0.80 g, 2.07 mmol) in anhydrous THF (10 ml) was added TFAA (0.87 g, 4.14 mmol) dropwise at 0°C, the reaction mixture was allowed to warm to room temperature and stirred for 1 hour. The reaction progress was monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was quenched with saturated NaHCO ₃ solution (10 ml) and extracted with EtOAc (3x15 ml). The organic layer was separated, washed with brine (2x15 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by flash column chromatography (silica gel 230-400 mesh, 20% EtOAc in n-hexane) to give the title intermediate 24 (0.70 g, 92%) as a pale yellow solid.

Analytical data: LCMS (ESI) m/z=369.90 [M+1] ⁺ ( ⁸¹ Br).

Intermediate 25. 5-(((1,4-dioxaspiro[4.5]decan-8-yl)methyl)sulfonyl)-2bromobenzonitrile

- 59 043371

To a stirred solution of intermediate 24 5-(((1,4-dioxaspiro[4.5]decan-8-yl)methyl)thio)-2-bromobenzonitrile (0.70 g, 1.90 mmol) in DCM (20 ml) 3-chloroperoxybenzoic acid (~70% in water) (0.94 g, 3.80 mmol) was added at 0°C in portions over 15 min, the reaction mixture was allowed to warm to room temperature and stirred for 12 h. reactions were monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was quenched with a saturated solution of NaHCO ₃ (10 ml). The organic layer was separated and the aqueous layer was extracted with DCM (3x15 ml). The combined organic layer was washed with brine (2x15 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by flash column chromatography (silica gel 230-400 mesh, 40% EtOAc in n-hexane) to give the title intermediate 25 (0.30 g, 39%) as an off-white solid.

Analytical data: LCMS (ESI) m/z=400.05 [M+1]+.

Intermediate 26: 2-bromo-5-(((4-oxocyclohexyl)methyl)sulfonyl)benzonitrile

A solution of intermediate 25 5-(((1,4-dioxaspiro[4.5]decan-8-yl)methyl)sulfonyl)-2bromobenzonitrile (0.20 g, 0.50 mmol) in 1 M HCl (20 ml) was heated at temperature 100°C for 12 hours. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 50% EtOAc in ngexane]. After completion of the reaction, the reaction mixture was cooled to room temperature, the pH was adjusted to 8 with saturated NaHCO ₃ solution (30 ml) and extracted with DCM (3x20 ml). The organic layer was separated, washed with brine (2x15 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the title intermediate 26 (0.12 g, crude) as a yellow solid, which was used in the next step without additional cleaning.

Analytical data: LCMS (ESI) m/z=399.05 [M+CH3CN+T ( ⁸¹ Br).

Intermediate 27: 2-bromo-5-(((4-oxocyclohexyl)methyl)sulfonyl)benzonitrile

3M methylmagnesium bromide (0.13 mL, 0. 40 mmol) at -20°C, the reaction mixture was allowed to warm to room temperature and stirred for 4 hours. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was quenched with saturated NH4C1 (20 mL) and extracted with EtOAc (3 x 20 mL). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the title intermediate 27 (0.10 g, crude) as a pale yellow solid, which was used in the next step without further purification.

Synthesized compound 11.

2',4'-difluoro-4-(((trans-4-hydroxy-4-methylcyclohexyl)methyl)sulfonyl)-[1,1'-biphenyl]-2carbonitrile

- 60 043371 (CHMSA-06-B)

HE

0=5=0 ^γ^ΟΝ

F

Synthesized compound 12.

2',4'-difluoro-4-(((cis-4-hydroxy-4-methylcyclohexyl)methyl)sulfonyl)-[1,1'-biphenyl]-2carbonitrile (CHMSA-06-A)

To a stirred solution of intermediate 27 2-bromo-5-(((4-hydroxy-4methylcyclohexyl)methyl)sulfonyl)benzonitrile (0.10 g, 0.27 mmol) and 2-(2,4-difluorophenyl)-4, 4,5,5tetramethyl-1,3,2-dioxaborolane (0.065 g, 0.27 mmol) in a mixture of dioxane:water (3:1, 8 ml) was added sodium carbonate (0.085 g, 0.81 mmol) and the reaction mixture degassed with argon for 20 min. Tetrakis(triphenylphosphine)palladium(0) (0.031 g, 0.027 mmol) was added to the mixture, the reaction mixture was degassed for 20 min and stirred at 100°C for 12 h in a sealed tube. The reaction progress was monitored by TLC [(silica gel TLC plate), 60% EtOAc in n-hexane] and LCMS. After completion of the reaction, the reaction mixture was cooled to room temperature, filtered through Celite®, washed with EtOAc (30 ml) and the filtrate was concentrated under reduced pressure. The resulting residue was diluted with water (20 ml) and extracted with EtOAc (3x20 ml). The organic layer was separated, washed with brine (2x15 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by flash column chromatography (silica gel 230-400 mesh, 40% EtOAc in n-hexane) to obtain a mixture of diastereomers. The crude material from this batch was mixed with another batch prepared in a similar manner (50 mg scale reaction). The combined crude material from both batches was purified by supercritical liquid chromatography (see details below) to obtain the title compounds synthesized 11 (0.007 g, 4%) and synthesized 12 (0.02 g, 12%) as off-white solids .

Analytical data (synthesized compound 11):

LCMS (ESI) m/z=388.15 [M-H ₂ O+1]+.

HPLC (see general method): retention time=8.23 min. Purity=96.29%. ^1H NMR (400 MHz, DMSO-d6) δ ppm: 8.53 (d, J=2.0 Hz, 1H), 8.29 (dd, J=8.0, 1.6 Hz, 1H), 7.91 (d, J=8.0 Hz, 1H), 7.72-7.66 (m, 1H), 7.58 - 7.53 (m, 1H), 7.36-7.32 ( m, 1H), 4.19 (s, 1H), 3.46 (d, J=6.4 Hz, 2H), 1.91 (lat s, 1H), 1.78 (lat s, 2H ), 1.51-1.47 (m, 2H), 1.36 - 1.20 (m, 4H), 1.07 (s, 3H).

Analytical data (synthesized compound 12):

LCMS (ESI) m/z=388.10 [M-H ₂ O+1]+.

HPLC (see general method): retention time=8.48 min. Purity=98.05%.

1H NMR (400 MHz, DMSO-d6) δ ppm: 8.54 (d, J=2.0 Hz, 1H), 8.29 (dd, J=8.0, 1.6 Hz, 1H), 7 .90 (d, J=8.4 Hz, 1H), 7.72 - 7.66 (m, 1H), 7.58 - 7.53 (m, 1H), 7.36 - 7.32 (m , 1H), 3.96 (s, 1H), 3.40 (d, J=5.6 Hz, 2H), 1.80 (lat s, 1H), 1.58 - 1.56 (m, 2H), 1.51-1.40 (m, 4H), 1.29 - 1.23 (m, 2H), 1.06 (s, 3H).

SKZHH conditions:

Mobile phases: A: CO ₂ ; B: 0.1% NH ₃ in MeOH.

Gradient: Started at 25% B, increased to 50% B over 5 min, held at 50% B for 4 min, decreased to 25% B over 1 min, and held at 25% B for 2 min.

Column: Chiralpak IG (250 mm x 4.6 mm, 5 µm). Wavelength: 258 nm. Flow rate: 3 ml/min.

- 61 043371

Synthesis scheme 8

NaH, DMF, from O C to 100 C compound 14 compound 13 □t -5 C to 75 C iii) KOH, EUN, Arr.COL.

Intermediate, conn. 3

Synthesized

Synthesized

Intermediate 28. 4-Bromo-3-chlorobenzenethiol

C.I.

To a stirred solution of 4-bromo-3-chloroaniline (5.00 g, 24.22 mmol) in concentrated HCl (6.67 mL) and ice (11.67 g) was added NaNO2 (1.74 g, 25.19 mmol ) and H2O (11.7 ml) at a temperature of 5°C and the resulting diazonium solution was added to a solution of potassium O-ethyl xanthate (7.76 g, 48.43 mmol) in H2O (11.7 ml). The resulting mixture was stirred at 75°C for 1.5 hours. The reaction mixture was cooled to room temperature, the pH was adjusted to 8 with saturated NaHCO ₃ solution (50 ml) and extracted with Et ₂ O (4x100 ml). The organic layer was separated, washed with water (200 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The resulting residue was added to a solution of KOH (6.21 g, 110.67 mmol) in EtOH (41.67 mL), and the resulting reaction mixture was refluxed for 17 hours. The reaction progress was monitored by TLC [(silica gel plate for TLC), 10% EtOAc in ngexane]. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain a crude residue. The resulting residue was diluted with water (50 ml) and washed with Et ₂ O (50 ml). The aqueous layer was acidified to pH 1-2 with 3N. H ₂ SO ₄ (80 ml) and extracted with DCM (3x150 ml). The combined organic layer was separated, washed with brine (200 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the title intermediate 28 (2.20 g, crude) as a pale brown oil, which was used as follows. stage without additional purification.

Intermediate 29. 8-(((4-bromo-3-chlorophenyl)thio)methyl)-1,4-dioxaspiro[4.5]decane

- 62 043371

Br

To a stirred solution of 4-bromo-3-chlorobenzenethiol intermediate 28 (3.42 g, 15.31 mmol) in DMF (20 ml) was added NaH (60% dispersion in mineral oil, 1.02 g, 25.52 mmol) at a temperature of 0°C and the said reaction mixture was stirred for 30 minutes. Intermediate 3 8-(bromomethyl)-1,4-dioxaspiro[4.5]decane (3.00 g, 12.76 mmol) dissolved in DMF (10 ml) was added dropwise to this and the reaction mixture was allowed to warm to room temperature , stirred for 30 min and then heated to a temperature of 100°C for 12 h. The reaction progress was monitored by TLC [(silica gel TLC plate), 20% EtOAc in n-hexane] and LCMS. After completion of the reaction, the reaction mixture was cooled to room temperature, quenched with water (30 ml) and extracted with EtOAc (3x30 ml). The combined organic layer was washed with brine (2x30 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The residue was purified by flash column chromatography (silica gel 230-400 mesh, 10% EtOAc in n-hexane) to give the title intermediate 29 (2.50 g) as a pale yellow solid.

Analytical data: LCMS (ESI) m/z=379.05 [M+1]+ ( ⁸¹ Br).

Intermediate 30. 8-(((4-bromo-3-chlorophenyl)sulfonyl)methyl)-1,4dioxaspiro [4.5] decane c-a o=s=o % Br

To a stirred solution of intermediate 29 8-(((4-bromo-3-chlorophenyl)thio)methyl)1,4-dioxaspiro[4.5]decane (2.00 g, 5.29 mmol) in AcOH (20 ml) and H2O (20 ml), potassium permanganate (2.51 g, 15.88 mmol) was added in portions at 0°C, the reaction mixture was allowed to reach room temperature and stirred for 8 hours. The progress of the reaction was monitored by TLC [( silica gel TLC plate), 20% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was basified to pH 8 with saturated NaHCO ₃ solution (20 ml) and extracted with EtOAc (3x30 ml). The organic layer was separated, washed with brine (50 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 20% EtOAc in n-hexane) to give the title intermediate 30 (1.20 g, 55%) as a colorless solid.

Analytical data: LCMS (ESI) m/z=411.05 [M+1] ⁺ ( ⁸¹ Br).

Intermediate 31. 4'-(((1,4-dioxaspiro[4.5]decan-8-yl)methyl)sulfonyl)-2-chloro[1,1'-biphenyl]-4-carbonitrile p?

o=s=o ό f CN

To a stirred solution of 8-(((4-bromo-3chlorophenyl)sulfonyl)methyl)-1,4-dioxaspiro[4.5]decane intermediate 30 (1.20 g, 2.93 mmol) in dioxane:water (3: 1, 20 mL), sodium carbonate (0.93 g, 8.79 mmol) and (4-cyanophenyl)boronic acid (0.43 g, 2.93 mmol) were added and the reaction mixture was degassed with argon for 30 min. Tetrakis(triphenylphosphine)palladium(0) (0.34 g, 0.29 mmol) was added to this mixture, the reaction mixture was degassed for 15 minutes and stirred at a temperature of 100°C for 12 hours in a sealed tube. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was cooled to room temperature, filtered through Celite® and the filtrate was concentrated under reduced pressure to obtain a crude residue. Water (10 ml) was added to the crude residue and extracted with EtOAc (3x20 ml). The organic layer was separated, washed with brine (15 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by flash column chromatography (silica gel 230-400 mesh, 10% EtOAc in n-hexane) to give the title intermediate 31 (0.50 g, 39%) as a pale yellow solid.

Analytical data: LCMS (ESI) m/z=432.10 [M+1]+.

Intermediate 32. 2-chloro-4'-(((4-oxocyclohexyl)methyl)sulfonyl)-[1,1'-biphenyl]-4carbonitrile

1^2^° o=s=o

CN

A solution of intermediate 31 4'-(((1,4-dioxaspiro[4.5]decan-8-yl)methyl)sulfonyl)-2'chloro-[1,1'-biphenyl]-4-carbonitrile (0.50 g , 1.16 mmol) in 0.5 M aqueous HCl solution (15 ml) was heated to a temperature of 100 ° C for 12 hours. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane ]. After completion of the reaction, the reaction mixture was cooled to 0°C, the pH was adjusted to 7 with 10% NaOH (10 ml) and extracted with EtOAc (3x15 ml). The organic layer was separated, washed with brine (20 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by flash column chromatography (silica gel 230-400 mesh, 20% EtOAc in n-hexane) to give the title intermediate 32 (0.43 g, 96%) as an off-white solid.

Analytical data: LCMS (ESI) m/z=429.25 [M+CH3CN+1X

Synthesized compound 13.

2'-chloro-4'-(((trans-4-hydroxy-4-methylcyclohexyl)methyl)sulfonyl)-[1,1'-biphenyl]-4carbonitrile (CHMSA-07-B)

Synthesized compound 14.

2'-chloro-4'-(((cis-4-hydroxy-4-methylcyclohexyl)methyl)sulfonyl)-[1,1'-biphenyl]-4-carbonitrile (CHMSA-07-A) o=s=o

A

Y^CI o

CN

To a stirred solution of intermediate 32 2'-chloro-4'-(((4oxocyclohexyl)methyl)sulfonyl)-[1,1'-biphenyl]-4-carbonitrile (0.43 g, 1.11 mmol) in anhydrous THF (10 ml) 3M methylmagnesium bromide (0.44 ml, 1.33 mmol) was added at -20°C, then the reaction mixture was allowed to warm to room temperature and stirred for 12 hours.

- 64 043371 reactions were monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was quenched with saturated NH4Cl solution (10 ml) and extracted with EtOAc (3x15 ml). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The crude material from this batch was mixed with another batch prepared in a similar manner (0.45 g scale reaction). The combined crude material from both batches was purified by supercritical liquid chromatography (see details below) to obtain the title compounds synthesized compound 13 (0.09 g, 10%) and synthesized compound 14 (0.15 g, 16%) as protein dry solids.

Analytical data (synthesized compound 13):

LCMS (ESI) m/z=386.10 [M-H2O+1]+.

HPLC (see general method): retention time=8.31 min. Purity=99.52%.

^1H NMR (400 MHz, DMSO- _d6 ) δ ppm: 8.12 (br s, 1H), 8.01 - 7.96 (m, 3H), 7.75 - 7.71 (m, 3H ), 4.19 (s, 1H), 3.43 (d, J=6.4 Hz, 2H), 1.89 (broad s, 1H), 1.83 - 1.75 (m, 2H) , 1.51 - 1.48 (m, 2H), 1.35 - 1.23 (m, 4H), 1.07 (s, 3H).

Analytical data (synthesized compound 14):

LCMS (ESI) m/z=386.15 [M-H ₂ O+1]+.

HPLC (see general method): retention time=8.57 min. Purity=97.31%.

1H NMR (400 MHz, DMSO-d ₆ ) δ ppm: 8.12 (d, J=2.0 Hz, 1H), 8.01 - 7.96 (m, 3H), 7.75-7.70 (m, 3H), 3.96 (s, 1H), 3.36 (d, J=6.4 Hz, 2H), 1.78 (br s, 1H), 1.59 - 1.57 ( m, 2H), 1.51 - 1.40 (m, 4H), 1.28 - 1.22 (m, 2H), 1.06 (s, 3H).

SKZHH conditions:

Mobile phases: A: CO ₂ ; B: 0.1% NH3 in MeOH.

Gradient: Started at 25% B, increased to 50% B over 5 min, held at 50% B for 4 min, decreased to 25% B over 1 min, and held at 25% B for 2 min.

Column: Chiralpak IG (250 mm x 4.6 mm, 5 µm). Wavelength: 256 nm. Flow rate: 3 ml/min.

Synthesis scheme 9

Intermediate, conn. 34

Intermediate, conn. 36

Intermediate, conn. 37

Synthesized Synthesized compound 15 compound 16

Intermediate 33: 4-bromo-2-(trifluoromethyl)benzenethiol

- 65 043371

To a stirred solution of 4-bromo-2-(trifluoromethyl)benzenesulfonyl chloride (2.00 g, 6.18 mmol) in toluene (20 ml), triphenylphosphine (4.86 g, 18.55 mmol) was added slowly at 0°C, the reaction mixture was then allowed to warm to room temperature and stirred for 16 hours. The reaction was monitored by TLC [(silica gel TLC plate), 20% EtOAc in nhexane].

After completion of the reaction, the reaction mixture was quenched with 1 N. HCl (5 ml) and concentrated under reduced pressure to obtain a crude residue. The resulting residue was made alkaline to pH ~ 10 with 10% KOH (~10 ml), the resulting solid was filtered, washed with water (10 ml) and the filtrate was extracted with Et ₂ O (2x25 ml). The aqueous layer was neutralized to pH 7 with 2N. HCl (~20 ml) and extracted with EtOAc (3x50 ml). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the title intermediate 33 (1.00 g, crude) as a brown liquid.

Analytical data: 1H NMR (400 MHz, CDC^) δ ppm: 7.73 (s, 1H), 7.46 (dd, J=8.4, 2.0 Hz, 1H),

7.24 (d, J=2.8 Hz, 1H), 3.75 - 3.73 (m, 1H).

Intermediate dioxaspiro[4.5]decane

34. 8-(((4-bromo-2-(trifluoromethyl)phenyl)thio)methyl)-1,4-

Br

To a stirred solution of 8-(bromomethyl)-1,4dioxaspiro[4.5]decane intermediate 3 (7.00 g, 29.77 mmol) and 4-bromo-2(trifluoromethyl)benzenethiol intermediate 33 (9.18 g, 35 .73 mmol) in acetone (100 ml), K ₂ CO ₃ (8.23 g, 59.54 mmol) was added and the reaction mixture was heated to a temperature of 60 ° C for 18 hours. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 30% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain a crude residue. The residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 1-20% EtOAc in n-hexane) to give the title intermediate 34 (6.00 g, 49%) as a brown oil. Analytical data: 1H NMR (400 MHz, CDCl ₃ ) δ ppm: 7.77 (br s, 1H), 7.60 - 7.58 (m, 1H), 7.32 (d, J=8.4 Hz , 1H), 3.96 (s, 4H), 2.89 (d, J=6.8 Hz, 2H), 1.94 - 1.92 (m, 2H), 1.79 - 1.76 ( m, 2H), 1.55 - 1.51 (m, 1H), 1.45 - 1.32 (m, 2H), 1.30 - 1.26 (m, 2H).

Intermediate 35. 8-(((4-bromo-2-(trifluoromethyl)phenyl)sulfonyl)methyl)-1,4dioxaspiro[4.5]decane

X o=s=o

Br

To a stirred solution of intermediate 34 8-(((4-bromo-2(trifluoromethyl)phenyl)thio)methyl)-1,4-dioxaspiro[4.5]decane (6.00 g, 14.59 mmol) in DCM (70 ml) 3-chloroperoxybenzoic acid (~60% in water) (12.58 g, 43.77 mmol) was added slowly at 0°C, the reaction mixture was allowed to warm to room temperature and stirred for 12 hours. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 30% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was cooled to 0°C, saturated NaHCO ₃ solution (50 ml) was slowly added and the layers were separated. The organic layer was washed with saturated Na ₂ S ₂ O ₃ solution (50 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 1-40% EtOAc in n-hexane) to give the title intermediate 35 (2.00 g, 31%) as a white solid.

Analytical data: LCMS (ESI) m/z=445.05 [M+1]+ ( ⁸¹ Br).

Intermediate 36. 8-(((2',4'-difluoro-3-(trifluoromethyl)-[1,1'-biphenyl]-4-66 043371 yl)sulfonyl)methyl)-1,4-dioxaspiro[4.5 ]dean

To a stirred solution of 8-(((4-bromo-2(trifluoromethyl)phenyl)sulfonyl)methyl)-1,4-dioxaspiro[4.5]decane intermediate 35 (1.00 g, 2.26 mmol) in dioxane: water (7:3, 10 ml) added sodium carbonate (0.72 g, 6.77 mmol), 2-(2,4-difluorophenyl)-4,4,5,5 tetramethyl-1,3,2-dioxaborolane (0.54 g, 2.26 mmol) and the reaction mixture was degassed with argon for 10 min. Tetrakis(triphenylphosphine)palladium(0) (0.26 g, 0.23 mmol) was added to the mixture, the reaction mixture was degassed for 10 min and stirred at 100°C for 12 h in a sealed tube. The reaction progress was monitored by TLC [(silica gel TLC plate), 30% EtOAc in n-hexane]. After completion of the reaction, the said reaction mixture was filtered through

Celite® and the filtrate were concentrated under reduced pressure to obtain a crude residue. The resulting residue was dissolved in EtOAc (30 ml) and washed with water (25 ml). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude residue. The residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 1-20% EtOAc in n-hexane) to give the title intermediate 36 (1.00 g, 93%) as a white solid.

Analytical data: LCMS (ESI) m/z=477.10 [M+1]+.

Intermediate 37. 4-(((2',4'-difluoro-3-(trifluoromethyl)-[1,1'-biphenyl]-4yl)sulfonyl)methyl)cyclohexan-1-one o=s=o Xa ^F 3 Q and F

A solution of intermediate 36 8-(((2',4'-difluoro-3-(trifluoromethyl)-[1,1'-biphenyl]-4yl)sulfonyl)methyl)-1,4-dioxaspiro[4.5]decane (1 .00 g, 2.10 mmol) in 0.5 M aqueous HCl solution (30 ml) was stirred at 70°C for 4 hours. The reaction progress was monitored by TLC [(silica gel TLC plate), 40% EtOAc in n-hexanes]. After completion of the reaction, the reaction mixture was cooled to 0°C, neutralized to pH 7 with 5% aqueous NaOH (~10 ml), stirred for 30 min and extracted with EtOAc (3x50 ml). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the title intermediate 37 (0.90 g, crude) as a white solid, which was used in the next step without further purification.

Analytical data: LCMS (ESI) m/z=433.10 [M+1] ⁺ .

Synthesized compound 15.

trans-4-(((2',4'-difluoro-3-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)sulfonyl)methyl)-1 methylcyclohexan-1-ol (CHMSA-08- B)

F

- 67 043371

Synthesized compound 16.

cis-4-(((2',4'-difluoro-3-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)sulfonyl)methyl)-1-methylcyclohexan1-ol (CHMSA-08-A )

To a stirred solution of intermediate 37 4-(((2',4'-difluoro-3-(trifluoromethyl)[1,1'-biphenyl]-4-yl)sulfonyl)methyl)cyclohexan-1-one (0.90 g, 2.08 mmol) in anhydrous THF (15 ml), 3M methylmagnesium bromide (0.83 ml, 2.50 mmol) was added dropwise at -20°C, the reaction mixture was allowed to warm to room temperature and stirred for 12 hours The reaction progress was monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was quenched with saturated aqueous NH ₄ Cl (20 ml) and extracted with EtOAc (3x50 ml). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by column chromatography (silica gel 230-400 mesh, gradient 1-40% EtOAc in n-hexane) to give 0.60 g of a mixture of diastereomers, which was purified by chiral preparative HPLC (see details below) to give the title compounds synthesized compound 15 (0.18 g, 19%) and synthesized compound 16 (0.06 g, 6%) as white solids.

Analytical data (synthesized compound 15):

LCMS (ESI) m/z=431.15 [M-H ₂ O+1]+.

HPLC (see general method): retention time=9.05 min. Purity=98.26%.

^1H NMR (400 MHz, DMSO-d6) δ ppm: 8.31 (d, J=8.8 Hz, 1H), 8.15 - 8.14 (m, 2H), 7.83-7.77 (m, 1H), 7.52 - 7.46 (m, 1H), 7.32 - 7.28 (m, 1H), 4.21 (s, 1H), 3.37 (d, J=6 ,4 Hz, 2H), 1.99 (lat s, 1H), 1.79 - 1.76 (m, 2H), 1.50 - 1.47 (m, 2H), 1.36 - 1, 20 (m, 4H), 1.07 (s, 3H).

Analytical data (synthesized compound 16):

LCMS (ESI) m/z=431.10 [M-H ₂ O+1]+.

HPLC (see general method): retention time=9.27 min. Purity=98.20%.

1H NMR (400 MHz, DMSO-d6) δ ppm: 8.31 (d, J=8.4 Hz, 1H), 8.13 (br s, 2H), 7.82 - 7.76 (m, 1H), 7.51 - 7.46 (m, 1H), 7.31 - 7.27 (m, 1H), 3.97 (s, 1H), 3.29 (d, J=6.4 Hz , 2H), 1.89 (br. s, 1H), 1.59 - 1.42 (m, 6H), 1.30 - 1.23 (m, 2H), 1.07 (s, 3H).

Chiral preparative HPLC method:

Column: YMC CHIRAL AMYLOSE-SA, 250 mm x 4.6 mm, 5 µm; mobile phases: A: n-hexane + 0.1% DEA and B: DCM:MeOH (1:1); flow rate: 1.0 ml/min; Isocratic mode: 15% B.

- 68 043371

Synthesis scheme 10

Synthesized Synthesized compound 17 compound 18

Intermediate 38 ,2-dioxaborolane

To a stirred solution of intermediate 35 8-(((4-bromo-2(trifluoromethyl)phenyl)sulfonyl)methyl)-1,4-dioxaspiro[4.5]decane (4.00 g, 9.02 mmol) in 1.4 -dioxane (40 ml), potassium acetate (2.66 g, 27.07 mmol) and bis(pinacolato)diboron (2.98 g, 11.73 mmol) were added at room temperature and the reaction mixture was degassed using argon in for 20 minutes. Bis(triphenylphosphine)palladium(II) dichloride (0.14 g, 0.198 mmol) was added to the mixture, the reaction mixture was degassed for 20 min and stirred at 100°C for 4 h in a sealed tube. The reaction progress was monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, the resulting residue was dissolved in EtOAc (150 ml) and washed with water (100 ml). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 1-30% EtOAc in n-hexane) to give the title intermediate 38 (3.80 g, 86%) as a white solid.

Analytical data: LCMS (ESI) m/z=409.10 [M+1]+ (corresponding boronic acid).

Intermediate 39. 2-(4-(((1,4-dioxaspiro[4.5]decan-8-yl)methyl)sulfonyl)-3(trifluoromethyl)phenyl)-3,5-difluoropyridine

- 69 043371

To a stirred solution of intermediate 38 2-(4-(((1,4-dioxaspiro[4.5]decane-8yl)methyl)sulfonyl)-3-(trifluoromethyl)phenyl)-4,4,5,5-tetramethyl-1 ,3,2-dioxaborolane (1.00 g, 2.04 mmol) and 2-bromo-3,5-difluoropyridine (0.40 g, 2.04 mmol) in dioxane:water (7:3, 10 ml ) sodium carbonate (0.65 g, 6.12 mmol) was added and the reaction mixture was degassed with argon for 15 min. Tetrakis(triphenylphosphine)palladium(0) (0.24 g, 0.20 mmol) was added to the mixture, the reaction mixture was degassed for 10 min and stirred at 100°C for 12 h in a sealed tube. The reaction progress was monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was filtered through Celite® and the filtrate was concentrated under reduced pressure to obtain a crude residue. The resulting residue was dissolved in EtOAc (100 ml) and washed with water (75 ml). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 1-50% EtOAc in n-hexane) to give the title intermediate 39 (0.80 g, 82%) as a white solid.

Analytical data: LCMS (ESI) m/z=478.10 [M+1] ⁺ .

Intermediate 40. 4-(((4-(3,5-difluoropyridin-2-yl)-2-(trifluoromethyl)phenyl)sulfonyl)methyl)cyclohexan-1-one

A solution of intermediate 39 2-(4-(((1,4-dioxaspiro[4.5]decan-8-yl)methyl)sulfonyl)3-(trifluoromethyl)phenyl)-3,5-difluoropyridine (0.80 g, 1 .68 mmol) in 0.5 M aqueous HCl solution (50 ml) was stirred at 100°C for 12 hours. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was cooled to 0°C, neutralized to pH 7 with 5% aqueous NaOH (~20 ml) and extracted with EtOAc (3x70 ml). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a white solid. The resulting residue was triturated with n-hexane (25 ml) to give the title intermediate 40 (0.70 g, 96%) as a white solid.

Analytical data: LCMS (ESI) m/z=474.10 [M+CH ₃ CN] ⁺ .

Synthesized compound 17.

trans-4-(((4-(3,5-difluoropyridin-2-yl)-2-(trifluoromethyl)phenyl)sulfonyl)methyl)-1methylcyclohexan-1-ol (CHMSA-09-B)

Synthesized compound 18.

cis-4-(((4-(3,5-difluoropyridin-2-yl)-2-(trifluoromethyl)phenyl)sulfonyl)methyl)-1

- 70 043371 methylcyclohexan-1-ol (CHMSA-09-A)

To a stirred solution of intermediate 40 4-(((4-(3,5-difluoropyridin-2yl)(trifluoromethyl)phenyl)sulfonyl)methyl)cyclohexan-1-one (0.70 g, 1.62 mmol) in THF ( 20 ml) 3M methylmagnesium bromide (0.65 ml, 1.95 mmol) was added at -20°C, the reaction mixture was allowed to warm to room temperature and stirred for 4 hours. The reaction progress was monitored by TLC [(silica gel plate for TLC), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was cooled to 0°C and quenched with saturated aqueous NH ₄ Cl (30 ml) and extracted with EtOAc (3x100 ml). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by column chromatography (silica gel 230-400 mesh, gradient 1-50% EtOAc in n-hexane) to give 0.50 g of a mixture of diastereomers, which was purified by SLC chromatography (see details below) to give the title compounds synthesized compound 17 (0.14 g, 19%) and synthesized compound 18 (0.15 g, 21%) as white solids.

Analytical data (synthesized compound 17):

LCMS (ESI) m/z=432.10 [M-H ₂ O+1]+.

HPLC (see general method): retention time=8.66 min. Purity=98.63%.

1H NMR (400 MHz, DMSO-66) δ ppm: 8.76-8.75 (m, 1H), 8.48-8.44 (m, 2H), 8.37 (d, J=8.4 Hz, 1H), 8.24 - 8.19 (m, 1H), 4.21 (s, 1H), 3.38 (d, J=6.4 Hz, 2H), 1.98 (lat. s , 1H), 1.79 - 1.75 (m, 2H), 1.50 - 1.46 (m, 2H), 1.37 - 1.20 (m, 4H), 1.07 (s, 3H ).

Analytical data (synthesized compound 18):

LCMS (ESI) m/z=432.10 [M-H2O+1L

HPLC (see general method): retention time=8.91 min. Purity=99.21%.

^1H NMR (400 MHz, DMSO-d6) δ ppm: 8.76 (br s, 1H), 8.48 - 8.44 (m, 2H), 8.38 (d, J=8.4 Hz , 1H), 8.24 - 8.20 (m, 1H), 3.97 (s, 1H), 3.30 (s, 1H), 1.88 (lat s, 1H), 1.59 - 1.56 (m, 2H), 1.51 - 1.42 (m, 4H), 1.30 - 1.24 (m, 2H), 1.07 (s, 3H). (1H combined with solvent peak).

SKZHH conditions:

Mobile phases: A: CO ₂ ; B: 0.1% NH ₃ in MeOH.

Gradient: Started at 10% B, increased to 40% B over 5 min, held at 40% B for 4 min, decreased to 10% B over 1 min, and held at 10% B for 2 min.

Column: Chiralpak IG (250 mm x 4.6 mm, 5 µm). Wavelength: 250 nm. Flow rate: 3 ml/min.

Synthesis scheme 11

Intermediate 41 carbonitrile

- 71 043371

To a stirred solution of intermediate 38 2-(4-(((1,4-dioxaspiro[4.5]decane-8yl)methyl)sulfonyl)-3-(trifluoromethyl)phenyl)-4,4,5,5-tetramethyl-1 ,3,2-dioxaborolane (1.00 g, 2.04 mmol) and 4-bromo-3-fluorobenzonitrile (0.49 g, 2.45 mmol) in dioxane:water (7:3, 10 ml) were added sodium carbonate (0.65 g, 6.12 mmol) and the reaction mixture was degassed with argon for 15 min. Tetrakis(triphenylphosphine)palladium(0) (0.24 g, 0.20 mmol) was added to the mixture, the reaction mixture was degassed for 10 min and stirred at 100°C for 12 h in a sealed tube. The reaction progress was monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane].

After completion of the reaction, the reaction mixture was filtered through Celite® and the filtrate was concentrated under reduced pressure to obtain a crude residue. The resulting residue was dissolved in EtOAc (50 ml) and washed with water (50 ml). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 1-35% EtOAc in n-hexane) to give the title intermediate 41 (0.90 g, 91%) as a white solid.

Analytical data: LCMS (ESI) m/z=484.15 [M+1]+.

Intermediate 42. 2-fluoro-4'-(((4-oxocyclohexyl)methyl)sulfonyl)-3'(trifluoromethyl)-[1,1'-biphenyl]-4-carbonitrile o=s=o

A.CF3 ST % CN

Solution of intermediate 41 4'-(((1,4-dioxaspiro[4.5]decan-8-yl)methyl)sulfonyl)-2fluoro-3'-(trifluoromethyl)-[1,1'-biphenyl]-4-carbonitrile (0.90 g, 1.86 mmol) in 0.5 M aq. HCl solution (30 ml) was stirred at 70°C for 12 hours. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was cooled to 0°C, neutralized to pH 7 with 5% aqueous NaOH (~20 ml) and extracted with EtOAc (3x50 ml). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the title intermediate 42 (0.85 g, crude) as a white solid, which was used in the next step without further purification.

Synthesized compound 19.

2-fluoro-4'-(((trans-4-hydroxy-4-methylcyclohexyl)methyl)sulfonyl)-3'-(trifluoromethyl)-[1,Hbiphenyl]-4-carbonitrile (CHMSA-02-B)

Synthesized compound 20.

2-fluoro-4'-(((cis-4-hydroxy-4-methylcyclohexyl)methyl)sulfonyl)-3'-(trifluoromethyl)-[1,1'biphenyl]-4-carbonitrile

- 72 043371

(CHMSA-02-A)

To a stirred solution of intermediate 42 2-fluoro-4'-(((4oxocyclohexyl)methyl)sulfonyl)-3'-(trifluoromethyl)-[1,1'-biphenyl]-4-carbonitrile (0.85 g, 1. 93 mmol) in anhydrous THF (10 ml), 3M methylmagnesium bromide (0.80 ml, 2.32 mmol) was added at -20°C, then the reaction mixture was allowed to warm to room temperature and stirred for 4 hours. During the reaction monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was cooled to 0°C and quenched with saturated aqueous NH ₄ Cl (~30 ml) and extracted with EtOAc (3x50 ml). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 1-50% EtOAc in n-hexane) to obtain 0.60 g of a mixture of diastereomers, which was purified by SLC chromatography (see details below) to obtain the indicated in the title compounds, synthesized compound 19 (0.09 g, 10%) and synthesized compound 20 (0.09 g, 10%) as white solids.

Analytical data (synthesized compound 19):

LCMS (ESI) m/z=438.10 [M-H ₂ O+1]+.

HPLC (see general method): retention time=8.77 min. Purity=99.07%. 1H NMR (400 MHz, DMSOL) δ ppm: 8.34 (d, J=8.4 Hz, 1H), 8.22 - 8.20 (m, 2H), 8.09 (d, J=10, 4 Hz, 1H), 7.96 - 7.88 (m, 2H), 4.21 (s, 1H), 3.38 (d, J=6.4 Hz, 2H), 1.99 (br. s, 1H), 1.82 - 1.73 (m, 2H), 1.50 - 1.47 (m, 2H), 1.37 - 1.20 (m, 4H), 1.07 (s, 3H).

Analytical data (synthesized compound 20):

LCMS (ESI) m/z=438.15 [M-H ₂ O+1]+.

HPLC (see general method): retention time=8.83 min. Purity=99.33%. 1H NMR (400 MHz, DMSO-d6) δ ppm: 8.35 (d, J=8.4 Hz, 1H), 8.22 - 8.19 (m, 2H), 8.09 (d, J= 10.0 Hz, 1H), 7.96 - 7.88 (m, 2H), 3.97 (s, 1H), 3.31 (d, J=6.4 Hz, 2H), 1.89 ( lat. s, 1H), 1.59 - 1.57 (m, 2H), 1.51 - 1.42 (m, 4H), 1.30 - 1.23 (m, 2H), 1.07 ( s, 3H).

SKZHH conditions:

Mobile phases: A: CO ₂ ; B: 0.1% NH ₃ in MeOH.

Gradient: Started at 10% B, increased to 40% B over 5 min, held at 40% B for 4 min, decreased to 10% B over 1 min, and held at 10% B for 2 min.

Column: Chiralpak IG (250 mm x 4.6 mm, 5 µm). Wavelength: 262 nm. Flow rate: 3 ml/min.

Synthesis scheme 12

Intermediate 43. 8-(((2',4'-dichloro-[1,1'-biphenyl]-4-yl)sulfonyl)methyl)-1,4- 73 043371 dioxaspiro[4.5]decane o-l © o=s=o

Cl

To a stirred solution of intermediate 6 2-(4-(((1,4-dioxaspiro[4.5]decane-8yl)methyl)sulfonyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (3.27 g, 7.74 mmol) to a dioxane:water mixture (3:1, 40 ml) was added sodium carbonate (2.46 g, 23.23 mmol) and 1-bromo-2,4-dichlorobenzene (1 .75 g, 7.74 mmol) and the reaction mixture was degassed with argon for 20 minutes. Tetrakis(triphenylphosphine)palladium(0) (0.89 g, 0.77 mmol) was added to the mixture, the reaction mixture was degassed for 10 min and stirred at 100°C for 12 h in a sealed tube. The reaction progress was monitored by TLC [(silica gel TLC plate), 30% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was filtered through Celite® and the filtrate was concentrated under reduced pressure to obtain a crude residue. The resulting residue was dissolved in EtOAc (150 ml) and washed with water (100 ml). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude residue. The residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 1-20% EtOAc in nhexane) to give the title intermediate 43 (3.00 g, 88%) as a white solid.

Analytical data: LCMS (ESI) m/z=441.10 [M+1]+.

Intermediate 44. 4-(((2',4'-dichloro-[1,1'-biphenyl]-4-yl)sulfonyl)methyl)cyclohexan-1 -one o=s=o tjr

Cl

A solution of intermediate 43 8-(((2',4'-dichloro-[1,1'-biphenyl]-4-yl)sulfonyl)methyl)-1,4dioxaspiro[4.5]decane (3.00 g, 6. 80 mmol) in 0.5 M aq. HCl (40 ml) was stirred at 70°C for 4 hours. The reaction was monitored by TLC [(silica gel TLC plate), 30% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was cooled to 0°C, neutralized to pH 7 with 5% aqueous NaOH, stirred for 30 min and extracted with EtOAc (3x50 ml). The combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude residue. The resulting residue was dissolved in DCM (10 ml), stirred at room temperature and n-hexane (50 ml) was slowly added to it. The resulting precipitate was filtered, washed with n-hexane (25 ml) and dried to give the title intermediate 44 (2.50 g, 93%) as a white solid. Analytical data: LCMS (ESI) m/z=397.10 [M+1]+.

Synthesized compound 21.

trans-4-(((2',4'-dichloro-[1,1'-biphenyl]-4-yl)sulfonyl)methyl)-1-methylcyclohexan-1-ol (CHMSA-11-B)

Synthesized compound 22.

cis-4-(((2',4'-dichloro-[1,1'-biphenyl]-4-yl)sulfonyl)methyl)-1-methylcyclohexan-1-ol

- 74 043371

To a stirred solution of intermediate 44 4-(((2',4'-dichloro-[1,G-biphenyl]-4yl)sulfonyl)methyl)cyclohexan-1-one (2.50 g, 6.29 mmol) in anhydrous THF (30 ml), 3M methylmagnesium bromide (2.52 ml, 7.55 mmol) was added dropwise at -20°C over 30 minutes, the reaction mixture was allowed to warm to room temperature and stirred for 12 hours. During the reaction monitored by TLC [(silica gel TLC plate), 40% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was quenched with saturated aqueous NH ₄ Cl (30 ml) and extracted with EtOAc (3x50 ml). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by column chromatography (silica gel 230-400 mesh, gradient 1-40% EtOAc in n-hexane) to obtain 1.00 g of a mixture of diastereomers, which was purified by SLC chromatography (see details below) to obtain the title compounds synthesized compound 21 (0.20 g, 8%) and synthesized compound 22 (0.20 g, 8%) as white solids.

Analytical data (synthesized compound 21):

LCMS (ESI) m/z=395.10 [M-H ₂ O+1] ⁺ .

HPLC (see general method): retention time=9.23 min. Purity=98.26%.

1H NMR (400 MHz, DMSO-d ₆ ) δ ppm: 8.00 (d, J=8.4 Hz, 2H), 7.81 - 7.80 (m, 1H), 7.71 (d, J =8.4 Hz, 2H), 7.57 (dd, J=8.4, 2.0 Hz, 1H), 7.51 (d, J=8.4 Hz, 1H), 4.18 (s , 1H), 1.85 (br. s, 1H), 1.77 - 1.73 (m, 2H), 1.49 1.46 (m, 2H), 1.33 - 1.15 (m, 4H), 1.06 (s, 3H) (2H combined at solvent peak).

Analytical data (synthesized compound 22):

LCMS (ESI) m/z=435.00 [M+Na] ⁺ .

HPLC (see general method): retention time=9.51 min. Purity=98.80%.

1H NMR (400 MHz, DMSO-d ₆ ) δ ppm: 8.00 (d, J=8.4 Hz, 2H), 7.80 (d, J=2.0 Hz, 1H), 7.71 ( d, J=8.4 Hz, 2H), 7.57 (dd, J=8.4, 2.0 Hz, 1H), 7.51 (d, J=8.4 Hz, 1H), 3, 94 (s, 1H), 3.27 (d, J=6.0 Hz, 2H), 1.76 (broad s, 1H), 1.56 - 1.39 (m, 6H), 1.26 - 1.18 (m, 2H), 1.06 (s, 3H).

SKZHH conditions:

Mobile phases: A: CO ₂ ; B: 0.1% NH3 in MeOH.

Gradient: Started at 10% B, increased to 40% B over 5 min, held at 40% B for 4 min, decreased to 10% B over 1 min, and held at 10% B for 2 min.

Column: Chiralpak IA (250 mm x 4.6 mm, 5 µm). Wavelength: 253 nm. Flow rate: 3 ml/min.

Synthesis scheme 13

- 75 043371

i) ZM MeMdBr.THF, from -20°C to RT ii) separation of diastereomers _F

Synthesized compound 24

Synthesized compound 23

Intermediate 45: 2-(4-(((1,4-dioxaspiro[4.5]decan-8-yl)methyl)sulfonyl)-2fluorophenyl)-3,5-difluoropyridine

To a stirred solution of intermediate 14 2-(4-(((1,4-dioxaspiro[4.5]decane-8yl)methyl)sulfonyl)-2-fluorophenyl)-4,4,5,5-tetramethyl-1,3, 2-dioxaborolane (3.00 g, 6.81 mmol) in a mixture of dioxane-water (3:1, 40 ml) was added sodium carbonate (2.17 g, 20.44 mmol) and 2-bromo-3,5- difluoropyridine (1.32 g, 6.81 mmol) and the reaction mixture was degassed using argon for 20 minutes. Tetrakis(triphenylphosphine)palladium(0) (0.79 g, 0.68 mmol) was added to the mixture, the reaction mixture was degassed for 10 min and heated at 100°C for 12 h in a sealed tube. The reaction progress was monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was cooled to room temperature, filtered through Celite® and the filtrate was concentrated under reduced pressure to obtain a crude residue. The resulting residue was dissolved in EtOAc (50 ml) and washed with water (50 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 30% EtOAc in n-hexane) to give the title intermediate 45 (2.50 g) as a colorless solid.

Analytical data: LCMS (ESI) m/z=428.15 [M+1] ⁺ .

Intermediate 46. 4-(((4-(3,5-difluoropyridin-2-yl)-3-fluorophenyl)sulfonyl)methyl)cyclohexan-1-one

Solution of intermediate 45 2-(4-(((1,4-dioxaspiro[4.5]decan-8-yl)methyl)sulfonyl)2-fluorophenyl)-3,5-difluoropyridine (2.50 g, 5.85 mmol ) in 0.5 M aqueous HCl solution (30 ml) was stirred at 70°C for 4 hours. The progress of the reaction was monitored by TLC [(silica gel TLC plate), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was brought to 0°C, neutralized to pH 7 with 5% aqueous NaOH (20 ml) and extracted with EtOAc (3x30 ml). The organic layer was separated, washed with brine (50 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified by flash column chromatography (silica gel 230-400 mesh, gradient 30% EtOAc in n-hexane) to give the title intermediate 46 (2.10 g, 94%) as a colorless oil. Analytical data: LCMS (ESI) m/z=384.10 [M+1]+.

Synthesized compound 23.

trans-4-(((4-(3,5-difluoropyridin-2-yl)-3-fluorophenyl)sulfonyl)methyl)-1-methylcyclohexan-1-ol

- 76 043371

Synthesized compound 24.

cis-4-(((4-(3,5-difluoropyridin-2-yl)-3-fluorophenyl)sulfonyl)methyl)-1-methylcyclohexan-1-ol (CHMSA-03-A) he

F

To a stirred solution of intermediate 46 4-(((4-(3,5-difluoropyridin-2-yl)-3fluorophenyl)sulfonyl)methyl)cyclohexan-1-one (2.10 g, 5.48 mmol) in anhydrous THF (20 ml) 3M methylmagnesium bromide (2.19 ml, 6.57 mmol) was added at -20°C, the reaction mixture was allowed to warm to room temperature and stirred for 4 hours. The reaction progress was monitored by TLC [(silica gel plate for TLC), 50% EtOAc in n-hexane]. After completion of the reaction, the reaction mixture was quenched with a saturated solution of NH4Cl (20 ml) and extracted with EtOAc (3x20 ml). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude residue. The resulting residue was purified using SLC chromatography (see details below) with obtaining the title compounds, synthesized compound 23 (0.10 g, 5%) and synthesized compound 24 (0.10 g, 5%) as white solids.

Analytical data (synthesized compound 23):

LCMS (ESI) m/z=382.10 [M-H ₂ O+1]+.

HPLC (see general method): retention time=7.91 min. Purity=99.42%.

^1H NMR (400 MHz, DMSO-06) δ ppm: 8.74 - 8.73 (m, 1H), 8.22 - 8.17 (m, 1H), 7.96 - 7.87 (m, 3H), 4.18 (s, 1H), 3.42 (d, J=6.4 Hz, 2H), 1.86 (br. s, 1H), 1.77 - 1.76 (m, 2H ), 1.49 - 1.46 (m, 2H), 1.33 - 1.20 (m, 4H), 1.06 (s, 3H).

Analytical data (synthesized compound 24):

LCMS (ESI) m/z=382.15 [M-H ₂ O+1] ⁺ .

HPLC (see general method): retention time=8.19 min. Purity=99.83%.

^1H NMR (400 MHz, DMSO-d ₆ ) δ ppm: 8.70 (d, J=2.0 Hz, 1H), 8.19 - 8.14 (m, 1H), 7.94 - 7, 84 (m, 3H), 3.92 (s, 1H), 3.33 (d, J=6.0 Hz, 2H), 1.72 (br. s, 1H), 1.53 - 1.36 (m, 6H), 1.24 - 1.17 (m, 2H), 1.02 (s, 3H).

SKZHH conditions:

Mobile phases: A: CO ₂ ; B: 0.1% NH ₃ in MeOH.

Gradient: Started at 10% B, increased to 40% B over 5 min, held at 40% B for 4 min, decreased to 10% B over 1 min, and held at 10% B for 2 min.

Column: Chiralpak IA (250 mm x 4.6 mm, 5 µm). Wavelength: 277 nm. Flow rate: 3 ml/min.

X-ray analysis.

Confirmation of the structure of CHMSA-04-B.

A crystal of suitable size was selected from the main sample CHMSA-04-B and single crystal X-ray diffraction analysis was performed on it at 100°K. The analysis confirmed the structure and showed a trans configuration of the hydroxyl group relative to the carbon substituent at position 4 of the cyclohexane ring.

Single crystal data were collected on a Rigaku Oxford Diffraction Supernova, Cu at Zero dual source Atlas CCD diffractometer equipped with an Oxford Cryosystems Cobra cooling unit. Data were collected using Cu Ka radiation as indicated in the experimental table. The structures were solved and refined using the Bruker AXS SHELXTL software package

- 77 043371 or crystallographic software OLEX ² . The reference diffraction pattern of the crystal structure was constructed according to C. F. a. Macrae, Mercury: visualization and analysis of crystal structures, J. Appl. Cryst., vol. 39, p. 453-457, 2006.

Table 1

X-ray analysis: sample and crystal data

Empirical formula C20H22F2O3S Mass according to formula 380.43 Temperature 100(2) °K Wavelength 1.54184 A Crystal size 0.200 x 0.050 x 0.030 mm Crystal habit Colorless, columnar singonia Monoclinic Space group P2i/c Unit cell dimensions a = 15.9704(3) A; a = 90° b = 11.9467(2) A; β = 107.047(2)° s = 9.9018(2) A; γ = 90° Volume 1806.19(7) A ³ Z 4 Density (calculation) 1.399 mg/ ^m3 Absorption coefficient 1.925 mm' ¹ F(000) 800 Diffractometer SuperNova, Dual, Si at zero, Atlas Radiation source SuperNova (C) X-ray Source, CuKa Data collection method omega scan Theta band for data acquisition from 4.700 to 74.582° Index ranges -19 >>? > 19;-14 > k < 13;-12 >> 12 Collected Reflections 33265 Independent reflections 3655 [R(int) = 0.0428] Coverage of independent reflections 99.9% Absorption correction Semi-empirical by equivalents Max, and min. transmission 1.00000 and 0.83197 Structure decoding technique direct methods Structure decoding program SHELXTL-2013 Refinement technique Full matrix least squares method on F ² Refinement program SHELXL-2013 Minimized function 1HH ² -g _s ² ) ² Data / restrictions / parameters 3655 /0/250 Degree of conformity to F ² 1.035 Δ/Omax 0.000 Final R-indexes (3246 data; 1>2o(1)) R1 = 0.0323, wR2 = 0.0817 Final R-indexes (all data) R1 = 0.0376, wR2 = 0.0855 Weight scheme w =1 I [σ ² (Fo ² )+(0.0451 P) ² +0.8328P] where P=(F0 ² +2F _c ² )/3 Largest diff. peak and hole 0.391 and -0.396 eA- ³

The following table shows the atomic coordinates and equivalent isotropic atomic displacement parameters (A ² ) (u(eq) is defined as one third of the trace of the orthogonalized tensor Uij). Fl is oriented 97% in one rotamer relative to the central aryl-aryl bond and 3% in a rotamer that is located at an angle of 180 degrees to the main rotamer.

-78043371

Atomic coordinates and equivalent isotropic atomic displacement parameters

Atom Ha y/b z/c U (eq.) S1 0.21867(2) 0.34174(3) 0.51209(3) 0.01580(10) F1A 0.55064(6) 0.39175(7) 0.25634(10) 0.0244(3) F1B 0.5486(19) 0.034(2) 0.447(3) 0.032(9) F2 0.73785(6) 0.11438(8) 0.18146(10) 0.0263(2) 01 -0.22081(6) 0.45131(8) 0.16221(11) 0.0176(2) 02 0.23618(7) 0.44273(9) 0.59611(11) 0.0245(2) OZ 0.18156(7) 0.24742(9) 0.56496(11) 0.0236(2) C1 -0.14088(10) 0.35565(13) 0.03519(16) 0.0237(3) C2 -0.13297(9) 0.41757(11) 0.17243(15) 0.0149(3) 03 -0.07428(9) 0.52098(11) 0.18838(16) 0.0184(3) C4 0.02201(9) 0.49109(12) 0.21008(16) 0.0185(3) C5 0.05785(8) 0.41229(11) 0.33642(15) 0.0146(3) About -0.00145(9) 0.30983(11) 0.32544(16) 0.0178(3) C7 -0.09762(9) 0.34201(11) 0.30111(15) 0.0170(3) C8 0.15142(8) 0.37854(11) 0.34119(14) 0.0156(3) C9 0.31699(9) 0.30016(12) 0.47931(14) 0.0153(3) SJ 0.32171(9) 0.19404(12) 0.42490(15) 0.0181(3) C11 0.39486(9) 0.16573(11) 0.38329(15) 0.0173(3) C12 0.46348(9) 0.24207(11) 0.39704(15) 0.0153(3) C13 0.45830(9) 0.34672(11) 0.45782(16) 0.0173(3) 014 0.38509(9) 0.37671(11) 0.49785(15) 0.0171(3) 015 0.53888(8) 0.21021(11) 0.34603(15) 0.0156(3) 016 0.57905(9) 0.28478(11) 0.27645(15) 0.0177(3) C17 0.64610(9) 0.25582(12) 0.22086(16) 0.0189(3) 018 0.67301(9) 0.14591(12) 0.23661(16) 0.0192(3) 019 0.63799(9) 0.06714(12) 0.30630(16) 0.0208(3) 020 0.57108(9) 0.10062(12) 0.36038(15) 0.0180(3)

The PRD X-ray diffraction pattern was calculated from the crystal structure (100°K) and compared with the experimentally obtained diffraction pattern of the crystals (taken at room temperature). The diffraction patterns are in good agreement and demonstrate that the resulting single crystal structure is representative of the supplied material.

Powder X-ray diffraction (XRD) patterns were obtained on a Bruker D8 diffractometer using Cu Ka radiation (40 kV, 40 mA) and a 0-20 goniometer equipped with a Ge monochromator. The incident beam passed through a 2.0 mm divergence slit, behind which there was a 0.2 mm anti-scatter slit and a knife collimator. The diffracted beam passed through an 8.0 mm receiving slit with 2.5° Soller slits, behind which the Lynxeye detector was located. Diffrac Plus XRD Commander and Diffrac Plus EVA software were used for data collection and analysis, respectively.

The PRD samples were tested under ambient conditions as flat plate samples using the compound in powder form. The sample was placed on a polished zero-background silicon substrate (510) by gentle pressing against a flat surface or placed in a cut-out cavity. The sample rotated in its own plane.

Parameters for collecting PDP data included:

Angular range: 2 to 42° 2θ;

Step size: 0.05° 2θ; And

- 79 043371

Data acquisition duration: 0.5 s/step (total acquisition time: 6.40 min).

Additional scaled-up synthesis methods Analytical gas chromatography (GC) Analyzes were performed on the following system:

System: Agilent 7890 series gas chromatograph or equivalent.

Column: HP-5, 30 mx0.32 mm, film thickness 0.25 μm (example: J&W, part number: 19091J413).

Oven program: 40°C (hold for 1 min), gradual increase at 10°C/min to 260°C (hold for 5 min).

Injector: 250°C.

Detector: 350°C FID.

Head Pressure: 10 psi (~68.9 kPa), constant pressure.

Carrier gas: Nitrogen.

Flow division: 10:1 (division).

Injection volume: 1 µl.

Liner: SGE FocusLiner with glass wool insert. Thinner: dichloromethane.

Analytical high performance liquid chromatography (HPLC) Analyzes were performed using the following system:

System: Agilent 1100 Series Liquid Chromatograph or equivalent.

Column: Acquity VEN Phenyl 4.6x30 mm; particle size 1.7 microns (eg: Waters No. 186004644).

Injection volume: 5 µl.

Flow rate: 2.0 ml/min.

Detection: Ultraviolet detection at 210 nm.

Column temperature: 40°C.

End of run: 2.3 min.

Solvents: A: water: TFA (100:0.03); B: acetonitrile:TFA (100:0.03)

Gradient: _________________________________

Time (min) A% IN% 0 95 5 5.2 5 95 5.7 5 95 5.8 95 5 6.2 95 5

Mass spectrometry conditions.

Analyzes were carried out on the following system:

System: Bruker Esquire 3000 Plus Ion Trap MS.

Ion polarity: Positive.

Type of ion source: ESI.

Nebulizer: 50 psi (~344.73 kPa).

Dry gas: 10 l/min.

Dry temperature: 350°C.

Target mass: 400 m/z.

Scan range: 50 m/z - 1000 m/z.

- 80 043371

Synthesis scheme 14

RRNZ, SVg4, DHM, , from 0°C to CT N

-----------* Вг vl J Intermediate, conn. 49

°C

Intermediate 47. 1-Methyl-2-oxabicyclo[2.2.2]octan-3-one

A 20 L flanged flask was charged with ethyl 4-oxocyclohexane-1-carboxylate (550 g, 3.23 mol) and THF (5060 ml) under N2. The reaction mixture was cooled to 0°C and 3M MeMgCl in THF (1078 ml, 3.23 mol) was added dropwise at 0-5°C for 30 minutes. The reaction mixture was warmed to room temperature and stirred for 2 hours. TLC (1% EtOAc/DCM) showed that no starting material remained. The reaction mixture was quenched with a mixture of a saturated solution of NH ₄ Cl (2.75 L) and water (5.5 L) at a temperature of <15°C. The product was extracted with EtOAc (5.5 L), the aqueous layer was separated and back-extracted with EtOAc (5.5 L). The organic layers were combined, dried over MgSO ₄ and concentrated in vacuo. 470.9 g of crude product were obtained, which was purified on silica gel (10 kg), eluting with 1% EtOAc/DCM. The pure fractions were concentrated in vacuo to give the title intermediate 47 (104.0 g, 23%) with a purity of >95% by NMR.

Analytical data:

^1H NMR (400 MHz, CDCl3) δ(ppm): 2.62-2.58 (m, 1H), 1.95 - 1.86 (m, 2H), 1.82 - 1.69 (m, 6H), 1.36 (s, 3H).

Intermediate 48. cis-4-(hydroxymethyl)-1-methylcyclohexan-1-ol ho^JLJ

Intermediate 47 1-methyl2-oxabicyclo[2.2.2]octan-3-one (104.0 g, 0.742 mol) and THF (1.5 L) were charged into a 5 L flanged flask under N ₂ atmosphere. The reaction mixture was cooled to 0°C and a solution of 3M LiAlH ₄ (739.8 ml, 2.219 mol) was added dropwise at 05°C for 30 minutes. This reaction mixture was warmed to room temperature and stirred for 1 hour. TLC (1% EtOAc/DCM) indicated that intermediate 47 1-methyl-2oxabicyclo[2.2.2]octan-3-one was consumed. The reaction mixture was quenched with a saturated aqueous solution of Rochelle's salt (0.75 L) to obtain a thick suspension. The suspension was partitioned between EtOAc (0.75 L) and water (1.5 L). The layers were separated and the aqueous layer was back-extracted with EtOAc (0.75 L). The organic layers were combined, dried over MgSO ₄ and concentrated in vacuo to give the title intermediate 48 (86.2 g, crude) >95% pure by NMR and

98.0% according to GC data.

Analytical data:

GC: retention time: 10.0 min; Purity: 98.0%.

¹ H NMR (400 MHz, CDCl ₃ ) δ (ppm): 3.48 (d, J=6.1 Hz, 2H), 1.75-1.57 (m, 4H), 1.47-1, 24 (m, 5H), 1.22 (s, 3H). The two exchangeable protons of this molecule do not appear in this NMR spectrum.

Intermediate 49. cis-4-(bromomethyl)-1-methylcyclohexan-1-ol r^Vi-one

A 2 L flange flask was charged with triphenylphosphine (334.7 g, 1.276 mol) and DCM (1 L) under N ₂ atmosphere. The reaction mixture was cooled to a temperature of 0°C and CBr ₄ (22.2 d, 0.670 mol) was charged in portions at a temperature of 0-5°C for 20 minutes. Intermediate 48 cis-4-(hydroxymethyl)-1methylcyclohexan-1-ol (92.0 g, 0.638 mol) was charged portionwise over 30 min at 05°C. The reaction mixture was warmed to room temperature and stirred for 3 hours, with NMR analysis indicating that ~13% of the starting material remained. Triphenylphosphine (33.5 g, 0.128 mol) was charged and the reaction mixture was stirred for 1 hour at room temperature. NMR analysis indicated that <1% of the starting material remained. Solids were removed by filtration and the filtrate was washed with water (1 L). The layers were separated and the aqueous layer was back-extracted with DCM (1 L). The organic layers were combined, dried over MgSO ₄ and concentrated in vacuo. The crude product was purified on silica (2 kg), loaded into 20% EtOAc/heptane, eluted with 25% EtOAc/heptane. The pure fractions were concentrated in vacuo to give the title intermediate 49 (96.1 g, 73%) with a purity of >95% by NMR and 98.3% by GC.

Analytical data:

GC: retention time: 11.0 min; Purity: 98.3%.

1H NMR (400 MHz, CDCl ₃ ) δ (ppm): 3.29 (d, J=6.7 Hz, 2H), 1.73 - 1.63 (m, 4H), 1.62 - 1.53 (m, 1H), 1.45-1.30 (m, 4H), 1.21 (s, 3H), 1.17 (s, 1H).

Intermediate 50. cis-4-{[(4-bromophenyl)sulfanyl]methyl}-1-methylcyclohexan-1-ol

VVoh /χΑχΤζ

GT

Intermediate 49 cis-4(bromomethyl)-1-methylcyclohexan-1-ol (42.3 g, 0.204 mol), 4-bromothiophenol (38.6 g, 0.204 mol) was charged into a 1 L three-neck flask under _N2 atmosphere. , cesium carbonate (133.1 g, 0.408 mol), MeOH (210 ml) and THF (420 ml). The reaction mixture was heated to 60°C for 2 hours, at which time HPLC showed that 2.2% of 4-bromothiophenol remained. Intermediate 49 cis-4-(bromomethyl)-1-methylcyclohexan-1-ol (0.86 g, 4.15 mmol) was charged into the reaction mixture and the reaction mixture was stirred for an additional 30 minutes at 60°C. HPLC showed that 0.3% 4-bromothiophenol remained. The reaction mixture was cooled to room temperature, filtered and the filtrate was concentrated in vacuo. The residue was partitioned between EtOAc (390 ml) and water (390 ml), the layers were separated and the aqueous layer was again extracted with EtOAc (195 ml). The organic layers were combined, dried over MgSO4 and concentrated in vacuo to give the title intermediate 50 (57.3 g, crude) with a purity of 99.4% by HPLC and >95% by NMR.

Analytical data:

HPLC: retention time: 3.5 min; Purity: 99.4%.

1H NMR (400 MHz, CDCl ₃ ) δ (ppm): 7.39 - 7.35 (m, 2H), 7.17 - 7.13 (m, 2H), 2.81 (d, J=6, 1 Hz, 2H), 1.75 - 1.70 (m, 2H), 1.67 - 1.62 (m, 2H), 1.49 - 1.40 (m, 1H), 1.39 - 1 .33 (m, 4H), 1.20 (s, 3H), 1.13 (s, 1H).

Intermediate 51. cis-4-[(4-bromobenzenesulfonyl)methyl]-1-methylcyclohexan-1-ol

A ₂ L flange flask was charged with intermediate 50 cis-4{[(4-bromophenyl)sulfanyl]methyl}-1-methylcyclohexan-1-ol (56.0 g, 0.178 mol) and DCM (560 ml). The reaction mixture was cooled to 0°C and meta-chloroperbenzoic acid (77%) (79.6 g, 0.355 mol) was charged in portions at 0-5°C for 15 minutes. The reaction mixture was warmed to room temperature and stirred for 8 hours. HPLC analysis indicated that 0.2% of the intermediate 50 cis-4-{[(4-bromophenyl)sulfanyl]methyl}-1-methylcyclohexan-1-ol remained. The reaction mixture was filtered and the filtrate was washed with saturated NaHCO ₃ solution (560 ml + 280 ml) and saturated Na ₂ S ₂ O ₃ solution (560 ml + 280 ml). The organic layers were separated, dried over MgSO ₄ and concentrated in vacuo to give the title intermediate 51 (57.3 g, crude) with a purity of 97.0% by HPLC and >95% by NMR.

Analytical data:

HPLC: retention time: 2.9 min; Purity: 97.0%.

'H NMR (400 MHz, CDCl ₃ ) δ (ppm): 7.78 - 7.74 (m, 2H), 7.71 - 7.67 (m, 2H), 2.99 (d, J=6 ,1 Hz, 2H), 1.98 - 1.85 (m, 1H), 1.76 - 1.68 (m, 2H), 1.64 - 1.58 (m, 2H), 1.52 - 1.33 (m, 4H), 1.19 (s, 3H), 1.10 (br s, 1H).

Intermediate 52. 1-Methyl-cis-4-{[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2yl)benzenesulfonyl]methyl}cyclohexan-1-ol

OH

A 1 L three-neck flask under _N2 atmosphere was charged with intermediate 51 cis-4[(4-bromobenzenesulfonyl)methyl]-1-methylcyclohexan-1-ol (46.5 g, 0.134 mol), potassium acetate (39.4 g , 0.402 mol), bis(pinacolato)diborane (44.2 g, 0.174 mol) and dioxane (460 ml). The flask was sealed and degassed by purging with nitrogen. Bis(triphenylphosphine)palladium(11) dichloride (1.42 g, 2.02 mmol) was charged into the reaction mixture. The flask was sealed and degassed by purging with nitrogen. The reaction mixture was refluxed (100°C) for 2 hours, at which time HPLC showed that 2.2% of intermediate 51 cis-4-[(4-bromobenzenesulfonyl)methyl]-1methylcyclohexan-1-ol remained. The reaction mixture was cooled to room temperature and filtered through celite. The filtrate was concentrated in vacuo to give the title intermediate 52 (98.2 g, 51.0 g active, crude) with a purity of 73.3% by HPLC and 52% by NMR.

Analytical data:

HPLC: retention time: 1.9 min; Purity: 73.3%.

Synthesized compound 25.

2-fluoro-4'-{[cis-4-hydroxy-4-methylcyclohexyl]methanesulfonyl}-[1,1'-biphenyl]-4-carbonitrile (CHMSA-01-A)

Intermediate 52 1-methylcis-4-{[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonyl]methyl was loaded into a 1-liter three-neck flask under _N2 atmosphere. }cyclohexan-1-ol (51.0 g, 0.129 mol), sodium carbonate (41.1 g, 0.388 mol), 4-bromo-3-fluorobenzonitrile (28.8 g, 0.144 mol), dioxane (500 ml) and water (153 ml). The flask was sealed and degassed by purging with nitrogen. Tetrakis(triphenylphosphine)palladium(0) (14.95 g, 0.013 mol) was charged into this reaction mixture. The flask was sealed and degassed by purging with nitrogen. The reaction mixture was refluxed (~90°C) for 11 hours, at which time HPLC showed that 0.9% of intermediate 52 1 -methyl-cis-4-{[4-(4,4,5, 5-tetramethyl-1,3,2-dioxaborolan)-2-yl)benzenesulfonyl]methyl}cyclohexan-1-ol. This reaction mixture was cooled to room temperature, filtered and concentrated in vacuo. The residue was dissolved in EtOAc (510 ml) and washed with water (510 mlx2). The aqueous layer was separated and back-extracted with EtOAc (510 ml). The organic layers were combined, dried over MgSO ₄ and concentrated in vacuo. 75.5 g of crude product were obtained, which was purified on silica gel (10 kg), eluting with 40-60% EtOAc/heptane. Pure fractions containing product were concentrated in vacuo to give the title intermediate 25 (26.1 g, 52%) with a purity of 97.9% by HPLC and >95% by NMR.

Analytical data:

HPLC: retention time: 3.1 min; Purity: 97.9%.

LCMS (ESI): m/z=370.10 [M-H ₂ O+1]+.

'H NMR (400 MHz, DMSO-d6) δ (ppm): 8.05 - 7.99 (m, 3H), 7.87 - 7.77 (m, 4H), 3.93 (s, 1H) , 3.25 (d, J=6.1 Hz, 2H), 1.77 - 1.63 (m, 1H), 1.56 - 1.48 (m, 2H), 1.48 - 1.33 (m, 4H), 1.23 - 1.12 (m, 2H), 1.01 (s, 3H).

- 83 043371

Intermediate methylcyclohexan-1-ol

Synthesis scheme 15

compound 53. cis-4-{[(4-bromo-3-fluorophenyl)sulfanyl]methyl}-1-

Intermediate 49 cis-4(bromomethyl)-1-methylcyclohexan-1-ol (35.5 g, 0.171 mol), 4-bromo-3-fluorobenzenethiol (35.5 g, 0.171 mol), cesium carbonate (111.7 g, 0.343 mol), MeOH (142 ml) and THF (284 ml). The reaction mixture was heated to 60°C for 2 hours, at which time HPLC showed that 3.5% of 4-bromo-3fluorobenzenethiol remained. Intermediate 49 cis-4-(bromomethyl)-1methylcyclohexan-1-ol (2.23 g, 10.77 mmol) was charged into the reaction mixture and the reaction mixture was stirred for an additional 30 minutes at 60°C. HPLC showed that 0.3% 4-bromo-3-fluorobenzenethiol remained. The reaction mixture was cooled to room temperature, filtered and the filtrate was concentrated in vacuo. The residue was partitioned between EtOAc (360 ml) and water (360 ml), the layers were separated and the aqueous layer was extracted again with EtOAc (180 ml). The organic layers were combined, dried over MgSO ₄ and concentrated in vacuo to give the title intermediate 53 (54.9 g, crude) with a purity of 97.6% by HPLC and >95% by NMR.

Analytical data:

HPLC: retention time: 3.6 min; Purity: 97.6%.

1H NMR (400 MHz, CDCl3) δ (ppm): 7.41 - 7.36 (m, 1H), 7.02 (dd, J=2.1, 9.5 Hz, 1H), 6.94- 6.90 (m, 1H), 2.82 (d, J=6.7 Hz, 2H), 1.74 - 1.69 (m, 2H), 1.68 - 1.62 (m, 2H) , 1.50 - 1.42 (m, 1H), 1.41 - 1.33 (m, 4H), 1.20 (s, 3H). The exchangeable protons of this molecule are not visible in this NMR spectrum.

Intermediate 54. cis-4-[(4-bromo-3-fluorobenzenesulfonyl)methyl]-1methylcyclohexan-1-ol

Intermediate 53 cis-4{[(4-bromo-3-fluorophenyl)sulfanyl]methyl}-1-methylcyclohexan-1-ol (54.0 g, 0.162 mol) was charged into a 2 L flanged flask under _N2 atmosphere. and DXM (540 ml). The reaction mixture was cooled to 0°C and meta-chloroperbenzoic acid (77%) (79.9 g, 0.356 mol) was charged in portions at 0-5°C for 15 minutes. The reaction mixture was warmed to room temperature and stirred for 8 hours. HPLC analysis showed that 0.9% of intermediate 53 cis-4-{[(4-bromo-3-fluorophenyl)sulfanyl]methyl}-1-methylcyclohexane- 1- 84 043371 ola. The reaction mixture was filtered and the filtrate was washed with saturated NaHCO ₃ solution (540 ml +

270 ml) and a saturated solution of Na ₂ S ₂ O ₃ (540 ml + 270 ml). The organic layers were separated and dried over

MgSO ₄ and concentrated in vacuo to give the title intermediate (50.1 g, crude) with a purity of 98.2% by HPLC and >95% by NMR.

Analytical data:

HPLC: retention time: 3.0 min; Purity: 98.2%.

^1H NMR (400 MHz, CDCl ₃ ) δ (ppm): 7.77 - 7.72 (m, 1H), 7.64 - 7.60 (m, 1H), 7.57 - 7.53 (m , 1H), 2.99 (d, J=6.7 Hz, 2H), 1.94 - 1.83 (m, 1H), 1.72 - 1.65 (m, 2H), 1.63 - 1.56 (m, 2H), 1.50 - 1.30 (m, 5H), 1.16 (s, 3H).

Intermediate 55. cis-4-{[3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2yl)benzenesulfonyl]methyl}-1-methylcyclohexan-1-ol

A 1 L three-neck flask under _N2 atmosphere was charged with intermediate 54 cis-4[(4-bromo-3-fluorobenzenesulfonyl)methyl]-1-methylcyclohexan-1-ol (45.0 g, 0.123 mol), potassium acetate ( 36.3 g, 0.370 mol), bis(pinacolato)diborane (40.7 g, 0.160 mol) and dioxane (450 ml). The flask was sealed and degassed by purging with nitrogen. Bis(triphenylphosphine)palladium(II) dichloride (1.3 g, 1.85 mmol) was charged into the reaction mixture. The flask was sealed and degassed by purging with nitrogen. The reaction mixture was refluxed (100°C) for 2 hours, with HPLC showing that 3.3% of intermediate 54 cis-4-[(4-bromo-3fluorobenzenesulfonyl)methyl]-1-methylcyclohexane-1 remained -ola. The reaction mixture was charged with bis(triphenylphosphine)palladium(II) dichloride (0.4 g, 0.57 mmol) before stirring at reflux (100°C) for 45 min, with no significant change in the HPLC profile. Reaction the mixture was cooled to room temperature and filtered through celite.The filtrate was concentrated in vacuo to give the title intermediate 55 (94.5 g, 35.9 g active, crude) with a purity of 90.0% by HPLC and 38% by NMR data.

Analytical data: HPLC: retention time: 1.9 min; Purity: 90.0%.

Synthesized compound 26.

cis-4-{[4-(3,5-difluoropyridin-2-yl)-3-fluorobenzenesulfonyl]methyl}-1-methylcyclohexan-1-ol (CHMSA-03-A)

Intermediate 55 cis4-{[3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonyl]methyl was charged into a 500 ml three-neck flask under _N2 atmosphere. } -1methylcyclohexan-1-ol (25.9 g, 0.063 mol), 2-bromo-3,5-difluoropyrdine (12.8 g, 0.066 mol), potassium carbonate (26.1 g, 0.189 mol), THF ( 260 ml) and water (78 ml). The flask was sealed and degassed by purging with nitrogen. (1,1'-bis(diphenylphosphino)ferrocene)dichloropalladium(II) (4.6 g, 6.3 mmol) was charged into the reaction mixture. The flask was sealed and degassed by purging with nitrogen. The reaction mixture was refluxed (65°C) for 1 hour, at which time HPLC showed that 0.9% of intermediate 55 cis-4-{[3-fluoro-4-(4,4,5,5 -tetramethyl-1,3,2dioxaborolan-2-yl)benzenesulfonyl]methyl}-1-methylcyclohexan-1-ol, and 2-bromo-3,5-difluoropyrdine were not observed. The reaction mixture was cooled to room temperature and filtered through a pad of celite. The filtrate was concentrated in vacuo to obtain 1.68 g of crude material. This crude material was combined with crude material from another similar reaction using 2 g of intermediate 55 cis-4-{[3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2yl )benzenesulfonyl]methyl}-1-methylcyclohexan-1-ol. The combined crude materials were purified on silica (50 eq.), eluting with 1% MeOH/DCM, to obtain 20.1 g of the title synthesized compound 26 with a purity of 97.0% by HPLC, but NMR of the isolated material showed some impurities, bound to cyclohexane and the remaining intermediate 55 cis-4-{[3-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonyl]methyl}-1methylcyclohexane- 1-olom. The material was combined with 6 g of crude material isolated from the mixed fractions and purified a second time on silica (50 eq.), eluting with 40-50% EtOAc/heptane. The pure fractions were concentrated in vacuo to give the title synthesized compound 26 (10.3 g, 38%) with a purity of 98.9% by HPLC and >95% by NMR.

Analytical data:

HPLC: retention time: 2.9 min; Purity: 98.9%.

LCMS (ESI): m/z=382.2 [M-H ₂ O+1]+.

1H NMR (400 MHz, DMSO-d ₆ ) δ (ppm): 8.70 (d, J=2.4 Hz, 1H), 8.19 - 8.12 (m, 1H), 7.95 - 7 .91 (m, 1H), 7.90 - 7.83 (m, 2H), 3.94 (s, 1H), 3.34 (d, J=6.7 Hz, 2H), 1.78 - 1.65 (m, 1H), 1.56 - 1.49 (m, 2H), 1.48 - 1.34 (m, 4H), 1.24 - 1.14 (m, 2H), 1, 02 (s, 3H).

Biological research

Biological study 1. Analysis of ATP release in monocytes.

The in vitro activity of test compounds was determined by incubation with human monocytic Thp1 cells followed by determination of adenosine triphosphate (ATP) levels using firefly luciferase.

ATP is present in all metabolically active cells. When cells lose integrity, their ability to synthesize ATP is quickly lost. Consequently, ATP concentration decreases when cells undergo necrosis or apoptosis, and its concentrations are commonly used as a marker of cell viability or cell proliferation. See, for example, Kang et al., 2015; Jiang et al., 2013. ATP levels can be monitored using a firefly (Photinus pyralis) luciferase-based system (see e.g. Auld et al., 2009) using commercially available kits. A system known as ATPIite™ was used to measure the effects of test compounds on cell viability in vitro. This one-step assay system is an adenosine triphosphate (ATP) monitoring system based on the light release caused by the reaction of ATP from cells with added luciferase and D-luciferin, as shown in the reaction diagram below:

MD ²⁺

Luciferin + ATP + O ₂ ------------* Oxyluciferin + AMP+

Luciferase CO2 + light

The light emitted is proportional to the ATP concentration.

Thp1 cells were seeded at 112,500 cells per well in 125 μl RPMI-1640 (without glucose) with 1% FBS in 96-well plates. Test compounds were prepared as 100 mM solutions in DMSO. These stock solutions were diluted in DMSO and then diluted 1000-fold in culture medium (RPMI) before adding directly to the wells to obtain the desired final compound concentration. After 24 hours incubation at 37°C/5% CO2, ATPLite™ (Perkin Elmer) (1:10 v/v, 10 μl) was added to each well. The plate was then incubated at room temperature for 5 min and the emitted light was quantified on a Viewlux with a measurement time of 0.3 s and a 4x4 sort.

The average results for each test compound were expressed as a percentage (%) of the control average; they reflected cell viability. The average values for the tested concentrations were then plotted and the IC ₅₀ was calculated by fitting the data to a four-parameter IC ₅₀ equation using software from Grafit (Erithacus Software). Each experiment was repeated twice and data are presented as the average IC ₅₀ value from both experiments.

The results are presented in the following table.

- 86 043371

Table 3

Analysis of ATP in Thpl monocytes

Compound 1С ₅ о (µM) NMS-S-01-A 0.63 ABD899 0.2 ABD900 1.1 CHMSA-01-A 0.7 CHMSA-01-B 4.4 CHMSA-02-A 0.1 CHMSA-02-B 0.3 CHMSA-03-A 0.7 CHMSA-03-B 5.4 CHMSA-04-A 0.98 CHMSA-04-B 2.4 CHMSA-05-A 0.8 CHMSA-05-B 2.7 CHMSA-06-A 1.8 CHMSA-06-B 4.5 CHMSA-07-A 1.3 CHMSA-07-B 2.5 CHMSA-08-A 0.2 CHMSA-08-B 0.4 CHMSA-09-A 0.3 CHMSA-09-B 0.5 CHMSA-10-A 1.63 CHMSA-10-B 1.96 CHMSA-11-A 0.5 CHMSA-11-B 0.8 CHMSA-12-A 1.8 CHMSA-12-B 3.2

⁽¹⁾ Obtained using a 9-point concentration range from 10 µM to 10 nM with n=2 parallel runs per concentration. Data represent the mean of 2 independent experiments.

The data demonstrate that many of the CHMSA compounds described herein, and in particular compounds CHMSA-02-A, CHMSA-02-B, CHMSA-08-A and CHMSA-09-A, exhibit high activity in the ATP assay in Thp1 monocytes, and also do not lose activity compared to reference compounds.

Biological study 2. Study of human and rat hepatocytes.

The metabolic stability of the test compounds was measured by determining the rate of disappearance of the compound when incubated in the presence of rat or human hepatocytes, the main source of the most important enzymes (cytochrome P450) involved in drug metabolism. Drug stability studies in the presence of primary hepatocytes are considered a valuable model to rapidly predict drug stability in vivo.

Rat or human hepatocytes were obtained from commercial sources and their viability was assessed using trypan blue solution before use. Test compounds (final concentration 1 μM, 0.1% DMSO, 0.9% acetonitrile) or marker (diclofenac or diltiazem, final assay concentration 1 μM, 0.1% DMSO, 0.9% acetonitrile) were incubated with pooled hepatocytes for 60 min, samples were collected at 6 time points and analyzed by LCMS/MS for the presence/amount of test compounds.

Each compound was incubated for 0, 5, 15, 30, 45, or 60 min. Reactions were stopped by adding methanol containing internal standard (1 μM tolbutamide) at appropriate times, stirred, and placed at −20°C for <1 h to quench and allow protein to precipitate. All samples were centrifuged (2500xg, 20 min, 4°C). Aliquots were analyzed by LCMS/MS. Reactions were carried out in duplicate at a temperature of 37°C.

The data was processed and the results were displayed as In (concentration) versus time. The elimination rate constant (slope of the regression line, k) was calculated using the following formula, where C(t) is the concentration at time t and C(0) is the initial concentration:

- 87 043371

In C(0) - In C(t) k = -------------t

The half-life ( _t1 / ₂ ) was calculated using the following formula:

In 2

C/2 “k

Intrinsic clearance (Clint) was calculated using the following formula, where [cell] is the concentration of hepatocytes in the analysis:

Cl _int = k

[cell]

The data is summarized in the following table.

Table 4 Hepatocyte stability

Compound ti/2 in rat (min) Clint in rat (µl/min/million cells) tl/2 У person (min) Clmt in humans (µl/min/million cells) NMS-S-01-A 7 188 154 7.6 ABD599 112 22 ABD899 24 57 149 9 ABD900 17.9 79 220 6.3 CHMSA-01-A 106 12.4 354.9 3.7 CHMSA-01-B 297.1 4.5 CHMSA-02-A 13 102.3 >412.2 <3.3 CHMSA-02-B 122.1 10.8 CHMSA-03-A 91.2 14.4 >460.0 <3.0 CHMSA-03-B >460.0 <3.0 CHMSA-04-A 47.1 37.4 27.5 40.6 CHMSA-04-B 31.4 51.3 >460 <3.0 CHMSA-05-A 396.8 3.3 CHMSA-05-B 227.4 6.5 CHMSA-08-A 73.2 18.0 CHMSA-08-B 138.0 9.8 CHMSA-09-A 112.3 12.1 CHMSA-09-B 25.0 52.9 CHMSA-10-A 10.7 149.2 37 30.8 CHMSA-10-B 4.8 336.2 195.1 5.8 CHMSA-11-A 172.3 7.8 CHMSA-11-B 99.8 13.3 CHMSA-12-A >436.5 <3.1 CHMSA-12-B >441.5 <3.1

The data demonstrate that many of the CHMSA compounds described herein demonstrated greater metabolic stability than the reference compounds, with compounds CHMSA-02-A, CHMSA-03-A, CHMSA-03-B, CHMSA-12-A and CHMSA-12-B showed exceptionally good stability.

Biological study 3. Water solubility.

Aqueous solubility was measured by equilibrating compounds in fasting artificial intestinal fluid (FaSSIF) and quantifying them by spectrophotometry.

FaSSIF was prepared as described below:

Preparation of empty FaSSIF: 0.21 g of sodium hydroxide (NaOH) granules, 1.97 g of sodium dihydrogen phosphate (NaH ₂ PO ₄ -H ₂ O) and 3.09 g of sodium chloride (NaCl) were dissolved in 400 ml of deionized water. The pH was adjusted to 6.5 with 1 M hydrochloric acid and additional deionized water was added to a final volume of 500 mL.

Preparation of FaSSIF: 0.056 g of SIF powder (containing sodium taurocholate and lecithin) (Phares AG) was dissolved in 25 ml of empty FaSSIF solution and stirred until the powder was completely dissolved. The solution was allowed to stand for 2 hours, during which it became opalescent; it was used in

- 88 043371 for 24 hours. The final composition of the solution was characterized as follows:

Sodium taurocholate: 3 mM.

Lecithin: 0.75 mM.

Osmolarity: 270±10 mOsmol.

pH: 6.5.

Water solubility was determined by adding a known concentration of test compound (dissolved in DMSO) to FaSSIF followed by incubation for 16 hours. Optical density was measured at the end of the incubation period for test compounds and a standard was used to determine solubility. Briefly, two samples were prepared for each determination: a reference sample consisting of a stock solution of the test compound in DMSO diluted with system solution (low absorbance phosphate-free buffer) and propanol; and a test sample (prepared in triplicate) consisting of 0.5 ml FaSSIF supplemented with 0.2 mM test compound. Each sample was incubated at room temperature for 16 h with constant shaking at 250 rpm. At the end of the incubation period, 0.3 ml of each sample was filtered through a pION filter plate (PION, Woburn, MA, USA), diluted 1:1 with propanol and scanned by UV spectrophotometry at λ _max (190–400 nM) using Spectra Max Plus version 2.1000 (Molecular Devices, Sunnyvale, CA, USA) with μSOL Explorer solubility software (pION, Woburn, MA, USA).

Solubility in FaSSIF was calculated using the following formula:

150

Solubility ^mg in FaSSIF ml

Sample OD * Cg * ^mog,ek U ^lya R ^naya

OD standard mass where OD is optical density;

Cr - standard concentration (33.4 µM); and the molecular weight is relative to the test compound (eg 381.44 for ABD735).

The data is summarized in the following table.

Table 5

Solubility in FaSSIF

Compound Solubility (mg/ml) ⁽¹⁾ Solubility (mg/ml) ⁽²⁾ NMS-S-01-A 0.06 ABD599 0.03 ABD735 0.02 ABD836 0.03 ABD899 0.06 0.13 ABD900 0.12 REF001 0.05 CHMSA-01-A 0.01 CHMSA-02-A 0.02 CHMSA-03-A 0.07 CHMSA-04-A >0.07 CHMSA-04-B 0.04

⁽¹⁾ Each study was carried out in three parallel experiments at pH 6.5.

⁽²⁾ Each study was carried out in two parallel experiments at pH 6.8.

The data demonstrated that the CHMSA compounds described herein have solubility equivalent to that of the reference compounds.

Biological study 4. Identification of metabolites.

To determine the propensity of compounds to form a biaryl metabolite, the formation of metabolites in rats was assessed.

Appropriate sulfonamide compounds (eg reference compound NMC-S-01-A) produce a sulfonamide metabolite that is persistent and has a long half-life. In addition, the biarylsulfonamide metabolite acts as a metabolic inducer in rats, which may make toxicity assessment in rodents difficult. Therefore, the lower the propensity to form a biarylsulfonamide metabolite, the greater the compound's potential suitability for development as a drug for use in humans.

The test compound was administered to Han Wistar rats (8-12 weeks old) and plasma samples were collected. Test compounds at a dose of 1 mg/kg were administered intravenously as a solution in 5% NMP, 5% Solutol HS and 90% saline. Animals were given free access to food throughout the study. Blood samples were collected under light isoflurane anesthesia at 12 time points.

- 89 043371 points after intravenous administration (0.033, 0.1, 0.167, 0.25, 0.5, 1, 2, 4, 6, 8, 12 and 24 hours). Blood samples were collected from a group of three rats at each time point and placed in a labeled microcentrifuge tube containing K2EDTA as an anticoagulant. Plasma samples were separated by whole blood centrifugation.

All samples were processed for protein precipitation analysis using acetonitrile and analyzed by a suitable LC-MS/MS method.

Upon completion of the study, the results were expressed in the form of metabolite detection and formation dynamics.

In fig. 1 shows a graph of plasma concentration (ng/ml) versus time after dosing (h) for the reference compound NMC-S-01-A (dark round marks) and the corresponding biarylsulfonamide metabolite (MET-001) (light round marks), obtained using the methods described herein. The metabolite was formed in large quantities and accumulated over time.

Table 6

Reference compound NMC-S-01-A and biarylsulfonamide metabolite (MET-001)

In fig. 2 is a plot of plasma concentration (ng/mL) versus time post-dose (h) for compound CHMSA-10-B (closed circle marks) and the corresponding biarylsulfonic acid metabolite (MET-002) (open circle marks), obtained with using the methods described herein. The metabolite was detected temporarily, 0.5 hours after administration.

Table 7

CHMSA-10-B and metabolite biarylsulfonic acid (MET-002)

While the reference compound HMC-C-01-A produced the biarylsulfonamide metabolite (MET001) in large quantities (which accumulated over time), the compound CHMSA-10B did not produce the biarylsulfonamide metabolite (MET001), and the biarylsulfonic acid metabolite (MET002) was detected only temporarily.

In summary, the data demonstrate that the CHMSA compounds described herein demonstrate significantly increased potential for development as a drug for human use compared to the reference compound (HMC-C-01-A).

Biological Study 5. hERG Ion Channel Assay.

Inhibition of the human cardiac specific potassium channel gene (hERG) ion channel mediates the repolarizing current IKr in the cardiac muscle action potential, indicating that it contributes to the electrical activity coordinating the heartbeat. When the ability of hERG to conduct electrical current across the cell membrane is suppressed or impaired, it can lead to a potentially fatal disorder called long QT syndrome. Because of this association between hERG and long QT syndrome, it is important to avoid hERG inhibition in drug development.

The activity of the compounds on the hERG ion channel was tested using two approaches: a binding assay and an automated patch-clamp system, the Qpatch method using stably transfected Chinese hamster ovary (hERG-CHO) cells. hERGCHO cells were cultured in Kaighn's Nutrient Mixture F12 (Invitrogen) + 10% FBS at 37°C for 1–3 days. Cells were maintained at 30°C for 24 to 48 h before applying the patch-clamp system to increase hERG current amplitude. Cells were subsequently harvested by trypsinization and stored in serum-free medium (SFM) in a Qpatch cell preparation compartment for up to 6 h at room temperature, then washed, resuspended in extracellular solution and transferred to a patch-clamp system for data recording.

Patch-clamp voltage protocol: After reaching the whole cell configuration, the cell was maintained at -80 mV. To measure the leakage current, a 50 ms pulse to -40 mV was applied and subtracted from the residual on-line current. The cell was then depolarized to +20 mV for 2 s, followed by a one-second pulse to −40 mV to detect residual hERG current. This pattern was applied every 5 s to monitor the current amplitude.

Extracellular solution: 137 mM NaCl, 4 mM KCl, 1.8 mM CaCl ₂ , 1 mM MgCl ₂ , 10 mM D(+)glucose, 10 mM HEPES buffer (pH adjusted to 7.4 with NaOH).

After the whole cell configuration was achieved, the extracellular solution (control) was first applied and the cell was stabilized for 2 min in the extracellular solution. The test compound was then applied cumulatively from low to high concentrations. The cell was incubated with each concentration tested for 5 min. During each incubation, the cell was repeatedly stimulated using the voltage protocol described above, and the amplitude of the residual current was continuously monitored.

Eligibility Criteria:

(1) Peak residual current >100 pA in control.

(2) Initial run-down <30% and stop before first application of test compound.

(3) Leakage currents <50% of reference residual current peaks at any time.

(4) rs <20 MΩ throughout the experiment.

The degree of inhibition (%) was obtained by measuring the amplitude of the residual current induced by a one-second test pulse to -40 mV after a two-second pulse to +20 mV, before and after incubation with the test compound. The difference in current was normalized to the control and multiplied by 100 to obtain the percentage inhibition.

The concentration (log) response curves were fitted with a logistic equation (three parameters assuming complete blocking of current at very high concentrations of the test compound) to obtain estimates of the 50% inhibitory concentration (IC ₅₀ ). The concentration-response relationship for each compound was plotted based on the percentage decrease in current amplitude at successive concentrations.

The results are presented in the following table.

Table 8

hERG ion channel activity

⁽¹⁾ IC ₅₀ was calculated using a four-parameter logistic equation automatically calculated in Grafit version 6.0.12 (Erithacus Software Ltd., Dr. Robin Leatherbarrow).

The data demonstrate that the CHMSA compounds described herein have the cardiac safety properties required for an orally active drug and have safety advantages over reference compounds such as ABD599 and ABD899, with CHMSA- 02-A and CHMSA-03-A show a particularly favorable profile.

Biological Study 6. Human Cytochrome P450 Inhibition Assay.

Inhibition of cytochrome P450 (CYP450) enzymes is a major cause of drug-drug interactions in clinical use and can complicate or halt the development of new drugs.

The ability of test compounds to inhibit the five most significant cytochrome P450 enzymes was measured by determining the activity of cytochrome P450 enzymes in recombinant cytochrome preparations called Bactosomes (Surech Ltd, Dundee, Scotland, UK DD2 1NH) in the presence of a specific marker substrate. Bactosomes are a highly efficient and economical source of recombinant CYP450s that have a higher specific enzyme activity compared to other sources such as liver microsomes. If a compound inhibits enzyme activity, the rate at which the marker substrate disappears is reduced. The following CYP450 isoforms were analyzed: CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4. Studies of the CYP450 inhibition potential of bactosomes have been accepted as a valuable model for rapidly predicting potential drug-drug interactions in vivo (see, e.g., Weaver et

- 91 043371 al., 2003).

Bactosomes were obtained from a commercial source (Sureh, Scotland, UK). Test compounds were incubated with bactosomes at 6 concentrations. Samples were incubated for 10 min, after which the reaction was stopped and samples were analyzed by LC-MS/MS multiple reaction monitoring (MRM) for the presence/amount of marker substrate.

CYP450 enzymes (final protein 75 pmol/ml for CYP1A2; 12.5 pmol/ml for CYP2C19; and 25 pmol/ml for CYP2C9, 2D6 and 3A4), 0.1 M phosphate buffer pH 7.4, marker and test compound (final concentration 50, 15.8, 5, 1.58, 0.5 and 0.158 μM; diluted from a 10 mM stock solution to give a final DMSO concentration of 1%) were preincubated at 37°C for 5 min. The reaction was initiated by adding 20 μl of 10 mM NADPH in phosphate buffer. The final incubation volume was 200 μl. For each CYP450 inhibition assay, the following control inhibitors were used: CYP1A2: α-naphthoflavone; CYP2C9: sulfaphenazole; CYP2C19: tranylcypromine; CYP2D6: quinidine; CYP3A4: ketoconazole.

Each compound was incubated for 10 min at 37°C. The reactions were stopped by the addition of methanol (final composition 1:1, aq:methanol). Incubation plates were shaken, cooled at 20°C for 2 hours, and centrifuged at 3500 rpm for 15 minutes at 4°C to pellet the protein. The supernatant was then transferred into vials for analysis using MC/MRM under the conditions shown in the following table.

Table 9

MS terms

HPLC: Waters Alliance 2790 MS/MS: Triple Quadrupole Quattro Ultima (Micromass, Manchester) Software: Analyst 1.5 Ionization mode: IER+ Scan mode: Multiple reaction monitoring (MRM) Column: Devosil C30 Column temperature (°C): 40 Phase A: 0.1% formic acid in water Phase B: 0.1% formic acid in methanol Gradient 97% A (0 - 0.3 min), 5% A (0.55 - 1.55 min), 97% A (1.6 min) Stop time 2.5 min Injection volume (µl): thirty Flow rate (ml/min): 1.2

IC ₅₀ values were determined by linear transformation in Microsoft Excel. The data is summarized in the following table.

Table 10 Human CYP450 inhibition

Compound IC50 (µM) CYP1A2 CYP2C9 CYP2C19 CYP2D6 CYP3A4 ABD899 >25 3.9 7.3 45.3 21.6 NMS-S-01-A 25 21 >25 16.6 >25 NMS-S-08-A 27 6.7 thirty 19 29 NMS-S-09-A 23 34 >50 >50 33 NMS-S-10-V >16 2.4 8.5 >16 9.2 NMS-S-11-A eleven 2.7 5.1 9.3 12 HMC-N-05-A 36 27 >50 >50 >50 CHMSA-01-A >50 39.4 >50 >50 >50 CHMSA-02-A >50 >50 >50 >50 >50 CHMSA-02-B >10 >10 >10 >10 >10 CHMSA-03-A >50 14.7 >50 >50 >50 CHMSA-05-A >50 7 36.6 >50 26.3 CHMSA-08-B >10 >25 >25 >10 ND CHMSA-09-A >25 >25 25 >25 >25 CHMSA-11-A >10 >10 25 >10 ND

ND: not defined.

- 92 043371

The data shows that the CHMSA compounds described herein exhibit reduced drug interaction potential compared to the reference compounds, with compounds CHMSA-01-A, CHMSA-02-A and CHMSA-03-A showing a particularly good profile .

Biological study 7. Collagen-induced arthritis in mice.

Seven to eight week old male DBA/1j mice were used for all procedures. Animals were housed in groups of 10 and maintained at 21°C ± 2°C with 12-hour light/dark cycles and free access to food and water. Complete Freund's adjuvant (CFA) was prepared by emulsifying bovine type II collagen at a concentration of 4 mg/ml with a 4 mg/ml suspension of Mycobacterium tuberculosis H37Ra in incomplete Freund's adjuvant (IFA) (0.85 ml paraffin oil and 0.15 ml mannid monooleate ) in a ratio of 1:1 (v/v). All mice were immunized subcutaneously with 200 μg of bovine type II collagen in CFA. 21 days later, all mice were immunized subcutaneously with 100 μg of bovine type II collagen in IFA. The mice began to develop signs and symptoms of arthritis after the booster immunization.

For macroscopic assessment of arthritis, the following signs were monitored on each paw of each mouse three times per week and summed to obtain the Arthritis Index (AI) (maximum AI for one animal is 16):

= no visible signs of arthritis.

= swelling and/or erythema of 1 finger.

= swelling and/or erythema of 2 fingers.

= swelling and/or erythema of more than 2 fingers.

= severe arthritis of the entire paw and toes.

Animals were assigned to treatment groups with a mean arthritis index of 2.5 and then administered once daily for 14 days with either test compounds via gavage or the positive control etanercept by subcutaneous injection at a dose of 10 mg/kg. After completion of the experiment, the mice were sacrificed.

Data were analyzed by obtaining the mean Arthritis Index score for each treatment group. The mean arthritis index was then compared to the arthritis index of control (untreated) animals using the following formula to obtain the percentage of disease inhibition.

- average arthritis index: animals % inhibition ₌ _ from the disease treatment group average arthritis index: animals not treated * 100

The data is summarized in the following table.

Table 11 ________Inhibition of arthritis

Compound Dose (mg/kg/day) Disease inhibition, % ABD899 10 77 NMS-S-01-A 10 40 HMC-N-01-A 10 45 NMS-S-02-A 10 61 HMC-N-02-A 10 36 NMS-S-01-V 10 26 HMC-N-01-B 10—1 (*) 38 CHMSA-01-A 10 63 CHMSA-03-A 10 62

(*) Reduced from 10 to 1 mg/kg/day due to mortality.

In fig. 3 is a graph of the mean arthritis index versus time (day of dosing) for test compound CHMSA-01-A at 10 mg/kg/day by gavage (light circles) and control (dark circles).

In fig. 4 is a graph of the mean arthritis index versus time (day of dosing) for test compound CHMSA-03-A at a dose of 10 mg/kg/day by gavage (light circles) and control (dark circles).

In fig. Figure 5 is a graph of the Arthritis Index versus time (day of dosing) for the reference compound ABD899 at 10 mg/kg/day (open circles, open squares), control (dark circles), and marketing-registered positive control. etanercept (triangular marks).

In fig. Figure 6 shows a graph of the arthritis index versus time (day of dosing) for the reference compound NMC-S-01-A at a dose of 10 mg/kg/day (light round marks) and control (dark round marks).

- 93 043371

These data demonstrate that the CHMSA compounds described herein, when administered orally, exhibit excellent in vivo activity in preventing the progression of established severe arthritis.

Reference connections.

The following reference compounds are mentioned above herein.

Table 12

Reference connections

The principles, preferred embodiments and modes of carrying out the present invention have been described above. However, the present invention should not be construed as limited to the specific embodiments discussed. To the contrary, the above-described embodiments should be considered illustrative and not limiting. It should be understood that changes to these embodiments may be made by those skilled in the art without departing from the scope of the present invention.

Bibliography

A number of publications are cited herein to more fully describe and disclose the invention and the prior art to which the invention relates. Full links to these publications are provided below.

Each of these publications is incorporated herein by reference in its entirety to the same extent as if each individual publication had been separately identified.

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Claims (18)

Steger etal., 2011, “Denosumab for the treatment of bone metastases in breast cancer: evidence and opinion,” Ther. Adv. Med. Oncol., Vol. 3, pp. 233243. Straub etal., 2010, “Energy regulation and neuroendocrine-immune control in chronic inflammatory diseases”, J. Intern. Med., Vol. 267, No. 6, pp. 543-560. Sun, 2010, “Mechanical loading, cartilage degradation and arthritis,” Annals of the New York Academy of Sciences, Vol. 1211, pp. 37-50. Takayanagi, 2009, “Osteoimmunology and the effects of the immune system on bone”, Nature Reviews Rheumatology, Vol. 5, pp. 667-676. Tanaka et al., 2003, “Signal transduction pathways regulating osteoclast differentiation and function”, J. Bone Miner. Metab., Vol. 21, pp. 123-133. Weinberg et al., 2010, “Mitochondrial metabolism and ROS generation are essential for Kras mediated tumorigenicity,” Proc. Natl. Acad. Sci. USA, Vol. 107, No. 19, pp. 87888793. Weaver, et al., 2003, “Cytochrome p450 inhibition using recombinant proteins and mass spectrometry/multiple reaction monitoring technology in a cassette incubation”, Drug Metabolism and Disposition, Vol. 31, No. 7, pp. 955-966. Weyandeta/., 2017a, “Immunometabolism in early and late stages of rheumatoid arthritis”, Nature Reviews Rheumatology, Vol. 13, pp. 291-301. Weyandeta/., 2017b, “Metabolic Signatures of T-cells and Macrophages in Rheumatoid Arthritis”, Curr. Opin. Immunol., Vol. 46, pp. 112-120. Wheaton et al., 2014, “Metformin inhibits mitochondrial complex I of cancer cells to reduce tumorigenesis,” eLife, Vol. 3, e02242. Yang et al., 2013, “Phosphofructokinase deficiency impairs ATP generation, autophagy, and redox balance in rheumatoid arthritis T cells,” J. Exp. Med., Vol. 210, pp. 21192134. CLAIM 1. Compound of the following formula: keel; keel; or a pharmaceutically acceptable salt thereof; where =X- independently represents -CH= or -N=; - R ¹ is independently H or -R ^1X ; - R ^1X is independently -F, -Cl or -CN; - R ² is independently -H or -R ^2X ; - R ^2x is independently -F, -Cl or -CN; - R ³ is independently -H or -R ^3X ; - R ^3X is independently -F, -Cl, -R ^3C , -R ^3F or -CN; - R ^3C independently represents a saturated straight or branched C _1-3 alkyl; - R ^3f is independently a saturated straight-chain or branched C _1-3 fluoro- - R ⁴ is independently -H or -R4X; - R4X is independently -F, -Cl, -R ^4C , -R ^4F or -CN; - R ^4C independently represents a saturated straight or branched C _1-3 alkyl; - R ^4f is independently a saturated straight-chain or branched C _1-3 fluoro- - R ⁵ represents -H; And - R ⁶ represents -H. 2. The compound according to claim 1, in which =X- represents -CH=. 3. The compound according to claim 1, in which =X- represents -N=. 4. The connection according to any one of claims 1-3, in which - R ^3C represents -CH ₃ ; - R ^3F represents -CF3; - R ^4C represents -CH3; And - R ^4F is -CF ₃ . - 98 043371 5. The connection according to any one of claims 1-4, in which -R ¹ represents -R ^1X ; And -R ² is -R ^2X . 6. The connection according to any one of claims 1-5, in which -R ³ is -R ^3X ; And -R ⁴ represents -H. 7. The connection according to claim 6, in which -R ^3X is independently -F, -Cl or -CN. 8. The connection according to any one of claims 1-5, in which -R ³ represents -H; And -R ⁴ is -R ^4X . 9. The connection according to claim 8, in which -R ^4X is -CF ₃ . 10. The connection according to any one of claims 1-5, in which -R ³ represents -H; And -R ⁴ represents -H. 11. A compound according to any one of claims 1 to 10, which is a compound of the following formula or a pharmaceutically acceptable salt thereof: 12. A compound according to any one of claims 1 to 10, which is a compound of the following formula or a pharmaceutically acceptable salt thereof: 13. A compound according to claim 1, which is a compound of one of the following formulas or a pharmaceutically acceptable salt thereof: - 99 043371 14. A compound according to claim 1, which is a compound of one of the following formulas or a pharmaceutically acceptable salt thereof: - 100 043371 - 101 043371 15. A compound according to claim 1, which is a compound of one of the following formulas or a pharmaceutically acceptable salt thereof: - 102 043371 16. A composition for treating a disorder, comprising a compound according to any one of claims 1 to 15 and a carrier, diluent or excipient; the disorder being inflammatory arthritis. 17. A composition for treating a disorder, comprising a compound according to any one of claims 1 to 15 and a carrier, diluent or excipient; wherein said disorder is rheumatoid arthritis; psoriatic arthritis; ankylosing spondylitis; spondyloarthritis; reactive arthritis; infectious arthritis; systemic lupus erythematosus; scleroderma; gout; acquired Still's disease; or juvenile idiopathic arthritis. 18. A composition for treating a disorder, containing a compound according to any one of claims 1 to 15 and wearing - 103 -