Tricyclic sulfone compounds as RORγ modulators

Abstract

As used herein, the term "compound of formula (I): TIFF2023511433000036.tif61153 (wherein all substituents are defined herein) or a stereoisomer or a pharmaceutically acceptable salt thereof.

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims priority to U.S. Provisional Application No. 62 / 965,206, filed January 24, 2020, the entire disclosure of which is incorporated herein by reference.

[0002] FIELD OF THE INVENTION The present invention relates to modulators of the retinoid-related orphan receptor RORγ and methods of using the modulators. The compounds described herein are particularly useful for treating a variety of diseases and disorders in humans and animals, including, but not limited to, psoriasis, rheumatoid arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, acute graft-versus-host disease, psoriatic arthritis, ankylosing spondylitis, systemic lupus erythematosus, lupus nephritis, Sjogren's syndrome, and multiple sclerosis. [Background technology]

[0003] Background of the Invention Retinoid-related orphan receptors, RORα, RORβ, and RORγ, play important roles in many biological processes, including organ development, immunity, metabolism, and circadian rhythms.See, for example, Dussault et al. in Mech. Dev. (1998) vol. 70, 147-153; Andre et al. in EMBO J. (1998) vol. 17, 3867-3877; Sun et al. in Science (2000) vol. 288, 2369-2373; and Jetten in Nucl. Recept. Signal. (2009) vol. 7, 1-32.

[0004] RORγ is expressed in several tissues, including the thymus, kidney, liver, and muscle. Two isoforms of RORγ, RORγ1 and RORγ2 (also known as RORγ and RORγt), have been identified. See, for example, Hirose et al. in Biochem. Biophys. Res. Commun. (1994) vol. 205, 1976-1983; Oritz et al. in Mol. Endocrinol. (1995) vol. 9, 1679-1691; and He et al. in Immunity (1998) vol. 9, 797-806. RORγt expression is restricted to lymphoid cell types, including CD4+CD8+ thymocytes, IL-17-producing helper T (Th17) cells, lymphoid tissue inducer (LTi) cells, and γδ cells. RORγt is essential for the development of lymph nodes and Peyer's patches and the normal differentiation of Th17, γδ, and LTi cells.See, for example, Sun et al. in Science (2000) vol.288, 2369-2373; Ivanov et al. in Cell (2006) vol.126, 1121-1133; Eberl et al. in Nat. Immunol. (2004) vol.5, 64-73; Ivanov et al. in Semin. Immunol. (2007) vol.19, 409-417; and Cua and Tato in Nat. Rev. Immunol. (2010) vol.10, 479-489.

[0005] Inflammatory cytokines (e.g., IL-17A (also called IL-17), IL-17F, and IL-22) produced by Th17 cells and other RORγ+ lymphocytes activate and direct immune responses to extracellular pathogens. See, e.g., Ivanov et al. in Semin. Immunol. (2007) vol. 19: 409-417; and Marks and Craft in Semin. Immunol. (2009) vol. 21, 164-171. RORγ directly suppresses IL-17 transcription, and disruption of RORγ in mice attenuates IL-17 production. See, e.g., Ivanov et al. in Cell (2006) vol. 126, 1121-1133.

[0006] Dysregulation of IL-17 production has been implicated in several human autoimmune and inflammatory diseases, including multiple sclerosis, rheumatoid arthritis, psoriasis, inflammatory bowel disease (IBD), and asthma. For example, Lock et al. in Nat. Med. (2002) vol. 8, 500-508; Tzartos et al. in Am. J. Pathol. (2008) vol. 172, 146-155; Kotake et al. in J. Clin. Invest. (1999) vol. 103, 1345-1352; Kirkham et al. in Arthritis Rheum. (2006) vol. 54, 1122-1131; Lowes et al. in J. Invest. Dermatol. (2008) vol. 128, 1207-1211; Leonardi et al. in N. Engl. J. Med. (2012) vol. 366, 1190-1199; Fujino et al. in Gut (2003) vol. 52, 65-70; Seiderer et al. in Inflamm. Bowel Dis. (2008) vol. 14, 437-445; Wong et al. in Clin. Exp. Immunol. (2001) vol. 125, 177-183; and Agache et al. in Respir. Med. (2010) 104: 1131-1137. In mouse models of these diseases, inhibition of IL-17 function with neutralizing antibodies or genetic disruption of IL-17 or the IL-17 receptor ameliorate the disease course or clinical symptoms. See, e.g., Hu et al. in Ann. NY Acad. Sci. (2011) vol. 1217, 60-76.

[0007] Disruption of RORγ in mice also attenuates disease progression or severity in animal models of autoimmunity and inflammation, including experimental autoimmune encephalomyelitis (EAE), imiquimod-induced psoriasis, colitis, and allergic airway disease. See, for example, Ivanov et al. in Cell (2006) vol. 126, 1121-1133; Yang et al. in Immunity (2008) vol. 28, 29-39; Pantelyushin et al. in J. Clin. Invest. (2012) vol. 122, 2252-2256; Leppkes et al. in Gastroenterology (2009) vol. 136, 257-267; and Tilley et al. in J. Immunol. (2007) vol. 178, 3208-3218.

[0008] Each of the references cited in this Background section is incorporated herein by reference in its entirety for all purposes. Summary of the Invention [Problem to be solved by the invention]

[0009] Although therapeutic agents exist for treating various inflammatory and autoimmune diseases, substantial unmet medical needs remain in these therapeutic areas. Based on the role of IL-17 in human disease and the validation of IL-17 and RORγ as targets in mouse disease models, compounds capable of modulating RORγt activity may offer therapeutic benefit in the treatment of many immune and inflammatory disorders. [Means for solving the problem]

[0010] In one embodiment, the present invention provides a compound of formula (I): [ka] [Wherein R is [ka] is] or a pharmaceutically acceptable salt thereof. The present invention also includes stereoisomers, solvates or prodrugs thereof.

[0011] In another aspect, the present invention provides (S)—N-((6aS,7R,9aS)-9a-((4-fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinolin-7-yl)-2-hydroxy-2-methyl-3-(methylsulfonyl)propanamide, formula (II): [ka] This includes compounds of the formula:

[0012] In another aspect, the present invention encompasses a pharmaceutical composition comprising a compound of formula (II) as described herein or a pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier, excipient, or diluent.

[0013] In another aspect, the present invention encompasses a pharmaceutical composition comprising a compound of formula (I) as described herein or a pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier, excipient, or diluent.

[0014] In another aspect, the invention encompasses a method of modulating RORy in a cell comprising contacting the cell with an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof as described herein. This aspect can be performed in vitro or in vivo.

[0015] In another aspect, the invention encompasses a method of treating a patient suffering from a disease or disorder modulated by RORy, comprising administering to the patient a therapeutically effective amount of a compound according to Formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described herein.

[0016] In another aspect, the invention encompasses a method of treating a disease or disorder in a patient selected from an inflammatory disease or disorder, an autoimmune disease or disorder, an allergic disease or disorder, a metabolic disease or disorder, and / or cancer, comprising administering to the patient a therapeutically effective amount of a compound according to Formula (I) or a pharmaceutically acceptable salt, or a pharmaceutical composition, as described herein.

[0017] Detailed Description of the Invention In one embodiment, the present invention provides a compound of formula (I): [ka] or a pharmaceutically acceptable salt or solvate thereof.

[0018] In another aspect, the present invention provides (S)—N-((6aS,7R,9aS)-9a-((4-fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinolin-7-yl)-2-hydroxy-2-methyl-3-(methylsulfonyl)propanamide, formula (II): [ka] This includes compounds of the formula:

[0019] In certain embodiments, the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt or solvate thereof.

[0020] In another embodiment, the present invention provides a method for preparing a compound of the present invention, or a pharmaceutically acceptable salt or solvate thereof.

[0021] In another embodiment, the present invention provides a compound of the present invention for use in therapy.

[0022] In another embodiment, the present invention provides a pharmaceutical combination of a compound of the present invention and another therapeutic agent for simultaneous, separate or sequential use in therapy.

[0023] In another embodiment, the present invention provides a compound of the present invention for use in the treatment of (or a method of treating) an inflammatory disease, including, but not limited to, psoriasis, rheumatoid arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, acute graft-versus-host disease, psoriatic arthritis, ankylosing spondylitis, systemic lupus erythematosus, lupus nephritis, Sjogren's syndrome, and multiple sclerosis.

[0024] The following are definitions of terms used in this specification and the appended claims: The first definition provided for a group or term herein applies to that group or term throughout the specification and claims, individually or as part of another group, unless otherwise stated.

[0025] Compounds of Formula I containing a carboxylic acid may exist in their free (non-ionized) form or may form salts, which are also within the scope of the present invention. Unless otherwise specified, reference to a compound of the invention is understood to include reference to both its free form and its salts. The term "salt" refers to base salts formed with inorganic and / or organic bases. Pharmaceutically acceptable (i.e., non-toxic and physiologically acceptable) salts are preferred, e.g., acceptable metal salts and amine salts, where the cation does not significantly contribute to the toxicity or biological activity of the salt. However, other salts may also be useful, e.g., in isolation or purification steps that may be used in manufacturing processes, and therefore, other salts are also considered within the scope of the present invention. Salts of compounds of Formula I may be formed, for example, by reacting a compound of Formula I with a certain amount of base (e.g., 1 equivalent) and precipitating the salt in a solvent, e.g., or by lyophilizing the aqueous solution.

[0026] Examples of base salts include ammonium salts, alkali metal salts (e.g., sodium, lithium, and potassium salts), alkaline earth metal salts (e.g., calcium and magnesium salts), barium, zinc, and aluminum salts, organic base salts such as organic amines (e.g., trialkylamines (e.g., triethylamine, procaine, dibenzylamine, N-benzyl-β-phenethylamine, ephenamine, N,N′-dibenzylethylene-diamine, dehydroabietylamine, N-ethylpiperidine, benzylamine, dicyclohexylamine), or similar pharmaceutically acceptable amines, and salts with amino acids (e.g., arginine, lysine), and the like.

[0027] As used herein, the phrase "pharmaceutically acceptable" refers to compounds, substances, compositions, and / or dosage forms that are, within the scope of ordinary medical judgment, suitable for contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, and that offer a reasonable benefit / risk ratio.

[0028] As used herein, "pharmaceutically acceptable salts" refers to derivatives of the compounds disclosed herein that are modified by preparing base salts of the parent compound. Examples of pharmaceutically acceptable salts include, but are not limited to, alkali salts or organic salts of carboxylic acids. Pharmaceutically acceptable salts include conventional non-toxic salts or quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic bases.

[0029] The pharmaceutically acceptable salts of the present invention can be synthesized from a parent compound containing an acidic moiety by conventional chemical methods. Generally, such salts are prepared by reacting the free acid form of the compound with a stoichiometric amount of an appropriate base in water, an organic solvent, or a mixture of the two. Non-aqueous solvents such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are generally preferred. A complete listing of suitable salts can be found in Remington's Pharmaceutical Sciences, 18th Edition, Mack Publishing Company, Easton, PA (1990), the entire contents of which are incorporated herein by reference.

[0030] Prodrugs and solvates of the compounds of the present invention are also contemplated. The term "prodrug" refers to a compound that, after administration to a patient, undergoes chemical conversion by metabolic or chemical processes to yield the compound of Formula I, and / or its salts and / or solvates. Any compound that is converted in the body to yield a bioactive agent (i.e., a compound of Formula I) is a prodrug within the spirit of the present invention. For example, the carboxy group of a compound of Formula I can form a physiologically hydrolyzable ester, which is hydrolyzed in the body to yield the compound of Formula I itself, which acts as a prodrug. Because hydrolysis often occurs primarily under the influence of digestive enzymes, such prodrugs are preferably administered orally. If the ester itself is active or hydrolysis occurs in the blood, it can be administered parenterally. Examples of physiologically hydrolyzable esters of a carboxylic acid-containing compound of Formula I include C 1-6 Alkylbenzyl, 4-methoxybenzyl, indanyl, phthalyl, methoxymethyl, C 1-6 Alkanoyloxy-C 1-6 alkyl (e.g., acetoxymethyl, pivaloyloxymethyl, or propionyloxymethyl), C 1-6 Alkoxycarbonyloxy-C 1-6Included are alkyl (e.g., methoxycarbonyloxymethyl or ethoxycarbonyloxymethyl, glycyloxymethyl, phenylglycyloxymethyl, (5-methyl-2-oxo-1,3-dioxolen-4-yl)-methyl), and other well-known physiologically hydrolyzable esters used, for example, in the penicillin and cephalosporin fields. Such esters may be prepared by conventional techniques known to those skilled in the art.

[0031] Various forms of prodrugs are well known in the art and are described in Rautio, J. et al., Nature Review Drug Discovery, 17, 559-587 (2018).

[0032] Another aspect of the present invention is a pharmaceutical composition comprising a compound of Formula I, or a pharmaceutical salt or solvate thereof, as described herein. The pharmaceutical compositions described herein generally comprise a combination of a compound described herein and a pharmaceutically acceptable carrier, diluent, or excipient. Such compositions are substantially free of pharmaceutically unacceptable ingredients, i.e., contain pharmaceutically unacceptable ingredients in amounts lower than those permitted by U.S. regulations at the time of filing this application. In some embodiments of this aspect, when the compound is dissolved or suspended in water, the composition may optionally further comprise another pharmaceutically acceptable carrier, diluent, or excipient. In other embodiments, the pharmaceutical compositions described herein are solid pharmaceutical compositions (e.g., tablets, capsules, etc.).

[0033] These compositions can be prepared by methods well known in the pharmaceutical arts and can be administered by a variety of routes, depending on whether local or systemic treatment is desired and which areas are to be treated. Administration can be topical (including delivery to the eye and mucous membranes, including the nose, vagina, and rectum), pulmonary (e.g., by inhalation or insufflation of powders or aerosols (e.g., nebulizers), intratracheal, intranasal, epithelial, and transdermal), intraocular, oral, or parenteral. Ocular delivery methods can include topical administration (eye drops), subconjunctival, peribulbar, or vitreous injection, or introduction via a balloon catheter or an ocular insert surgically placed in the conjunctival sac. Parenteral administration can include intravenous, intraarterial, subcutaneous, intraperitoneal, or intramuscular injection or infusion, or intracranial (e.g., intrathecal or intraventricular) administration. Parenteral administration can be in the form of a single bolus dose or, for example, via a continuous infusion pump. Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.

[0034] Pharmaceutical compositions may also contain the compounds described above as active ingredients in combination with one or more pharmaceutically acceptable carriers. In preparing the compositions described herein, the active ingredients are generally mixed with excipients, diluted by the excipients, and enclosed in a carrier, such as a capsule, a sachet, a paper container, or other container. When the excipient serves as a diluent, it can be a solid, semi-solid, or liquid substance that acts as a vehicle, carrier, or medium for the active ingredient. Thus, the composition can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as solids or in liquid media), for example, ointments containing up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injection solutions, and sterile packaged powders.

[0035] In the preparation of the formulation, the active compound can be milled before being combined with other ingredients to provide a suitable particle size.If the active compound is substantially insoluble, it can be milled to a particle size of less than 200 mesh.If the active compound is substantially water-soluble, its particle size can be adjusted by milling to be substantially uniformly dispersed in the formulation, for example, about 40 mesh.

[0036] Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginic acid, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup and methylcellulose.Preparation can also include lubricant (for example, talc, magnesium stearate and mineral oil); wetting agent; emulsifier and suspending agent; preservative (for example, methyl- and propylhydroxy-benzoate); sweetener; and flavoring agent.The composition described herein can be formulated by using methods known in the art so that active ingredient is released quickly, sustained or delayed after being administered to patients.

[0037] The active compounds are effective over a wide dosage range and are generally administered in a pharmaceutically effective amount. However, it will be understood that the amount of compound actually administered will typically be determined by a physician depending on the relevant circumstances, including the condition to be treated, the selected route of administration, the compound actually administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, etc.

[0038] For the preparation of solid compositions such as tablets, the primary active ingredient is mixed with pharmaceutical excipients to form a solid preformulation composition containing a homogeneous mixture of the compounds described herein. When these preformulation compositions are considered homogeneous, the active ingredient is generally dispersed evenly throughout the composition so that the composition can be readily subdivided into equally effective unit dosage forms (e.g., tablets, pills, and capsules). This preformulation solid is then subdivided into unit dosage forms of the type described above, containing, for example, 0.1 to about 500 mg of the active ingredient of the compounds described herein.

[0039] Tablet or pill can be coated or mixed to provide the dosage form that produces sustained action effect.For example, tablet or pill can comprise an inner preparation and an outer preparation component, and the latter is in the form of coating the former.These two components can be separated by an enteric layer that resists disintegration in the stomach and allows the inner component to reach the duodenum intact or be delayed release.Various materials can be used for the enteric layer or coating, and these materials include many polymeric acids and the mixture of polymeric acids with materials such as shellac, cetyl alcohol and cellulose acetate.

[0040] Liquid forms into which the compounds and compositions can be taken for oral or injectable administration include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and emulsions flavored with edible oils (e.g., cottonseed oil, sesame oil, coconut oil, or peanut oil), as well as elixirs and similar pharmaceutical vehicles.

[0041] Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents, or mixtures thereof, and powders.Liquid or solid compositions can contain suitable pharmaceutically acceptable excipients as described above.In some embodiments, the compositions are administered orally or nasally for local or systemic effect.The compositions can be administered by nebulizer using an inert gas.The solutions administered by nebulizer can be inhaled directly from the nebulizing device, or the nebulizing device can be attached to a face tent mask or intermittent positive pressure respirator.Solution, suspension, or powder compositions can be administered orally or nasally from a device that delivers the formulation in an appropriate manner.

[0042] The amount of compound or composition administered to a patient will vary depending on what is being administered, the purpose of administration (e.g., prophylaxis or treatment), the condition of the patient, the method of administration, etc. When used therapeutically, the composition may be administered to a patient already suffering from a disease in an amount sufficient to cure or at least partially halt the symptoms of the disease and its complications. The effective dose will depend on the disease state being treated and factors such as the severity of the disease, the patient's age, weight, and general health, etc., as determined by the attending physician.

[0043] The compositions administered to a patient may be in the form of pharmaceutical compositions described above. These compositions may be sterilized by conventional sterilization techniques or may be sterile filtered. Aqueous solutions may be packaged for immediate use or lyophilized, and the lyophilized preparation may be combined with a sterile aqueous carrier prior to administration. The pH of the compound formulations is generally 3 to 11, more preferably 5 to 9, and most preferably 7 to 8. It is understood that the use of certain of the aforementioned excipients, carriers, or stabilizers will result in the formation of pharmaceutically acceptable salts.

[0044] The therapeutic dosage of a compound can vary depending, for example, on the particular application for which the treatment is being administered, the method of administration of the compound, the condition and status of the patient, and the judgment of the prescribing physician. The proportion or concentration of a compound described herein in a pharmaceutical composition can vary depending on numerous factors, including dosage, chemical characteristics (e.g., hydrophobicity), and route of administration. For example, for parenteral administration, the compounds described herein can be provided in an aqueous physiological buffer solution containing about 0.1 to about 10% w / v of the compound. A typical dosage range is about 1 μg / kg to about 1 g / kg of body weight per day. In some embodiments, the dosage range is about 0.01 mg / kg to about 100 mg / kg of body weight per day. The dosage will likely depend on variables such as the type and progression of the disease or disorder, the overall health of the particular patient, the biological availability of the compound, the formulation of excipients, and its route of administration. Effective doses can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0045] The compounds of the present invention are useful for the prevention, diagnosis, and treatment of various medical disorders in humans or animals. The compounds are used to inhibit or reduce one or more activities associated with RORγ receptors compared to the RORγ receptors in the absence of the compounds. Thus, in one embodiment of the present invention, a method for treating a disease or disorder in a patient selected from an autoimmune disease or disorder, asthma, an allergic disease or disorder, a metabolic disease or disorder, and cancer comprises administering to the patient a therapeutically effective amount of a compound of formula (I) described herein, or a pharmaceutically acceptable salt, solvate, or pharmaceutical composition. See, for example, LA Solt et al., "Action of RORs and their ligands in (patho)physiology," Trends Endocrinol. Metab. 2012, 23 (12): 619-627; MS Maddur et al., "Th17 cells: biology, pathogenesis of autoimmune and inflammatory diseases, and therapeutic strategies," Am. J. Pathol. 2012 Jul;181(1):8-18; and AM Jetten, "Retinoid-related orphan receptors (RORs): critical roles in development, immunity, circadian rhythm, and cellular metabolism," Nucl. Recept. Signal. 2009;7:e003, the contents of each of which are incorporated herein by reference in their entirety, as well as the references cited in the Background section. In some embodiments, the autoimmune disease or disorder is selected from rheumatoid arthritis, ankylosing spondylitis, psoriasis and psoriatic arthritis, multiple sclerosis, inflammatory bowel disease, Sjogren's syndrome and systemic lupus erythematosus.In some embodiments, the allergic disease or disorder is selected from allergic rhinitis and dermatitis.In some embodiments, the metabolic disease or disorder is selected from obesity, obesity-induced insulin resistance and type II diabetes.

[0046] In certain embodiments, the disease or disorder is rheumatoid arthritis.

[0047] In other embodiments, the disease or disorder is multiple sclerosis.

[0048] In other embodiments, the disease or disorder is ankylosing spondylitis.

[0049] In other embodiments, the disease or disorder is inflammatory bowel disease.

[0050] In other embodiments, the disease or disorder is lupus.

[0051] In other embodiments, the disease or disorder is Sjogren's syndrome.

[0052] In other embodiments, the disease or disorder is psoriasis.

[0053] In other embodiments, the disease or disorder is psoriatic arthritis.

[0054] In other embodiments, the disease or disorder is graft-versus-host disease (GVHD).

[0055] In other embodiments, the disease or disorder is autoimmune uveitis.

[0056] In other embodiments, the disease or disorder is obesity and / or insulin resistance.

[0057] In other embodiments, the disease or disorder is melanoma.

[0058] In some embodiments, the medical disorder diagnosed, treated, or prevented by the use of the compounds of the present disclosure may be, for example, an autoimmune disorder. In other embodiments, the disorder diagnosed, treated, or prevented by the use of the compounds of the present disclosure may be an inflammatory disorder. For example, in some embodiments, the disorder is selected from arthritis, diabetes, multiple sclerosis, uveitis, rheumatoid arthritis, psoriasis, asthma, bronchitis, allergic rhinitis, chronic obstructive pulmonary disease, atherosclerosis, Helicobacter pylori infection, and inflammatory bowel disease. In other embodiments, the disorder is selected from Crohn's disease, ulcerative colitis, sprue, and food allergy. In other embodiments, the disorder is autoimmune encephalomyelitis, imiquimod-induced psoriasis, colitis, or allergic airway disease.

[0059] As used herein, the phrase "therapeutically effective amount" refers to the amount of an active compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue, system, animal, individual, or human that is desired by a researcher, veterinarian, physician, or other clinician.

[0060] In certain embodiments, a therapeutically effective amount can be an amount adequate for (1) disease prevention; e.g., preventing a disease, condition, or disorder in an individual who may be predisposed to the disease, condition, or disorder, but who has not yet experienced or manifested symptoms or signs of the disease; (2) disease inhibition; e.g., inhibiting a disease, condition, or disorder in an individual who is experiencing or manifesting symptoms or signs of the disease, condition, or disorder; or (3) disease amelioration; e.g., improving a disease, condition, or disorder (i.e., improving the condition and / or symptoms), such as reducing the severity of the disease, in an individual who is experiencing or manifesting symptoms or signs of the disease, condition, or disorder.

[0061] As used herein, the terms "treatment" and "treating" refer to (i) the amelioration of the aforementioned condition; e.g., the improvement of a disease, condition, or disorder (i.e., the reversal or amelioration of the condition and / or symptoms), such as a reduction in the severity of the disease in an individual experiencing or manifesting a symptom or sign of the disease, condition, or disorder; (ii) the elicitation of a biological or medicinal response in a tissue, system, animal, individual, or human that is sought by a researcher, veterinarian, medical doctor, or other clinician; or (iii) the prevention of the aforementioned condition; e.g., the prevention of a disease, condition, or disorder in an individual experiencing or manifesting a symptom or sign of the disease, condition, or disorder. These methods are for illustrative purposes only and

[0062] (Manufacturing method) The compounds of the present invention can be synthesized by many methods available to those skilled in the art of organic chemistry. Methods for preparing the intermediates and compounds of the present invention are described in the following examples. These methods are for illustrative purposes and are not intended to limit the techniques that those skilled in the art may practice to prepare the intermediates and compounds disclosed in the present invention. Various methods for preparing the compounds of the present invention will be readily apparent to those skilled in the art. Chemists skilled in the synthetic arts will frequently devise desirable alternative preparation methods based on one or more considerations (e.g., shorter reaction times, cheaper starting materials or reagents, ease of operation and purification, higher yields, catalyst compatibility, avoidance of toxic reagents, availability of specialized equipment, and reduced number of steps).

[0063] Preparation of homochiral example compounds can be carried out by techniques known to those skilled in the art. For example, homochiral compounds can be prepared by separating racemic products or diastereomers, for example, by chiral-phase preparative HPLC. Alternatively, example compounds can be prepared by known methods to obtain enantiomerically or diastereomerically enriched products.

[0064] The reactions and techniques described for preparing the intermediates and compounds of the present invention are carried out in solvents appropriate to the reagents and materials used and are suitable for the transformations being effected. It should be understood, and will be readily recognized by those skilled in the art, that all reaction conditions (such as selection of solvent, reaction atmosphere, reaction temperature, experimental time, and workup method) are selected to be conditions appropriate for the reaction. Those skilled in the art of organic synthesis will understand that functional groups present on various portions of the molecule must be compatible with the proposed reagents and reactions. Those skilled in the art will readily recognize that there are limitations on the substituents compatible with the reaction conditions, and that alternatives will be required if the substituents present are not suitable. The reactions may require the determination to change the order of synthetic steps or to select a different course of action for certain reactions to obtain the desired compounds of the present invention. It is also recognized that another important consideration in planning any synthetic route in this field is the judicious selection of protecting groups used to protect reactive functional groups present in the compounds described in this invention. For the experienced experimenter, an authoritative reference describing many protecting group alternatives is Greene's Protective Groups in Organic Synthesis, Fourth Edition, by Wuts and Greene, Wiley and Sons (2007). [Example]

[0065] The following examples illustrate specific and preferred embodiments of the present invention and are not intended to limit the scope of the invention. Chemical and scientific abbreviations and symbols have their common and accustomed meanings unless otherwise specified. Alternative abbreviations used in the examples and elsewhere in this application are defined below. Common intermediates are generally useful in the preparation of more than one example and are identified sequentially by the intermediate number and step in their preparation (e.g., Intermediate 1, Step A), or by the intermediate number only if the compound is a title compound. Example compounds are identified by the example number and preparation step (e.g., Example 1, Step A) if the compound is an intermediate, or by the example number alone if the compound is the title compound of the example. A chemical engineer skilled in the art of synthesis may devise alternative preparation procedures that may be desirable based on one or more considerations, such as shorter reaction times, less expensive starting materials, ease of handling or isolation, improved yields, compatibility with catalysis, avoidance of toxic reagents, availability of specialized equipment, or a reduced number of linear steps. Starting materials and intermediates not explicitly prepared are either commercially available or known from the literature.

[0066] Organic solutions were dried over anhydrous sodium sulfate or anhydrous magnesium sulfate, followed by decantation or filtration to remove residual water. Solvent removal was achieved by concentration under reduced pressure. Column chromatography was generally performed using a CombiFlash® automated chromatography system (Teledyne Isco) with pre-packed silica gel cartridges eluted with the indicated solvent or solvent mixture.

[0067] Analytical HPLC was performed under the following conditions: Column - XBridge TM C 18, 2.1 x 50 mm, 1.7 μm (Waters Corp.); temperature 50 °C; mobile phase A - 5:95 MeCN-water (containing 0.1% TFA); mobile phase B - 95:5 MeCN-gradient; gradient, 0 to 100% B over 3 min, then 100% B for 0.75 min; flow rate 1 mL / min; detection by MS and UV (220 nm).

[0068] Preparative HPLC was performed under the following conditions: Column - XBridge TM C 18 19 x 200 mm, 5 μm (Waters Corp.); Mobile Phase A - 5:95 MeCN-water (containing 10 mM ammonium acetate); Mobile Phase B - 95:5 MeCN-water (containing 10 mM ammonium acetate); Gradient: increasing B followed by isocratic gradient; Flow rate 20 mL / min. Separation of enantiomers or diastereomers by chiral supercritical fluid chromatography was carried out using the conditions specified individually. Mass spectrometry data were obtained by liquid chromatography-mass spectrometry using electrospray ionization.

[0069] Compound names were determined using ChemBioDraw Ultra version 14.0.0.126 (PerkinElmer Inc.). The following abbreviations are used: [Table 1]

[0070] Intermediate 1 (6aS,7R,9aS)-9a-((4-Fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinolin-7-amine, HCl salt [ka] Process A: 7,8-Dihydroquinoline-2,5(1H,6H)-dione [ka] A mixture of 3-amino-2-cyclohexen-1-one (12.0 g, 108 mmol) and ethyl propionate (14.6 mL, 144 mmol) was heated to 100°C overnight. The temperature was then increased to 170°C for 2 hours and then cooled. The dark brown mixture was treated dropwise with MeOH (35 mL) over 20 minutes, gradually forming a suspension, which was stirred at 70°C for 2 hours. The mixture was cooled to 0°C and stirred for 1 hour. The precipitate was collected by filtration, washed with a 2:1 mixture of heptane and DCM (30 mL), then MeOH (10 mL), and air-dried to give 7,8-dihydroquinoline-2,5(1H,6H)-dione as a tan solid (4.4 g). LCMS m / z 163.9 (M+H) + . 1 H NMR (499 MHz, DMSO-d6) δ 12.08 (br s, 1H), 7.77 (d, J=9.5 Hz, 1H), 6.24 (d, J=9.5 Hz, 1H), 2.79 (t, J=6.2 Hz, 2H), 2.44 - 2.40 (m, 2H), 2.00 (quin, J=6.4 Hz, 2H).

[0071] Process B: 2-((2,6-dichlorobenzyl)oxy)-7,8-dihydroquinolin-5(6H)-one [ka] A mixture of 7,8-dihydroquinoline-2,5(1H,6H)-dione (8.65 g, 53.0 mmol), 2-(bromomethyl)-1,3-dichlorobenzene (15.3 g, 63.6 mmol), and CsCO (17.3 g, 53.0 mmol) in MeCN (200 ml) was stirred at room temperature for 22 hours. The precipitate was removed by filtration, and the filtrate was concentrated. The residue was partitioned between EtOAc and water, and the organic phase was washed with brine, dried over MgSO, and concentrated. The residue was purified by silica gel column chromatography (220 g) eluting with EtOAc-hexane (5-10% gradient) to give 2-((2,6-dichlorobenzyl)oxy)-7,8-dihydroquinolin-5(6H)-one as a white solid (14.5 g). LCMS m / z 322.0, 324.0 (M+H) + . 1 H NMR (499 MHz, CDCl3) δ 8.19 (d, J=8.6 Hz, 1H), 7.39 - 7.36 (m, 2H), 7.28 - 7.24 (m, 1H), 6.69 (d, J=8.6 Hz, 1H), 5.68 (s, 2H), 3.08 (t, J=6.2 Hz, 2H), 2.69 - 2.60 (m, 2H), 2.19 (quin, J=6.4 Hz, 2H).

[0072] Process C: 2-((2,6-Dichlorobenzyl)oxy)-7,8-dihydroquinolin-5-yl trifluoromethanesulfonate [ka] A solution of 2-((2,6-dichlorobenzyl)oxy)-7,8-dihydroquinolin-5(6H)-one (8.24 g, 25.6 mmol) and N,N-bis(trifluoromethylsulfonyl)aniline (11.9 g, 33.2 mmol) in anhydrous THF was cooled to −78° C. and treated with 1 M potassium bis(trimethylsilyl)amide (1.0 M in toluene; 34.5 mL, 34.5 mmol) over approximately 10 min. After stirring for 80 min, the mixture was treated with water and allowed to warm to room temperature. Saturated aqueous NaHCO3 was added, and the mixture was extracted with ether. The organic phase was washed with brine, dried over Na2SO4, and concentrated. The residue was purified by silica gel column chromatography (220 g) eluting with EtOAc-hexane (0-5% gradient) to give 2-((2,6-dichlorobenzyl)oxy)-7,8-dihydroquinolin-5-yl trifluoromethanesulfonate as a colorless syrup (7.29 g). LCMS m / z 454.0, 456.0 (M+H). + . 1 H NMR (499 MHz, CDCl) δ 7.54 (d, J = 8.5 Hz, 1H), 7.39 (d, J = 8.1 Hz, 2H), 7.27–7.21 (m, 1H), 6.68 (d, J = 8.5 Hz, 1H), 5.94 (t, J = 4.8 Hz, 1H), 5.62 (s, 2H), 3.04 (t, J = 8.6 Hz, 2H), 2.63 (td, J = 8.6, 4.8 Hz, 2H). An additional portion of the impurity 2-((2,6-dichlorobenzyl)oxy)-7,8-dihydroquinolin-5-yl trifluoromethanesulfonate was also obtained as another colorless syrup (4.39 g).

[0073] Process D: 2-((2,6-dichlorobenzyl)oxy)-5-((4-fluorophenyl)thio)-7,8-dihydroquinoline [ka] A mixture of 2-((2,6-dichlorobenzyl)oxy)-7,8-dihydroquinolin-5-yl trifluoromethanesulfonate (20.2 g, 44.5 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthine (1.29 g, 2.22 mmol), tris(dibenzylideneacetone)dipalladium(0) (1.63 g, 1.78 mmol), and DIEA (14.0 mL, 80 mmol) in 1,4-dioxane (127 mL) in a pressure-resistant flask was treated with 4-fluorobenzenethiol (8.5 mL, 80 mmol). The mixture was purged with nitrogen for 5 minutes, and the vessel was sealed under a nitrogen atmosphere and heated at 115 °C for 3 hours. The mixture was cooled to room temperature, filtered through a Celite pad, and the solid was washed with ether. The filtrate was partitioned between saturated aqueous NaHCO3 and ether. The organic phase was washed with brine, dried over MgSO4, and concentrated. The residue was purified by silica gel column chromatography (750 g) eluting with EtOAc-hexane (0-25% gradient) to give 2-((2,6-dichlorobenzyl)oxy)-5-((4-fluorophenyl)thio)-7,8-dihydroquinoline (16.0 g) as a pale yellow syrup. LCMS m / z 432.0, 434.0 (M+H). + . 1 H NMR (499 MHz, CDCl3) δ 7.65 (d, J=8.5 Hz, 1H), 7.36 (d, J=8.0 Hz, 2H), 7.26 - 7.21 (m, 3H), 6.96 (t, J=8.7 Hz, 2H), 6.52 (d, J=8.5 Hz, 1H), 6.40 (t, J=4.6 Hz, 1H), 5.58 (s, 2H), 3.01 (t, J=8.4 Hz, 2H), 2.57 (td, J=8.4, 4.6 Hz, 2H).

[0074] Process E: 2-((2,6-dichlorobenzyl)oxy)-5-((4-fluorophenyl)sulfonyl)-7,8-dihydroquinoline [ka] A solution of 2-((2,6-dichlorobenzyl)oxy)-5-((4-fluorophenyl)thio)-7,8-dihydroquinoline (16.0 g, 37.0 mmol) in DCM (463 mL) was cooled on an ice-water bath and treated with mCPBA (19.9 g, 89.0 mmol) in small portions. The resulting suspension was stirred at 0 °C for 2 h, then filtered, and the collected solid was washed with DCM. The filtrate was treated with saturated aqueous NaHCO3, and the mixture was stirred for 10 min. The organic phase was separated and washed successively with saturated aqueous NaHCO3, 10% aqueous Na2S2O3, and brine, dried over MgSO4, and concentrated. The residue was dried in vacuo to give 2-((2,6-dichlorobenzyl)oxy)-5-((4-fluorophenyl)sulfonyl)-7,8-dihydroquinoline as an off-white solid (16.3 g). LCMS m / z 464.2, 466.1 (M+H) + . 1 H NMR (499 MHz, CDCl3) δ 8.08 (d, J=8.7 Hz, 1H), 7.98 - 7.89 (m, 2H), 7.37 - 7.34 (m, 2H), 7.31 (t, J=4.9 Hz, 1H), 7.26 - 7.22 (m, 1H), 7.18 (t, J=8.6 Hz, 2H), 6.58 (d, J=8.7 Hz, 1H), 5.56 (s, 2H), 3.00 - 2.86 (m, 2H), 2.67 (td, J=8.4, 4.9 Hz, 2H).

[0075] Process F: Ethyl 3-((2,6-dichlorobenzyl)oxy)-9a-((4-fluorophenyl)sulfonyl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinoline-7-carboxylate (mixture of diastereomers) [ka] A solution of 2-((2,6-dichlorobenzyl)oxy)-5-((4-fluorophenyl)sulfonyl)-7,8-dihydroquinoline (17.1 g, 36.8 mmol) and ethyl 4-chlorobutanoate (21.1 mL, 147 mmol) in THF (184 mL) was cooled to −78° C. and treated with lithium bis(trimethylsilyl)amide (1.0 M in THF; 147 mL, 147 mmol) over 30 minutes. The resulting mixture was stirred at −78° C. for 2 hours and then allowed to warm slowly to room temperature overnight. After 18 hours, the mixture was cooled to 0° C. and treated with saturated aqueous NH4Cl. The mixture was concentrated in vacuo and then partitioned between ether and saturated aqueous NH4Cl. The organic phase was washed successively with saturated aqueous NH4Cl and brine, dried over MgSO4, and concentrated. The resulting light brown syrup was triturated with ether to form a solid. The suspension was stirred at 0° C. for 30 minutes, and the solid was collected by filtration, washed twice with cold ether, and dried under vacuum to give a diastereomeric mixture of ethyl 3-((2,6-dichlorobenzyl)oxy)-9a-((4-fluorophenyl)sulfonyl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinoline-7-carboxylate as a light brown solid (7.83 g). LCMS m / z 578.5, 580.5 (M+H) + . 1H NMR (499 MHz, CDCl3) δ 7.68 (d, J=8.6 Hz, 1H), 7.41 - 7.34 (m, 4H), 7.29 - 7.23 (m, 1H), 7.05 (t, J=8.6 Hz, 2H), 6.67 (d, J=8.7 Hz, 1H), 5.64 - 5.49 (m, 2H), 4.23 (q, J=7.2 Hz, 2H), 3.28 (td, J=8.7, 5.9 Hz, 1H), 3.18 (ddd, J=13.9, 7.6, 3.6 Hz, 1H), 2.73 - 2.65 (m, 1H), 2.58 (dt, J=16.9, 5.2 Hz, 1H), 2.37 (dtd, J=12.5, 9.8, 7.6 Hz, 1H), 2.16 (ddd, J=13.9, 10.0, 7.2 Hz, 1H), 2.11 - 2.01 (m, 2H), 1.96 (ddd, J=16.9, 9.8, 4.4 Hz, 1H), 1.45 (dtd, J=13.8, 9.3, 4.6 Hz, 1H), 1.32 (t, J=7.2 Hz, 3H). The filtrate was concentrated, and the residue was purified by column chromatography on silica gel (330 g) eluting with EtOAc-hexane (5-25% gradient) to give another ethyl acetate compound of slightly reduced purity. 3-((2,6-Dichlorobenzyl)oxy)-9a-((4-fluorophenyl)sulfonyl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinoline-7-carboxylate was obtained as an off-white glassy solid (7.68 g).

[0076] Process G: Ethyl 9a-((4-fluorophenyl)sulfonyl)-3-hydroxy-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinoline-7-carboxylate (mixture of diastereomers) [ka] A diastereomeric mixture of ethyl 3-((2,6-dichlorobenzyl)oxy)-9a-((4-fluorophenyl)sulfonyl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinoline-7-carboxylate (17.0 g, 29.4 mmol) in DCE (147 mL) was treated with HCl (4.0 M in 1,4-dioxane; 44.1 mL, 176 mmol) at room temperature. The mixture was heated at 50° C. for 18 h and then concentrated. The light brown syrup residue was dissolved in DCM (20 mL) and added slowly to stirred ether (400 mL). The resulting suspension was stirred for 30 minutes, and the solid was collected by filtration, washed three times with ether, and dried under vacuum to give a diastereomeric mixture of ethyl 9a-((4-fluorophenyl)sulfonyl)-3-hydroxy-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinoline-7-carboxylate (12.3 g). LCMS m / z 420.4 (M+H). + . 1 H NMR (499 MHz, MeOH-d4) δ 7.82 (d, J=9.3 Hz, 1H), 7.69 - 7.63 (m, 2H), 7.35 (t, J=8.8 Hz, 2H), 6.68 (d, J=9.4 Hz, 1H), 4.18 (q, J=7.2 Hz, 2H), 3.20 (td, J=8.4, 5.8 Hz, 1H), 2.97 (ddd, J=13.8, 7.5, 4.4 Hz, 1H), 2.76 (q, J=8.5 Hz, 1H), 2.63 - 2.55 (m, 1H), 2.24 - 2.16 (m, 1H), 2.15 - 2.03 (m, 3H), 2.02 - 1.94 (m, 1H), 1.62 - 1.46 (m, 1H), 1.27 (t, J=7.2 Hz, 3H).

[0077] Process H: Ethyl 9a-((4-fluorophenyl)sulfonyl)-3-(((trifluoromethyl)sulfonyl)oxy)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinoline-7-carboxylate (mixture of diastereomers) [ka] A solution of ethyl 9a-((4-fluorophenyl)sulfonyl)-3-hydroxy-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinoline-7-carboxylate (12.3 g, 29.3 mmol) in DCM (147 mL) was treated with pyridine (4.74 mL, 58.6 mmol). The solution was cooled in an ice-water bath and treated dropwise with trifluoromethanesulfonic anhydride (7.40 mL, 44.0 mmol) over 30 min. The mixture was stirred at 0-5 °C for 1 h and then treated again with additional pyridine (1.19 mL, 14.7 mmol) and trifluoromethanesulfonic anhydride (1.48 mL, 8.79 mmol). The mixture was stirred at 0-5 °C for 1.5 h and then treated with saturated aqueous NH4Cl (2 mL). The mixture was diluted with EtOAc, washed successively with water and brine, dried over Na2SO4, and concentrated to give a crude diastereomeric mixture of ethyl 9a-((4-fluorophenyl)sulfonyl)-3-(((trifluoromethyl)sulfonyl)oxy)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinoline-7-carboxylate (15.5 g), which was used without purification. LCMS m / z 552.4 (M+H) + .

[0078] Process I: Ethyl 3-bromo-9a-((4-fluorophenyl)sulfonyl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinoline-7-carboxylate (mixture of diastereomers) [ka] A vigorously stirred solution of the crude diastereomeric mixture of ethyl 9a-((4-fluorophenyl)sulfonyl)-3-(((trifluoromethyl)sulfonyl)oxy)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinoline-7-carboxylate (15.5 g, 28.1 mmol) in DCM (20 mL) and toluene (160 mL) was treated with LiBr (11.0 g, 126 mmol) followed by portionwise p-toluenesulfonic acid (5.88 g, 30.9 mmol). The mixture was stirred at 50 °C for 16 h, then cooled to rt and diluted with DCM and water. The organic phase was separated, washed with brine, dried over NaSO, and concentrated. The residue was purified by silica gel column chromatography (120 g) eluting with EtOAc-hexane (0-30% gradient) to give a diastereomeric mixture of ethyl 3-bromo-9a-((4-fluorophenyl)sulfonyl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinoline-7-carboxylate (10.2 g). LCMS m / z 482.4, 484.4 (M+H). + . 1 H NMR (499 MHz, CDCl3) δ 7.67 (d, J=8.3 Hz, 1H), 7.43 - 7.36 (m, 3H), 7.15 - 7.09 (m, 2H), 4.22 (q, J=7.2 Hz, 2H), 3.30 (td, J=8.8, 5.9 Hz, 1H), 3.21 (ddd, J=13.8, 7.6, 3.2 Hz, 1H), 2.70 - 2.63 (m, 2H), 2.36 (dtd, J=12.4, 10.0, 7.4 Hz, 1H), 2.17 - 1.99 (m, 4H), 1.46 - 1.37 (m, 1H), 1.31 (t, J=7.2 Hz, 3H).

[0079] Process J: Ethyl 9a-((4-fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinoline-7-carboxylate (diastereomeric mixture) [ka] Activated copper was prepared by adding zinc powder (24.6 g, 376 mmol) in small portions over 10 minutes to a solution of CuSO4 tetrahydrate (45.1 g, 283 mmol) in water (250 mL) with stirring. The mixture was stirred for 10 minutes, and then the suspension was decanted to separate the red precipitate. This was washed twice by decantation with water and then stirred with 1 M aqueous HCl (400 mL) for 2.5 hours. The supernatant was decanted, and the precipitate was stirred with fresh water and then washed repeatedly by decantation until the pH of the supernatant was approximately 7. The solid was stored under water and an inert atmosphere (argon or nitrogen). For use, the solid was washed twice by decantation with MeOH, then twice by decantation with diethyl ether, and dried in vacuo.

[0080] A sample of dry activated copper powder (15.8 g, 248 mmol) in a pressure flask was treated with a solution of a diastereomeric mixture of ethyl 3-bromo-9a-((4-fluorophenyl)sulfonyl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinoline-7-carboxylate (10.2 g, 21.2 mmol) in dry DMF (141 mL). The mixture was flushed with nitrogen for 2 minutes and then treated with 1,1,1,2,3,3,3-heptafluoro-2-iodopropane (12.0 mL, 85.0 mmol). The vessel was sealed under nitrogen and heated at 120 °C for 2.5 hours. The mixture was cooled to room temperature, mixed with water (300 mL) and ether (350 mL), and filtered through a pad of Celite. The solid was washed three times with ether. The combined filtrate was separated, and the organic phase was washed successively with saturated aqueous NaHCO3, 10% aqueous LiCl, and brine, dried over MgSO4, and concentrated to give the crude diastereomeric mixture of ethyl 9a-((4-fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinoline-7-carboxylate as a brown sticky solid (12.0 g), which was used without purification. LCMS m / z 572.2 (M+H) +. 1 H NMR (499 MHz, CDCl3) δ 8.01 (d, J=8.2 Hz, 1H), 7.60 (dd, J=8.2, 2.6 Hz, 1H), 7.23 - 7.17 (m, 2H), 7.04 - 6.98 (m, 2H), 4.28 - 4.23 (m, 2H), 3.40 - 3.32 (m, 2H), 2.74 - 2.63 (m, 2H), 2.53 - 2.43 (m, 1H), 2.25 - 2.14 (m, 3H), 1.85 - 1.77 (m, 2H), 1.33 (t, J=7.2 Hz, 3H).

[0081] Process K: 9a-((4-Fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinoline-7-carboxylic acid (diastereomeric mixture) [ka] The crude diastereomeric mixture of ethyl 9a-((4-fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinoline-7-carboxylate (12.0 g, 21.0 mmol) was combined with LiOH monohydrate (3.52 g, 84.0 mmol) in THF-EtOH-water (3:1:1, 175 mL) and stirred at rt for 18 h. The mixture was concentrated in vacuo, and the residue was partitioned between EtOAc (250 mL) and 0.5 M aqueous HCl (181 mL). The organic phase was washed with brine, dried over MgSO4, and concentrated to give a diastereomeric mixture of 9a-((4-fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinoline-7-carboxylic acid as a light yellow solid (11.4 g). LCMS m / z 544.3 (M+H) + . 11H NMR (499 MHz, DMSO-d6) δ 12.67 - 12.06 (m, 1H), 8.80 (s, 1H), 7.93 (d, J=8.2 Hz, 1H), 7.77 (dd, J=8.3, 2.9 Hz, 1H), 7.32 - 7.27 (m, 3H), 3.27 - 3.20 (m, 1H), 3.05 (ddd, J=14.1, 6.8, 2.2 Hz, 1H), 2.80 - 2.71 (m, 1H), 2.61 (dt, J=16.4, 3.9 Hz, 1H), 2.30 (s, 1H), 2.20 - 2.13 (m, 1H), 2.13 - 1.99 (m, 3H), 1.42 (br s, 1H).

[0082] Project L: 2-(Trimethylsilyl)ethyl (9a-((4-fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinolin-7-yl)carbamate (diastereomeric mixture)

Chem.

[0083] Process M: 2-(Trimethylsilyl)ethyl (9a-((4-fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinolin-7-yl)carbamate, four separate diastereomers [ka] A diastereomeric mixture of 2-(trimethylsilyl)ethyl (9a-((4-fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinolin-7-yl)carbamate (1.2 g) was dissolved in MeOH (17 mL) and separated by SFC (column: Whelko-RR, 5 x 50 cm, 10 μm (Phenomenex Inc.); pressure: 100 bar; temperature: 35 °C; mobile phase: CO2-MeOH (85:15); flow rate: 300 mL / min; 1 mL injection, 3.5 min cycle time). Three fractions were collected.

[0084] The first eluted fraction was concentrated to give a single diastereomer of 2-(trimethylsilyl)ethyl (9a-((4-fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinolin-7-yl)carbamate as a white solid (0.396 g). LCMS m / z 659.0 (M+H) + . 1 H NMR (499 MHz, CDCl3) δ 8.09 (d, J=8.2 Hz, 1H), 7.62 (dd, J=8.2, 2.6 Hz, 1H), 7.15 - 7.08 (m, 2H), 6.97 (t, J=8.5 Hz, 2H), 5.60 (br d, J=8.9 Hz, 1H), 4.25 - 4.16 (m, 2H), 4.16 - 4.08 (m, 1H), 3.32 (dt, J=14.2, 7.0 Hz, 1H), 2.75 - 2.64 (m, 2H), 2.31 (dt, J=14.5, 7.3 Hz, 1H), 2.26 - 2.19 (m, 1H), 2.18 - 2.09 (m, 2H), 1.69 - 1.60 (m, 1H), 1.37 (qd, J=12.8, 3.6 Hz, 1H), 1.07 - 1.00 (m, 2H), 0.07 (s, 9H). The second eluted fraction was concentrated to give the second single diastereomer of 2-(trimethylsilyl)ethyl (9a-((4-fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinolin-7-yl)carbamate as a white solid (0.375 g). LCMS m / z 659.0 (M+H) + . 1 H NMR (499 MHz, CDCl3) δ 8.09 (d, J=8.2 Hz, 1H), 7.62 (dd, J=8.3, 2.6 Hz, 1H), 7.17 - 7.08 (m, 2H), 6.97 (t, J=8.5 Hz, 2H), 5.60 (br d, J=8.7 Hz, 1H), 4.26 - 4.06 (m, 3H), 3.32 (dt, J=14.2, 7.0 Hz, 1H), 2.77 - 2.60 (m, 2H), 2.31 (dt, J=14.6, 7.4 Hz, 1H), 2.26 - 2.19 (m, 1H), 2.19 - 2.07 (m, 2H), 1.70 - 1.59 (m, 1H), 1.37 (qd, J=12.8, 3.4 Hz, 1H), 1.07 - 1.00 (m, 2H), 0.07 (s, 9H). The third elution fraction was concentrated, and the residue was dissolved in MeOH (17 mL) and further separated by SFC (column: cellulose-4, 3 x 25 cm, 5 μm; pressure: 100 bar; temperature: 35 °C; mobile phase: CO2-MeOH (85:15) (containing 0.1% NH4OH); flow rate: 180 mL / min; 1 mL injection, 1.1 min cycle time). Two further fractions were collected.

[0085] The first eluted fraction from the second separation was concentrated to give the third single diastereomer of 2-(trimethylsilyl)ethyl (9a-((4-fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinolin-7-yl)carbamate as a white solid (46 mg). LCMS m / z 659.0 (M+H) + . 1 H NMR (499 MHz, CDCl3) δ 8.27 (br d, J=8.3 Hz, 1H), 7.66 (dd, J=8.2, 2.5 Hz, 1H), 7.13 (br s, 2H), 6.95 (t, J=8.6 Hz, 2H), 4.68 - 4.56 (m, 2H), 4.26 - 4.14 (m, 2H), 3.21 (ddd, J=14.0, 11.0, 8.2 Hz, 1H), 3.12 - 2.97 (m, 1H), 2.70 (br d, J=16.1 Hz, 1H), 2.44 - 2.22 (m, 2H), 1.89 (br dd, J=7.9, 3.9 Hz, 1H), 1.72 - 1.62 (m, 1H), 1.55 - 1.48 (m, 1H), 1.20 - 1.08 (m, 1H), 1.05 - 0.96 (m, 2H), 0.06 (s, 9H).

[0086] The second eluting fraction from the second separation was concentrated to give the fourth single diastereomer of 2-(trimethylsilyl)ethyl (9a-((4-fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinolin-7-yl)carbamate as a white solid (46 mg). LCMS m / z 659.0 (M+H) + . 1H NMR (499 MHz, CDCl3) δ 8.27 (br d, J=8.2 Hz, 1H), 7.66 (dd, J=8.3, 2.6 Hz, 1H), 7.14 (br s, 2H), 6.95 (t, J=8.5 Hz, 2H), 4.68 - 4.56 (m, 2H), 4.30 - 4.12 (m, 2H), 3.28 - 3.14 (m, 1H), 3.13 - 3.00 (m, 1H), 2.70 (br d, J=17.0 Hz, 1H), 2.39 - 2.23 (m, 2H), 1.89 (br dd, J=8.2, 3.8 Hz, 1H), 1.66 (br dd, J=11.3, 8.7 Hz, 1H), 1.56 - 1.48 (m, 1H), 1.21 - 1.08 (m, 1H), 1.00 (br t, J=8.3 Hz, 2H), 0.06 (s, 9H).

[0087] Process N: (6aS,7R,9aS)-9a-((4-Fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinolin-7-amine, HCl salt [ka] A solution of the second diastereomer of 2-(trimethylsilyl)ethyl (9a-((4-fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinolin-7-yl)carbamate (0.375 g, 0.569 mmol) obtained from Step M of Intermediate 1 in DCE (4.8 mL) was cooled on an ice-water bath and treated with HCl (4.0 M in 1,4-dioxane; 1.42 mL, 5.69 mmol), and the resulting mixture was stirred with heating at 40° C. for 6 hours. The mixture was cooled to room temperature and concentrated to give (6aS,7R,9aS)-9a-((4-fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinolin-7-amine, HCl salt as a white solid (0.33 g). LCMS m / z 514.9 (M+H) + . 1H NMR (499 MHz, MeOH-d4) δ 8.01 (d, J=8.3 Hz, 1H), 7.71 (dd, J=8.2, 2.7 Hz, 1H), 7.31 (dd, J=8.5, 4.9 Hz, 2H), 7.17 (t, J=8.7 Hz, 2H), 3.62 - 3.52 (m, 1H), 3.37 (dd, J=7.1, 4.8 Hz, 1H), 3.15 - 3.06 (m, 1H), 2.72 (dt, J=16.7, 3.9 Hz, 1H), 2.46 (ddd, J=14.7, 9.8, 7.3 Hz, 1H), 2.38 - 2.25 (m, 3H), 2.00 (ddd, J=16.7, 12.9, 3.8 Hz, 1H), 1.53 - 1.41 (m, 1H). The absolute configuration was determined from the anomalous dispersion signals using the Flack method by single crystal X-ray analysis (Acta Cryst. B, 2013, 69, 249).

[0088] Intermediate 2 (S)-2-Hydroxy-2-methyl-3-(methylsulfonyl)propanoic acid [ka] Process A: Methyl 2-hydroxy-2-methyl-3-(methylthio)propanoate (racemic) [ka] A solution of racemic methyl 2-methyloxirane-2-carboxylate (5.00 g, 42.6 mmol) and acetic acid (3.05 mL, 53.3 mmol) in anhydrous MeOH (125 mL) was stirred in an ice-water bath under a nitrogen atmosphere. When the internal temperature reached approximately 5 °C, sodium methanethiolate (11.0 g, 149 mmol) was added dropwise over approximately 5 minutes, resulting in an exotherm to approximately 20 °C. After 10 minutes, the internal temperature was approximately 12 °C, and the cooling bath was removed and stirring continued. Two hours after the addition was complete, the mixture was cooled in an ice-water bath and treated with acetic acid (6.1 mL, 107 mmol). The mixture was concentrated in vacuo, and the remaining slug was partitioned between ether (300 mL) and water (50 mL). The aqueous phase was extracted twice with ether (2x100 mL) and the combined organic phases were washed successively with saturated aqueous NaHCO3 and brine, dried over Na2SO4 and concentrated in vacuo to give racemic methyl 2-hydroxy-2-methyl-3-(methylthio)propionate as a pale yellow liquid (6.87 g). LCMS m / z 186.9 (M+Na). + . 1 H NMR (499 MHz, CDCl3) δ 3.85 - 3.78 (m, 3H), 3.48 (s, 1H), 2.97 (d, J=14.1 Hz, 1H), 2.77 (d, J=13.9 Hz, 1H), 2.18 (s, 3H), 1.49 (s, 3H).

[0089] Process B: Methyl 2-hydroxy-2-methyl-3-(methylsulfonyl)propanoate (racemic) [ka] A solution of racemic methyl 2-hydroxy-2-methyl-3-(methylthio)propanoate (8.00 g, 48.7 mmol) in DCM (400 mL) was stirred in an ice-water bath and treated with mCPBA (33.6 g, 146 mmol) in several portions over approximately 5 min. After 5 min more, additional mCPBA (11.2 g, 48.7 mmol) was added. Stirring was continued for another 5 min, after which the cooling bath was removed and the white suspension was stirred at rt. After 1.75 h, the mixture was filtered and the white solid was washed twice with DCM. The combined filtrate was cooled in an ice-water bath and treated with aqueous NaSO (20 g in 125 mL) in several portions, resulting in an exotherm. After thorough mixing, the layers were separated and the aqueous phase was extracted with DCM. The combined organic phases were washed successively with saturated aqueous NaHCO3 and water, and the two aqueous phases were combined and extracted again with DCM. The combined organic phases were dried over Na2SO4, filtered, and concentrated to give racemic methyl 2-hydroxy-2-methyl-3-(methylsulfonyl)propanoate as a white solid (6.98 g). LCMS m / z 219.1 (M+Na). + . 1 H NMR (499 MHz, CDCl3) δ 3.88 (s, 3H), 3.61 (dd, J=15.1, 1.0 Hz, 1H), 3.44 (d, J=15.0 Hz, 1H), 3.06 (s, 3H), 1.52 (s, 3H).

[0090] All aqueous phases were combined, treated with solid NaCl, and concentrated to a concentrated sludge. The solid was collected by filtration and stirred with DCM while still damp. The mixture was filtered, and the collected solid was washed with additional DCM. The layers of the filtrate were separated, the aqueous phase was extracted with DCM, and the combined organic phase was washed with saturated aqueous NaHCO3, dried over Na2SO4, and concentrated to give additional racemic methyl 2-hydroxy-2-methyl-3-(methylsulfonyl)propanoate as a pale yellow solid (2.23 g).

[0091] Process C: Methyl (R)-2-hydroxy-2-methyl-3-(methylsulfonyl)propanoate and Methyl (S)-2-hydroxy-2-methyl-3-(methylsulfonyl)propanoate [ka] Racemic methyl 2-hydroxy-2-methyl-3-(methylsulfonyl)propanoate (25.3 g) was dissolved in DCM (40 mL) and MeOH (250 mL) and separated by SFC (column: Chiralpak AD-H, 5 x 25 cm, 5 μm; pressure: 100 bar; temperature: 40 °C; mobile phase: CO2-MeOH (82:18); flow rate: 290 mL / min; 0.7 mL injection with a cycle time of 1.2 min). Two fractions were collected. The first eluted fraction was concentrated to give methyl (R)-2-hydroxy-2-methyl-3-(methylsulfonyl)propanoate as a white solid (12.3 g). LCMS m / z 197.2 (M+H) + . 1 H NMR (499 MHz, CDCl3) δ 3.88 (s, 3H), 3.80 (s, 1H), 3.60 (dd, J=15.1, 1.0 Hz, 1H), 3.43 (d, J=15.0 Hz, 1H), 3.06 (s, 3H), 1.51 (s, 3H). The second eluted fraction was concentrated to give methyl (S)-2-hydroxy-2-methyl-3-(methylsulfonyl)propanoate as a white solid (11.9 mg). LCMS m / z 197.0 (M+H) + . 1 H NMR (499 MHz, chloroform-d) δ 3.88 (s, 3H), 3.80 (s, 1H), 3.60 (dd, J=15.1, 1.0 Hz, 1H), 3.43 (d, J=15.0 Hz, 1H), 3.06 (s, 3H), 1.51 (s, 3H). The absolute configuration of this enantiomer was determined from the anomalous dispersion signals by the Flack method using single crystal X-ray analysis in Example 1.

[0092] Process D: (S)-2-Hydroxy-2-methyl-3-(methylsulfonyl)propanoic acid [ka] A solution of methyl (S)-2-hydroxy-2-methyl-3-(methylsulfonyl)propanoate (2.00 g, 10.2 mmol) in THF (51 mL) and MeOH (17 mL) was stirred in an ice-water bath and treated with a solution of LiOH hydrate (0.684 g, 16.3 mmol) in water (17 mL). The cold bath was removed, and the slightly cloudy solution was stirred at room temperature. After 90 minutes, the mixture was stirred in an ice-water bath, treated with 1 M aqueous HCl (16.3 mL, 16.3 mmol), and concentrated in vacuo to remove most of the organic solvent. The aqueous residue was frozen on dry ice-acetone and lyophilized to give (S)-2-hydroxy-2-methyl-3-(methylsulfonyl)propanoic acid as a white solid (2.75 g) containing LiCl and residual water; estimated purity 67%. This material was used without further purification. LCMS m / z 181.1 (M−H) - . 1 H NMR (499 MHz, DMSO-d6) δ 3.53 - 3.37 (m, 2H), 3.00 (s, 3H), 1.37 (s, 3H).

[0093] Intermediate 3 (R)-2-Hydroxy-2-methyl-3-(methylsulfonyl)propanoic acid [ka] Following the procedure used, intermediate 2, methyl (R)-2-hydroxy-2-methyl-3-(methylsulfonyl)propanoate (89 mg, 0.454 mmol) was prepared and converted to (R)-2-hydroxy-2-methyl-3-(methylsulfonyl)propanoic acid. LCMS m / z 183.0 (M+H) + .

[0094] Example 1 (S)-N-((6aS,7R,9aS)-9a-((4-fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinolin-7-yl)-2-hydroxy-2-methyl-3-(methylsulfonyl)propanamide [ka] A mixture of (6aS,7R,9aS)-9a-((4-fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinolin-7-amine, HCl salt (Intermediate 1; 9.90 g, 18.0 mmol) in DMF (150 mL) was treated with (S)-2-hydroxy-2-methyl-3-(methylsulfonyl)propanoic acid (Intermediate 2; 6.50 g, 24.3 mmol), HATU (10.6 g, 27.9 mmol), and DIEA (12.6 mL, 71.9 mmol) at 0 °C, and the mixture was stirred at rt for 1.5 h. The mixture was cooled in an ice-water bath and treated with water (20 mL). Partitioned between EtOAc and saturated aqueous NaHCO3, the organic phase was washed successively with 10% aqueous NaHCO3 and brine, and dried over MgSO4. The residue was purified by silica gel column chromatography (220 g) eluting with EtOAc-hexane (0-100% gradient) to give (S)-N-((6aS,7R,9aS)-9a-((4-fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinolin-7-yl)-2-hydroxy-2-methyl-3-(methylsulfonyl)propanamide as a white solid (12.0 g). LCMS m / z 679.3 (M+H). + . 1H NMR (499 MHz, MeOH-d4) δ 8.15 (d, J=8.3 Hz, 1H), 7.73 (dd, J=8.3, 2.7 Hz, 1H), 7.34 (dd, J=8.4, 5.1 Hz, 2H), 7.16 (t, J=8.7 Hz, 2H), 4.20 - 4.09 (m, 1H), 3.66 (d, J=1.0 Hz, 1H), 3.50 (d, J=14.9 Hz, 1H), 3.29 - 3.21 (m, 1H), 3.12 (s, 3H), 3.07 - 3.01 (m, 1H), 2.71 (dt, J=16.7, 4.0Hz, 1H), 2.45 (dt, J=14.0, 6.8 Hz, 1H), 2.26 - 2.14 (m, 2H), 2.08 - 1.99 (m, 1H), 1.94 (ddd, J=16.7, 12.3, 4.1 Hz, 1H), 1.55 (s, 3H), 1.49 - 1.38 (m, 1H). The absolute configuration was determined from the anomalous dispersion signals by single-crystal X-ray analysis using the Flack method.

[0095] The following additional examples were prepared from Intermediate 1 and the appropriate carboxylic acid using the method of Example 1 and purified by preparative HPLC. [Table 2]

[0096] (General RORγ Gal4 reporter assay) The inverse agonist activity of potential ligands for RORy was measured by luminescence inhibition in a Gal4-luciferase reporter assay in Jurkat cells. Jurkat cells stably overexpressing the RORγ receptor, Jurkat pEx / Gal / hRORγCLBD / HYGpG5luc / blast, were seeded at 10,000 cells / well in 384-well solid white cell culture plates (Perkin Elmer #6007899) in assay buffer RPMI 1640 (Gibco 11875-085 1 L) containing 0.1% BSA, 100× HEPES (Gibco 15360-080), 100 mM sodium pyruvate (Gibco 11360-040), 50 mg / mL hygromycin B (Invitrogen 10687-010), and 10 mg / mL blasticidin (Invitrogen R210-01). Three-fold serial dilutions of test compounds (100 nL) at final concentrations ranging from 40 mM to 0.67 nM were added to the cells, which were then incubated overnight.

[0097] The next day, cells were lysed with 10 μL of Steady-Glo Luciferase Assay System (Promega Cat. No. EZ550) and immediately analyzed. 50 The IC value was determined. 50 Values were defined as the test compound concentration required to reduce luciferase activity by 50% and were calculated using a four-parameter logistic equation to fit the normalized data.

[0098] IC of compounds in the RORγ Gal4 reporter assay 50 The values are shown below. [Table 3]

[0099] (Human whole blood assay) Compounds were diluted in dimethyl sulfoxide (DMSO) and dispensed (60 nL / well) into clear, V-bottom 384-well plates (Matrix Technologies) using ECHO acoustic liquid handling technology. Human whole blood samples (30 μL) were added to each well using a CyBio FeliX liquid dispensing device, and the plate was shaken on a plate shaker for 3 minutes and incubated at 37°C for 1 hour. The wells were then treated with CD3+CD28 at a final concentration of 1 μg / mL in 30 μL per well of AIM-V medium. The plate was shaken on a plate shaker for 3 minutes, and the reaction mixture was then incubated at 37°C for 20 hours. Plasma was collected from each sample by centrifugation (450 g, 5 minutes, ambient temperature). Aliquots of plasma samples (4 μL) were then dispensed into individual wells of a white, shallow 384-well ProxiPlate (PerkinElmer) using a FeliX liquid handling device, and IL17A content was measured using AlphaLISA technology according to the manufacturer's instructions.

[0100] Using proprietary BMS data analysis software, the EC 50 values were determined; where baseline was established using the mean value of DMSO and 100% induction was established using the value of the highest concentration of reference compound tested.

[0101] Patch clamp assay protocol for sodium channel recording Cardiac sodium ion channel assays were performed using human embryonic kidney cells (HEK 293) stably expressing the cloned human sodium channel gene SCN5A ion channel.

[0102] In the sodium channel assay, compounds were evaluated at 10 μM, and efficacy was calculated by measuring the inhibition of peak inward current. Two stimulation frequencies, 1 Hz and 4 Hz (compound A), were used to examine the rate-dependent effects of compounds on sodium channels. All results are reported as mean ± SEM. DMSO was used as a vehicle, and the final concentration of DMSO did not exceed 0.1%.

[0103] Membrane current recordings were performed using a whole-cell modification of the conventional patch clamp technique with a Multiclamp 700 series miniature patch clamp amplifier (Axon Instruments, Foster City, California). Cells expressing the SCN5A sodium ion channel were placed in a Plexiglas bath chamber, mounted on the stage of an inverted microscope, and continuously perfused with bath solution.

[0104] The sodium current bath solution contained (mM): 140 NaCl, 4 KCl, 1.8 CaCl, 1 MgCl, 10 glucose, 10 HEPES (pH 7.4, NaOH). The filling solution for patch pipettes used for sodium channel experiments contained (mM): 130 KCl, 1 MgCl, 1 CaCl, 5 ATP-K, 10 EGTA, 10 HEPES (pH 7.2, KOH).

[0105] To determine steady-state inhibition, sodium currents were elicited every 5 seconds using the following voltage protocol (0.2 Hz): Cells were held at a potential of -90 mV and stepped to -20 mV over 45 ms. Maximal sodium currents in response to the depolarizing step to -20 mV were monitored in control buffer and after test substance application until a new steady state was achieved in the presence of the test substance. To assess rate-dependent inhibition of sodium currents, trains of voltage steps (30 voltage sweeps each) at frequencies of 1 Hz and 4 Hz were applied to cells before test substance application (control) and after steady-state inhibition with test substance, as determined at a frequency of 0.2 Hz. The voltage waveforms used in rate-dependent experiments were the same as those used to assess steady-state inhibition at a 0.2 Hz stimulation frequency. Rate-dependent inhibition was calculated by comparing the average of the last three of the 30 voltage sweeps in the presence of the test substance with the average of the last three of the 30 voltage sweeps under control conditions at each frequency tested.

[0106] Each compound was tested in duplicate at a concentration of 10 μM with 1 Hz stimulation unless otherwise noted (compound A was tested in triplicate with both 1 Hz and 4 Hz stimulation). Currents were sampled at a rate greater than twice the low-pass filter rate. The flow rate was held constant throughout the experiment. All currents were recorded at room temperature to 25°C.

[0107] The IC50 values of Compound 1 and reference Compound A in the RORγGal4 reporter assay, human whole blood assay and Na channel assay are provided below.

[0108] Compound A is disclosed and claimed in U.S. Patent No. 9,815,859. Compound 1 was found to be twice as potent in a human whole blood assay and essentially inactive in a Na channel assay, whereas Compound A exhibited an IC50 of 8.6 μM in this assay. These differences made Compound 1 a superior candidate for further development.

Table 4

Claims

1. (S)—N-((6aS,7R,9aS)-9a-((4-fluorophenyl)sulfonyl)-3-(perfluoropropan-2-yl)-6,6a,7,8,9,9a-hexahydro-5H-cyclopenta[f]quinolin-7-yl)-2-hydroxy-2-methyl-3-(methylsulfonyl)propanamide, formula (II): 【Chemistry 3】 Compound.

2. 10. A pharmaceutical composition comprising a compound of claim 1 and a pharmaceutically acceptable carrier or diluent.

3. 10. A pharmaceutical composition comprising the compound of claim 1 for treating a disease or disorder selected from an autoimmune disease or disorder, asthma, an allergic disease or disorder, a metabolic disease or disorder, and cancer.

4. 4. The pharmaceutical composition of claim 3, wherein the autoimmune disease or disorder is selected from psoriasis, rheumatoid arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, acute graft-versus-host disease, psoriatic arthritis, ankylosing spondylitis, systemic lupus erythematosus, lupus nephritis, Sjogren's syndrome, and multiple sclerosis.

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