Triazinone derivatives as NLRP3 inhibitors

Abstract

The present invention relates to a novel compound having the formula: 6-[[(3R)-1-ethyl-3-piperidinyl] amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1, 2, 4-triazine-5-one or 6-[[(3R)-1-ethyl-3-piperidinyl] amino]-3-(2-hydroxy-3-bicyclo [4.2. 0] oct-1, 3, 5-trialkenyl)-4-methyl-1, 2, 4-triazine-5-one, and a pharmaceutically acceptable salt thereof, and to a pharmaceutical composition comprising the same. Comprising the compound; and methods of using the compounds. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to organic compounds that can be used for the treatment and / or prevention in mammals, and particularly to compounds that modulate NLRP3 inhibition.

[0002] The present invention provides novel compounds selected from the following

[0003] 6-[[(3R)-1-Ethyl-3-piperidinyl]amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazin-5-one; and

[0004] 6-[[(3R)-1-Ethyl-3-piperidinyl]amino]-3-(2-hydroxy-3-bicyclo[4.2.0]octa-1,3,5-trienyl)-4-methyl-1,2,4-triazin-5-one;

[0005] 6-[[(3R)-1-Ethyl-3-piperidinyl]-methyl-amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazin-5-one; and

[0006] or a pharmaceutically acceptable salt thereof. Background Art

[0007] The NOD-like receptor (NLR) family (NLRP3 inflammasome containing a pyrin domain) is a component of the inflammatory process, and its abnormal activity is pathogenic in genetic disorders such as cryopyrin-associated periodic syndromes (CAPS) and complex diseases such as multiple sclerosis, type 2 diabetes, Alzheimer's disease, and atherosclerosis.

[0008] NLRP3 is an intracellular signaling molecule that can sense many pathogen-derived, environmental, and host-derived factors. Upon activation, NLRP3 binds to apoptosis-associated speck-like protein containing a caspase activation and recruitment domain (ASC). Then, ASC polymerizes to form large aggregates called ASC specks. The polymeric ASC then interacts with the cysteine protease caspase-1 to form a complex called the inflammasome. This leads to the activation of caspase-1, which cleaves the precursor forms of the pro-inflammatory cytokines IL-1β and IL-18 (called pro-IL-1β and pro-IL-18, respectively), thereby activating these cytokines. Caspase-1 also mediates a type of inflammatory cell death called pyroptosis. ASC specks can also recruit and activate caspase-8, which can process pro-IL-1β and pro-IL-18 and trigger apoptotic cell death.

[0009] Caspase-1 cleaves pro-IL-1β and pro-IL-18 into their active forms, which are secreted by cells. Active Caspase-1 also cleaves gasdermin-D to trigger pyroptosis. Caspase-1 can also mediate the release of alarm protein molecules such as IL-33 and high-mobility group box 1 (HMGB1) by controlling the pyroptotic cell death pathway. Caspase-1 also cleaves intracellular IL-1R2, leading to its degradation and the release of IL-1α. In human cells, Caspase-1 can also control the processing and secretion of IL-37. Many other substrates of Caspase-1, such as components of the cytoskeleton and glycolytic pathways, may contribute to Caspase-1-dependent inflammation.

[0010] NLRP3-dependent ASC specks are released into the extracellular environment, where they can activate Caspase-1, induce the processing of Caspase-1 substrates, and propagate inflammation.

[0011] The active cytokines derived from NLRP3 inflammasome activation are important drivers of inflammation and interact with other cytokine pathways to form an immune response to infection and injury. For example, IL-1β signaling induces the secretion of the pro-inflammatory cytokines IL-6 and TNF. IL-1β and IL-18 act in concert with IL-23 to induce memory CD4 Th17 cells and γδ T cells to produce IL-17 in the absence of T cell receptor engagement. IL-18 and IL-12 also act together to induce IFN-γ production from memory T cells and NK cells, driving a Th1 response.

[0012] The hereditary CAPS diseases Muckle-Wells syndrome (MWS), familial cold autoinflammatory syndrome (FCAS), and neonatal-onset multisystem inflammatory disease (NOMID) are caused by gain-of-function mutations in NLRP3, thus defining NLRP3 as a key component of the inflammatory process. NLRP3 is also associated with the pathogenesis of many complex diseases, which particularly include metabolic disorders such as type 2 diabetes, atherosclerosis, obesity, and gout.

[0013] The role of NLRP3 in central nervous system diseases is emerging, and lung diseases have also been shown to be affected by NLRP3. NLRP3 is also thought to play a role in many central nervous system diseases, including Parkinson's disease (PD), Alzheimer's disease (AD), dementia, Huntington's disease, cerebral malaria, and brain injury caused by pneumococcal meningitis (Walsh et al., Nature Reviews, 15:84-97, 2014 and Dempsey et al. Brain. Behav. Immun. 2017 61:306-316). NLRP3 has also been shown to play a role in many lung diseases, including chronic obstructive pulmonary disease (COPD), asthma (including steroid-resistant asthma), asbestosis, and silicosis (De Nardo et al., Am. J. Pathol., 184:42-54, 2014 and Kim et al. Am J Respir Crit Care Med. 2017 196(3):283-97). In addition, NLRP3 has a role in the development of liver disease, kidney disease, and aging. Many of these associations have been defined using Nlrp3 - / - mice, but there are also insights into the specific activation of NLRP3 in these diseases. In type 2 diabetes (T2D), the deposition of islet amyloid polypeptide in the pancreas activates NLRP3 and IL-1β signaling, leading to cell death and inflammation.

[0014] Several small molecules have been shown to inhibit the NLRP3 inflammasome. Glibenclamide inhibits IL-1β production at micromolar concentrations in response to the activation of NLRP3 but not NLRC4 or NLRP1. Other previously characterized weak NLRP3 inhibitors include parthenolide, 3,4-methylenedioxy-β-nitrostyrene, and dimethyl sulfoxide (DMSO), although these agents have limited potency and are non-specific.

[0015] Current treatments for NLRP3-related diseases include biologic agents that target IL-1. These are the recombinant IL-1 receptor antagonist anakinra, the neutralizing IL-1β antibody canakinumab, and the soluble decoy IL-1 receptor rilonacept. These approaches have been shown to successfully treat CAPS, and these biologic agents have been used in clinical trials for other IL-1β-related diseases.

[0016] There is a need to provide compounds having improved pharmacological and / or physiological and / or physicochemical properties, and / or compounds that provide useful alternatives to known compounds. In particular, compounds having good activity and improved in vitro stability. SUMMARY OF THE INVENTION

[0017] The present invention provides novel compounds selected from the following

[0018] 6-[[(3R)-1-Ethyl-3-piperidinyl]amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazin-5-one; and

[0019] 6-[[(3R)-1-Ethyl-3-piperidinyl]amino]-3-(2-hydroxy-3-bicyclo[4.2.0]octa-1,3,5-trienyl)-4-methyl-1,2,4-triazin-5-one;

[0020] 6-[[(3R)-1-Ethyl-3-piperidinyl]-methyl-amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazin-5-one; and

[0021] or a pharmaceutically acceptable salt thereof.

[0022] The term "pharmaceutically acceptable salt" refers to those salts that retain the biological effects and properties of the free base or free acid and are not otherwise undesirable biologically or otherwise. These salts are formed with inorganic acids such as trifluoroacetic acid, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid (especially hydrochloric acid) and organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, N-acetylcysteine. Additionally, these salts can be prepared by adding an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium salts. Salts derived from organic bases include, but are not limited to, salts of the following substances: primary amines, secondary amines and tertiary amines, including substituted amines of naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, lysine, arginine, N-ethylpiperidine, piperidine, polyamine resins. The compounds of this formula can also exist in the form of zwitterions. Particularly preferred pharmaceutically acceptable salts of the compounds of this formula are salts formed with formic acid and salts formed with hydrochloric acid, yielding hydrochloride, dihydrochloride or trihydrochloride.

[0023] The abbreviation uM means micromole and is equivalent to the symbol μM.

[0024] The abbreviation uL means microliter and is equivalent to the symbol μL.

[0025] The abbreviation ug means microgram and is equivalent to the symbol μg.

[0026] Another embodiment of the present invention provides compounds and their pharmaceutically acceptable salts or esters according to those as described herein, particularly provides compounds and their pharmaceutically acceptable salts according to those as described herein, and more particularly provides compounds according to those as described herein.

[0027] Particular examples of compounds as described herein are selected from

[0028] 6-[[(3R)-1-ethyl-3-piperidinyl]amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazin-5-one;

[0029] 6-[[(3R)-1-ethyl-3-piperidinyl]amino]-3-(2-hydroxy-3-bicyclo[4.2.0]octan-1,3,5-tri

[0030] 4-methyl-1,2,4-triazine-5-one;

[0031] and pharmaceutically acceptable salts thereof.

[0032] Another particular example of a compound as described herein is 6-[[(3R)-1-ethyl-3-piperidinyl]-methyl-amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazin-5-one or a pharmaceutically acceptable salt thereof.

[0033] One embodiment of the present invention provides a compound of formula 6-[[(3R)-1-ethyl-3-piperidinyl]amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazine-5-one or a pharmaceutically acceptable salt thereof.

[0034] One embodiment of the present invention provides a compound of formula 6-[[(3R)-1-ethyl-3-piperidinyl]amino]-3-(2-hydroxy-3-bicyclo[4.2.0]octa-1,3,5-trienyl)-4-methyl-1,2,4-triazine-5-one or a pharmaceutically acceptable salt thereof.

[0035] Another embodiment of the present invention provides a pharmaceutical composition or medicine containing the compound of the present invention and a therapeutic inert carrier, diluent or excipient, and a method for preparing such compositions and medicines using the compound of the present invention. In one example, the compound can be formulated as a galenical administration form by mixing with a physiologically acceptable carrier (i.e., a carrier that is nontoxic to the recipient at the dosage and concentration used) at ambient temperature at an appropriate pH and at a desired purity. The pH of the formulation depends primarily on the specific use and concentration of the compound, but is preferably in the range of about 3 to about 8. In one example, the compound of the formula is prepared in an acetate buffer of pH 5. In another embodiment, the compound of the formula is sterile. The compound can be, for example, stored as a solid or amorphous composition, as a lyophilized formulation or as an aqueous solution.

[0036] The compositions are formulated, dosed and administered in a manner consistent with good medical practice. Factors to be considered in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the timing of administration, and other factors known to the practicing physician.

[0037] The compounds of the invention can be administered by any suitable means, including orally, topically (including buccal and sublingual), rectally, vaginally, transdermally, parenterally, subcutaneously, intraperitoneally, intraluminally, intradermally, intrathecally and epidurally and intranasally, and (if required for local treatment) intralesionally. Parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration.

[0038] The compounds of the invention can be administered in any convenient dosage form, for example, tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions can contain conventional components in pharmaceutical formulations, for example, diluents, carriers, pH regulators, sweeteners, fillers and other active agents.

[0039] Conventional formulations are prepared by mixing the compounds of the invention with a carrier or excipient. Suitable carriers and excipients are well known to those skilled in the art and are described in detail, for example, in Ansel, Howard C. et al., Ansel’s Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams and Wilkins, 2004; Gennaro, Alfonso R. et al. Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams and Wilkins, 2000; and Rowe, Raymond C. Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005. The formulations can also contain one or more buffering agents, stabilizers, surfactants, wetting agents, lubricants, emulsifying agents, suspending agents, preservatives, antioxidants, opacifying agents, glidants, processing aids, colorants, sweeteners, flavorants, diluents and other known additives to provide an aesthetic presentation of the medicament (e.g., the compounds of the invention or their pharmaceutical compositions) or to facilitate the preparation of the pharmaceutical product (e.g., the drug).

[0040] The compounds and their pharmaceutically acceptable salts can be processed together with pharmaceutically inert inorganic or organic auxiliaries for the production of tablets, coated tablets, dragees, hard gelatin capsules, injection solutions or topical preparations. For example, lactose, corn starch or its derivatives, talc, stearic acid or its salts, etc. can be used as such auxiliaries for tablets, dragees and hard gelatin capsules.

[0041] Suitable auxiliaries for soft gelatin capsules are, for example, vegetable oils, waxes, fats, semi-solid substances and liquid polyols, etc.

[0042] Suitable auxiliaries for the preparation of solutions and syrups are, for example, water, polyols, sucrose, invert sugar, glucose, etc.

[0043] Suitable auxiliaries for injection solutions are, for example, water, alcohols, polyols, glycerol, vegetable oils, etc.

[0044] Suitable auxiliaries for suppositories are, for example, natural or hardened oils, waxes, fats, semi-solid or liquid polyols, etc.

[0045] Suitable auxiliaries for topical ophthalmic preparations are, for example, cyclodextrins, mannitol or many other carriers and excipients known in the art.

[0046] In addition, pharmaceutical preparations can contain preservatives, solubilizers, thickening substances, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, fragrances, salts for altering the osmotic pressure, buffer masking agents or antioxidants. They can also contain other therapeutically valuable substances.

[0047] The dosage can vary within a wide range and will of course be suitable for the various requirements in each specific case. Generally speaking, in the case of oral administration, a daily dosage of about 0.1 mg to 20 mg per kg body weight, preferably about 0.5 mg to 4 mg per kg body weight (e.g., about 300 mg per person), should be appropriate, which is preferably divided into 1 to 3 separate doses (which can consist of, for example, the same amount). In the case of topical administration, the formulation can contain 0.001% to 15% by weight of the drug, and the required dosage can be 0.1 mg to 25 mg, administered once a day or once a week, or several times a day (2 to 4 times), or several times a week. However, it is obvious that when shown to be applicable, the upper or lower limits given in this text can be exceeded.

[0048] One embodiment of the present invention is a compound as described herein according to the present invention, which is used as a therapeutically active substance.

[0049] One embodiment of the present invention is a compound as described herein according to the present invention, which is used for treating or preventing a disease, disorder or condition, wherein the disease, disorder or condition responds to NLRP3 inhibition.

[0050] One embodiment of the present invention is a compound as described herein according to the present invention for treating or preventing a disease, disorder or condition, wherein the disorder or condition responds to NLRP3 inhibition.

[0051] As used herein, the term "NLRP3 inhibition" refers to a complete or partial reduction in the level of NLRP3 activity and includes, for example, inhibiting active NLRP3 and / or inhibiting the activation of NLRP3.

[0052] There is evidence that NLRP3-induced IL-1 and IL-18 play a role in the inflammatory responses associated with or caused by a variety of different disorders (Menu et al., Clinical and Experimental Immunology, 166:1-15, 2011; Strowig et al., Nature, 481:278-286, 2012).

[0053] In one embodiment, the disease, disorder or condition is selected from:

[0054] (i) Inflammation;

[0055] (ii) Autoimmune diseases;

[0056] (iii) Cancer;

[0057] (iv) Infection;

[0058] (v) Central nervous system diseases;

[0059] (vi) Metabolic diseases;

[0060] (vii) Cardiovascular diseases;

[0061] (viii) Respiratory diseases;

[0062] (ix) Liver diseases;

[0063] (x) Kidney diseases;

[0064] (xi) Eye diseases;

[0065] (xii) Skin diseases;

[0066] (xiii) Lymphatic disorders;

[0067] (xiv) Psychological disorders;

[0068] (xv) Graft-versus-host disease;

[0069] (xvi) Allodynia;

[0070] (xvii) Conditions associated with diabetes; and

[0071] (xviii) Any disease in which an individual has been determined to carry a germline or somatic non-silent mutation in NLRP3

[0072] In another embodiment, the disease, disorder or condition is selected from:

[0073] (i) Cancer;

[0074] (ii) Infection;

[0075] (iii) Central nervous system diseases;

[0076] (iv) Cardiovascular diseases;

[0077] (v) Liver diseases;

[0078] (vi) Eye diseases; or

[0079] (vii) Skin diseases.

[0080] In yet another exemplary embodiment of the invention, the disease, disorder or condition is inflammation. Examples of inflammation that can be treated or prevented include inflammatory responses associated with or caused by the following diseases:

[0081] (i) Skin diseases such as contact hypersensitivity, bullous pemphigoid, sunburn, psoriasis, atopic dermatitis, contact dermatitis, allergic contact dermatitis, seborrheic dermatitis, lichen planus, scleroderma, pemphigus, epidermolysis bullosa, urticaria, erythema or alopecia;

[0083] (ii) Arthropathies such as osteoarthritis, systemic juvenile idiopathic arthritis, adult-onset Still's disease, relapsing polychondritis, rheumatoid arthritis, juvenile chronic arthritis, gout or seronegative spondyloarthropathies (e.g., ankylosing spondylitis, psoriatic arthritis or Reiter's disease);

[0084] Ter's disease);

[0085] (iii) Muscle diseases such as polymyositis or myasthenia gravis;

[0086] (iv) Gastrointestinal diseases such as inflammatory bowel disease (including Crohn's disease and ulcerative colitis), colitis, gastric ulcer, celiac disease, proctitis, pancreatitis, eosinophilic gastroenteritis, mastocytosis, antiphospholipid syndrome, or food-related allergies that may have effects outside the gut (e.g., migraine, rhinitis or eczema);

[0087] (e.g., migraine, rhinitis or eczema);

[0088] (v) Respiratory diseases, such as chronic obstructive pulmonary disease (COPD), asthma (including eosinophilic asthma, bronchial asthma, allergic asthma, endogenous asthma, exogenous asthma or dust asthma, and especially chronic or refractory asthma, such as late-stage asthma and airway hyperresponsiveness), bronchitis, rhinitis (including acute rhinitis, allergic rhinitis, atrophic rhinitis, chronic rhinitis, caseous rhinitis, hypertrophic rhinitis, pustular rhinitis, dry rhinitis, drug-induced rhinitis, membranous rhinitis, seasonal rhinitis such as hay fever and vasomotor rhinitis), sinusitis, idiopathic pulmonary fibrosis (IPF), sarcoidosis, farmer's lung, silicosis, asbestosis, inflammation caused by volcanic ash, adult respiratory distress syndrome, allergic pneumonia or idiopathic

[0089] interstitial pneumonia;

[0090] (vi) Vascular diseases, such as atherosclerosis, Behcet's disease, vasculitis or Wegener's granulomatosis;

[0091] granulomatosis;

[0092] (vii) Autoimmune diseases, such as systemic lupus erythematosus, Sjogren's syndrome, systemic sclerosis, Hashimoto's thyroiditis, type I diabetes, idiopathic thrombocytopenic purpura or Graves' disease;

[0093] Graves' disease;

[0094] (viii) Eye diseases, such as uveitis, allergic conjunctivitis or vernal conjunctivitis;

[0095] (ix) Neurological diseases, such as multiple sclerosis or encephalomyelitis;

[0096] (x) Infections or infection-related diseases, such as acquired immunodeficiency syndrome (AIDS), acute or chronic bacterial infections, acute or chronic parasitic infections, acute or chronic viral infections, acute or chronic fungal infections, meningitis, hepatitis (hepatitis A, B or C or other viral hepatitis), peritonitis, pneumonia, epiglottitis, malaria, dengue hemorrhagic fever, leishmaniasis, streptococcal myositis, Mycobacterium tuberculosis (including co-infection with Mycobacterium tuberculosis and HIV), Mycobacterium avium intracellulare, Pneumocystis carinii pneumonia, orchitis / epididymitis, Legionella, Lyme disease, influenza A, Epstein-Barr virus infection, viral encephalitis / aseptic meningitis or pelvic

[0097] inflammatory diseases;

[0098] (xi) Kidney diseases, such as mesangial proliferative glomerulonephritis, nephrotic syndrome, nephritis, glomerulonephritis, obesity-related glomerulopathy, acute renal failure, acute kidney injury, uremia, nephrotic syndrome, renal fibrosis (including chronic crystal nephropathy) or renal hypertension;

[0099] (xii) Lymphatic diseases, such as Castleman disease;

[0100] (xiii) Diseases of the immune system or diseases involving the immune system, such as hyper-IgE syndrome, leprosy, familial hemophagocytic lymphohistiocytosis or graft-versus-host disease;

[0101] (xiv) Liver diseases, such as chronic active hepatitis, non-alcoholic steatohepatitis (NASH), alcoholic hepatitis, non-alcoholic fatty liver disease (NAFLD), alcoholic fatty liver disease (AFLD), alcoholic steatohepatitis (ASH), primary biliary cirrhosis, fulminant hepatitis, liver fibrosis or liver failure;

[0102] (xv) Cancers, including those listed above;

[0103] (xvi) Burns, trauma, injury, bleeding or stroke;

[0104] (xvii) Radiation exposure;

[0105] (xviii) Metabolic diseases, such as type 2 diabetes (T2D), atherosclerosis, obesity, gout or pseudogout; and / or

[0106] (xix) Pain, such as inflammatory hyperalgesia, pelvic pain, allodynia, neuropathic pain or bone pain caused by cancer.

[0107] One embodiment of the present invention is a compound according to the present invention as described herein for treating or preventing a disease, disorder or condition selected from:

[0108] (i) Inflammation;

[0109] (ii) Autoimmune diseases;

[0110] (iii) Cancers;

[0111] (iv) Infections;

[0112] (v) Central nervous system diseases;

[0113] (vi) Metabolic diseases;

[0114] (vii) Cardiovascular diseases;

[0115] (viii) Respiratory diseases;

[0116] (ix) Liver diseases;

[0117] (x) Kidney diseases;

[0118] (xi) Eye diseases;

[0119] (xii) Skin diseases;

[0120] (xiii) Lymphatic disorders;

[0121] (xiv) Psychological disorders;

[0122] (xv) Graft-versus-host disease;

[0123] (xvi) Allodynia;

[0124] (xvii) Conditions associated with diabetes; and

[0125] (xviii) Any disease in which an individual has been determined to carry a germline or somatic non-silent mutation in NLRP3.

[0126] One embodiment of the present invention is the use of a compound as described herein according to the present invention in the treatment or prevention of a disease, disorder or condition, wherein the disease, disorder or condition responds to NLRP3 inhibition.

[0127] One embodiment of the present invention is the use of a compound as described herein according to the present invention in the treatment or prevention of a disease, disorder or condition selected from Alzheimer's disease and Parkinson's disease.

[0128] One embodiment of the present invention is the use of a compound as described herein according to the present invention for the treatment or prevention of a disease, disorder or condition selected from asthma or COPD.

[0129] One embodiment of the present invention is a compound as described herein according to the present invention for use in the treatment or prevention of a disease, disorder or condition selected from Alzheimer's disease and Parkinson's disease.

[0130] One embodiment of the present invention is a compound as described herein according to the present invention for use in the treatment or prevention of a disease, disorder or condition selected from asthma or COPD.

[0131] One embodiment of the present invention is the use of a compound as described herein according to the present invention for the preparation of a medicament for the treatment or prevention of a disease, disorder or condition selected from Alzheimer's disease and Parkinson's disease.

[0132] One embodiment of the present invention is the use of a compound as described herein according to the present invention for the preparation of a medicament for the treatment or prevention of a disease, disorder or condition selected from asthma or COPD.

[0133] One embodiment of the present invention is a method for the treatment or prevention of a disease, disorder or condition selected from Alzheimer's disease and Parkinson's disease, the method comprising administering an effective amount of a compound as described herein according to the present invention.

[0134] One embodiment of the present invention is a method for treating or preventing a disease, disorder or condition selected from asthma or COPD, the method comprising administering an effective amount of a compound according to the present invention as described herein.

[0135] One embodiment of the present invention relates to a method for inhibiting NLRP3, the method comprising administering an effective amount of a compound according to the present invention as described herein.

[0136] Another embodiment of the present invention is a compound of the formula as described herein, which is manufactured according to any one of the said methods.

[0137] One embodiment of the present invention is a pharmaceutical composition comprising a compound according to the present invention as described herein and a pharmaceutically inert carrier.

[0138] Determination Procedure

[0139] NLRP3 and Pyroptosis

[0140] It is well known that the activation of NLRP3 leads to pyroptosis, and this feature plays an important role in the manifestation of clinical diseases (Yang-gang Liu et al., Cell Death & Disease, 2017, 8(2), e2579; Alexander Wree et al., Hepatology, 2014, 59(3), 898-910; Alex Baldwin et al., Journal of Medicinal Chemistry, 2016, 59(5), 1691-1710; Ema Ozaki et al., Journal of Inflammation Research, 2015, 8, 15-27; Zhen Xie and Gang Zhao, Neuroimmunology Neuroinflammation, 2014, 1(2), 60-65; Mattia Cocco et al., Journal of Medicinal Chemistry, 2014, 57(24), 10366-10382; T. Satoh et al., Cell Death & Disease, 2013, 4, e644). Therefore, it is expected that inhibitors of NLRP3 will prevent pyroptosis and the release of pro-inflammatory cytokines (such as IL-1β) from cells.

[0141] THP-1 Cells: Culture and Preparation

[0142] Grow THP-1 cells (ATCC#TIB-202) in RPMI containing L-glutamine (Gibco#11835) supplemented with 1 mM sodium pyruvate (Sigma#S8636) and penicillin (100 units / ml) / streptomycin (0.1 mg / ml) (Sigma#P4333) in 10% fetal bovine serum (FBS) (Sigma#F0804). Cells are passaged routinely and grown to confluence (about 10 6 cells / ml). On the day of the experiment, harvest THP-1 cells and resuspend them in RPMI medium (without FBS). Then count the cells and check viability (>90%) by trypan blue (Sigma#T8154). Make appropriate dilutions to obtain a concentration of 625,000 cells / ml. Add LPS (Sigma#L4524) to the diluted cell solution to obtain a final assay concentration (FAC) of 1 μg / ml. Aliquot 40 μl of the final preparation into each well of a 96-well plate. The plate thus prepared is used for compound screening.

[0143] THP-1 Cell Pyroptosis Assay

[0144] Compound screening is carried out in a stepwise assay according to the following method.

[0145] 1. Seed THP-1 cells (25,000 cells / well) containing 1.0 μg / ml LPS in 40 μl RPMI medium (without FBS) in a 96-well, black-wall, clear-bottom cell culture plate coated with poly-D-lysine (VWR

[0146] #734-0317).

[0147] 2. Add 5 μl of the compound (8-point half-log dilution, 10 μM highest dose) or vehicle (DMSO 0.1% FAC) to the appropriate wells.

[0148] 3. Incubate at 37 °C, 5% CO2 for 3 hours.

[0149] 4. Add 5 μl of nigericin (Sigma#N7143) (FAC 5 μM) to all wells.

[0150] 5. Incubate at 37 °C, 5% CO2 for 1 hr.

[0151] 6. At the end of the incubation period, spin the plate at 300 x g for 3 minutes and remove the supernatant.

[0152] 7. Then add 50 μl of resazurin (Sigma#R7017) (FAC 100 μM resazurin, dissolved in RPMI medium without FBS), and incubate the plate at 37 °C and 5% CO2 for an additional 1 - 2 hours.

[0153] 8. Read the plate in the Envision reader at Ex 560 nm and Em 590 nm

[0154] 9. IC 50 The data fit a non - linear regression equation (log inhibitor vs. response variable slope 4 - parameter)

[0155] The results of the pyroptosis assay are summarized in Table 1 below, as THP IC 50 .

[0156] Human Whole Blood IL-1β Release Assay

[0157] For systemic delivery, the ability to inhibit NLRP3 when the compound is present in the bloodstream is very important. For this reason, the NLRP3 inhibitory activity of multiple compounds in human whole blood was investigated according to the following protocol.

[0158] Human whole blood in lithium heparin tubes was obtained from healthy donors from a panel of volunteer donors.

[0159] 1. Place 80 μl of whole blood containing 1 μg / ml LPS into a 96 - well clear - bottom cell culture plate (Corning #3585)

[0160] 2. Add 10 μl of the compound (8 - point half - log dilution, 10 μM highest dose) or vehicle (DMSO 0.1% FAC) to the appropriate wells

[0161] 3. Incubate at 37 °C, 5% CO2 for 3 hours

[0162] 4. Add 10 μl of nigericin (Sigma #N7143) (10 μM FAC) to all wells

[0163] 5. Incubate at 37 °C, 5% CO2 for 1 hr

[0164] 6. At the end of the incubation period, spin the plate at 300 xg for 5 minutes to pellet the cells and remove 20 μl of the supernatant, and add it to a 96 - well V - bottom plate for IL - 1β analysis (Note: These plates containing the supernatant can be stored at - 80 °C for later analysis)

[0165] 7. IL - 1β was measured according to the manufacturer's protocol (Perkin Elmer - AlphaLisa IL - 1 Kit AL220F - 5000)

[0166] 8. IC 50 The data fit a non - linear regression equation (log inhibitor vs. response variable slope 4 - parameter)

[0167] The results of the human whole blood assay are summarized in Table 1 below as HWB IC 50 .

[0168] hERG Screening Assay

[0169] During the development of small molecule drugs, one of the most common adverse side effects leading to drug failure is arrhythmia. This failure is usually related to the ability of the drug to inhibit the human ether-à-go-go related gene (hERG) cardiac potassium channel. Therefore, no inhibition or low inhibition of the hERG cardiac potassium channel is considered beneficial.

[0170] Cells

[0171] The CHO crelox hERG cell line (ATCC reference number PTA-6812, female Chinese hamster cells) was generated and validated at Roche. Ready-to-use frozen CHO-hERG cells were cryopreserved at Evotec (Germany) and used directly in the experiments.

[0172] Experimental solutions

[0173] The extracellular solution contained (in mM): NaCl 150; KCl 4; CaCl2 1; MgCl2 1; HEPES 10; pH 7.2 - 7.4 with NaOH, osmolarity 290 - 330 mOsm. The internal solution contained (in mM): KCl, 10; KF, 100; NaCl, 10; HEPES, 10; EGTA, 20; pH = 7.0 - 7.4 with KOH, osmolarity 260 - 300 mOsm.

[0174] Electrophysiology

[0175] The effect of the compound on hERG K+-current parameters will be evaluated in at least 4 cells at 2 concentrations.

[0176] The hERG test was performed using an automated patch clamp system 384 (Nanion Technologies GmbH, Germany). The K+ current was measured using the patch voltage clamp technique in the whole cell configuration at 35 - 37 °C.

[0177] The cells were held at a resting voltage of -80 mV and stimulated by the In Figure 1 voltage pattern shown (pulse pattern to elicit outward K + current) to activate the hERG channel and conduct the IKhERG current outward, with a stimulation frequency of 0.1 Hz (6 bpm)

[0178] Data analysis

[0179] The amplitudes of IKhERG at each drug concentration were recorded and compared with the vehicle control value (set at 100%) to define the fractional blockade. The concentration-response data conformed to the following relationship

[0180]

[0181] The concentration-response curves were fitted using the EworkBook suite (ID Business Solutions Ltd, UK) by non-linear regression analysis. Data fitting was performed using a 4-parameter Logistic model (fit = (A+(B / (1+((x / C)^D)))), where A = 0 and B = 100).

[0182] The results of the hERG assays are summarized in Table 2 below as hERG IC 20 。

[0183] Transcellular P-gp Assay:

[0184] For general assays, transfected LLC-PK1 cells (porcine kidney epithelial cells) overexpressing human or mouse P-gp were used and cultured on 96-well semi-permeable filter plates, where they formed a polarized monolayer with tight junctions and served as a barrier between the apical and basolateral compartments.

[0185] P-gp was expressed in the apical membrane of the monolayer.

[0186] The tightness of the cell monolayer and the functional activity of P-gp were confirmed by adding the cell-impermeable tracer fluorescein and the reference P-gp substrate edoxaban, respectively.

[0187] PAMPA :

[0188] PAMPA (Parallel Artificial Membrane Permeability Assay) was the first-line permeability screening for candidate drugs. The PAMPA assay used an artificial phospholipid membrane to simulate transcellular absorption conditions. This assay determined the permeability values that could be used for compound optimization and ranking purposes and the input parameters for computer models to predict intestinal absorption.

[0189] The donor concentration was measured at t-start (reference) and compared with the donor and acceptor concentrations after a certain time (t-end) to calculate the extent of passage of the compound through the membrane.

[0190] Microsomal Stability :

[0191] On a TECAN (Tecan Group Ltd, Switzerland) automated liquid handling system, incubation was carried out in a 96-well plate at 37 °C in microsomes with a test compound (0.5 mg / mL) at 1 μM and cofactor NADPH. After a 10-minute pre-incubation step of the test compound with microsomes, the enzymatic reaction was initiated by adding the cofactor. At 1, 3, 6, 9, 15, 25, 35, and 45 minutes, aliquots of the culture were taken and quenched with 1:3 (v / v) acetonitrile containing an internal standard. The samples were then cooled and centrifuged, and the supernatant was subsequently analyzed by LC-MS / MS2.

[0192] Hepatocyte Metabolic Stability :

[0193] Assay description:

[0194] Biomaterials. Cryopreserved hepatocytes [mouse, rat, rabbit, monkey, and human (male and female; mixed)] were obtained. Throughout the study, the viability of the reconstituted hepatocytes was at least 80%. Ready-to-use rat / human cultures [long-term hepatocyte co-cultures; pooled (for humans, n = 5 for males and n = 5 for females)] with stromal mouse fibroblasts (negative control; pooled) and the plates for incubation were used to obtain application medium and maintenance medium.

[0195] Metabolism was carried out by suspending hepatocytes. Primary pooled cryopreserved hepatocytes were reconstituted in pre-warmed William's E medium containing 10% FCS, 0.05 mg / mL streptomycin, 50 U / mL penicillin, and 0.4 mM L-glutamine; as well as 0.01 mg / mL gentamicin, 0.048 mg / mL hydrocortisone, and 0.004 mg / mL insulin, with a final suspension density of 1 × 106 cells / mL. Incubation was carried out fully automatically using a liquid handling system (Tecan) equipped with a CO2 incubator and an orbital shaker. After adding a test compound such as 1 μM (1 × 105 cells / well) to the wells, the 96-well hepatocyte suspension culture plate was incubated at 37 °C in 5% CO2. At the designated time points, the samples were quenched by adding acetonitrile (including the internal standard) to the incubation wells for up to 2 hours.

[0196] Metabolism was carried out by Incubation of the test substances (e.g., 1 μM, 0.1% v / v DMSO) carried out in the suspension assay was performed in a 96-well plate containing a co-culture of adherent hepatocytes and mouse fibroblast control cells or separate control cells (5% CO2 atmosphere and 37 °C). Human The incubation medium in [it] was the same as that in the suspension of hepatocytes. At defined time points (2, 18, 26, 48, 72, and 96 hours), all wells were quenched with ice-cold acetonitrile containing an internal standard.

[0197] The samples were then centrifuged appropriately and the supernatant was analyzed by LC-MS / MS. The incubation was performed with n = 1 or 2.

[0198] Pharmacokinetic Characteristics of the Test Substance in Miniature Swine:

[0199] After intravenous and oral administrations, the pharmacokinetics of the test substance were determined in minipigs. The experimental design consisted of three male minipigs, and each animal received a single intravenous bolus dose and a single oral dose of the test item. The intravenous dose was administered at a nominal dose volume of 1 mL / kg. The oral dose was administered by gavage at a nominal dose volume of 5 mL / kg. There was a washout period of at least 7 days between the last sampling of the same animal and the next administration. The content of all formulations was within the required range of 85% to 115% of the nominal content. After administration, blood samples (1 mL) were withdrawn from the saphenous vein (via cannula) or jugular vein of each animal at pre-dose 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 24 hours after the intravenous dose and at pre-dose 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 24 hours, 48 hours after the oral dose. Hematocrit was measured at all time points. The blood:plasma partition factor was measured at 2 hours and 4 hours time points, and urine was collected as a single sample 24 hours after the administration. Blood samples (nominal 1 mL) were withdrawn from the saphenous vein (via cannula) or jugular vein of each animal into polypropylene tubes containing K2EDTA anticoagulant and centrifuged (1500 g, 10 minutes, 4 °C) to prepare plasma for analysis. The residual blood cells were discarded. The plasma vials were capped and stored on wet ice for no more than 60 minutes, then transferred to <-50 °C storage (usually -80 °C), and then analyzed using a specific LC-MS method.

[0200] Toxicity Evaluation of the Test Substance in Miniature Swine

[0201] The maximum tolerated dose (MTD) of the test item was determined after once-daily oral (by gavage) administration to minipigs. Then the toxicity of repeated daily administration for 14 consecutive days was evaluated. In addition, the toxicokinetic characteristics of the test item were characterized. Obtained from Ellegaard in Dalmose, Denmark sufficient specially bred Miniature pigs (animals: in the range of 2 to 3 months of age and a body weight range of 4 to 6 kg). At the start of dosing, the animals are within 4 to 5 months of age and the body weight range is within 6 to 9.5 kg. A dose volume of 10 mL / kg is used. The individual dose volume is based on the most recent body weight of each animal to achieve target dose levels of 30, 100, and 300 mg / kg / day or other dose levels depending on non-MTD results. Blood samples are collected on Day 1 and Day 14 to determine the drug concentration in plasma and derived pharmacokinetic parameters. The animals are not fed on the day of the scheduled necropsy. Each animal is anesthetized by intramuscular injection of a Zoletil mixture and then sacrificed by exsanguination. All tissues are preserved in an appropriate fixative. Further analyses include food consumption, body weight, clinical pathology, and complete histopathological examination of the target organs.

[0202] Table 1: NLRP3 inhibitory activity

[0203]

[0204] Table 2: hERG inhibition assay

[0205]

[0206] Table 3: Metabolic stability in human hepatocytes

[0207]

[0208] The present invention will now be illustrated by the following examples, which are not limiting.

[0209] If the preparation examples are obtained in the form of a mixture of enantiomers or diastereomers, the pure enantiomers or diastereomers can be obtained by the methods described herein or methods known to those skilled in the art such as, for example, chiral chromatography or crystallization.

[0210] Experimental Methods

[0211] Abbreviations:

[0212]

[0213]

[0214] Examples

[0215] If not otherwise stated, all examples and intermediates are prepared under a nitrogen atmosphere.

[0216] Intermediates

[0217] Intermediate 1: 3-Chloro-6-[[(3R)-1-ethyl-3-piperidinyl]amino]-4-methyl-1,2,4-triazin-5-one

[0218]

[0219] Step A: 6-Bromo-2-[(4-methoxyphenyl)methyl]-4-methyl-1,2,4-triazine-3,5-dione

[0220] 6-Bromo-4-methyl-2H-1,2,4-triazine-3,5-dione (CAS# 15870-75-4, 13.8 g, 63.1 mmol, 1.0 eq) and potassium carbonate (4.84 g, 31.5 mmol, 0.50 eq) were suspended in dry DMF (125 mL) and 4-methoxybenzyl chloride (10.3 mL, 75.7 mmol, 1.2 eq) was added. The reaction mixture was stirred at room temperature for 24 h. The reaction mixture was diluted with EtOAc (50 mL) and washed with 10 wt% aqueous LiCl solution (2 x 30 mL), dried using a phase separator and concentrated in vacuo. The resulting residue was purified by silica gel chromatography (0-50% EtOAc / isohexane) to afford the title compound (15.9 g, 77% yield), which was a white solid. 1 1H NMR (500 MHz, DMSO-d6) [ppm]: δ 7.33 - 7.25 (m, 2H), 6.97 - 6.89 (m, 2H), 5.00 (s, 2H), 3.74 (s, 3H), 3.20 (s, 3H).

[0221] Step B: 6-[[(3R)-1-Ethyl-3-piperidinyl]amino]-2-[(4-methoxyphenyl)methyl]-4-methyl-1,2,4-triazine-3,5-dione

[0222] (3R)-1-Ethylpiperidin-3-amine (6.0 g, 46.9 mmol, 1.53 eq) and the aforementioned 6-bromo-2-[(4-methoxyphenyl)methyl]-4-methyl-1,2,4-triazine-3,5-dione (10.0 g, 30.7 mmol, 1.0 eq) and cesium carbonate (20 g, 61.3 mmol, 2.0 eq) were dissolved in DMSO (125 mL), and the mixture was degassed (N2) for 5 minutes. The reaction vessel was evacuated and backfilled with N2 (3x), then (rac)-BINAP Pd G3 (1 g, 1.01 mmol, 0.030 eq) was added, and the reaction mixture was placed under N2 and then stirred at 95 °C for 24 hours. The reaction mixture was partitioned between EtOAc (500 mL) and water (500 mL). The organic phase was separated, washed with brine (3 x 300 mL), dried using a phase separator, and concentrated in vacuo. The resulting residue was purified by silica gel chromatography (0 - 7% in DCM (0.7 N ammonia in MeOH)) to afford the title compound (10.4 g, 86% yield), which was an orange oil. LCMS m / z 374.2 [M+H] + , ESI pos.

[0223] Step C: 6-[[(3R)-1-Ethyl-3-piperidinyl]amino]-4-methyl-2H-1,2,4-triazine-3,5-dione; trifluoromethanesulfonate

[0224] The aforementioned 6-[[(3R)-1-ethyl-3-piperidinyl]amino]-2-[(4-methoxyphenyl)methyl]-4-methyl-1,2,4-triazine-3,5-dione (10.4 g, 25.1 mmol, 1.0 eq) was dissolved in DCM (75 mL). Trifluoromethanesulfonic acid (3.33 mL, 37.7 mmol, 1.5 eq) was added to the reaction. The resulting solution was stirred at room temperature for 24 hours. Another portion of trifluoromethanesulfonic acid (3.33 mL, 37.7 mmol, 1.5 eq) was added and the reaction mixture was stirred for a further 3 hours. The reaction mixture was concentrated in vacuo and the resulting residue was purified by silica gel chromatography (0 - 10% (0.7 N ammonia in MeOH / DCM)) to afford the title compound (17.04 g, 84% yield), which was a yellow oil. LCMS m / z 254.5 [M+H] + , ESI pos.

[0225] Step D: 3-Chloro-6-[[(3R)-1-ethyl-3-piperidinyl]amino]-4-methyl-1,2,4-triazin-5-one

[0226] The aforementioned 6-[[(3R)-1-ethyl-3-piperidinyl]amino]-4-methyl-2H-1,2,4-triazine-3,5-dione; trifluoromethanesulfonate (17.04 g, 21.1 mmol, 1.0 eq) was dissolved in phosphorus oxychloride (75.0 mL, 804.6 mmol, 38.1 eq). The reaction mixture was stirred at 120 °C for 72 h. A 15 mL aliquot of the reaction mixture was concentrated in vacuo, and the resulting residue was diluted with EtOAc (200 mL) and washed with 1:1 brine:saturated aqueous NaHCO3 (200 mL). The organic phase was separated, and the aqueous phase was back-extracted with EtOAc (200 mL). The combined organic extracts were dried (MgSO4) and concentrated in vacuo to afford the title compound (1.03 g, 17% yield), which was a brown oil. The remaining reaction mixture was subjected to the same work-up conditions, scaled up, and afforded the title compound (5.06 g, 79% yield), which was a brown oil. LCMS m / z 274.4 ([37Cl]M+H) + , ESI pos.

[0227] Intermediate 2: 5-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)indan-4-ol

[0228]

[0229] A mixture of commercially available 5-bromoindan-4-ol (CAS# 575504-23-3) (950 mg, 4.46 mmol, 1.0 eq), bis(pinacolato)diboron (5.66 g, 22.3 mmol, 5.0 eq), Pd(dppf)Cl2·DCM complex (364.1 mg, 0.45 mmol, 0.1 eq), and potassium acetate (1.31 g, 13.4 mmol, 3.0 eq) in MeCN (40 mL) was degassed with nitrogen for 5 min, after which the reaction mixture was heated to 80 °C and stirred at this temperature for 16 h. The reaction mixture was cooled and concentrated in vacuo. The residue was purified by silica gel chromatography (0-100% EtOAc / isohexane) to afford the title compound (211.8 mg, 17%), which was a colorless oil. LCMS: no ionization. 1 1H NMR (500 MHz, CDCl3) [ppm]: δ 7.91 (s, 1H), 7.43 (d, 1H), 6.81 (d, 1H), 2.94 - 2.86 (m, 4H), 2.07 (d, 2H), 1.35 (s, 12H).

[0230] Intermediate 3: 3-Bromobicyclo[4.2.0]octa-1(6),2,4-triene-2-ol

[0231]

[0232] Step A: 5 - Benzyloxybicyclo[4.2.0]octa - 1(6),2,4 - trien - 7 - ol

[0233] At 0 °C under N2, a 2.5 M solution of butyllithium in hexanes (38.0 mL, 95.01 mmol, 5.0 eq) was added dropwise to stirred THF (100 mL). The reaction mixture was allowed to return to room temperature and then stirred for about 16 h. In another flask, a 2.5 M solution of butyllithium in hexanes (15.2 mL, 38.0 mmol, 2.0 eq) was added dropwise to a solution of 2,2,6,6 - tetramethylpiperidine (6.41 mL, 38.0 mmol, 2.0 eq) in THF (60 mL) at 0 °C, and the reaction mixture was stirred for 30 min. Then the first flask was cooled to - 78 °C and 3 - benzyloxybromobenzene (5000 mg, 19.0 mmol, 1.0 eq; CAS# 53087 - 13 - 1) in THF (25 mL) was added. Then a solution of lithium 2,2,6,6 - tetramethylpiperidide was added dropwise to the reaction mixture via cannula, and the reaction was stirred for 1 h. The reaction was quenched by the addition of NH4Cl (200 mL, saturated aqueous solution) and allowed to return to room temperature. The reaction was diluted with water (100 mL) and extracted with EtOAc (3 x 150 mL). The combined organic layers were dried over MgSO4, concentrated in vacuo and purified by silica gel column chromatography (0 - 10% EtOAc / isohexane) to afford the title compound (3.22 g, 74% yield), which was a white solid. LCMS: m / z = 225.3 [M - H] - , ESI neg.

[0234] Step B : 2 - Benzyloxy - 8 - bromo - bicyclo[4.2.0]octa - 1(6),2,4 - triene

[0235] At room temperature, carbon tetrabromide (1.76 g, 5.3 mmol, 1.2 eq) and triphenylphosphine (2.09 g, 7.96 mmol, 1.8 eq) were added to a stirred solution of the aforementioned 5 - benzyloxybicyclo[4.2.0]octa - 1(6),2,4 - trien - 7 - ol (1.0 g, 4.42 mmol, 1.0 eq) in Et2O (40 mL), and the reaction was stirred for 2 days. The solution was filtered, the solid was washed with diethyl ether (40 mL), and the filtrate was concentrated in vacuo. The crude material was purified by silica gel column chromatography (0 - 20% EtOAc / isohexane) to afford the title compound (1186 mg, 4.1 mmol, 89% yield), which was a colorless oil. LCMS: m / z not observed. 11H NMR (500 MHz, DMSO-d6) [ppm]: δ 7.48–7.43 (m, 2H), 7.43–7.38 (m, 2H), 7.37–7.28 (m, 2H), 6.87 (d, 1H), 6.77 (d, 1H), 5.74 (dd, 1H), 5.32 (d, 1H), 5.27 (d, 1H), 3.94–3.81 (m, 1H), 3.40–3.34 (m, 1H).

[0236] Step C : Bicyclo[4.2.0]octa-1(6),2,4-triene-2-ol

[0237] A mixture of the aforementioned 2-benzyloxy-8-bromo-bicyclo[4.2.0]octa-1(6),2,4-triene (1180.0 mg, 4.08 mmol, 1.0 eq) and 5 wt% palladium on carbon, type R434, 50 wt% water (173.7 mg, 0.04 mmol, 0.01 eq) in ethanol (25 mL) was stirred under H2 (5 bar) for 3 h. The reaction mixture was filtered and diluted with DCM (200 mL), then washed with sodium thiosulfate (1 x 50 mL, 10% aqueous solution), sodium bicarbonate (1 x 50 mL, saturated aqueous solution) and brine (1 x 50 mL). The organic phase was dried over MgSO4, concentrated in vacuo and purified by silica gel column chromatography (0 - 10% EtOAc / isohexane) to afford the title compound (471.7 mg, 3.93 mmol, 91% yield), which was a pale grey crystalline solid. LCMS: m / z = 121.1 [M+H] + , ESI pos.

[0238] Step D : 3-Bromobicyclo[4.2.0]octa-1(6),2,4-triene-2-ol

[0239] At 0 °C, N-bromosuccinimide (628.34 mg, 3.53 mmol, 0.95 eq) in DCM (10 mL) was added portionwise to a stirred solution of the aforementioned bicyclo[4.2.0]octa-1(6),2,4-triene-2-ol (470.0 mg, 3.72 mmol, 1.0 eq) and diisopropylamine (52.08 μL, 0.37 mmol, 0.1 eq) in DCM (40 mL), and the reaction mixture was stirred for 1 h. The residue was concentrated in vacuo and purified by silica gel column chromatography (0 - 10% EtOAc / isohexane) to afford the title compound (522.4 mg, 2.62 mmol, 67% yield), which was a white solid. LCMS: m / z not observed. 11H NMR (500 MHz, CDCl3) [ppm]: δ 7.32 (d, 1H), 6.55 (d, 1H), 5.43 (s, 1H), 3.20–3.13 (m, 2H), 3.13–3.06 (m, 2H).

[0240] Examples:

[0241] Example 1: 6-[[(3R)-1-Ethyl-3-piperidinyl]amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazin-5-one

[0242]

[0243] The mixture of the aforementioned 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indan-4-yl intermediate 2 (60.0 mg, 0.23 mmol, 1.25 eq), 3-chloro-6-[[(3R)-1-ethyl-3-piperidinyl]amino]-4-methyl-1,2,4-triazin-5-one intermediate 1 (50.0 mg, 0.18 mmol, 1.0 eq), meCgPPh Pd G3 (CAS#. 2230788-58-4) (12.2 mg, 0.02 mmol, 0.1 eq) and potassium carbonate (76.29 mg, 0.55 mmol, 3.0 eq) in a mixture of 1,4-dioxane (2 mL) and water (0.5 mL) was degassed with nitrogen for 5 minutes, and then the reaction mixture was heated to 90 °C for 2 hours. The reaction mixture was cooled and concentrated in vacuo. The residue was purified by silica gel chromatography (24 g column, 0-10% (0.7 M NH3) MeOH / DCM, then 12 g column, 0-7% (0.7 M NH3) MeOH / DCM) to give the title compound (19.6 mg, 28%), which was an off-white solid. LCMS m / z = 370.2 [M+H] + , ESI pos.

[0244] Example 2: 6-[[(3R)-1-Ethyl-3-piperidinyl]amino]-3-(2-hydroxy-3-bicyclo[4.2.0]octa-1,3,5-trienyl)-4-methyl-1,2,4-triazin-5-one

[0245]

[0246] The mixture of 3-bromobicyclo[4.2.0]octa-1(6),2,4-triene-2-ol intermediate 3 (100 mg, 0.50 mmol, 1.0 eq), bis(pinacolato)diboron (640.0 mg, 2.52 mmol, 5.0 eq), Pd(dppf)Cl2·DCM complex (50.0 mg, 0.06 mmol, 0.12 eq; CAS# 95464-05-4) and potassium acetate (150.0 mg, 1.53 mmol, 3.0 eq) in 1,4-dioxane (5 mL) was degassed with N2 for 5 minutes and then heated to 90 °C and stirred for 6 hours. The reaction mixture was cooled to room temperature, then K3PO4 (2 mL, 10% aqueous solution) was added and the reaction mixture was stirred for 30 minutes. The reaction mixture was diluted with water (30 mL) and extracted with DCM (30 mL). The aqueous layer was acidified to ~pH 3 - 4 by dropwise addition of HCl (1 M aqueous solution) and then extracted again with DCM (5 x 30 mL). The combined organic extracts were dried over MgSO4 and concentrated in vacuo. The resulting residue was dissolved in 1,4-dioxane (4 mL) and 3-chloro-6-[[(3R)-1-ethyl-3-piperidinyl]amino]-4-methyl-1,2,4-triazin-5-one intermediate 1 (140.0 mg, 0.36 mmol, 0.72 eq), Xphos Pd G3 (22.0 mg, 0.03 mmol, 0.05 eq), potassium carbonate (210.0 mg, 1.52 mmol, 3.02 eq) and water (1 mL) were added. The resulting mixture was degassed with N2 for 5 minutes and then heated to 90 °C for 4 hours. The reaction mixture was cooled to room temperature, concentrated in vacuo and purified by silica gel column chromatography (0 - 10% (0.7 M NH3)MeOH / DCM) to afford 58 mg of the desired compound with a purity of about 93%. The material was further purified by reverse phase chromatography (0.1% ammonium hydroxide, 5 - 35% MeCN / water) to give the title compound (33.8 mg, 18% yield), which was a white solid. LCMS: m / z = 356.2 [M+H] + , ESIpos.

[0247] Example 3: 6-[[(3R)-1-Ethyl-3-piperidinyl]-methyl-amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazin-5-one

[0248]

[0249] Step A: 3-(4-Benzyloxyindan-5-yl)-6-[[(3R)-1-ethyl-3-piperidinyl]amino]-4-methyl-1,2,4-triazin-5-one

[0250] 2-(4-benzyloxyindan-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (737.0 mg, 2.1 mmol, 1.0 eq, CAS# 2878443-82-2), 3-chloro-6-[[(3R)-1-ethylpiperidin-3-yl]amino]-4-methyl-1,2,4-triazin-5-one intermediate 1 (603.57 mg, 2.0 mmol, 0.95 eq), XPhos Pd G3 (178.32 mg, 0.21 mmol, 0.1 eq) and saturated aqueous sodium carbonate (3.0 mL, 2.1 mmol, 1.0 eq) were dissolved in MeCN (15 mL) and the mixture was degassed with nitrogen and then stirred at 80 °C for 20 h. The mixture was diluted with EtOAc (100 mL) and washed with brine (100 mL). The organic phase was separated, dried using a phase separator and concentrated in vacuo. The resulting residue was purified by flash silica chromatography (24 g column, 0 - 10% (0.7N ammonia in MeOH) / DCM) to afford the title compound (675.0 mg, 1.47 mmol, 67% yield), which was a yellow solid. LCMS: m / z = 460.3 [M+H] + , ESIpos.

[0251] Step B: 3-(4-benzyloxyindan-5-yl)-6-[[(3R)-1-ethylpiperidin-3-yl]methyl-amino]-4-methyl-1,2,4-triazin-5-one

[0252] 3-(4-Benzyloxyindan-5-yl)-6-[[(3R)-1-ethylpiperidin-3-yl]amino]-4-methyl-1,2,4-triazin-5-one (250.0 mg, 0.54 mmol, 1.0 eq) was dissolved in NMP (2 mL), and sodium hydride (60% in mineral oil, 108.79 mg, 2.72 mmol, 5.0 eq) was added. The mixture was stirred for 15 minutes, then iodomethane (0.04 mL, 0.65 mmol, 1.2 eq) was added. The mixture was stirred for 18 hours, then additional sodium hydride (60% in mineral oil, 108.79 mg, 2.72 mmol, 5.0 eq) and iodomethane (0.04 mL, 0.65 mmol, 1.2 eq) were added, and the mixture was stirred for a further 2 hours. The mixture was quenched with MeOH (5 mL), then water (10 mL) and diluted with EtOAc (50 mL). The organic phase was washed with brine (3 x 30 mL), dried using a phase separator, and concentrated in vacuo. The resulting residue was purified by flash silica chromatography (4 g column, 0 - 7% (0.7N NH3 in MeOH) / DCM) to give the title compound (13.0 mg, 4% yield), which was a pale yellow solid. LCMS: m / z = 474.5 [M+H] + , ESI pos.

[0253] Step B: 6-[[(3R)-1-Ethyl-3-piperidinyl]-methyl-amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazin-5-one

[0254] 3-(4-Benzyloxyindan-5-yl)-6-[[(3R)-1-ethylpiperidin-3-yl]-methyl-amino]-4-methyl-1,2,4-triazin-5-one (13.0 mg, 0.02 mmol, 1.0 eq) was dissolved in EtOH (1 mL), and Pd(OH)2 (carbon supported, 10 wt%, 7.53 mg, 0.0 mmol, 0.2 eq) was added. The mixture was then stirred under H2 (2 bar) for 24 hours, then the reaction mixture was filtered through a plug of celite and the filtrate was concentrated in vacuo. The resulting residue was purified by flash silica chromatography (4 g column, 0 - 10% (0.7N NH3 in MeOH / DCM) to give the title compound (7.0 mg, 73% yield), which was an off-white solid. LCMS: m / z = 384.3 [M+H] + , ESI pos.

[0255] Example A

[0256] The compounds of this formula can be used as active ingredients for producing tablets composed of the following in a manner known per se:

[0257]

[0258] Example B

[0259] The compounds of this formula can be used as active ingredients for producing capsules composed of the following in a manner known per se:

[0260]

[0261]

Claims

1. A compound selected from 6-[[(3R)-1-Ethyl-3-piperidinyl]amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazin-5-one; 6-[[(3R)-1-Ethyl-3-piperidinyl]amino]-3-(2-hydroxy-3-bicyclo[4.2.0]octa-1,3,5-trienyl)-4-methyl-1,2,4-triazin-5-one; 6-[[(3R)-1-Ethyl-3-piperidinyl]-methyl-amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazin-5-one; or a pharmaceutically acceptable salt thereof.

2. The compound according to claim 1, wherein the compound is 6-[[(3R)-1-Ethyl-3-piperidinyl]amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazin-5-one or a pharmaceutically acceptable salt thereof.

3. The compound according to claim 1, wherein the compound is 6-[[(3R)-1-Ethyl-3-piperidinyl]amino]-3-(2-hydroxy-3-bicyclo[4.2.0]octa-1,3,5-trienyl)-4-methyl-1,2,4-triazin-5-one or a pharmaceutically acceptable salt thereof.

4. The compound according to claim 1, wherein the compound is 6-[[(3R)-1-Ethyl-3-piperidinyl]-methyl-amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazin-5-one or a pharmaceutically acceptable salt thereof.

5. The compound according to any one of claims 1 to 4, which is used as a therapeutic active substance.

6. The compound according to any one of claims 1 to 4, which is used for treating or preventing a disease, disorder or condition, wherein the disease, disorder or condition responds to NLRP3 inhibition.

7. A pharmaceutical composition comprising the compound according to any one of claims 1 to 4 and a therapeutically inert carrier.

8. Use of the compound according to any one of claims 1 to 4 for treating or preventing a disease, disorder or condition, wherein the disease, disorder or condition responds to NLRP3 inhibition.

9. The compound according to any one of claims 1 to 4, which is used for treating or preventing a disease, disorder or condition selected from asthma or COPD.

10. The compound according to any one of claims 1 to 4, which is used for treating or preventing a disease, disorder or condition selected from Alzheimer's disease and Parkinson's disease.

11. Use of the compound according to any one of claims 1 to 4 in treating or preventing a disease, disorder or condition selected from asthma or COPD.

12. Use of the compound according to any one of claims 1 to 4 in treating or preventing a disease, disorder or condition selected from Parkinson's disease or Alzheimer's disease.

13. Use of the compound according to any one of claims 1 to 4 for preparing a medicament for treating or preventing a disease, disorder or condition selected from asthma or COPD.

14. Use of a compound according to any one of claims 1 to 4 for the manufacture of a medicament for the treatment or prevention of a disease, disorder or condition selected from Parkinson's disease or Alzheimer's disease.

15. A method of inhibiting NLRP3, the method comprising administering an effective amount of a compound as claimed in any one of claims 1 to 4 to inhibit NLRP3.

16. A method of treating or preventing a disease, disorder or condition, the method comprising administering an effective amount of a compound according to any one of claims 1 to 4, wherein the disease, disorder or condition is selected from asthma or COPD.

17. A method of treating or preventing a disease, disorder or condition, the method comprising administering an effective amount of a compound according to any one of claims 1 to 4, wherein the disease, disorder or condition is selected from Parkinson's disease or Alzheimer's disease.

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