Triazine derivatives as NLRP3 inhibitors

Abstract

TIFF2026522012000019.tif36165 The present invention relates to general formula (I) (wherein A, R 1 , R 2 The present invention relates to novel compounds having (and n as described herein), compositions comprising the compound, and methods for using the compound.

Description

[Technical Field]

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

[0002] This invention relates to formula I: [ka] (In the formula, A is -O- or CH2, R 1 is a hydroxyalkyl or acetyl, R 2 It is an alkyl, n is either 0 or 1. (If n is 0, then A is CH2.) This invention provides novel compounds and pharmaceutically acceptable salts thereof.

[0003] Furthermore, the present invention includes all racemic mixtures, all their corresponding enantiomers and / or optical isomers. [Background technology]

[0004] The pyrin domain-containing protein 3 (NLRP3) inflammasome of the NOD-like receptor (NLR) family is a component of inflammatory processes, 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.

[0005] NLRP3 is an intracellular signaling molecule that senses many pathogen-derived, environmental, and host-derived factors. When activated, NLRP3 binds to apoptosis-related speckled proteins (ASCs) that contain caspase activation and recruitment domains. The ASCs then polymerize to form large aggregates known as ASC specks. The polymerized ASCs then interact with the cysteine ​​protease caspase-1 to form a complex called the inflammasome. This results in the activation of caspase-1, which cleaves the precursor forms of the pro-inflammatory cytokines IL-1β and IL-18 (referred to as pro-IL-1β and pro-IL-18, respectively), thereby activating these cytokines. Caspase-1 also mediates a type of inflammatory cell death known as pyroptosis. ASC specks can also recruit and activate caspase-8, which can process pro-IL-1β and pro-IL-18 to induce apoptotic cell death.

[0006] Caspase-1 cleaves pro-IL-1β and pro-IL-18, converting them to their active forms which are then secreted from cells. Activated caspase-1 also cleaves gasdelmin-D, inducing pyroptosis. Through its control of the pyroptotic cell death pathway, caspase-1 also mediates the release of alarmin molecules such as IL-33 and high-mobility groupbox-1 protein (HMGB1). Caspase-1 also cleaves intracellular IL-1R2, leading to its degradation and allowing IL-1α to be released. In human cells, caspase-1 can also regulate the processing and secretion of IL-37. Many other caspase-1 substrates, such as components of the cytoskeleton and glycolysis pathway, can contribute to caspase-1-dependent inflammation.

[0007] NLRP3-dependent ASC specks are released into the extracellular environment, where they activate caspase-1, induce caspase-1 substrate processing, and can propagate inflammation.

[0008] Active cytokines derived from NLRP3 inflammasome activation are key drivers of inflammation and interact with other cytokine pathways to form immune responses to infection and injury. For example, IL-1β signaling induces the secretion of pro-inflammatory cytokines IL-6 and TNF. IL-1β and IL-18 synergistically with IL-23 to induce IL-17 production by memory-CD4 Th17 cells and γδ T cells in the absence of T cell receptor binding. IL-18 and IL-12 also act synergistically to induce IFN-γ production from memory T cells and NK cells, promoting the Th1 response.

[0009] Because Mackle-Wells syndrome (MWS), familial cold autoinflammatory syndrome (FCAS), and neonatal-onset multiorgan inflammatory disease (NOMID), all hereditary CAPS disorders, are caused by gain-of-function mutations in NLRP3, NLRP3 is defined as a key component of the inflammatory process. NLRP3 is also involved in the pathogenesis of many complex diseases, particularly metabolic disorders such as type 2 diabetes, atherosclerosis, obesity, and gout.

[0010] The role of NLRP3 in central nervous system disorders is becoming clearer, and it has also been shown that lung diseases are affected by NLRP3. NLRP3 has been suggested to be involved in several central nervous system pathologies, including Parkinson's disease (PD), Alzheimer's disease (AD), dementia, Huntington's disease, cerebral malaria, and brain injury due to 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 be involved in several 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). Furthermore, NLRP3 is involved in the development of liver disease, kidney disease, and aging. Many of these associations are linked to NLRP3. - / - Although defined using mice, insights into the specific activation of NLRP3 in these diseases also exist. In type 2 diabetes (T2D), the deposition of islet amyloid polypeptides in the pancreas activates NLRP3 and IL-1β signaling, leading to cell death and inflammation.

[0011] Several small molecules have been shown to inhibit the NLRP3 inflammasome. Glybrid inhibits IL-1β production at micromolar concentrations in response to NLRP3 activation, but does not respond to NLRC4 or NLRP1 activation. Other previously characterized weak NLRP3 inhibitors include parthenolide, 3,4-methylenedioxy-β-nitrostyrene, and dimethyl sulfoxide (DMSO), but these drugs have limited efficacy and are nonspecific.

[0012] Current treatments for NLRP3-related diseases include biological 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 proven successful in the treatment of CAPS, and these biological agents are being used in clinical trials for other IL-1β-related diseases.

[0013] There is a need to provide compounds with improved pharmacological and / or physiological and / or physicochemical properties, and / or compounds that provide useful alternatives to known compounds.

[0014] Furthermore, when developing NLRP3 inhibitors for treating peripheral indications, it is advantageous to minimize the exposure of NLRP3 inhibitory compounds in the brain compared to systemic exposure, since central exposure does not provide a therapeutic benefit. Furthermore, this approach minimizes the risk of potential side effects in the central nervous system (CNS), and thus provides an opportunity to administer higher doses if necessary. The compounds of formula I achieve this by showing increased efflux and / or reduced passive permeability in a transcellular assay expressing the active P-gp transporter without compromising systemic distribution. P-gp (P-glycoprotein) is an important transporter expressed in capillary endothelial cells that form the blood-brain barrier and the blood-testis barrier, returning foreign substances back to the capillaries and limiting exposure to the brain.

Brief Description of the Drawings

[0015] [Figure 1] It is a diagram showing the voltage pattern in the hERG test.

Summary of the Invention

[0016] The present invention relates to formula I:

Chemical

[0017] The term "acetyl" refers to a group of the formula -C(=O)-R' (where R' is an alkyl group). An example of acetyl is -C(=O)CH3.

[0018] The term "alkyl" refers to a monovalent linear or branched saturated hydrocarbon group consisting of 1 to 6 carbon atoms. In some embodiments, unless otherwise stated, alkyl refers to a group consisting of 1 to 6 carbon atoms (C 1-6 -alkyl) or 1 to 4 carbon atoms (C 1-4 -Contains alkyl) C 1-6 Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, and pentyl. A specific alkyl group is methyl.

[0019] The term "hydroxy" refers to the -OH group.

[0020] The term "hydroxyalkyl" refers to an alkyl group in which at least one hydrogen atom of the alkyl group is replaced by a hydroxyl group. Examples of hydroxyalkyl groups include hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxymethylethyl, hydroxymethylpropyl, and dihydroxypropyl. An example of a hydroxyalkyl group is hydroxyethyl.

[0021] The term "pharmaceutically acceptable salt" refers to a salt that retains the biological efficacy and properties of a free base or free acid and is not biologically or otherwise undesirable. Salts are formed from 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, and N-acetylcysteine. In addition, these salts can be prepared by adding an inorganic base or organic base to a free acid. Salts derived from inorganic bases include, but are not limited to, salts of sodium, potassium, lithium, ammonium, calcium, and magnesium. Salts derived from organic bases include, but are not limited to, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and salts of basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, lysine, arginine, N-ethylpiperidine, piperidine, and polyamine resins. Compounds of formula I can also exist in zwitterionic form. Particularly preferred pharmaceutically acceptable salts of compounds of formula I are salts formed with formic acid, and salts formed with hydrochloric acid that yield hydrochloride, dihydrochloride, or trihydrochloride.

[0022] The abbreviation uM stands for micromolar and is equivalent to the symbol μM.

[0023] The abbreviation uL stands for microliter and is equivalent to the symbol μL.

[0024] The abbreviation ug stands for microgram and is equivalent to the symbol μg.

[0025] The compounds of formula I may contain several asymmetric centers and can exist in the form of optically pure enantiomers, mixtures of enantiomers, such as racemates, etc., optically pure diastereoisomers, mixtures of diastereoisomers, racemates of diastereoisomers or mixtures of racemates of diastereoisomers.

[0026] According to the Cahn-Ingold-Prelog rules, an asymmetric carbon atom can have the configuration of "R" or "S".

[0027] Also, one embodiment of the present invention provides a compound according to formula I described herein and a pharmaceutically acceptable salt or ester thereof, particularly a compound according to formula I described herein and a pharmaceutically acceptable salt thereof, more specifically, a compound according to formula I described herein.

[0028] One embodiment of the present invention provides a compound according to formula I described herein in which R 1 is hydroxyalkyl.

[0029] One embodiment of the present invention provides a compound according to formula I described herein in which R 2 is methyl.

[0030] Specific examples of the compounds of formula I described herein are 3-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-6-[[(3R)-1-(2-hydroxyethyl)-3-piperidyl]amino]-4-methyl-1,2,4-triazin-5-one, 6-[[(3R)-1-(2-hydroxyethyl)-3-piperidyl]amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazin-5-one, and pharmaceutically acceptable salts thereof.

[0031] Other specific examples of the compounds of formula I described herein are 3-(4-hydroxyindan-5-yl)-6-[[(3R)-1-(3-hydroxypropyl)-3-piperidyl]amino]-4-methyl-1,2,4-triazine-5-one, 3-(2-hydroxy-3-bicyclo[4.2.0]octa-1(6),2,4-trienyl)-6-[[(3R)-1-(2-hydroxyethyl)-3-piperidyl]amino]-4-methyl-1,2,4-triazine-5-one, Selected from the pharmaceutically acceptable salts thereof.

[0032] Another specific example of the compound of formula I described herein is 6-[[(3R)-1-acetyl-3-piperidyl]amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazine-5-one, or a pharmaceutically acceptable salt thereof.

[0033] Another embodiment of the present invention provides a pharmaceutical composition or drug containing the compound of the present invention and a therapeutically inert carrier, diluent, or excipient, as well as a method of using the compound of the present invention to prepare such compositions and drugs. In one example, the compound of formula I can be formulated into a gallenical dosage form by mixing it with a physiologically acceptable carrier, i.e., a carrier that is nontoxic to the recipient at the dose and concentration used, at ambient temperature, at a suitable pH, and to a desired degree of purity. The pH of the formulation depends primarily on the specific application and the concentration of the compound, but is preferably in the range of about 3 to about 8. In one example, the compound of formula I is formulated in an acetate buffer at pH 5. In another embodiment, the compound of formula I is sterile. The compound can be stored, for example, as a solid or amorphous composition, as a lyophilized formulation, or as an aqueous solution.

[0034] The composition is formulated, administered, and given in a manner consistent with medical practice. Factors to be considered in this context include the specific disorder being treated, the specific mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of drug delivery, the method of administration, the administration scheduling, and other factors known to the medical practitioner.

[0035] The compounds of the present invention may be administered by any suitable means, including orally, topically (including buccal and sublingual), rectally, vaginally, percutaneously, parenterally, subcutaneously, intraperitoneally, intrapulmonaryly, intradermally, intrathecally, and epidurally, as well as intranasally, and, if desired for topical treatment, intrafocal administration. Parenteral administrations include intramuscular, intravenous, intra-arterial, intraperitoneal, or subcutaneous administration.

[0036] The compounds of the present invention can be administered in any convenient dosage form, such as tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, and patches. Such compositions may contain common components in pharmaceutical preparations, such as diluents, carriers, pH adjusters, sweeteners, fillers, and further active agents.

[0037] Typical formulations are prepared by mixing the compound of the present 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, Ansel, Howard C., et al., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, Alfonso R. et al., Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C., Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005. The formulation may also contain one or more buffers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, flow enhancers, processing aids, colorants, sweeteners, fragrances, flavorings, diluents, and other known additives for providing a drug (i.e., the compound of the present invention or its pharmaceutical composition) in an appealing manner or for assisting in the manufacture of a pharmaceutical product (i.e., a drug).

[0038] Compounds of formula I and pharmaceutically acceptable salts thereof can be treated with pharmaceutically inert, inorganic or organic adjuvants for the production of tablets, coated tablets, sugar-coated tablets, hard gelatin capsules, injectable solutions, or topical formulations. Lactose, corn starch or its derivatives, talc, stearic acid or its salts, etc., can be used as such adjuvants for tablets, sugar-coated tablets, and hard gelatin capsules, for example.

[0039] Suitable adjuvants for soft gelatin capsules include, for example, vegetable oils, waxes, fats, semi-solid substances, and liquid polyols.

[0040] Suitable adjuvants for the production of solutions and syrups include, for example, water, polyols, sucrose, invert sugar, and glucose.

[0041] Suitable adjuvants for injection solutions include, for example, water, alcohol, polyol, glycerol, and vegetable oil.

[0042] Suitable adjuvants for suppositories include, for example, natural or hydrogenated oils, waxes, fats, semi-solid or liquid polyols.

[0043] Suitable adjuvants for topical ophthalmic formulations include, for example, cyclodextrin, mannitol, or many other carriers and excipients known in the art.

[0044] Furthermore, pharmaceutical preparations may contain preservatives, solubilizers, viscosity-enhancing substances, stabilizers, humectants, emulsifiers, sweeteners, colorants, flavoring agents, salts for altering osmotic pressure, buffers, masking agents, or antioxidants. These may also contain other substances of therapeutic value.

[0045] Dosages can vary over a wide range and, of course, are tailored to the individual requirements of each specific case. Generally, for oral administration, a daily dose of approximately 0.1 mg to 20 mg per kg of body weight, preferably approximately 0.5 mg to 4 mg per kg of body weight (e.g., approximately 300 mg per person), is preferably divided into 1 to 3 individual doses, which, if appropriate, may consist of equal amounts, for example. For topical administration, the formulation may contain 0.001% to 15% by weight of the drug, and the required dose, which may be between 0.1 and 25 mg, can be administered by a single dose per day or per week, by multiple doses per day (2 to 4 times), or by multiple doses per week. However, where indicated, it will be clear that the upper or lower limits given herein may be exceeded.

[0046] One embodiment of the present invention is a compound according to formula I described herein, for use as a therapeutically active substance.

[0047] One embodiment of the present invention is a compound according to Formula I described herein for use in the treatment or prevention of a disease, disorder, or condition that is responsive to NLRP3 inhibition.

[0048] One embodiment of the present invention is a compound according to Formula I described herein for the treatment or prevention of a disease, disorder, or pathological condition in which the disorder or pathological condition is responsive to NLRP3 inhibition.

[0049] As used herein, the term "NLRP3 inhibition" refers to a complete or partial reduction of the activity level of NLRP3, including, for example, inhibition of active NLRP3 and / or inhibition of NLRP3 activation.

[0050] There is evidence regarding the roles of NLRP3-induced IL-1 and IL-18 in inflammatory responses associated with or resulting from numerous different disorders (Menu et al., Clinical and Experimental Immunology, 166:1-15, 2011; Strowig et al., Nature, 481:278-286, 2012).

[0051] In one embodiment, the disease, disorder, or pathological condition is selected from the following: (i) inflammation; (ii) autoimmune diseases; (iii) cancer; (iv) infectious diseases; (v) metabolic disease; (vi) cardiovascular disease; (vii) respiratory diseases; (viii) Liver disease, (ix) renal disease; (x) eye diseases; (xi) skin diseases; (xii) Lymphatic system pathology, (xiii) Graft-versus-host disease, (xiv) allodynia; (xv) Conditions related to diabetes, and (xvi) Any disease in which an individual is determined to have germline or somatic non-silent mutations in NLRP3.

[0052] In another embodiment, the disease, disorder, or condition is selected from the following: (i) Cancer, (ii) Infectious diseases, (iii) Cardiovascular disease, (iv) Liver disease, (v) Eye diseases, and (vi) Skin diseases.

[0053] In a further typical embodiment of the present invention, the disease, disorder, or pathological condition is inflammation. Examples of inflammation that can be treated or prevented include inflammatory responses related to or resulting from the following: (i) Skin conditions 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. (ii) Joint conditions such as osteoarthritis, systemic juvenile idiopathic arthritis, adult-onset Still's disease, relapsing polychondritis, rheumatoid arthritis, juvenile chronic arthritis, gout, or seronegative spondyloarthropathy (e.g., ankylosing spondylitis, psoriatic arthritis, or Reiter's disease), (iii) Muscle conditions such as polymyositis or myasthenia gravis, (iv) Gastrointestinal conditions 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), (v) respiratory conditions such as chronic obstructive pulmonary disease (COPD), asthma (eosinophilic, bronchial, allergic, endogenous, exogenous or dust-induced asthma, especially chronic or refractory asthma, e.g., delayed-onset asthma and airway hyperresponsiveness), bronchitis, rhinitis (acute rhinitis, allergic rhinitis, atrophic rhinitis, chronic rhinitis, caseous rhinitis, hypertrophic rhinitis, suppurative rhinitis (rhinitis pumlenta), xerotic rhinitis, drug-induced rhinitis, membranous rhinitis, seasonal rhinitis, e.g., hay fever and vasomotor rhinitis), sinusitis, idiopathic pulmonary fibrosis (IPF), sarcoidosis, farmer's lung, silicosis, asbestosis, ash-induced inflammation, adult respiratory distress syndrome, hypersensitivity pneumonitis, or idiopathic interstitial pneumonia. (vi) Vascular conditions such as atherosclerosis, Behçet's disease, vasculitis, or Wegener's granulomatosis, (vii) Autoimmune conditions such as systemic lupus erythematosus, Sjögren's syndrome, systemic sclerosis, Hashimoto's thyroiditis, type 1 diabetes, idiopathic thrombocytopenic purpura, or Graves' disease. (viii) Eye conditions such as uveitis, allergic conjunctivitis, or vernal conjunctivitis, (ix) Infections or infection-related conditions, 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 (A, B, or C, or other viral hepatitis), peritonitis, pneumonia, epiglottitis, malaria, hemorrhagic dengue fever, leishmaniasis, streptococcal myositis, Mycobacterium tuberculosis (including co-infection with Mycobacterium tuberculosis and HIV), Mycobacterium avium intracellulare, Pneumocystis jirovecii pneumonia, orchitis / epididymitis, Legionella, Lyme disease, influenza A, Epstein-Barr virus infection, viral encephalitis / aseptic meningitis, or pelvic inflammatory disease. (x) Renal conditions such as mesangial proliferative glomerulonephritis, nephrotic syndrome, nephritis, glomerulonephritis, obesity-related glomerulopathy, acute renal failure, acute kidney injury, uremia, nephritis syndrome, renal fibrosis including chronic crystalline nephropathy, or renal hypertension. (xi) Lymphatic system disorders such as Castleman disease, (xii) Immune system or related conditions, e.g., hyper-IgE syndrome, multibacterial leprosy, familial hemophagocytic lymphohistiocytosis, or graft-versus-host disease. (xiii) Liver conditions such as chronic active hepatitis, non-alcoholic steatohepatitis (NASH), alcohol-induced hepatitis, non-alcoholic fatty liver disease (NAFLD), alcoholic fatty liver disease (AFLD), alcoholic steatohepatitis (ASH), primary biliary cirrhosis, fulminant hepatitis, hepatic fibrosis, or liver failure. (xiv) Cancers including the cancers listed above, (xv) radiation exposure; (xvi) Type 2 diabetes (T2D), atherosclerosis, obesity, metabolic disorders such as gout or pseudogout, and / or

[0054] One embodiment of the present invention is a compound according to Formula I described herein for the treatment or prevention of a disease, disorder, or condition selected from the following: inflammation, autoimmune diseases, cancer, infectious disease, metabolic diseases, cardiovascular disease, respiratory diseases, Liver disease, kidney disease, eye diseases, skin diseases, Lymphatic system pathology, Graft-versus-host disease Allodynia, Conditions related to diabetes, and Any disease in which an individual is determined to have a germline or somatic non-silent mutation in NLRP3.

[0055] One embodiment of the present invention is the use of a compound according to Formula I described herein for the treatment or prevention of a disease, disorder, or condition selected from asthma and COPD.

[0056] One embodiment of the present invention is the use of a compound according to Formula I described herein for the treatment or prevention of cardiovascular disease, disorder, or condition.

[0057] One embodiment of the present invention is the use of a compound according to Formula I described herein for the treatment or prevention of cardiovascular metabolic diseases, disorders, or conditions.

[0058] One embodiment of the present invention is the use of a compound according to Formula I described herein for the treatment or prevention of a disease, disorder, or condition selected from cryopyrin-associated periodic syndromes.

[0059] One embodiment of the present invention is a compound according to Formula I described herein for the treatment or prevention of a disease, disorder, or condition selected from asthma and COPD.

[0060] One embodiment of the present invention is a compound according to Formula I described herein for the treatment or prevention of cardiovascular disease, disorder, or condition.

[0061] One embodiment of the present invention is a compound according to Formula I described herein for the treatment or prevention of cardiovascular metabolic diseases, disorders, or conditions.

[0062] One embodiment of the present invention is a compound according to Formula I described herein for the treatment or prevention of a disease, disorder, or condition selected from cryopyrin-associated periodic syndromes.

[0063] One embodiment of the present invention is the use of a compound according to Formula I described herein for the preparation of a pharmaceutical product for the treatment or prevention of a disease, disorder, or condition selected from asthma and COPD.

[0064] One embodiment of the present invention is the use of a compound according to Formula I described herein for the preparation of a pharmaceutical product for the treatment or prevention of cardiovascular disease, disorder, or condition.

[0065] One embodiment of the present invention is the use of a compound according to Formula I described herein for the preparation of a pharmaceutical product for the treatment or prevention of cardiovascular metabolic diseases, disorders, or conditions.

[0066] One embodiment of the present invention is the use of a compound according to Formula I described herein for the preparation of a pharmaceutical product for the treatment or prevention of a disease, disorder, or condition selected from cryopyrin-associated periodic syndromes.

[0067] One embodiment of the present invention is a method for treating or preventing a disease, disorder, or condition selected from asthma and COPD, comprising administering an effective amount of a compound according to Formula I described herein.

[0068] One embodiment of the present invention is a method for treating or preventing a cardiovascular disease, disorder, or condition, comprising administering an effective amount of a compound according to Formula I described herein.

[0069] One embodiment of the present invention is a method for treating or preventing a cardiovascular metabolic disease, disorder, or condition, comprising administering an effective amount of a compound according to Formula I described herein.

[0070] One embodiment of the present invention is a method for treating or preventing a disease, disorder, or condition selected from cryopyrin-associated periodic syndromes, comprising administering an effective amount of a compound according to Formula I described herein.

[0071] One embodiment of the present invention relates to a method for inhibiting NLRP3, comprising administering an effective amount of a compound according to Formula I described herein.

[0072] Furthermore, one embodiment of the present invention is a compound of formula I described herein, when manufactured according to any one of the processes described.

[0073] One embodiment of the present invention is a pharmaceutical composition comprising a compound according to Formula I described herein and a therapeutically inert carrier.

[0074] Assay procedure NLRP3 and pyroptosis It is well established that NLRP3 activation leads to cellular pyroptosis, and that this characteristic plays a crucial role in the manifestation of clinical diseases (Yan-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 & 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, inhibitors of NLRP3 are expected to block pyroptosis and the release of pro-inflammatory cytokines (e.g., IL-1β) from cells.

[0075] THP-1 cells: culture and preparation THP-1 cells (ATCC#TIB-202) were grown 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). The cells were periodically passaged until confluence (approximately 10) 6Cells were grown to a concentration of 625,000 cells / ml. On the day of the experiment, THP-1 cells were collected and resuspended in RPMI medium (without FBS). The cells were then counted, and viability (>90%) was confirmed by trypan blue (Sigma#T8154). Appropriate dilution was performed to obtain a concentration of 625,000 cells / ml. LPS (Sigma#L4524) was added to this diluted cell solution to obtain a final assay concentration (FAC) of 1 μg / ml. 40 μl of the final preparation was dispensed into each well of a 96-well plate. The plates prepared in this way were used for compound screening.

[0076] THP-1 cell pyroptosis assay For compound screening, we followed the stepwise assay method described below. THP-1 cells (25,000 cells / well) containing 1.0 μg / ml LPS are seeded into 40 μl of RPMI medium (without FBS) in a 96-well black-walled, transparent-bottom cell culture plate (VWR#734-0317) coated with poly-D-lysine. Add 5 μl of the compound (8-point semi-logarithmic dilution using the highest dose of 10 μM) or the vehicle (DMSO 0.1% FAC) to the appropriate well. Incubate at 37°C and 5% CO2 for 3 hours. Add 5 μl of nigericin (Sigma #N7143) (FAC 5 μM) to all wells. Incubate at 37°C and 5% CO2 for 1 hour. At the end of the incubation period, rotate the plate at 300xg for 3 minutes and remove the supernatant. Next, add 50 μl of resazurin (Sigma #R7017) (100 μM resazurin in RPMI medium without FAC FBS) and incubate the plate at 37°C and 5% CO2 for a further 1-2 hours. The plate was read with an Envision reader at Ex 560nm and Em 590nm. I C 50 The data is fitted to a nonlinear regression equation (log-inhibitor versus response variable gradient, 4 parameters).

[0077] The results of the pyroptosis assay were obtained from THP IC. 50 This is summarized in Table 1 below.

[0078] Human whole blood IL-1β release assay For systemic delivery, the ability of a compound to inhibit NLRP3 when present in the bloodstream is crucial. Therefore, the NLRP3 inhibitory activity of numerous compounds in human whole blood was investigated according to the following protocol. Human whole blood in Li-heparin tubules was obtained from healthy donors selected from a volunteer donor panel. Plate out 80 μl of whole blood containing 1 μg / ml of LPS into a 96-well clear-bottom cell culture plate (Corning #3585). Add 10 μl of the compound (8-point semi-logarithmic dilution using the highest dose of 10 μM) or the vehicle (DMSO 0.1% FAC) to the appropriate wells. Incubate at 37°C and 5% CO2 for 3 hours. Add 10 μl of nigericin (Sigma# N7143) (10 μM FAC) to all wells. Incubate at 37°C and 5% CO2 for 1 hour. At the end of the incubation period, rotate the plate at 300×g for 5 minutes to pellet the cells, remove 20 μl of supernatant, and add it to a 96-well v-bottom plate for IL-1β analysis. (Note: These plates, including the supernatant, can be stored at -80°C for analysis on a later date.) IL-1β was measured according to the manufacturer's protocol (Perkin Elmer-AlphaLisa IL-1 Kit AL220F-5000). I C 50 The data is fitted to a nonlinear regression equation (log-inhibitor versus response variable gradient, 4 parameters).

[0079] Results of human whole blood assays, HWB IC 50 This is summarized in Table 1 below.

[0080] Microsomal stability: Microsomes (0.5 mg / mL) and 1 μM of the test compound in cofactor NADPH were incubated in a 96-well plate at 37°C using a TECAN (Tecan Group Ltd, Switzerland) automated liquid handling system. After a 10-minute pre-incubation step with the microsomes and test compound, the enzymatic reaction was initiated by adding the cofactor. Aliquots of the incubation were taken at 1, 3, 6, 9, 15, 25, 35, and 45 minutes and quenched with 1:3 (v / v) acetonitrile containing an internal standard. The samples were then cooled, centrifuged, and the supernatant was analyzed by LC-MS / MS2.

[0081] Metabolic stability in hepatocytes: Assay description: Biological materials. Obtain cryopreserved hepatocytes [mouse, rat, rabbit, monkey, and human (mixed sexes)]. The viability of reconstituted hepatocytes should be at least 80% throughout the entire experiment. Obtain ready-to-use rat / human HepatoPac® culture systems [long-term hepatocyte co-culture medium, pooled (human male n=5, and female n=5)], stromal mouse fibroblasts (negative control, pooled), incubation plates, application medium, and maintenance medium.

[0082] Metabolism by suspended hepatocytes. Primary pooled cryopreserved hepatocytes were reconstituted in pre-warmed William's E medium containing 10% FCS, 0.05 mg / mL streptomycin and 50 U / mL penicillin, 0.4 mM L-glutamine, as well as 0.01 mg / mL gentamicin, 0.048 mg / mL hydrocortisone, and 0.004 mg / mL insulin, to a final suspension density of 1 × 10⁶ cells / mL. Incubation was performed fully automated using a Liquid Handling System (Tecan) equipped with an orbital shaker and CO₂ incubator. After adding, for example, 1 μM of the test compound to each well (1 × 10⁵ cells / well), the 96-well hepatocyte suspension culture plate was incubated at 5% CO₂ and 37°C. The sample was quenched at specified time points up to 2 hours by adding acetonitrile (containing the internal standard) to the incubation wells.

[0083] Metabolism by HepatoPac®. The test substance used in the suspension assay (e.g., 1 μM, 0.1% v / v DMSO) is incubated in a 96-well plate containing either adherent hepatocytes and mouse fibroblast control cells, or control cells alone (5% CO2 atmosphere, 37°C). The incubation medium for human HepatoPac® is the same as that used in the suspended hepatocytes. At predetermined time points (2, 18, 26, 48, 72, and 96 hours), all wells are quenched with ice-cold acetonitrile containing an internal standard.

[0084] Subsequently, the sample is properly centrifuged, and the supernatant is analyzed by LC-MS / MS. Incubation is performed with n=1 or n=2.

[0085] hERG Screening Assay In the development process of small molecule drugs, arrhythmias are one of the most frequent side effects that lead to drug failure. Such failures are often related to the drug's ability to inhibit human ether-a-go-go related gene (hERG) cardiac potassium channels. Therefore, no or low inhibition of hERG cardiac potassium channels is considered beneficial.

[0086] cell A CHO clerox hERG cell line (ATCC reference Nr. PTA-6812, female Chinese hamster cells) was generated and validated using Roche. Ready-to-use cryopreserved CHO-hERG cells were cryopreserved at Evotec (Germany) and used directly in experiments.

[0087] Experimental solution The extracellular solution contains the following (in mM): NaCl 150; KCl 4; CaCl21; MgCl21; HEPES 10; pH 7.2-7.4 with NaOH, osmolality 290-330 mOsm. The internal solution contains the following (in mM): KCl 10; KF 100; NaCl 10; HEPES 10; EGTA 20; pH 7.0-7.4 with KOH, osmolality 260-300 mOsm.

[0088] Electrophysiology The effect of the compound on the hERG K+ current parameter will be evaluated at two concentrations in at least four cells.

[0089] The hERG test is performed using the SynchroPatch® 384 automated patch clamp system (Nanion Technologies GmbH, Germany). The K+ current is measured at 35-37°C in a whole cell composition using patch voltage clamp technology.

[0090] The cells were held at a quiescent voltage of -80mV and stimulated with the voltage pattern shown in Figure 1 (outward K at 35-37°C). +The pulse pattern used to extract the current activated the hERG channel with a stimulation frequency of 0.1 Hz (6 bpm), conducting an outward IKhERG current.

[0091] Data Analysis The amplitude of the IKhERG was recorded at each drug concentration and fractional blocks were defined by comparing it to the vehicle control value (set as 100%). The concentration-response data were fitted according to the following relationship: [Table 1]

[0092] The concentration-response curves were fitted using nonlinear regression analysis with the EworkBook suite (ID Business Solutions Ltd, UK). Data fitting was performed using a four-parameter logistic model (fit=(A+(B / (1+((x / C)^D)))), where A=0 and B=100).

[0093] hERG assay results, hERG IC 20 This is summarized in Table 3 below.

[0094] Transcellular P-gp assay: A typical assay uses transfected LLC-PK1 cells (porcine kidney epithelial cells) overexpressing human P-gp or mouse P-gp, cultured on a 96-well semipermeable filter membrane plate, where these cells form a polarized monolayer with tight junctions, acting as a barrier between the apical and basal-lateral compartments.

[0095] P-gp is expressed in the monolayer apical membrane.

[0096] The adhesion of the cell monolayer and the functional activity of P-gp are confirmed by the addition of Lucifer Yellow, a cell impermeability marker, and edoxaban, a reference P-gp substrate, respectively.

[0097] PAMPA: PAMPA (Parallel Microwave Permeability Assay) is a first-line permeability screening method for drug candidates. The PAMPA assay uses an artificial phospholipid membrane to mimic transcellular absorption conditions. This assay determines permeability values ​​that can be used for compound optimization and ranking, as well as for input parameters in in silico models predicting intestinal absorption.

[0098] The donor concentration is measured at t-start (reference) and compared with the donor and acceptor concentrations after a certain period of time (t-end) to calculate the extent to which the compound passes through the membrane.

[0099] Bacterial reverse mutagenesis test (AMES): The compound testing is carried out as outlined in these guidelines. Test No. 471: Bacterial Reverse Mutation Test | OECD Guidelines for Testing Chemicals, Part 4: Health Effects | OECD iLibrary (oecd-ilibrary.org)

[0100] Bacterial culture: The bacterial strains used were TA98, TA100, TA1535, TA97a, and TA102. Batch of each strain was maintained as frozen stock. The vials were thawed and used for inoculation into cultures in nutrient broth. The cultures were placed in an incubator set to 37°C with agitation for approximately 10 hours to obtain working cultures of at least 10⁸ cells / mL.

[0101] To ensure that the cultures are at the appropriate stage of growth and culture density, samples are taken from each culture at the end of the incubation period and the culture density is evaluated by either viable cell count plating or OD650 evaluation.

[0102] treatment: Each compound and positive control concentration is included in a 3-series, and each vehicle control in a 6-series.

[0103] The formulation is prepared using DMSO and allows for exposure up to the solubility limit or a maximum of 1000 μg / well for the freely soluble test substance. This concentration is equivalent to 5000 μg / plate used in a typical plate-integrated Ames assay.

[0104] Concentrations are typically set at semi-logarithmic intervals in a single experiment. For soluble compounds, the concentrations are 0, 3.2, 10, 32, 100, 320, and 1000 μg / well.

[0105] The positive controls used are as follows: Abbreviation Name Stock used 2NF 2-Nitrofluorene TA98-S-9 NaN3 Sodium azide TA100 and TA1535-S-9 AAC 9-aminoacridin TA97a-S-9 MMC Mitomycin C TA102-S-9 B[a]P benzo[a]pyrene TA98+S-9 AAN Aminoanthracene TA100, TA1535, TA97a and TA102+S-9

[0106] Plating is achieved by the following series of additions to 400 μL of replenished molten agar at 45 ± 1°C: 20 μL bacterial culture • 20 μL of test substance solution / vehicle control / positive control • 100 μL of 10% S-9 mix or buffer solution

[0107] Then, mix quickly and pour into the mutation plate (wells).

[0108] Once solidified, invert the plate and incubate it in a dark incubator set to 37°C for 2-3 days.

[0109] toxicity: Toxicity is detected by the following parameters: • Reduction of background loans • Significant reduction in revertant mutants compared to the simultaneous vehicle control. • Reduction of mutagenic response.

[0110] Scoring: Bacterial colony scoring is performed manually or electronically using an automated colony counter.

[0111] In vitro mammalian cell micronucleus test: The testing of the compound is carried out as outlined in these guidelines. Test number 487: In vitro mammalian cell micronucleus test | OECD Guidelines for the Testing of Chemicals, Section 4: Health Effects | OECD iLibrary (oecd-ilibrary.org)

[0112] Cell culture: The cultures are maintained in a tissue culture flask containing GlutaMAX-1-supplemented HEPES-buffered RPMI 1640 medium with 10% (v / v) heat-inactivated fetal bovine serum and 100 units / mL / 100 μg / mL penicillin / streptomycin, in a humidified incubator with 5% (v / v) CO2 in air set to 37°C. Cells are subcultured at low to medium density at least once before treatment.

[0113] The day before the procedure, the cells are subcultured at a density of approximately 7 × 10⁴ cells / mL. The cells are maintained in a humidified environment before the procedure at 37°C and 5% (v / v) CO₂ in the air.

[0114] treatment: Cultured human lymphoblastoid TK6 cells were exposed to a compound for 3 hours in the presence of S-9, followed by a 24-hour recovery period. Furthermore, a continuous 27-hour treatment in the absence of S-9 was included, as several chemicals have been reported to only exhibit a positive effect after prolonged treatment. This corresponds to approximately 1.5–2.0 times the average doubling time (cell cycle approximately 15 hours) of TK6 cells used in this laboratory. All cultures were sampled 27 hours after the start of treatment.

[0115] Prepare a diluent in DMSO that allows for maximum exposure up to the solubility limit, whichever is lower: 1 mM or 500 μg / mL.

[0116] Typically, at least 12 concentrations are spaced 0.7 times apart from the upper limit (for soluble compounds with MW ≥ 500, the concentrations are 9.887, 14.12, 20.18, 28.82, 41.18, 58.82, 84.04, 120.1, 171.5, 245, 350, and 500 μg / mL). The final concentration of DMSO is 1% v / v. Positive controls are noscapine in the absence of S-9 and cyclophosphamide in the presence of S-9. Two replicates are set up for each compound concentration, and multiple concurrent vehicle controls and positive controls are included for each treatment. The cultures are incubated in 96-well plates at 37°C and 5% (v / v) CO2 for the treatment time. The 3-hour treated cultures are washed once and re-incubated in fresh medium for 24 hours.

[0117] Collection: At the specified sampling time, aliquots of cell suspension from the designated culture will be taken for cell counting using a Coulter Counter. The designated culture for analysis will be centrifuged at approximately 200 g for 5 minutes. After resuspending the cells in 0.075 M KCl, they will be fixed with fresh cold methanol / glacial acetic acid (7:1 v / v). The fixed cells will be stored in the fixative at 2–8°C before slide preparation.

[0118] Air-dry the slides before staining, then immerse them in 12.5 μg / mL acridine orange in phosphate-buffered saline (PBS), pH 6.8, for about 10 minutes, followed by washing with PBS for a few seconds (while stirring).

[0119] Cytotoxicity readout and concentration selection: Toxicity is expressed as population doubling (PD) compared to the vehicle control. PD is calculated for each concentration as follows: PD = [log(N / X0)] / log2 In the formula, N = average final cell number at each concentration / culture X0 = Start (baseline) count

[0120] The maximum concentration for micronucleus analysis should not exceed (approximately) 50% cytotoxicity, be the highest concentration tested, or be the lowest precipitate concentration observed by eye at the end of the treatment incubation period.

[0121] Slides from the highest selective concentration and at least two lower concentrations will be analyzed to cover the range of cytotoxicity from maximum to almost none, or none at all, as needed. A minimum of 1000 mononuclear cells (2000 per concentration) from each culture will be analyzed for micronuclei.

[0122] Evaluation criteria: The compound is thought to induce chromosomal abnormalities and / or aneuploidy events in the following cases: -A statistically significant increase in the frequency of MNMON cells is observed at concentrations of 1 or more. - The incidence of cells with such a concentration of micronuclei exceeds the normal range in both replicates. - A concentration-related increase in the proportion of cells with micronuclei is observed (positive trend test).

[0123] If all of the above criteria are met, the compound is considered positive in this assay.

[0124] If none of the above criteria are met, the compound is considered negative in this assay.

[0125] Results that only partially meet the above criteria are treated on a case-by-case basis, but in the context of screening studies, they are concluded as either positive, negative, or inconclusive. Evidence of concentration-related effects is considered useful, but is not essential for evaluating positive results. Biological relevance should take into account, for example, the consistency of responses within and between concentrations, as well as between experiments (where applicable), or effects that occur only at highly toxic concentrations.

[0126] Pharmacokinetic profiles of test substances in miniature pigs: The pharmacokinetics of the test substance were determined in miniature pigs after intravenous and oral administration. The experimental design consisted of three male miniature pigs, each receiving a single intravenous bolus and a single oral dose of the test substance. The intravenous dose was administered at a nominal dose volume of 1 mL / kg. The oral dose was administered via gastric administration at a nominal dose volume of 5 mL / kg. There was a washout period of at least 7 days between the last sampling and the next administration to the same animal. The content of all formulations was within the desired range of 85–115% of the nominal content. After administration, blood samples (1 mL) were collected from the saphenous vein (via cannula) or jugular vein of each animal before administration, and at 5, 15, 30 minutes, 1, 2, 4, 8, and 24 hours after IV administration, as well as before administration and at 15, 30 minutes, 1, 2, 4, 6, 8, 24, and 48 hours after oral administration. Hematocrit was calculated at all time points. Blood:plasma partition coefficients were determined at 2 and 4 hours, and urine was collected as a single sample over 24 hours post-dose administration. Blood samples (nominal 1 mL) were collected from the saphenous vein (via cannula) or jugular vein of each animal into polypropylene tubes containing K2EDTA anticoagulant, and centrifuged (1500 g, 10 min, 4°C) to prepare plasma for analysis. Remaining blood cells were discarded. Plasma vials were capped and stored on wet ice for 60 minutes or less, then transferred to storage at <-50°C (nominal -80°C) before analysis by specific LC-MS methods.

[0127] Toxicity evaluation of test substances in miniature pigs The maximum tolerated dose (MTD) of the test substance is determined after once-daily oral (gastric) administration to miniature pigs. The toxicity of repeated daily administrations for 14 days is then evaluated. Furthermore, the toxicological profile of the test substance is characterized. Sufficiently reared Gottingen miniature pigs are obtained from Ellegaard Gottingen, Dalmose, Denmark (animals: 2-3 months old and 4-6 kg body weight range). At the start of administration, the animals are 4-5 months old and weigh in the 6-9.5 kg range. A dose volume of 10 mL / kg is used. Individual dose volumes are calculated from each animal's current body weight to 30, 100, and 300 mg / kg / day or other target dose levels, depending on the MTD result. Blood samples are taken on days 1 and 14 to determine the drug concentration in plasma and the derived toxicological parameters. Animals are not fed on the scheduled necropsy day. Each animal is anesthetized by intramuscular injection of a zoletyl mixture and then euthanized by bleeding. All tissues are preserved in an appropriate fixative. Further analysis includes food consumption, body weight, clinicopathology, and complete histopathological examination of the target organ. [Table 2] [Table 3] [Table 4]

[0128] The present invention will now be described by the following examples, which do not have a limiting effect.

[0129] If the preparation is obtained as a mixture of enantiomers or diastereoisomers, the pure enantiomers or diastereoisomers can be obtained by the methods described herein or by methods known to those skilled in the art, such as chiral chromatography or crystallization.

[0130] Experimental method [Table 5]

[0131] Preparation of intermediates: Intermediate 1: 2-(4-benzyloxy-2,3-dihydrobenzofuran-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (CAS:2923540-31-0) [ka] Process A: 4-benzyloxy-5-bromo-2,3-dihydrobenzofuran To a solution of 5-bromocoumaran-4-ol (CAS No. 2279149-27-6, 4.59 g, 20.26 mmol, 1.00 equivalent) in acetonitrile (40 mL), potassium carbonate (5.6 g, 40.51 mmol, 2.00 equivalent), followed by benzyl bromide (4.89 g, 3.4 mL, 28.57 mmol, 1.41 equivalent) was added. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was extracted with ethyl acetate and water. The aqueous layer was back-extracted with ethyl acetate. The organic layers were washed with water and brine. The combined organic layers were dried over sodium sulfate, filtered, and concentrated under vacuum. The crude product was adsorbed onto ISOLUTE HM-N and purified by flash chromatography (silica gel, 220 g, ethyl acetate gradient of 0% to 10% heptane) to obtain the title compound (6.17 g, yield 95%) as a colorless oil. LCMS: m / z 305.1 / 307.0 [M+H] + ,ESI pos.

[0132] Step B: 2-(4-benzyloxy-2,3-dihydrobenzofuran-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane To a solution of 4-benzyloxy-5-bromo-2,3-dihydrobenzofuran (Example 1, Step A) (6.16 g, 19.18 mmol, 1.00 equivalent) and 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (CAS No. 61676-62-8, 5.47 g, 6.0 mL, 29.41 mmol, 1.53 equivalents) in tetrahydrofuran (80 mL), a 1.6 M solution of n-butyllithium in hexane (19 mL, 30.4 mmol, 1.59 equivalents) was added dropwise within 40 minutes at -76°C. The mixture was stirred at -76°C for 2.5 hours. The reaction mixture was heated to -60°C, quenched at -60°C with saturated NH4Cl aqueous solution, heated to room temperature, and then extracted with ethyl acetate and saturated NH4Cl aqueous solution. The aqueous layer was back-extracted with ethyl acetate. The organic layers were washed with brine. The combined organic layers were dried over sodium sulfate, filtered, and concentrated under vacuum. The crude product was adsorbed onto ISOLUTE HM-N and purified by flash chromatography (silica gel, 120 g, ethyl acetate gradient of 0% to 10% heptane) to obtain the title compound (5.78 g, yield 81%) as a colorless oil. LCMS: m / z 353.1[M+H] + ESI pos.

[0133] Intermediate 2 6-[[(3R)-1-benzyl-3-piperidyl]amino]-3-chloro-4-methyl-1,2,4-triazine-5-one [ka] Process A: 6-bromo-2-[(4-methoxyphenyl)methyl]-4-methyl-1,2,4-triazine-3,5-dione 6-Bromo-4-methyl-2H-1,2,4-triazine-3,5-dione (CAS No. 15870-75-4, 13.8 g, 63.1 mmol, 1.00 equivalent) and potassium carbonate (4.84 g, 31.5 mmol, 0.50 equivalent) were suspended in dry DMF (125 mL), and 4-methoxybenzyl chloride (10.3 mL, 75.7 mmol, 1.2 equivalents) was added. The reaction mixture was stirred at room temperature for 24 hours. The reaction mixture was diluted with HCl (50 mL), washed with 10 wt% LiCl aqueous solution (2 × 30 mL), dried using a phase separator, and concentrated under vacuum. The residue was purified by silica gel chromatography (0-50% HCl / isohexane) to obtain the title compound (15.9 g, yield 77%) as a white solid. 1 H NMR(500 MHz,DMSO-d6)δ7.33-7.25(m,2H),6.97-6.89(m,2H),5.00(s,2H),3.74(s,3H),3.20(s,3H).

[0134] Process B: 6-[[(3R)-1-benzyl-3-piperidyl]amino]-2-[(4-methoxyphenyl)methyl]-4-methyl-1,2,4-triazine-3,5-dione The mixture of 6-bromo-2-[(4-methoxyphenyl)methyl]-4-methyl-1,2,4-triazine-3,5-dione (5.00 g, 15.33 mmol, 1.00 equivalent), (R)-3-amino-1-benzylpiperidine (CAS No. 168466-84-0, 3.40 g, 17.87 mmol, 1.17 equivalents), cesium carbonate (9.99 g, 30.66 mmol, 2.00 equivalents), Pd(OAc)2 (172 mg, 0.77 mmol, 0.05 equivalents), and xanthophos (444 mg, 0.77 mmol, 0.05 equivalents) in MeCN (100 mL) was degassed with N2 for 5 minutes, then heated to 50°C and stirred for 1 hour. The reaction mixture was then heated to 80°C and stirred for 18 hours. After cooling to room temperature, the mixture was partitioned into toluene (250 mL) and water (100 mL). The organic phase was isolated, washed with brine (50 mL), dried using a phase separator, and concentrated under vacuum. The residue was purified by chromatography on silica gel (0-7% in DCM (0.7N ammonia in MeOH)) to obtain the title compound (5.50 g, 70% yield) as a brown solid. LCMS m / z 436.3[M+H] + ESI pos.

[0135] Process C: 6-[[(3R)-1-benzyl-3-piperidyl]amino]-4-methyl-2H-1,2,4-triazine-3,5-dione Trifluoromethanesulfonic acid (2.91 mL, 32.83 mmol, 2.6 equivalents) was added dropwise at room temperature to a stirred solution of 6-[[(3R)-1-benzyl-3-piperidyl]amino]-2-[(4-methoxyphenyl)methyl]-4-methyl-1,2,4-triazine-3,5-dione (6.47 g, 12.63 mmol, 1.00 equivalent) in DCM (40 mL) and MeCN (20 mL). The reaction system was heated to 35 °C and stirred for 5 days. The reaction system was cooled to room temperature, and then K3PO4 (100 mL, 50% aqueous solution) was added. The reaction mixture was extracted with DCM (3 × 100 mL), and the combined organic layers were dried over MgSO4 and concentrated under vacuum. The crude product was purified by column chromatography on silica gel (0-10% MeOH (0.7M NH3) / DCM) to obtain the title compound (4.1 g, 96% yield) as a light brown solid. LCMS m / z 315.9[M+H] + ESI pos.

[0136] Process D: 6-[[(3R)-1-benzyl-3-piperidyl]amino]-3-chloro-4-methyl-1,2,4-triazine-5-one 6-[[(3R)-1-benzyl-3-piperidyl]amino]-4-methyl-2H-1,2,4-triazine-3,5-dione (4.8 g, 12.94 mmol, 1.00 equivalent) was suspended in phosphorus oxychloride (54.4 mL, 583.63 mmol, 45.11 equivalents), and the reaction mixture was heated to 100°C and stirred for 4 days. The reaction mixture was cooled to room temperature and concentrated under vacuum. The resulting residue was dissolved in MeCN (20 mL) and added in portions to a vigorously stirred mixture of pharmaceutically acceptable phosphate (150 mL) and K3PO4 (250 mL, 50% aqueous solution). The aqueous material was separated and extracted again with pharmaceutically acceptable phosphate (2 × 150 mL). The combined organic layers were dried over MgSO4, concentrated under vacuum, and purified by column chromatography (0-5% MeOH (0.7 M NH3) / DCM) on silica gel to obtain the title compound (3.1 g, 50% yield) as a light brown solid. LCMS m / z 334.3 / 336.3[M+H] + ESI pos.

[0137] Intermediate 3 2-(2-benzyloxy-3-bicyclo[4.2.0]octa-1,3,5-trienyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane [ka] Step A: 2-benzyloxy-3-bromovicyclo[4.2.0]octa-1,3,5-triene Potassium carbonate (458.26 mg, 3.32 mmol, 2.0 equivalents) was added at room temperature to a stirred solution of 3-bromovicyclo[4.2.0]octa-1(6),2,4-trien-2-ol (330.0 mg, 1.66 mmol, 1.0 equivalent, CAS No. 2676864-45-0) in MeCN (5 mL), and the reaction mixture was stirred for 5 minutes. Bromomethylbenzene (0.24 mL, 1.99 mmol, 1.2 equivalents) was then added, and the reaction mixture was stirred for a further 16 hours. The reaction mixture was diluted with water (50 mL) and ethyl acetate (50 mL), and the layers were separated. The aqueous layer was extracted again with ethyl acetate (2 × 50 mL), and the combined organic extract was washed with brine (1 × 50 mL), dried over MgSO4, and concentrated under vacuum. The crude product was purified by column chromatography (0-10% siRNA / heptane) on silica gel to obtain the title compound (440.9 mg, 91% yield) as a colorless oil. This was allowed to stand and crystallize to obtain a white solid. LCMS m / z 289.2 / 291.3[M+H] + ESI pos.

[0138] Step B: 2-(2-benzyloxy-3-bicyclo[4.2.0]octa-1,3,5-trienyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane The mixture of 2-benzyloxy-3-bromovicyclo[4.2.0]octa-1,3,5-triene (0.34 g, 1.17 mmol, 1.0 equivalent), bis(pinacorato)diborone (0.59 g, 2.34 mmol, 2.0 equivalent), cesium carbonate (0.76 g, 2.34 mmol, 2.0 equivalent), Pd(OAc)2 (26.24 mg, 0.12 mmol, 0.1 equivalent), and tris(4-methoxy-3,5-dimethylphenyl)phosphane (51.02 mg, 0.12 mmol, 0.1 equivalent) in 1,4-dioxane (8 mL) was degassed for about 5 minutes and then heated at 85°C for 1 hour. The reaction mixture was cooled to room temperature and then diluted with water (50 mL) and ethyl acetate (50 mL). The organic layer was separated, and the aqueous layer was extracted again with toluene (2 × 50 mL). The combined organic layers were washed with brine (1 × 50 mL), dried over MgSO4, and concentrated under vacuum. The crude material was purified by column chromatography on silica gel (0-5% toluene / heptane) to obtain the title compound (257.5 mg, yield 64%) as a white solid. LCMS m / z 336.3[M+H] + ESI pos.

[0139] Preparation of the example: Example 1: 3-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-6-[[(3R)-1-(2-hydroxyethyl)-3-piperidyl]amino]-4-methyl-1,2,4-triazine-5-one [ka] Step A: 3-(4-benzyloxy-2,3-dihydrobenzofuran-5-yl)-6-[[(3R)-1-benzyl-3-piperidyl]amino]-4-methyl-1,2,4-triazine-5-one A mixture of 2-(4-benzyloxy-2,3-dihydrobenzofuran-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (intermediate 1, 928.53 mg, 2.64 mmol, 1.1 equivalent, CAS number 2923540-31-0), 6-[[(3R)-1-benzyl-3-piperidyl]amino]-3-chloro-4-methyl-1,2,4-triazine-5-one (intermediate 2, 1.0 g, 2.4 mmol, 1.00 equivalent), saturated sodium carbonate aqueous solution (4.5 mL), and Xphos Pd G3 (101.55 mg, 0.12 mmol, 0.05 equivalent) in MeCN (18 mL) was degassed with nitrogen for 5 minutes, and then heated at 80°C for 2 hours. The reaction mixture was cooled to room temperature, then diluted with ELISA (25 mL), and acidified to approximately pH 2 with 1 M HCl (approximately 40 mL). The layers were separated, and the organic phase was further extracted with 1 M HCl (2 x 25 mL). The combined aqueous layer was basicized with solid NaOH until the pH was approximately 10-12, and then extracted with DCM (4 x 50 mL). The combined organic layers were dried over MgSO4, filtered, and concentrated under vacuum to obtain the title compound (1.10 g, yield 61%) as a brown foamy solid, which was used in the next step without further purification. LCMS m / z 524.3[M+H] + ESI pos.

[0140] Step B: 3-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-4-methyl-6-[[(3R)-3-piperidyl]amino]-1,2,4-triazine-5-one Carbon-supported palladium (5R87L) (319.88 mg, 0.15 mmol, 0.1 equivalent) was added to a stirred solution of 3-(4-benzyloxy-2,3-dihydrobenzofuran-5-yl)-6-[[(3R)-1-benzyl-3-piperidyl]amino]-4-methyl-1,2,4-triazine-5-one (1.10 g, 1.47 mmol, 1.0 equivalent) in ethanol (15 mL), and the reaction mixture was stirred under H2 (2 bar) for approximately 16 hours. The reaction mixture was filtered (GF / F paper), concentrated under vacuum, and the crude substance was then purified by column chromatography on silica gel (40 g cartridge, 0-10% MeOH (0.7 M NH3) / DCM) to obtain the title compound (401.5 mg, yield 71.69%) as a grayish-white solid. LCMS m / z 344.2[M+H] + ESI pos.

[0141] Step C: 3-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-6-[[(3R)-1-(2-hydroxyethyl)-3-piperidyl]amino]-4-methyl-1,2,4-triazine-5-one 1,4-dioxane-2,5-diol (124.65 mg, 1.04 mmol, 1.1 equivalents) in EtOH (2 mL) was added dropwise to a stirred solution of 3-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-4-methyl-6-[[(3R)-3-piperidyl]amino]-1,2,4-triazine-5-one (360.0 mg, 0.94 mmol, 1.0 equivalent) in ethanol (12 mL). Then, sodium triacetoxyborohydride (499.95 mg, 2.36 mmol, 2.5 equivalents) was added in portions, and the reaction mixture was stirred for 2 hours. Further 1,4-dioxane-2,5-diol (56.66 mg, 0.47 mmol, 0.5 equivalents) and sodium triacetoxyborohydride (249.97 mg, 1.18 mmol, 1.25 equivalents) were added, and the reaction mixture was stirred for 3 hours. The reaction mixture was diluted with MeOH (20 mL) and then concentrated under vacuum. The resulting residue was dissolved in 1 M aqueous HCl (10 mL), washed with DCM (3 × 10 mL), and the aqueous layer was then basicized to approximately pH 9-10 using solid K3PO4 and extracted with DCM (3 × 20 mL). The combined organic layers were dried over MgSO4, concentrated under vacuum, and purified by RP chromatography. The impurity-containing substance was dissolved in 1.94 mL of DMSO, filtered, and purified by reverse-phase preparative HPLC (Waters 2767 Sample Manager, Waters 2545 Binary Gradient Module, Waters Systems Fluidics Organiser, Waters 515 ACD pump, Waters 515 Makeup pump, Waters 2998 Photodiode Array Detector, Waters QDa) using a Waters XBridge BEH C18 ODB preparative column, 130 Å, 5 μm, 30 mm × 100 mm, flow rate 40 mL / min, eluting over 17.5 minutes with 0.3% ammonia in a water-MeCN gradient using UV across all wavelengths with PDA, QDA, and ELS detectors. The at-column dilution pump supplied 2 mL / min of methanol throughout the process, which is included in the following MeCN percentage.Gradient information: 0.0-0.5 min, 5% MeCN; 0.5-15.5 min, gradient from 5% MeCN to 17.5% MeCN; 15.5-15.6 min, gradient from 17.5% MeCN to 100% MeCN; 15.6-17.5 min, held at 100% MeCN. The clean fraction was evaporated using a freeze-dryer. The recovered material was dissolved in DCM (5 mL) and water (5 mL), the organic layer was separated, and washed with water (2 × 5 mL). The combined aqueous layer was adjusted to pH 7 (using 1 M HCl aqueous solution and saturated NaHCO3 aqueous solution) and extracted with DCM (4 × 5 mL). The combined organic layer was dried over MgSO4 and concentrated under vacuum to obtain the title compound (15.7 mg, yield 4%) as a grayish-white solid. LCMS m / z 388.2 [M + H]. + ESI pos.

[0142] Example 2: 6-[[(3R)-1-(2-hydroxyethyl)-3-piperidyl]amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazine-5-one [ka] Process A: 3-(4-benzyloxyindan-5-yl)-6-[[(3R)-1-benzyl-3-piperidyl]amino]-4-methyl-1,2,4-triazine-5-one A mixture of 2-(4-benzyloxyindan-5-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (CAS No. 2878443-82-2, 782 mg, 2.23 mmol, 1.10 equivalents), 6-[[(3R)-1-benzyl-3-piperidyl]amino]-3-chloro-4-methyl-1,2,4-triazine-5-one (intermediate 2, 850 mg, 2.04 mmol, 1.0 equivalent), saturated sodium carbonate aqueous solution (4 mL), and Xphos Pd G3 (87 mg, 0.1 mmol, 0.05 equivalents) in MeCN (16 mL) was degassed with nitrogen for 5 minutes and then heated at 80°C for 1 hour. The reaction mixture was cooled to room temperature, then diluted with ELISA (25 mL), and acidified to approximately pH 2 with 1 M HCl (approximately 30 mL). The layers were separated, and the organic phase was further extracted with 1 M HCl (2 × 20 mL). The combined aqueous phase was basicized with solid NaOH until the pH was approximately 10, and extracted with SiO2 (4 × 50 mL). The combined organic phase was dried over MgSO4, filtered, and concentrated to obtain the title compound (980 mg, yield 69%) as a dark brown foam. LCMS m / z 522.3 [M + H] + ESI pos.

[0143] Process B: 3-(4-hydroxyindan-5-yl)-4-methyl-6-[[(3R)-3-piperidyl]amino]-1,2,4-triazine-5-one Carbon-supported palladium (5R87L) (320 mg, 0.15 mmol, 0.10 equivalents) was added to a stirred solution of 3-(4-benzyloxyindan-5-yl)-6-[[(3R)-1-benzyl-3-piperidyl]amino]-4-methyl-1,2,4-triazine-5-one (980 mg, 1.50 mmol, 1.00 equivalent) in ethanol (15 mL), and the reaction mixture was stirred under H2 (2 bar) for approximately 16 hours. Further carbon-supported palladium (320 mg, 0.15 mmol, 0.10 equivalents) was added, and the reaction mixture was stirred under H2 (2 bar) for a further 3 hours, then filtered (GF / F paper) and concentrated under vacuum. The crude substance was purified by column chromatography on silica gel (40 g cartridge, 0-10% MeOH (0.7 M NH3) / DCM) to obtain the title compound (402 mg, 70% yield) as a grayish-white solid. LCMS m / z 342.2[M+H] + ESI pos.

[0144] Process C: 6-[[(3R)-1-(2-hydroxyethyl)-3-piperidyl]amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazine-5-one EtOH (2 mL) containing 1,4-dioxan-2,5-diol (93 mg, 0.77 mmol, 1.20 equivalents) was added dropwise at 0°C to a stirred solution of 3-(4-hydroxyindan-5-yl)-4-methyl-6-[[(3R)-3-piperidyl]amino]-1,2,4-triazine-5-one (250 mg, 0.64 mmol, 1.00 equivalent) in ethanol (12 mL). Then, sodium triacetoxyborohydride (341 mg, 1.61 mmol, 2.50 equivalents) was added in portions, and the reaction mixture was stirred at 0°C for 30 minutes. The reaction mixture was allowed to return to room temperature, stirred for 1 hour, and then quenched by adding water (20 mL). The reaction mixture was further diluted with NaHCO3 (50 mL), and the aqueous layer was extracted with DCM (3 × 50 mL). The combined organic layers were dried over MgSO4, concentrated under vacuum, and then purified by column chromatography on silica gel (40g cartridge, 0-10% MeOH (0.7M NH3) / DCM). The impurity-containing material was dissolved in 3.7 mL of DMSO, filtered, and purified by reverse-phase preparative HPLC (Waters 2767 Sample Manager, Waters 2545 Binary Gradient Module, Waters Systems Fluidics Organiser, Waters 515 ACD pump, Waters 515 Makeup pump, Waters 2998 Photodiode Array Detector, Waters QDa) using a Waters XBridge BEH C18 ODB preparative column, 130 Å, 5 μm, 30 mm × 100 mm, flow rate 40 mL / min, eluting over 12.5 minutes with 0.3% ammonia in a water-MeCN gradient using UV across all wavelengths with PDA, QDA, and ELS detectors. The at-column dilution pump dispensed 2 mL / min of methanol throughout the entire method, which is included in the following percentages of MeCN. Gradient information: 0.0–0.5 min, 5% MeCN; 0.5–10.5 min, sloped from 5% MeCN to 32.5% MeCN; 10.5–10.6 min, sloped from 32.5% MeCN to 100% MeCN; 10.6–12.5 min, held at 100% MeCN.The fraction containing the product was evaporated using Genevac, and the impurity-containing material was then dissolved in 1.87 mL of DMSO, filtered, and purified by reverse-phase preparative HPLC (Waters 2767 Sample Manager, Waters 2545 Binary Gradient Module, Waters Systems Fluidics Organiser, Waters 515 ACD pump, Waters 515 Makeup pump, Waters 2998 Photodiode Array Detector, Waters QDa) using a Waters XBridge BEH C18 ODB preparative column, 130 Å, 5 μm, 30 mm × 100 mm, flow rate 40 mL / min, with UV across all wavelengths using PDA, QDA, and ELS detectors, eluting with 0.3% ammonia in a water-MeCN gradient over 17.5 minutes. The at-column dilution pump supplied 2 mL / min of methanol throughout the process, which is included in the following MeCN percentage. Gradient information: 0.0-0.5 min, 5% MeCN; 0.5-15.5 min, gradient from 5% MeCN to 25% MeCN; 15.5-15.6 min, gradient from 25% MeCN to 100% MeCN; 15.6-17.5 min, held at 100% MeCN. The clean fraction was evaporated using a freeze-dryer. The recovered material was dissolved in DCM (5 mL) and water (5 mL), the organic layer was separated, and washed with water (2 × 5 mL). The combined aqueous layer was adjusted to pH 7 (using 1 M HCl aqueous solution and saturated NaHCO3 aqueous solution) and extracted with DCM (4 × 5 mL). The combined organic layer was dried over MgSO4 and concentrated under vacuum to obtain the title compound (39.3 mg, yield 16%) as a white solid. LCMS m / z 386.3 [M + H]. + ESI pos.

[0145] Example 3 3-(4-hydroxyindan-5-yl)-6-[[(3R)-1-(3-hydroxypropyl)-3-piperidyl]amino]-4-methyl-1,2,4-triazine-5-one; formic acid [ka] 3-iodopropanol (16.1 mg, 0.09 mmol, 0.99 equivalents) in DMF (0.300 mL) was added dropwise at room temperature to a stirred solution in DMF (0.500 mL) of 3-(4-hydroxyindan-5-yl)-4-methyl-6-[[(3R)-3-piperidyl]amino]-1,2,4-triazine-5-one (30.0 mg, 0.09 mmol, 1.0 equivalent; Example 2, Step B) and DIPEA (30.0 μL, 0.17 mmol, 1.96 equivalents), and the reaction mixture was stirred overnight (28 hours). The mixture was transferred to a separation funnel and rinsed with ELISA (50 mL) and 1 M HCl (50 mL). The separated organic layer was further extracted with 1 M HCl (50 mL). The combined aqueous layer was basicized with saturated NaHCO3 aqueous solution until a pH of approximately 8 was achieved, and then extracted with Â15 (3 × 25 mL). The combined organic layer was dried over sodium sulfate, filtered, and concentrated. The crude product was then purified by column chromatography on C18 silica gel (26 g cartridge, 0 → 100% gradient using 0.1% formic acid / 0.1% formic acid aqueous solution in MeCN) to obtain the title compound (19.0 mg, yield 54%) as a grayish-white solid. LCMS m / z 400.0 [M + H] + ESI pos.

[0146] Example 4: 3-(2-hydroxy-3-bicyclo[4.2.0]octa-1(6),2,4-trienyl)-6-[[(3R)-1-(2-hydroxyethyl)-3-piperidyl]amino]-4-methyl-1,2,4-triazine-5-one [ka] Step A: 3-(2-benzyloxy-3-bicyclo[4.2.0]octa-1(6),2,4-trienyl)-4-methyl-6-[[(3R)-1-benzyl-3-piperidyl]amino]-1,2,4-triazine-5-one A mixture of 2-(2-benzyloxy-3-bicyclo[4.2.0]octa-1,3,5-trienyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane intermediate 3 (110.8 mg, 0.33 mmol, 1.1 equivalents), 3-chloro-4-methyl-6-[[(3R)-1-benzyl-3-piperidyl]amino]-1,2,4-triazine-5-one intermediate 2 (100.0 mg, 0.3 mmol, 1.0 equivalent), saturated sodium carbonate aqueous solution (95.25 mg, 0.9 mmol, 3.0 equivalents), and XPhos Pd G3 (12.69 mg, 0.01 mmol, 0.05 equivalents) in MeCN (3 mL) was degassed with nitrogen for 5 minutes, and then heated at 80°C for 1.5 hours. The reaction mixture was cooled to room temperature, then diluted with toluene (10 mL), and acidified to approximately pH 2 with 1 M HCl (approximately 10 mL). The layers were separated, and the organic layer was further extracted with 1 M HCl (2 × 10 mL). The combined aqueous layer was basicized with solid K3PO4 until the pH was approximately 10, and then extracted with DCM (4 × 20 mL). The combined organic layer was dried over MgSO4, filtered, and concentrated under vacuum to obtain the title compound (161.3 mg, yield 85%) as a yellow viscous oil, which was used in the next step without further purification. LCMS m / z 508.4[M+H] + ESI pos.

[0147] Step B: 3-(2-hydroxy-3-bicyclo[4.2.0]octa-1(6),2,4-trienyl)-4-methyl-6-[[(3R)-3-piperidyl]amino]-1,2,4-triazine-5-one Carbon-supported palladium (62.89 mg, 0.03 mmol, 0.1 equivalent) (a 1:1 mixture of 87 L and R39) was added to a stirred solution of 3-(2-benzyloxy-3-bicyclo[4.2.0]octa-1(6),2,4-trienyl)-4-methyl-6-[[(3R)-1-benzyl-3-piperidyl]amino]-1,2,4-triazine-5-one (150.0 mg, 0.3 mmol, 1.0 equivalent) in ethanol (5 mL). The reaction mixture was stirred under H2 (2 bar) for approximately 16 hours. The reaction mixture was filtered (GF / F paper), concentrated under vacuum, and the crude substance was then purified by column chromatography on silica gel (0-10% MeOH (0.7 M NH3) / DCM) to obtain the title compound (40.8 mg, yield 42%) as a white solid. LCMS m / z 328.2[M+H] + ESI pos.

[0148] Step C: 3-(2-hydroxy-3-bicyclo[4.2.0]octa-1(6),2,4-trienyl)-6-[[(3R)-1-(2-hydroxyethyl)-3-piperidyl]amino]-4-methyl-1,2,4-triazine-5-one 2-iodoethanol (16.47 mg, 0.1 mmol, 0.95 equivalents) in DMF (0.1 mL) was added dropwise to a stirred solution in DMF (0.400 mL) of 3-(2-hydroxy-3-bicyclo[4.2.0]octa-1(6),2,4-trienyl)-4-methyl-6-[[(3R)-3-piperidyl]amino]-1,2,4-triazine-5-one (33.0 mg, 0.1 mmol, 1.0 equivalent) and DIPEA (26.34 μL, 0.15 mmol, 1.5 equivalents) at room temperature, and the reaction mixture was stirred for approximately 24 hours. The reaction mixture was diluted with HCl (10 mL) and 1 M aqueous HCl (10 mL), and the layers were separated. The organic layer was washed with 1 M aqueous HCl (2 × 5 mL), and then the combined aqueous layer was washed with HCl (3 × 10 mL). The aqueous layer was neutralized to approximately pH 8 with saturated NaHCO3 aqueous solution, and then extracted with DCM (3 × 20 mL). The combined organic layers were dried over MgSO4, concentrated under vacuum, and purified by column chromatography on silica gel (0-10% MeOH (0.7 M NH3) / DCM) to obtain the title compound (11.5 mg, yield 30%) as a white solid. LCMS m / z 372.3 [M + H] + ESI pos.

[0149] Example 5 6-[[(3R)-1-acetyl-3-piperidyl]amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazine-5-one [ka] HATU (31.01 mg, 0.13 mmol, 1.5 equivalents) was added in portions at room temperature to a stirred solution of 3-(4-hydroxyindan-5-yl)-4-methyl-6-[[(3R)-3-piperidyl]amino]-1,2,4-triazine-5-one (30.0 mg, 0.09 mmol, 1.0 equivalent; Example 2, Step B), acetic acid (5.8 mg, 0.1 mmol, 1.1 equivalents), and DIPEA (22.96 μL, 0.13 mmol, 1.5 equivalents) in DMF (1 mL), and the reaction mixture was stirred for 1 hour. The reaction mixture was diluted with water (15 mL), and the aqueous mixture was extracted with DCM (3 × 15 mL). The combined organic layers were washed with brine (1 × 20 mL), dried over MgSO4, and concentrated under vacuum. The crude substance was purified by column chromatography on silica gel (0-5% MeOH (0.7M NH3) / DCM) to obtain the title compound (6.8 mg, 18% yield) as a white solid. LCMS m / z 384.2[M+H] + ESI pos.

[0150] Example RE-A: 6-[[(3R)-1-ethyl-3-piperidyl]amino]-3-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-4-methyl-1,2,4-triazine-5-one [ka] RE-A is Example 2 of International Publication No. WO2022238347A1, and was synthesized according to the procedure described therein.

[0151] Example A The compound of formula I can be used as an active ingredient in a manner known to itself to produce tablets of the following composition: per tablet Active ingredient 200mg Microcrystalline cellulose 155mg Corn starch 25mg Talc 25mg Hydroxypropylmethylcellulose 20mg 425mg

[0152] Example B The compound of formula I can be used as an active ingredient in known methods to produce capsules of the following composition: per capsule Active ingredient: 100.0 mg Corn starch 20.0 mg Lactose 95.0 mg Talc 4.5mg Magnesium stearate 0.5mg 220.0 mg

Claims

1. Formula I: 【Chemistry 1】 (In the formula, A is -O- or CH ２ And, R １ is a hydroxyalkyl or acetyl, R ２ It is an alkyl, n is either 0 or 1, If n is 0, then A is CH ２ (is) Compounds of and pharmaceutically acceptable salts thereof.

2. R １ The compound according to claim 1, wherein is a hydroxyalkyl group.

3. R ２ The compound according to claim 1 or 2, wherein is methyl.

4. below: 3-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-6-[[(3R)-1-(2-hydroxyethyl)-3-piperidyl]amino]-4-methyl-1,2,4-triazine-5-one, 6-[[(3R)-1-(2-hydroxyethyl)-3-piperidyl]amino]-3-(4-hydroxyindan-5-yl)-4-methyl-1,2,4-triazine-5-one, Compounds according to any one of claims 1 to 3, selected from, and pharmaceutically acceptable salts thereof.

5. below: 3-(4-hydroxyindan-5-yl)-6-[[(3R)-1-(3-hydroxypropyl)-3-piperidyl]amino]-4-methyl-1,2,4-triazine-5-one, 3-(2-hydroxy-3-bicyclo[4.2.0]octa-1(6),2,4-trienyl)-6-[[(3R)-1-(2-hydroxyethyl)-3-piperidyl]amino]-4-methyl-1,2,4-triazine-5-one, Compounds according to any one of claims 1 to 3, selected from, and pharmaceutically acceptable salts thereof.

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

7. A pharmaceutical composition comprising a compound according to any one of claims 1 to 6 and a therapeutically inert carrier.

8. A compound according to any one of claims 1 to 6, for use as a therapeutically active substance.

9. A compound according to any one of claims 1 to 6 for use in the treatment or prevention of a disease, disorder, or pathological condition, wherein the disease, disorder, or pathological condition is responsive to NLRP3 inhibition.

10. A compound according to any one of claims 1 to 6, for the treatment or prevention of a disease, disorder, or condition selected from asthma or COPD.

11. Use of a compound according to any one of claims 1 to 6 for the treatment or prevention of a disease, disorder or condition, wherein the disease, disorder or condition is responsive to NLRP3 inhibition.

12. Use of the compound according to any one of claims 1 to 6 in the treatment or prevention of a disease, disorder, or condition selected from asthma or COPD.

13. Use of the compound according to any one of claims 1 to 6 for the preparation of a pharmaceutical product for the treatment or prevention of a disease, disorder, or condition selected from asthma or COPD.

14. A method for inhibiting NLRP3, comprising administering an effective amount of a compound according to any one of claims 1 to 6 to inhibit NLRP3.

15. A method for treating or preventing a disease, disorder, or condition, wherein the method comprises administering an effective amount of a compound according to any one of claims 1 to 6, wherein the disease, disorder, or condition is selected from asthma or COPD.

16. The invention as described prior to this specification.

Images