Crystalline forms, chemical processes and chemical compounds of NLRP3 inflammasome inhibitors

Abstract

This specification generally relates to crystalline forms of inhibitors of the NLRP3 inflammasome, pharmaceutical compositions containing such crystalline forms, chemical processes useful for preparing inhibitors of the NLRP3 inflammasome, and intermediate compounds useful in such processes. Such crystalline forms and pharmaceutical compositions are useful for inhibiting NLRP3 inflammasome activity and may be useful as therapeutic agents.

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims the benefit of U.S. Provisional Patent Application No. 63 / 477,516, filed December 28, 2022, the entire contents of which are incorporated herein by reference.

[0002] FIELD OF THE INVENTION Described herein are crystalline forms of inhibitors of the NLRP3 inflammasome, pharmaceutical compositions comprising such crystalline forms, chemical processes useful for preparing inhibitors of the NLRP3 inflammasome, and intermediate compounds useful in such processes. [Background technology]

[0003] The NLRP3 inflammasome is a multiprotein complex consisting of the NLR family pyrin domain-containing 3 (NLRP3) protein, the PYD and CARD domain-containing (ASC, also known as PYCARD), and caspase 1 (CASP1), a stress-sensing pathway that leads to inflammatory responses (Swanson KV et al. Nat Rev Immunol. 2019 Aug;19(8):477-489). Upon activation, these three proteins condense into a large multiprotein complex, the speck.

[0004] The NLRP3 protein consists of three domains: PYD, NACHT, and LRR (Sharif H et al. Nature. 2019 Jun;570(7761):338-343). The amino-terminal PYD domain is thought to be important for binding of NLRP3 to the PYD domain of ASC. The NACHT domain possesses ATPase activity, which is suggested to regulate oligomerization, potentially via conformational changes in the LRR domain. The LRR domain is thought to induce autoinhibition by folding onto the NACHT domain. The activity of the NLRP3 protein is further regulated by numerous post-translational modifications, including phosphorylation and ubiquitination.

[0005] Numerous cellular stressors, including pathogen-associated molecular patterns (PAMPs), endogenous danger signals (DAMPs), and environmental irritants, have been shown to lead to the condensation of inflammasomes into specks. Inflammasome activation is thought to require two steps (McKee CM et al. J Leukoc Biol. 2020 Sep;108(3):937-952). The initial priming step, which acts to increase the levels of inflammasome components, can be initiated, for example, by lipopolysaccharide (LPS, a common PAMP). LPS is detected via toll-like receptors, leading to NF-kB-driven transcription of NLRP3 and IL1B. Secondary insults initiate the rapid oligomerization of inflammasome components into specks, producing activated caspase-1.

[0006] In addition to this two-step process, very high induction of NLRP3 transcription has been shown to drive inflammasome activation in a single step, typically through prolonged LPS exposure.

[0007] The downstream effects of activated NLRP3 inflammasomes are further amplified through caspase-1-mediated cleavage and subsequent activation of gasdermin D. Upon activation, gasdermin D forms large pores, leading to a regulated form of lytic cell death called pyroptosis (Kovacs SB et al. Trends Cell Biol. 2017 Sep;27(9):673-684). Indeed, pyroptosis amplifies inflammation through the release of cellular contents, followed by the recruitment and influx of additional immune cells.

[0008] Dysregulated inflammasome drive, even at low levels over several years, can likely lead to tissue damage and chronic disease, as has been demonstrated for cryopyrin-associated periodic syndromes 1, 2, and 3, in which a causative genetic lesion in NLRP3 has been identified (Kacar M et al. Rheumatology (Oxford). 2019 Nov 1;58(Suppl 6):vi31-vi43).

[0009] NLRP3 inflammasome activation has been associated with multiple indications (as discussed herein) with often demonstrated presence or activity in affected tissues, and thus, inhibition of the NLRP3 inflammasome resolves unwanted inflammation.

[0010] The NLRP3 inflammasome can regulate both acute kidney injury (AKI) and chronic kidney disease (CKD), and mice lacking NLRP3 inflammasome components and their downstream mediators are protected from renal injury in experimental models of both AKI and CKD (Hutton HL et al. Nephrology. 2016 21(9):736-744). Inflammation plays a key role in the pathogenesis of AKI. After an initial ischemic, sepsis, or nephrotoxic trigger, the release of inflammatory cytokines and chemokines by renal endothelial cells and tubular epithelium can lead to leukocyte recruitment and subsequent kidney injury. The role of the inflammasome in this process has been demonstrated in both biomarker studies and experimental models of AKI (Andersen K et al. Kidney Int. 2014 Nov;86(5):965-78). Increasing evidence from clinical and experimental studies indicates that both systemic and local renal inflammation play important roles in the development and progression of diabetic kidney disease (DKD) (Tang SCW et al. Nat Rev Nephrol. 2020 Apr;16(4):206-222). Specifically, the NLRP3 inflammasome links sensing metabolic stress in the diabetic kidney to activation of a pro-inflammatory cascade via induction of IL-1β and IL-18, leading to chronic injury and renal dysfunction in CKD / DKD (Shahzad K et al. J Am Soc Nephrol. 2016 Aug;27(8):2270-5).

[0011] Studies have implicated the NLRP3 inflammasome in cardiovascular disease (An N et al. Front Immunol. 2019 Jul 10;10:1592). The relationship between the NLRP3 inflammasome and coronary atherosclerotic heart disease and vascular injury via cholesterol crystals / monosodium glutamate and downstream factors has been well documented (Jin Y et al. J Am Heart Assoc. 2019 Jun 18;8(12):e012219). In addition, the NLRP3 inflammasome may also be involved in the pathological mechanisms of cardiomyopathy, including myocardial infarction (MI), cardiac remodeling, and cardiac hypertrophy (An N et al. Front Immunol. 2019 Jul 10;10:1592).

[0012] Nonalcoholic fatty liver disease (NAFLD) is defined as excess hepatic fat accumulation (steatosis) exceeding 5% caused by causes other than alcohol consumption. Fatty liver progresses to nonalcoholic steatohepatitis (NASH) with or without fibrosis in a variable proportion of individuals, ultimately leading to cirrhosis, liver failure, and hepatocellular carcinoma in susceptible individuals (Friedman et al. Nat Med. 2018 Jul;24(7):908-922). Inflammation involving the NLRP3 inflammasome contributes to the pathogenesis of most acute and chronic liver diseases, including NAFLD, NASH, alcoholic steatohepatitis, chronic hepatitis C virus (HCV) infection, ischemia-reperfusion injury, and paracetamol-induced liver injury (Szabo et al. Nat Rev Gastroenterol Hepatol 2015;12:387-400). Hepatic NLRP3 and downstream target mRNA levels are increased in NASH and correlate with liver collagen expression levels in humans. In addition, NLRP3-induced activation increases liver fibrosis in mice, and NLRP3 knockout mice are protected from experimentally induced NASH, including liver inflammation and fibrosis (Wree et al. J Mol Med, 2014, DOI: 10.1007 / s00109-014-1170-1). Inhibition of the NLRP3 inflammasome using a small molecule inhibitor (MCC950) reduces liver inflammation and fibrosis in experimental NASH models in which mice are fed a high-fat diet or a methionine- and choline-deficient diet (Mridha et al. Journal of Hepatology, 2017, DOI: 10.1016 / j.jhep.2017.01.022). Therefore, NLRP3 inflammasome inhibition can protect against liver diseases, including NAFLD and NASH.

[0013] Several hyperactivating mutations in NLRP3 are associated with autoinflammatory disorders, resulting in the inappropriate release of inflammatory cytokines, including IL-1β, and inflammatory symptoms. Cryopyrin-associated periodic syndromes (CAPS) include familial cold autoinflammatory syndrome (FCAS), Muckle-Wells syndrome (MWS), chronic infantile neurologic cutaneous articular (CINCA) syndrome, and neonatal-onset multisystem inflammatory disease (NOMID) (Booshehri ML et al. J Clin Immunol. 2019 Apr;39(3):277-286).

[0014] The NLRP3 inflammasome has also been implicated in gout and pseudogout, as monosodium urate (MSU) and calcium pyrophosphate dihydrate (CPPD), both crystals found in gout, are activators of the NLRP3 inflammasome (Martinon F et al. Nature 440:237-241, 2006). In sarcoidosis, the NLRP3 inflammasome has been identified as one of the key cellular pathways (Riteau N et al. Eur Respir J. 2020;55(3):2000149), and increased activity has been shown in the lungs of sarcoid patients.

[0015] Evidence suggests that inflammasomes play a role in autoimmune diseases and that inhibition of the NLRP3 inflammasome may have positive effects in rheumatoid arthritis (RA), multiple sclerosis (MS), Addison's disease, celiac disease, systemic lupus erythematous (SLE), and vitiligo (Shaw PJ et al. Trends Mol Med. 2011 Feb;17(2):57-64).

[0016] In inflammatory skin diseases, NLRP3 inflammasome activation has been shown in acne vulgaris (Li ZJ et al. J Invest Dermatol. 2014 Nov; 134(11):2747-2756) and hidradenitis suppurativa (Kelly G et al. Br J Dermatol. 2015 Dec; 173(6):1431-9).

[0017] Emerging evidence suggests that persistent activation of NLRP3 may be involved in the progression of several chronic lung diseases, including idiopathic pulmonary fibrosis (IPF), chronic obstructive pulmonary disease (COPD), and asthma (De Nardo D. et al. Am J Pathol. 2014 Jan;184(1):42-54).

[0018] In inflammatory bowel disease (IBD), evidence suggests that inflammasome-driven IL-1β and IL-18 play a role in IBD pathology, and that NLRP3 inflammasome inhibitors may be effective in ulcerative colitis (UC) and Crohn's disease (Zhen Y et al. Front Immunol. 2019 Feb 28;10:276).

[0019] Therefore, inhibitors of the NLRP3 inflammasome may be useful in treating the diseases and conditions described herein that are associated with NLRP3 inflammasome activation. However, to date, no small molecule synthetic inhibitors of the NLRP3 inflammasome have been approved for medical use.

[0020] Small molecule inhibitors of the NLRP3 inflammasome have been previously discussed, for example, in WO 2020 / 234715(A1), WO 2022 / 135567(A1), and WO 2022 / 216971(A1); notwithstanding the foregoing, there continues to be a need for additional compounds that are inhibitors of the NLRP3 inflammasome, and processes for making such compounds, which may therefore be particularly promising for development as therapeutic agents.

[0021] Compounds 1 and 2 described herein are described in International Application No. PCT / EP2022 / 068292 and are inhibitors of the NLRP3 inflammasome. Stable crystalline forms of such compounds, which may be useful in the commercial manufacture of pharmaceutical compositions, and pharmaceutical compositions comprising such stable crystalline forms, are desirable. Furthermore, efficient processes for making such compounds are desirable, along with novel chemical intermediates useful in such synthetic processes. Summary of the Invention

[0022] This specification describes, in part, a crystalline form of (2S)-3-[[4-[2-hydroxy-4-(trifluoromethyl)phenyl]phthalazin-1-yl]amino]propane-1,2-diol (Compound 1).

[0023] This specification also describes, in part, a crystalline form of Compound 1, which is crystalline form A.

[0024] This specification also describes, in part, a crystalline form of 2-(4-(((1R,2R)-2-hydroxycyclohexyl)amino)pyrido[3,4-d]pyridazin-1-yl)-5-(trifluoromethyl)phenol (Compound 2), hydrochloride salt.

[0025] This specification also describes, in part, pharmaceutical compositions comprising the crystalline forms disclosed herein and a pharmaceutically acceptable excipient.

[0026] This specification also describes, in part, the crystalline forms or pharmaceutical compositions disclosed herein for use in therapy.

[0027] This specification also describes, in part, a crystalline form or pharmaceutical composition disclosed herein for use in treating a subject having a disease or condition involving NLRP3 inflammasome activity.

[0028] The present specification also describes, in part, a method for inhibiting NLRP3 inflammasome activity in a subject in need thereof, the method comprising administering to the subject a crystalline form or pharmaceutical composition disclosed herein.

[0029] The present specification also describes, in part, a process for the preparation of a compound of formula (Ic) or a salt thereof, comprising:

[0030] [ka] (a) reacting 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran with an alkyllithium, optionally n-BuLi, to form (2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)lithium; (b) optionally adding magnesium chloride or magnesium bromide to the reaction product of (a); (c) reacting the product of (a) or (b) with a compound of formula (Ia):

[0031] [ka] forming a compound of formula (Ib) or a salt thereof;

[0032] [ka] (d) reacting a compound of formula (Ib) or a salt thereof with hydrazine, optionally in the presence of an acid such as acetic acid, to form a compound of formula (Ic) or a salt thereof; (e) isolating the compound of formula (Ic) or a salt thereof, The process is described in formula (Ia), formula (Ib) and formula (Ic) where X=CH or N.

[0033] This specification also describes, in part, a process for the preparation of 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof. (a) reacting 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran with an alkyllithium, optionally n-BuLi, to form (2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)lithium; (b) optionally adding magnesium chloride or magnesium bromide to the reaction product of (a); (c) reacting the product of (a) or (b) with phthalic anhydride to form 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof; (d) reacting 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof with aqueous acid to form 2-[2-hydroxy-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof; (e) reacting 2-[2-hydroxy-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof with hydrazine to form 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof; (f) isolating 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof.

[0034] This specification also describes, in part, a process for the preparation of 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof, comprising: (a) reacting 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof with a chlorinating agent such as POCl to form 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof; (b) isolating 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof.

[0035] This specification also describes, in part, a process for the preparation of Compound 1 or a salt thereof, comprising: (a) reacting 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran with an alkyllithium, optionally n-BuLi, to form (2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)lithium; (b) optionally adding magnesium chloride or magnesium bromide to the reaction product of (a); (c) reacting the product of (a) or (b) with phthalic anhydride to form 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof; (d) reacting 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof with aqueous acid to form 2-[2-hydroxy-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof; (e) reacting 2-[2-hydroxy-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof with hydrazine to form 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof; (f) reacting 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof with a chlorinating agent such as POCl to form 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof; (g) reacting 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof with (1) (2S)-3-amino-1,2-propanediol or a salt thereof to form Compound 1 or a salt thereof, or (2) diol-protected (2S)-3-amino-1,2-propanediol or a salt thereof, followed by deprotection of the resulting product to form Compound 1 or a salt thereof; (h) isolating Compound 1 or a salt thereof.

[0036] This specification also describes, in part, a process for the preparation of Compound 1 or a salt thereof, comprising: (a) reacting 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran with an alkyllithium, optionally n-BuLi, to form (2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)lithium; (b) optionally adding magnesium chloride or magnesium bromide to the reaction product of (a); (c) reacting the product of (a) or (b) with tert-butyl methyl phthalate to form tert-butyl 2-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)benzoyl)benzoate; (d) reacting tert-butyl 2-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)benzoyl)benzoate with hydrazine, optionally in the presence of an acid such as acetic acid, to form 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof; (e) reacting 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof with diol-protected (2S)-3-amino-1,2-propanediol or a salt thereof in the presence of a coupling reagent, followed by removal of the THP and diol-protecting groups from the resulting product by reaction with an acid to form compound 1 or a salt thereof; (f) isolating Compound 1 or a salt thereof.

[0037] This specification also describes, in part, a process for the preparation of Compound 2 or a salt thereof, comprising: (a) reacting 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran with an alkyllithium, optionally n-BuLi, to form (2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)lithium; (b) optionally adding magnesium chloride or magnesium bromide to the reaction product of (a); (c) reacting the product of (a) or (b) with 3-(tert-butyl) 4-methylpyridine-3,4-dicarboxylate to form tert-butyl 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)benzoyl)nicotinate or a salt thereof; (d) reacting tert-butyl 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)benzoyl)nicotinate or a salt thereof with hydrazine, optionally in the presence of an acid such as acetic acid, to form 1-[2-tetrahydropyran-2-yloxy-4-(trifluoromethyl)phenyl]-3H-pyrido[3,4-d]pyridazin-4-one or a salt thereof; (e) reacting 1-[2-tetrahydropyran-2-yloxy-4-(trifluoromethyl)phenyl]-3H-pyrido[3,4-d]pyridazin-4-one with (1R,2R)-2-aminocyclohexan-1-ol or a salt thereof in the presence of a coupling reagent to form (1R,2R)-2-((1-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)pyrido[3,4-d]pyridazin-4-yl)amino)cyclohexan-1-ol or a salt thereof; (f) removing the THP protecting group from (1R,2R)-2-((1-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)pyrido[3,4-d]pyridazin-4-yl)amino)cyclohexan-1-ol or a salt thereof by reaction with an acid to form compound 2 or a salt thereof; (g) isolating compound 2 or a salt thereof.

[0038] This specification also describes, in part:

[0039] [ka] or a salt thereof.

[0040] This specification also describes, in part:

[0041] [ka] or a salt thereof.

[0042] This specification also describes, in part:

[0043] [ka] or a salt thereof.

[0044] This specification also describes, in part:

[0045] [ka] or a salt thereof.

[0046] This specification also describes, in part:

[0047] [ka] or a salt thereof, wherein X is CH or N.

[0048] This specification also describes, in part:

[0049] [ka] or a salt thereof, wherein X is CH or N.

[0050] This specification also describes, in part:

[0051] [ka] or a salt thereof, wherein X is CH or N.

[0052] This specification also describes, in part:

[0053] [ka] or a salt thereof.

[0054] This specification also describes, in part:

[0055] [ka] or a salt thereof.

[0056] This specification also describes, in part:

[0057] [ka] or a salt thereof.

[0058] This specification also describes, in part:

[0059] [ka] or a salt thereof.

[0060] Further aspects of the present disclosure will become apparent to those skilled in the art upon reading this specification. [Brief explanation of the drawings]

[0061] [Figure 1] 1 is an X-ray powder diffraction pattern of Compound 1 Form A. [Figure 2] 1 shows the TG / DTA traces from the analysis of Compound 1 Form A. DETAILED DESCRIPTION OF THE INVENTION

[0062] As used herein, "Compound 1" is a compound having the chemical name (2S)-3-[[4-[2-hydroxy-4-(trifluoromethyl)phenyl]phthalazin-1-yl]amino]propane-1,2-diol and the following chemical structure:

[0063] [ka]

[0064] As used herein, "Compound 2" is a compound having the chemical name 2-(4-(((1R,2R)-2-hydroxycyclohexyl)amino)pyrido[3,4-d]pyridazin-1-yl)-5-(trifluoromethyl)phenol and the following chemical structure:

[0065] [ka]

[0066] As described in the Examples herein, Compound 1 and Compound 2 are inhibitors of the NLRP3 inflammasome and may therefore be useful in treating disease states in which inhibition of the NLRP3 inflammasome is beneficial.

[0067] Terms not specifically defined herein should be understood to have the meaning that would be given to them by one of ordinary skill in the art in light of this disclosure and the context.

[0068] "About" can generally mean an acceptable degree of error for the quantity measured, given the nature or precision of the measurement. Exemplary degrees of error are within a percent (%), typically within 10%, and more typically within 5% of a given value or range of values.

[0069] Embodiments provided herein as "comprising" one or more features may also be considered to disclose corresponding embodiments "consisting of" such features.

[0070] Concentrations, amounts, volumes, percentages, and other numerical values ​​may be presented herein in a range format, with it being understood that such range format is used merely for convenience and brevity and should be interpreted flexibly to include not only the numerical values ​​expressly stated as the limits of the range, but also all individual numerical values ​​or subranges subsumed within that range, as if each numerical value and subrange were expressly written.

[0071] The chemical names of the compounds described herein were generated using ChemDraw® Professional version 19.0.0.22 from PerkinElmer®. Those skilled in the art will understand that different chemical naming software may generate different chemical names for a particular compound. When a compound described herein is shown in the form of a chemical name and as a formula, the formula shall control in the event of any discrepancy.

[0072] The compounds and salts described herein can exist in solvated and unsolvated forms. For example, a solvated form can be a hydrated form, such as a hemi-hydrate, mono-hydrate, di-hydrate, tri-hydrate, or an alternative amount thereof. All such solvated and unsolvated forms of the compounds described herein are encompassed herein.

[0073] The atoms of the compounds and salts described herein may exist as their isotopes. All compounds described herein in which an atom is replaced by one or more of its isotopes (e.g., where one or more carbon atoms are 11 C or 13 C carbon isotope or one or more hydrogen atoms 2 H or 3 Compounds described herein in which the H isotope is included herein.

[0074] The compounds described herein may exist in one or more geometric, optical, enantiomeric, and diastereomeric forms, including, but not limited to, cis- and trans-forms, E- and Z-forms, and R-, S-, and meso-forms. Unless otherwise specified, reference to a particular compound encompasses all such isomeric forms, including racemates and other mixtures. If desired, such isomers can be separated from their mixtures by the application or adaptation of known methods (e.g., chromatographic and recrystallization techniques). If desired, such isomers can be prepared by the application or adaptation of known methods.

[0075] The compounds described herein may contain one or more chiral centers. Unless a structure or chemical name herein indicates chirality, the structure or name is intended to encompass any single stereoisomer corresponding to that structure or name, as well as any mixture of stereoisomers (e.g., racemates). The structures herein are represented by solid and dashed wedges (i.e.,

[0076] [ka] When including bonds depicted as ), the solid and dashed wedges are intended to indicate the absolute configuration of that chiral center.

[0077] It is well known in the art how such optically active forms can be separated. For example, a single stereoisomer can be obtained by isolation from a mixture of isomers (e.g., a racemate) using, for example, chiral chromatographic separation. In other embodiments, a single stereoisomer can be obtained, for example, by direct synthesis from chiral starting materials.

[0078] According to one embodiment, the compounds described herein are provided as a single enantiomer in an enantiomeric excess (% ee) of 95% or more, 98% or more, or 99% or more. Advantageously, the single enantiomer is present in an enantiomeric excess of 99% or more.

[0079] According to one embodiment, the compounds described herein are provided as single enantiomers in enantiomeric excess (% ee) in the range of 95-100%.

[0080] The compounds described herein may exist in one or more tautomeric forms, including, but not limited to, keto and enol forms. A reference to a particular compound includes all tautomeric forms, including mixtures thereof. Thus, a structure shown herein as one tautomer is intended to include other tautomers as well.

[0081] Crystalline morphology In formulating a drug composition, it is important that the drug substance be in a form that can be conveniently handled and processed, not only from the standpoint of obtaining a commercially viable manufacturing process, but also from the standpoint of the subsequent manufacture of pharmaceutical formulations (e.g., oral dosage forms such as tablets) containing the active compound.

[0082] The different physical properties of crystalline forms with respect to each other and with respect to the amorphous state can significantly affect the chemical and pharmaceutical processing of the compounds, especially when the compounds are prepared or used on an industrial scale.

[0083] Furthermore, it is important in the manufacture of oral drug compositions that they provide a reliable and reproducible plasma concentration profile of the drug after administration to a patient. Patient-to-patient variability in the absorption profile of a drug in the stomach, intestine, or bloodstream can affect the safety and efficacy of the drug.

[0084] The chemical stability, solid-state stability, and "shelf life" of the active ingredient are also very important factors: drug substances and compositions containing them should be able to be effectively stored for a significant period of time without exhibiting significant changes in the physicochemical properties of the active ingredient (e.g., its chemical composition, density, hygroscopicity, and solubility).

[0085] Additionally, it is important to be able to provide drugs in as chemically pure a form as possible.

[0086] Amorphous materials can present problems in this regard: for example, such materials are typically difficult to handle and formulate, provide unreliable solubility, and are often found to be unstable and chemically impure.

[0087] Those skilled in the art will appreciate that the above problems can be solved if the drug can be readily obtained in a stable crystalline form.

[0088] Therefore, in the manufacture of commercially viable and pharmaceutically acceptable drug compositions, it is important to provide the drug in a crystalline and stable form whenever possible.

[0089] However, it should be noted that this goal is not always achievable. In fact, it is typically not possible to predict from molecular structure alone what the crystallization behavior of a compound, either by itself or in salt form, will be. This can only be determined experimentally.

[0090] In one embodiment, Compound 1 can be prepared in a crystalline form. This crystalline form can be characterized as a particular polymorphic form. When an embodiment is described as relating to a crystalline form, the degree of crystallinity can be greater than about 60%, optionally greater than about 80%, greater than about 90%, or greater than about 95%. In one embodiment, the degree of crystallinity is greater than about 98%.

[0091] Certain crystalline forms described herein provide X-ray powder diffraction patterns substantially similar to those shown in the figures, with various 2-theta values ​​as described herein. It will be understood that the 2-theta values ​​of X-ray powder diffraction patterns may vary slightly from machine to machine or sample to sample, and therefore the quoted values ​​should not be construed as absolute.

[0092] It is known that an X-ray powder diffraction pattern may have one or more measurement errors depending on the measurement conditions (such as the device or machine used). In particular, it is generally known that the intensities in an X-ray powder diffraction pattern may vary depending on the measurement conditions. Therefore, it should be understood that the crystalline forms described herein are not limited to crystals that provide X-ray powder diffraction patterns identical to those shown in the figures, and that any crystals that provide X-ray powder diffraction patterns substantially identical to those shown in the figures fall within the scope of the embodiments described herein. Those skilled in the art of X-ray powder diffraction can judge the substantial identity of X-ray powder diffraction patterns.

[0093] Those skilled in the art of X-ray powder diffraction will recognize that the relative intensities of peaks can be affected, for example, by particles greater than 30 μm in size and non-unitary aspect ratios, which can affect the analysis of the sample. Those skilled in the art will also recognize that the position of reflections can be affected by the exact height at which the sample is positioned within the diffractometer and the zero calibration of the diffractometer. The surface flatness of the sample can also have a small effect. Therefore, the diffraction pattern data presented should not be interpreted as absolute values. (Jenkins, R & Snyder, RL, "Introduction to X-Ray Powder Diffractometry," John Wiley & Sons 1996; Bunn, CW (1948), "Chemical Crystallography," Clarendon Press, London; Klug, HP & Alexander, LE (1974), X-Ray Diffraction Procedures). Peak intensities are described herein as vs (very strong), s (strong), m (medium), and w (weak), corresponding to relative intensity percentages (based on the most intense peak) of 25-100%, 10-25%, 3-10%, and 1-3%, respectively. Relative intensities are obtained from diffractograms measured with a fixed slit.

[0094] Generally, the measurement error in diffraction angles in X-ray powder diffractograms is about 5% or less, particularly plus or minus 0.2° 2-theta, and such degree of measurement error should be taken into account when considering the X-ray powder diffraction patterns in the figures herein and when reading the data presented herein. Furthermore, it should be understood that intensities may vary depending on experimental conditions and sample preparation (preferred orientation).

[0095] Useful crystalline polymorphic forms of Compound 1 were produced using the conditions described in Example 1. In all of the embodiments of the crystalline forms listed herein, the peaks in the X-ray diffraction pattern are Cu K α It is measured using radiation.

[0096] Thus, in one embodiment, Compound 1 is provided in crystalline form.

[0097] In one embodiment, a crystalline form of Compound 1 is provided, wherein the crystalline form is Form A.

[0098] Thus, in one embodiment, there is provided polymorphic Form A of Compound 1. This polymorphic form is Cu K α It may be characterized by providing at least one of the following 2θ values ​​measured using radioactivity: 13.0, 19.3, 19.8, 22.7, and 24.1 degrees.

[0099] Polymorphic Form A of Compound 1 is characterized by providing an X-ray powder diffraction pattern substantially as shown in FIG.

[0100] In one embodiment, polymorphic Form A of Compound 1 is provided, having an X-ray powder diffraction pattern with at least one particular peak at about 2-theta=19.3°.

[0101] In one embodiment, polymorphic Form A of Compound 1 is provided, having an X-ray powder diffraction pattern with at least one particular peak at about 2-theta=13.0°.

[0102] In one embodiment, polymorphic Form A of Compound 1 is provided, having an X-ray powder diffraction pattern with at least two specific peaks at about 2-theta=13.0 and 19.3°.

[0103] In one embodiment, polymorphic Form A of Compound 1 is provided, having an X-ray powder diffraction pattern with at least five specific peaks at about 2-theta = 13.0, 19.3, 19.8, 22.7, and 24.1 degrees.

[0104] In one embodiment, polymorphic Form A of Compound 1 is provided, having an X-ray powder diffraction pattern with particular peaks at about 2-theta = 10.7, 11.6, 13.0, 16.1, 19.3, 19.8, 22.7, 24.1, 26.3, 29.5, and 30.9 degrees.

[0105] In one embodiment, polymorphic Form A of Compound 1 is provided, having an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in FIG.

[0106] In one embodiment, polymorphic Form A of Compound 1 is provided, having an X-ray powder diffraction pattern with at least one particular peak at 2-theta=19.3°±0.2° 2-theta.

[0107] In one embodiment, polymorphic Form A of Compound 1 is provided, having an X-ray powder diffraction pattern with at least one particular peak at 2-theta=13.0°±0.2° 2-theta.

[0108] In one embodiment, polymorphic Form A of Compound 1 is provided, having an X-ray powder diffraction pattern with at least two specific peaks at 2-theta=13.0 and 19.3°, each of which is ±0.2° 2-theta.

[0109] In one embodiment, polymorphic Form A of Compound 1 is provided, having an X-ray powder diffraction pattern with at least five specific peaks at 2-theta=13.0, 19.3, 19.8, 22.7, and 24.1 degrees, each of which is ±0.2 degrees 2-theta.

[0110] In one embodiment, polymorphic Form A of Compound 1 is provided, having an X-ray powder diffraction pattern with particular peaks at 2-theta=10.7, 11.6, 13.0, 16.1, 19.3, 19.8, 22.7, 24.1, 26.3, 29.5, and 30.9 degrees, each of which is ±0.2 degrees 2-theta.

[0111] In one embodiment, crystalline Form A of Compound 1 is unsolvated.

[0112] Useful crystalline polymorphic forms of the hydrochloride salt of Compound 2 have been produced using the conditions described in Example 3. Thus, in one embodiment, a crystalline form of the hydrochloride salt of Compound 2 is provided.

[0113] Pharmaceutical Composition Because Compound 1 is an inhibitor of the NLRP3 inflammasome, pharmaceutical compositions comprising crystalline Compound 1, particularly polymorphic Form A of Compound 1 (described herein), and a pharmaceutically acceptable excipient are expected to be useful for treating disease states in which inhibition of the NLRP3 inflammasome is beneficial.

[0114] Thus, in one embodiment, there is provided a pharmaceutical composition comprising Compound 1 in a crystalline form and a pharmaceutically acceptable excipient.

[0115] In one embodiment, a pharmaceutical composition is provided comprising Compound 1 in a crystalline form and a pharmaceutically acceptable excipient, wherein the crystalline form is Form A.

[0116] In one embodiment, a pharmaceutical composition is provided comprising polymorphic Form A of Compound 1 and a pharmaceutically acceptable excipient.

[0117] In one embodiment, a pharmaceutical composition is provided comprising polymorphic Form A of Compound 1, having an X-ray powder diffraction pattern with at least one particular peak at about 2-theta=19.3°, and a pharmaceutically acceptable excipient.

[0118] In one embodiment, a pharmaceutical composition is provided comprising polymorphic Form A of Compound 1, having an X-ray powder diffraction pattern with at least one particular peak at about 2-theta=13.0°, and a pharmaceutically acceptable excipient.

[0119] In one embodiment, a pharmaceutical composition is provided comprising polymorphic Form A of Compound 1, having an X-ray powder diffraction pattern with at least two specific peaks at about 2-theta=13.0 and 19.3°, and a pharmaceutically acceptable excipient.

[0120] In one embodiment, a pharmaceutical composition is provided comprising polymorphic Form A of Compound 1 having an X-ray powder diffraction pattern with at least five specific peaks at about 2-theta=13.0, 19.3, 19.8, 22.7, and 24.1°, and a pharmaceutically acceptable excipient.

[0121] In one embodiment, there is provided a pharmaceutical composition comprising polymorphic Form A of Compound 1 having an X-ray powder diffraction pattern with particular peaks at about 2-theta = 10.7, 11.6, 13.0, 16.1, 19.3, 19.8, 22.7, 24.1, 26.3, 29.5, and 30.9 degrees, and a pharmaceutically acceptable excipient.

[0122] In one embodiment, there is provided a pharmaceutical composition comprising polymorphic Form A of Compound 1, having an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in FIG. 1, and a pharmaceutically acceptable excipient.

[0123] In one embodiment, there is provided a pharmaceutical composition comprising polymorphic Form A of Compound 1 having an X-ray powder diffraction pattern with at least one particular peak at 2-theta=19.3°±0.2° 2-theta, and a pharmaceutically acceptable excipient.

[0124] In one embodiment, there is provided a pharmaceutical composition comprising polymorphic Form A of Compound 1 having an X-ray powder diffraction pattern with at least one particular peak at 2-theta=13.0°±0.2° 2-theta, and a pharmaceutically acceptable excipient.

[0125] In one embodiment, there is provided a pharmaceutical composition comprising polymorphic Form A of Compound 1 having an X-ray powder diffraction pattern with at least two specific peaks at 2-theta=13.0 and 19.3°, each of which is ±0.2° 2-theta, and a pharmaceutically acceptable excipient.

[0126] In one embodiment, there is provided a pharmaceutical composition comprising polymorphic Form A of Compound 1 having an X-ray powder diffraction pattern with at least five specific peaks at 2-theta=13.0, 19.3, 19.8, 22.7, and 24.1 degrees, each of which is ±0.2 degrees 2-theta, and a pharmaceutically acceptable excipient.

[0127] In one embodiment, there is provided a pharmaceutical composition comprising polymorphic Form A of Compound 1 having an X-ray powder diffraction pattern with particular peaks at 2-theta=10.7, 11.6, 13.0, 16.1, 19.3, 19.8, 22.7, 24.1, 26.3, 29.5, and 30.9 degrees, each of which is ±0.2 degrees 2-theta, and a pharmaceutically acceptable excipient.

[0128] Because Compound 2 is an inhibitor of the NLRP3 inflammasome, pharmaceutical compositions comprising crystalline Compound 2, particularly the hydrochloride salt of Compound 2, and a pharmaceutically acceptable excipient are expected to be useful for treating disease states in which inhibition of the NLRP3 inflammasome is beneficial.

[0129] Thus, in one embodiment, there is provided a pharmaceutical composition comprising the hydrochloride salt of Compound 2 in crystalline form and a pharmaceutically acceptable excipient.

[0130] The pharmaceutical compositions described herein may contain one or more pharmaceutically acceptable excipients. The excipients selected for inclusion in a particular composition depend on factors such as the mode of administration and the form of the composition provided. Suitable pharmaceutically acceptable excipients are well known to those skilled in the art and are described, for example, in "Handbook of Pharmaceutical Excipients," Sixth Edition, Pharmaceutical Press, edited by Rowe, Ray C; Sheskey, Paul J; and Quinn, Marian. Pharmaceutically acceptable excipients can function, for example, as adjuvants, diluents, carriers, stabilizers, flavoring agents, coloring agents, fillers, binders, disintegrants, lubricants, glidants, thickeners, and coating agents. As those skilled in the art will understand, a particular pharmaceutically acceptable excipient can perform more than one function, and can perform different functions, depending on how much of the excipient is present in the composition and what other excipients are present in the composition.

[0131] In one embodiment, the amount of pharmaceutically acceptable excipient in the pharmaceutical composition is 1 mg or more. In a further embodiment, the amount of pharmaceutically acceptable excipient in the pharmaceutical composition is 10 mg or more. In a further embodiment, the amount of pharmaceutically acceptable excipient in the pharmaceutical composition is 100 mg or more.

[0132] The pharmaceutical compositions may be in a form suitable for oral use (e.g., as tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, or dispersible powders or granules), topical use (e.g., as creams, ointments, or aqueous or oily suspensions), administration by inhalation (e.g., as a finely divided powder), administration by insufflation (e.g., as a finely divided powder), or as a suppository for rectal administration. The compositions may be obtained by conventional procedures well known in the art. Compositions intended for oral use may contain additional ingredients, such as one or more colorants, sweeteners, flavoring agents, and / or preservatives.

[0133] The term "pharmaceutically acceptable" is used to specify that an object (e.g., an excipient) is suitable for use in patients. An exemplary list of pharmaceutically acceptable salts can be found in "Handbook of Pharmaceutical Salts: Properties, Selection and Use", P.H. Stahl and C.G. Wermuth, editors, Weinheim / Zurich: Wiley-VCH / VHCA, 2002.

[0134] Therapeutic Uses of Crystalline Forms and Pharmaceutical Compositions As a result of their NLRP3 inflammasome inhibitory activity, the crystalline forms and pharmaceutical compositions described herein comprising Compound 1 or Compound 2 are expected to be useful in therapy.

[0135] The term "therapy" is intended to have its ordinary meaning of addressing a disease or condition to completely or partially alleviate one, some, or all of the symptoms of the disease or condition, or to correct or compensate for an underlying pathology. The term "therapy" also includes "prophylaxis," unless specifically indicated to the contrary. The terms "therapeutic" and "therapeutically" are to be construed in corresponding ways.

[0136] The term "prevent" is intended to have its ordinary meaning and includes primary prevention, to prevent the onset of a disease or condition, and secondary prevention, where a disease or condition has already occurred and the patient is temporarily or permanently protected from exacerbation or worsening of the disease or condition, or from the onset of new symptoms associated with the disease or condition.

[0137] The term "treatment" is used synonymously with "therapy." Similarly, the term "treating" can be considered as "applying therapy," as "therapy" is defined herein.

[0138] In one embodiment, a method is described for treating a disease or condition involving NLRP3 inflammasome activity in a subject in need of such treatment, the method comprising administering to the subject a therapeutically effective amount of a crystalline form or pharmaceutical composition described herein.

[0139] In one embodiment, the present invention relates to kidney diseases such as acute kidney injury, chronic kidney disease, and diabetic kidney disease; cardiovascular diseases such as coronary atherosclerotic heart disease, cardiomyopathy, myocardial infarction, cardiac hypertrophy, and ischemia-reperfusion injury; liver diseases such as non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, alcoholic steatohepatitis, chronic hepatitis C virus infection, and paracetamol-induced liver injury; inflammatory diseases such as autoinflammatory disorders, cryopyrin-associated periodic syndrome, familial cold autoinflammatory syndrome (FCAS), Muckle-Wells syndrome (MWS), chronic infantile neurological, cutaneous, and articular (CINCA) syndrome, and neonatal-onset multisystem inflammatory disease (NOMID); rheumatoid arthritis (rheumatoid arthritis), and rheumatoid arthritis (rheumatoid arthritis). inflammatory skin diseases, such as acne, hidradenitis suppurativa, and hidradenitis suppurativa; inflammatory bowel diseases, such as ulcerative colitis (UC) and Crohn's disease; autoimmune diseases, such as gout, pseudogout, rheumatoid arthritis (RA), multiple sclerosis (MS), Addison's disease, celiac disease, systemic lupus erythematosus (SLE), and vitiligo; and respiratory diseases, such as chronic lung disease, idiopathic pulmonary fibrosis (IPF), chronic obstructive pulmonary disease (COPD), and asthma, in a subject in need of such treatment, the method comprising administering to the subject a therapeutically effective amount of a crystalline form or pharmaceutical composition described herein.

[0140] In one embodiment, the present invention relates to a method for treating acute kidney injury, chronic kidney disease, diabetic kidney disease, coronary atherosclerotic heart disease, cardiomyopathy, myocardial infarction, cardiac hypertrophy, ischemia-reperfusion injury, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, alcoholic steatohepatitis, chronic hepatitis C virus infection, paracetamol-induced liver injury, autoinflammatory disorders, cryopyrin-associated periodic syndrome, familial cold autoinflammatory syndrome (FCAS), Muckle-Wells syndrome (MWS), chronic infantile neurological, cutaneous, and articular (CINCA) syndrome, neonatal-onset multisystem inflammatory disease (NOMID), Methods are provided for treating a disease or condition selected from acne vulgaris, hidradenitis suppurativa, ulcerative colitis (UC), Crohn's disease, gout, pseudogout, rheumatoid arthritis (RA), multiple sclerosis (MS), Addison's disease, celiac disease, systemic lupus erythematosus (SLE), vitiligo, chronic lung disease, idiopathic pulmonary fibrosis (IPF), chronic obstructive pulmonary disease (COPD), and asthma in a subject in need of such treatment, comprising administering to the subject a therapeutically effective amount of a crystalline form or pharmaceutical composition described herein.

[0141] In one embodiment, there is provided a crystalline form or pharmaceutical composition described herein for use in therapy.

[0142] In one embodiment, there is provided a crystalline form or pharmaceutical composition described herein for use in a method for treating a disease or condition described herein.

[0143] In one embodiment, there is provided the use of a crystalline form or pharmaceutical composition described herein in the manufacture of a medicament for a disease or condition described herein.

[0144] The term "therapeutically effective amount" refers to an amount of a crystalline form or pharmaceutical composition described in any of the embodiments herein that is effective to provide "therapy" in a subject or to "treat" a disease or condition in a subject. A therapeutically effective amount may cause any of the observable or measurable changes in a subject, as described above in the definitions of "therapy," "treatment," and "prevention." As will be recognized by those skilled in the art, an effective amount may vary depending on the route of administration, the use of excipients, and the use in combination with other agents. For example, when a combination therapy is used, the amount of a crystalline form or pharmaceutical composition described herein and the amount of another pharmaceutically active agent, when combined, are jointly effective to treat the target disorder or condition in a subject. In this context, the combined amount is a "therapeutically effective amount" if, when combined, it is sufficient to reduce the symptoms of a disease or condition that responds to inhibition of the NLRP3 inflammasome, as described above. Typically, such an amount can be determined by one skilled in the art.

[0145] A "subject" includes, for example, mammals, such as humans.

[0146] Process for the preparation of Compound 1 and Compound 2 The synthetic routes to compounds 1 and 2 from our previous work used Li-halogen exchange of benzyl-, PMB-, or methoxy-protected bromophenolic compounds followed by reaction of the lithiated compound with an ester (see, e.g., Scheme 1 and Comparative Example 1). Similar routes have also been used in the synthesis of other known inhibitors of the NLRP3 inflammasome (see, e.g., U.S. Pat. No. 11,319,319). Such lithium-halogen exchange requires cryogenic temperatures due to the nature of the reaction and the instability of the resulting lithiated compounds, and such cryogenic conditions are undesirable for larger-scale syntheses. Furthermore, the use of brominated starting materials is undesirable from a commercial perspective, given the bromination step required to prepare such materials.

[0147] [ka]

[0148] Alternatively, compounds 1 and 2 can be prepared by Suzuki-Miyaura coupling with an appropriate boronic acid or boronic ester (see, for example, the synthesis described in Scheme 2 and Example 1). Similar routes have also been used in the synthesis of other known inhibitors of the NLRP3 inflammasome (see, for example, WO 2022 / 135567). However, such synthetic procedures require an additional step to synthesize a boronic acid / boronic ester intermediate, and the starting materials for the synthesis of boronic acids / boronic esters tend to be halogenated, which is undesirable given the halogenation step required to prepare such starting materials. Furthermore, the use of palladium catalysts in pharmaceutical synthesis is undesirable because such residual palladium may contaminate the reaction product, and the level of residual palladium in pharmaceuticals must be controlled.

[0149] [ka]

[0150] Therefore, improved processes for the synthesis of inhibitors of the NLRP3 inflammasome, such as compounds 1 and 2, that are more amenable to large-scale production and offer improved efficiency are desirable.

[0151] It has been discovered that the use of tetrahydropyran (THP)-protected phenols allows for efficient ortho-lithiation at non-cryogenic temperatures, thus avoiding the use of bromine-containing starting materials. Reaction of the lithiated intermediate with tert-butyl methyl phthalate or 3-(tert-butyl)4-methylpyridine-3,4-dicarboxylate (formula (Ia)) provides an intermediate (formula (Ib)), which can be cyclized with hydrazine to form an intermediate (formula (Ic)) useful in the synthesis of compounds 1 and 2. (See Scheme 3.)

[0152] [ka]

[0153] Thus, in one embodiment, there is provided a process for the preparation of a compound of formula (Ic) or a salt thereof, comprising: (a) reacting 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran with an alkyllithium, optionally n-BuLi, to form (2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)lithium; (b) optionally adding magnesium chloride or magnesium bromide to the reaction product of (a); (c) reacting the product of (a) or (b) with a compound of formula (Ia) to form a compound of formula (Ib) or a salt thereof; (d) reacting a compound of formula (Ib) or a salt thereof with hydrazine, optionally in the presence of an acid such as acetic acid, to form a compound of formula (Ic) or a salt thereof; (e) isolating the compound of formula (Ic) or a salt thereof, In the formula (Ia), (Ib) and (Ic), X=CH or N is provided.

[0154] In one embodiment, X=CH. In one embodiment, X=N.

[0155] Steps (a)-(c): Suitable solvents for steps (a)-(c) include ethers, such as THF. The alkyllithium may be n-BuLi or t-BuLi, optionally n-BuLi. Steps (a)-(c) may be carried out at a temperature of -80°C to 20°C, e.g., -80°C to 10°C, -80°C to 0°C, -40°C to 10°C, and -20°C to 5°C. Optionally, step (c) is carried out at a temperature of -80°C to -60°C, e.g., -80°C to -70°C, or about -78°C. Optionally, steps (a)-(c) may be carried out in the presence of a coordinating ligand, such as N,N,N',N'-tetramethyl-1,2-ethanediamine. In one embodiment, magnesium chloride is added in step (b).

[0156] Step (d): In one embodiment, the reaction mixture from step (c) is used directly in step (d) without further purification. The hydrazine used in step (d) may be in the form of hydrazine hydrate. Suitable solvents for step (d) include THF and EtOH. Step (d) may be carried out at a temperature of -40°C to 40°C, for example, -30°C to 30°C, -30°C to 10°C, and -30°C to 0°C. The acid used in step (d) may be a carboxylic acid, such as acetic acid.

[0157] Step (e): In one embodiment, the compound of formula (Ic) or a salt thereof may be isolated by precipitation and filtration.

[0158] Surprisingly, it has been found that the cyclization step with hydrazine to form compounds of formula (Ic) (see Scheme 3) can be carried out without protection of the phenol (such as with a THP group) and using a carboxylic acid group instead of the tert-butyl ester present in compounds of formula (Ib), as shown in Scheme 4, thus providing 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof (which is used in the synthesis of compound 1) in a more efficient manner.

[0159] [ka]

[0160] Thus, in one embodiment, there is provided a process for the preparation of 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof, comprising: (a) reacting 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran with an alkyllithium, optionally n-BuLi, to form (2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)lithium; (b) optionally adding magnesium chloride or magnesium bromide to the reaction product of (a); (c) reacting the product of (a) or (b) with phthalic anhydride to form 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof; (d) reacting 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof with aqueous acid to form 2-[2-hydroxy-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof; (e) reacting 2-[2-hydroxy-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof with hydrazine to form 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof; (f) isolating the 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof.

[0161] Steps (a)-(c): Suitable solvents for steps (a)-(c) include ethers, such as THF. The alkyllithium may be n-BuLi or t-BuLi, optionally n-BuLi. Steps (a)-(c) may be carried out at a temperature of -80°C to 20°C, e.g., -80°C to 10°C, -80°C to 0°C, -40°C to 10°C, and -20°C to 5°C. Optionally, step (c) is carried out at a temperature of -80°C to -60°C, e.g., -80°C to -70°C, or about -78°C. Optionally, steps (a)-(c) may be carried out in the presence of a coordinating ligand, such as N,N,N',N'-tetramethyl-1,2-ethanediamine. In one embodiment, magnesium chloride is added in step (b).

[0162] Step (d): A suitable solvent for step (d) is THF. Optionally, the aqueous acid is hydrochloric acid. Step (d) can be carried out at a temperature of 0°C to 70°C, for example, 10°C to 60°C, 20°C to 50°C, and 30°C to 40°C.

[0163] Step (e): The hydrazine used in step (e) may be in the form of hydrazine hydrate. Suitable solvents for step (e) include THF. Step (e) may be carried out at a temperature of 0°C to 80°C, for example, 20°C to 70°C, and 30°C to 60°C.

[0164] Step (f): In one embodiment, the compound of formula (Ic) or a salt thereof can be isolated by precipitation, such as precipitation from a mixture of THF and heptane, optionally 2:1 THF / heptane, and filtration.

[0165] Alternatively, the order of hydrazine cyclization and removal of the THP protecting group can be reversed, as shown in Scheme 5.

[0166] [ka]

[0167] Thus, in one embodiment, there is provided a process for the preparation of 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof, comprising: (a) reacting 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran with an alkyllithium, optionally n-BuLi, to form (2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)lithium; (b) optionally adding magnesium chloride or magnesium bromide to the reaction product of (a); (c) reacting the product of (a) or (b) with phthalic anhydride to form 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof; (d) reacting 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof with hydrazine to form 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof; (e) reacting 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof with aqueous acid to form 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof; (f) isolating the 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof.

[0168] Furthermore, it was found that a phenol protecting group is not required for the chlorination of 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one with a suitable chlorinating agent such as POCl. This is useful in the commercial synthesis of NLRP3 inhibitors such as compounds 1 and 2, as the THP phenol protecting group can undergo side reactions with chlorinating agents (such as POCl) and result in undesirable alkyl chloride impurities in such commercial processes.

[0169] Accordingly, there is provided a process for the preparation of 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof, comprising the steps of: (a) reacting 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof with a chlorinating agent such as POCl to form 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof; (b) isolating the 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof.

[0170] Step (a): A suitable solvent for step (a) is MeCN. The reaction of step (a) can be carried out at a temperature of 0°C to 80°C, for example, 20°C to 70°C, and 30°C to 60°C.

[0171] In one embodiment, the 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof used in step (a) is prepared by a process described herein.

[0172] Step (b): In one embodiment, 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof is isolated by precipitation, such as precipitation from a mixture of MeCN and water.

[0173] Also provided are processes for the preparation of compounds 1 and 2, or salts thereof, utilizing efficient syntheses of the intermediates described herein.

[0174] Thus, in one embodiment, there is provided a process for the preparation of Compound 1 or a salt thereof, comprising: (a) reacting 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran with an alkyllithium, optionally n-BuLi, to form (2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)lithium; (b) optionally adding magnesium chloride or magnesium bromide to the reaction product of (a); (c) reacting the product of (a) or (b) with phthalic anhydride to form 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof; (d) reacting 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof with aqueous acid to form 2-[2-hydroxy-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof; (e) reacting 2-[2-hydroxy-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof with hydrazine to form 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof; (f) reacting 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof with a chlorinating agent such as POCl to form 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof; (g) reacting 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof with (1) (2S)-3-amino-1,2-propanediol or a salt thereof to form Compound 1 or a salt thereof, or (2) diol-protected (2S)-3-amino-1,2-propanediol or a salt thereof, followed by deprotection of the resulting product to form Compound 1 or a salt thereof; (h) isolating Compound 1 or a salt thereof.

[0175] Steps (a)-(c): Suitable solvents for steps (a)-(c) include ethers, such as THF. The alkyllithium may be n-BuLi or t-BuLi, optionally n-BuLi. Steps (a)-(c) may be carried out at a temperature of -80°C to 20°C, e.g., -80°C to 10°C, -80°C to 0°C, -40°C to 10°C, and -20°C to 5°C. Optionally, step (c) is carried out at a temperature of -80°C to -60°C, e.g., -80°C to -70°C, or about -78°C. Optionally, steps (a)-(c) may be carried out in the presence of a coordinating ligand, such as N,N,N',N'-tetramethyl-1,2-ethanediamine. In one embodiment, magnesium chloride is added in step (b).

[0176] Step (d): A suitable solvent for step (d) is THF. Optionally, the aqueous acid is hydrochloric acid. Step (d) can be carried out at a temperature of 0°C to 70°C, for example, 10°C to 60°C, 20°C to 50°C, and 30°C to 40°C.

[0177] Step (e): The hydrazine used in step (e) may be in the form of hydrazine hydrate. Suitable solvents for step (e) include THF. Step (e) may be carried out at a temperature of 0°C to 80°C, for example, 20°C to 70°C, and 30°C to 60°C.

[0178] Step (f): A suitable solvent for step (f) is MeCN. The reaction of step (f) can be carried out at a temperature of 0°C to 80°C, for example, 20°C to 70°C, and 30°C to 60°C.

[0179] Step (g): Suitable solvents for step (g) are NMP or DMF, optionally a polar aprotic solvent such as NMP. The reaction of step (g) can be carried out at a temperature of 20°C to 150°C, for example, 60°C to 140°C, and 100°C to 140°C.

[0180] Diol-protected (2S)-3-amino-1,2-propanediol refers to a compound in which the two alcohol groups of (2S)-3-amino-1,2-propanediol are protected using known protecting groups suitable for 1,2-diols. Such protection can be provided by forming a five-membered ketal, such as a five-membered acetal, from the 1,2-diol. Thus, in one embodiment, the diol-protected (2S)-3-amino-1,2-propanediol is (S)-(2,2-dimethyl-1,3-dioxolan-4-yl)methanamine.

[0181] Removal of the diol protecting group can be carried out according to known methods. In one embodiment, the diol protecting group is removed using aqueous or alcoholic acid. In one embodiment, the diol protecting group is removed using hydrochloric acid in an alcohol solvent such as isopropyl alcohol.

[0182] Step (h): Isolation of Compound 1 or a salt thereof can be carried out by adding water to a solution of Compound 1 or a salt thereof to precipitate Compound 1 or a salt thereof. For example, water is added to a solution of Compound 1 or a salt thereof in NMP to precipitate Compound 1 or a salt thereof.

[0183] Alternatively, the order of hydrazine cyclization and removal of the THP protecting group may be reversed. Thus, in one embodiment, there is provided a process for the preparation of compound 1 or a salt thereof, comprising: (a) reacting 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran with an alkyllithium, optionally n-BuLi, to form (2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)lithium; (b) optionally adding magnesium chloride or magnesium bromide to the reaction product of (a); (c) reacting the product of (a) or (b) with phthalic anhydride to form 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof; (d) reacting 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof with hydrazine to form 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof; (e) reacting 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof with aqueous acid to form 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof; (f) reacting 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof with a chlorinating agent such as POCl to form 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof; (g) reacting 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof with (1) (2S)-3-amino-1,2-propanediol or a salt thereof to form Compound 1 or a salt thereof, or (2) diol-protected (2S)-3-amino-1,2-propanediol or a salt thereof, followed by deprotection of the resulting product to form Compound 1 or a salt thereof; (h) isolating Compound 1 or a salt thereof.

[0184] In one embodiment, there is provided a process for the preparation of Compound 1 or a salt thereof, comprising: (a) reacting 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran with an alkyllithium, optionally n-BuLi, to form (2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)lithium; (b) optionally adding magnesium chloride or magnesium bromide to the reaction product of (a); (c) reacting the product of (a) or (b) with tert-butyl methyl phthalate to form tert-butyl 2-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)benzoyl)benzoate; (d) reacting tert-butyl 2-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)benzoyl)benzoate with hydrazine, optionally in the presence of an acid such as acetic acid, to form 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof; (e) reacting 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof with diol-protected (2S)-3-amino-1,2-propanediol or a salt thereof in the presence of a coupling reagent, followed by removal of the THP and diol-protecting groups from the resulting product by reaction with an acid to form compound 1 or a salt thereof; (f) isolating Compound 1 or a salt thereof.

[0185] Steps (a)-(c): Suitable solvents for steps (a)-(c) include ethers, such as THF. The alkyllithium may be n-BuLi or t-BuLi, optionally n-BuLi. Steps (a)-(c) may be carried out at a temperature of -80°C to 20°C, e.g., -80°C to 10°C, -80°C to 0°C, -40°C to 10°C, and -20°C to 5°C. Optionally, step (c) is carried out at a temperature of -80°C to -60°C, e.g., -80°C to -70°C, or about -78°C. Optionally, steps (a)-(c) may be carried out in the presence of a coordinating ligand, such as N,N,N',N'-tetramethyl-1,2-ethanediamine. In one embodiment, magnesium chloride is added in step (b).

[0186] Step (d): In one embodiment, the reaction mixture from step (c) is used directly in step (d) without further purification. The hydrazine used in step (d) may be in the form of hydrazine hydrate. Suitable solvents for step (d) include THF and EtOH. Step (d) may be carried out at a temperature of -40°C to 40°C, for example, -30°C to 30°C, -30°C to 10°C, and -30°C to 0°C. The acid used in step (d) may be a carboxylic acid, such as acetic acid.

[0187] Step (e): The coupling reagent is a suitable reagent for activating a carboxyl group for amide synthesis. Such coupling agents are known in the art and include, for example, ((1H-benzo[d][1,2,3]triazol-1-yl)oxy)tri(pyrrolidin-1-yl)phosphonium hexafluorophosphate (V). Optionally, the reaction of step (e) can be carried out in the presence of a base such as DBU. Optionally, the reaction of step (e) can be carried out in the presence of a molecular sieve, for example, a 0.3 nm molecular sieve, to trap water. Suitable solvents for step (e) include polar protic solvents such as MeCN. Step (e) can be carried out at a temperature of 20°C to 70°C, for example, 30°C to 60°C.

[0188] Removal of the THP and diol protecting groups can be carried out according to known methods. In one embodiment, the THP and diol protecting groups are removed using aqueous or alcoholic acid. In one embodiment, the THP and diol protecting groups are removed using hydrochloric acid in an alcoholic solvent such as isopropyl alcohol.

[0189] In one embodiment, there is provided a process for the preparation of Compound 2 or a salt thereof, comprising: (a) reacting 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran with an alkyllithium, optionally n-BuLi, to form (2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)lithium; (b) optionally adding magnesium chloride or magnesium bromide to the reaction product of (a); (c) reacting the product of (a) or (b) with 3-(tert-butyl) 4-methylpyridine-3,4-dicarboxylate to form tert-butyl 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)benzoyl)nicotinate or a salt thereof; (d) reacting tert-butyl 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)benzoyl)nicotinate or a salt thereof with hydrazine, optionally in the presence of an acid such as acetic acid, to form 1-[2-tetrahydropyran-2-yloxy-4-(trifluoromethyl)phenyl]-3H-pyrido[3,4-d]pyridazin-4-one or a salt thereof; (e) reacting 1-[2-tetrahydropyran-2-yloxy-4-(trifluoromethyl)phenyl]-3H-pyrido[3,4-d]pyridazin-4-one with (1R,2R)-2-aminocyclohexan-1-ol or a salt thereof in the presence of a coupling reagent to form (1R,2R)-2-((1-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)pyrido[3,4-d]pyridazin-4-yl)amino)cyclohexan-1-ol or a salt thereof; (f) removing the THP protecting group from (1R,2R)-2-((1-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)pyrido[3,4-d]pyridazin-4-yl)amino)cyclohexan-1-ol or a salt thereof by reaction with an acid to form compound 2 or a salt thereof; (g) isolating compound 2 or a salt thereof.

[0190] Steps (a)-(c): Suitable solvents for steps (a)-(c) include ethers, such as THF. The alkyllithium may be n-BuLi or t-BuLi, optionally n-BuLi. Steps (a)-(c) may be carried out at a temperature of -80°C to 20°C, e.g., -80°C to 10°C, -80°C to 0°C, -40°C to 10°C, and -20°C to 5°C. Optionally, step (c) is carried out at a temperature of -80°C to -60°C, e.g., -80°C to -70°C, or about -78°C. Optionally, steps (a)-(c) may be carried out in the presence of a coordinating ligand, such as N,N,N',N'-tetramethyl-1,2-ethanediamine. In one embodiment, magnesium chloride is added in step (b).

[0191] Step (d): In one embodiment, the reaction mixture from step (c) is used directly in step (d) without further purification. The hydrazine used in step (d) may be in the form of hydrazine hydrate. Suitable solvents for step (d) include THF and EtOH. Step (d) may be carried out at a temperature of -40°C to 40°C, for example, -30°C to 30°C, -30°C to 10°C, and -30°C to 0°C. The acid used in step (d) may be a carboxylic acid, such as acetic acid.

[0192] Step (e): The coupling reagent is a suitable reagent for activating a carboxyl group for amide synthesis. Such coupling agents are known in the art and include, for example, ((1H-benzo[d][1,2,3]triazol-1-yl)oxy)tri(pyrrolidin-1-yl)phosphonium hexafluorophosphate (V). Optionally, the reaction of step (e) can be carried out in the presence of a base such as DBU. Optionally, the reaction of step (e) can be carried out in the presence of a molecular sieve, for example, a 0.3 nm molecular sieve, to trap water. Suitable solvents for step (e) include polar protic solvents such as MeCN. Step (e) can be carried out at a temperature of 20°C to 70°C, for example, 30°C to 60°C.

[0193] Step (f): Removal of the THP protecting group can be carried out according to known methods. In one embodiment, the THP protecting group is removed using aqueous or alcoholic acid. In one embodiment, the THP protecting group is removed using hydrochloric acid in an alcohol solvent such as isopropyl alcohol.

[0194] Chemical Intermediates As described herein, provided herein are novel chemical intermediates that are useful for the synthesis of inhibitors of the NLRP3 inflammasome, such as Compound 1 and Compound 2.

[0195] Thus, in one embodiment,

[0196] [ka] or a salt thereof.

[0197] In one embodiment,

[0198] [ka] or a salt thereof.

[0199] In one embodiment,

[0200] [ka] or a salt thereof.

[0201] In one embodiment,

[0202] [ka] or a salt thereof.

[0203] In one embodiment,

[0204] [ka] or a salt thereof, wherein X is CH or N.

[0205] In one embodiment,

[0206] [ka] or a salt thereof, wherein X is CH or N.

[0207] In one embodiment,

[0208] [ka] or a salt thereof, wherein X is CH or N.

[0209] In one embodiment,

[0210] [ka] or a salt thereof.

[0211] In one embodiment,

[0212] [ka] or a salt thereof.

[0213] In one embodiment,

[0214] [ka] or a salt thereof.

[0215] In one embodiment,

[0216] [ka] or a salt thereof.

[0217] In one embodiment, there is provided a chemical intermediate or a salt thereof described in the Examples herein.

[0218] In one embodiment, there is provided Compound 1, or a salt thereof, obtainable by any of the processes described herein.

[0219] In one embodiment, there is provided Compound 1, or a salt thereof, obtained by any of the processes described herein.

[0220] In one embodiment, there is provided Compound 2, or a salt thereof, obtainable by any of the processes described herein.

[0221] In one embodiment, there is provided Compound 2, or a salt thereof, obtained by any of the processes described herein.

[0222] Salts of the compounds described herein may be, for example, acid addition salts or base addition salts. Acid addition salts of the compounds described herein can be formed by contacting the compounds with a suitable inorganic or organic acid under conditions known to those skilled in the art. Acid addition salts can be formed using, for example, inorganic acids selected from the group consisting of hydrochloric acid, hydrobromic acid, sulfuric acid, and phosphoric acid. Acid addition salts can also be formed using organic acids selected from the group consisting of trifluoroacetic acid, citric acid, maleic acid, oxalic acid, acetic acid, formic acid, benzoic acid, fumaric acid, succinic acid, tartaric acid, lactic acid, pyruvic acid, methanesulfonic acid, benzenesulfonic acid, and para-toluenesulfonic acid.

[0223] Base addition salts of the compounds described herein can be formed by contacting the compounds with a suitable inorganic or organic base under conditions known to those skilled in the art. For example, it may be possible to prepare alkali metal (such as sodium, potassium, or lithium) or alkaline earth metal (such as calcium) salts by treating the compounds with an alkali metal or alkaline earth metal hydroxide or alkoxide (e.g., ethoxide or methoxide) or a suitable basic organic amine (e.g., choline or meglumine) in an aqueous medium.

[0224] In the processes described herein, it may be necessary to protect reactive functional groups (e.g., hydroxy, dihydroxy) in intermediates described in the synthesis herein to avoid their undesired participation in reactions leading to the formation of the desired product. Conventional protecting groups, such as those described in P.G.M.Wuts, "Greene's Protective Groups in Organic Synthesis," Fifth Edition, John Wiley & Sons Inc., 2014, can be used. For example, if a phenolic hydroxy group is protected as a methyl ether, the protecting group can be removed using BBr3 in dichloromethane. Benzyl protecting groups can be removed by hydrogenation over a palladium catalyst, and para-methoxybenzyl groups can be removed using HCl in alcohol. Acetal protecting groups of diols can be removed by treatment with acid (e.g., AcOH / HO or HCl in 1,4-dioxane). [Example]

[0225] The crystalline forms, chemical processes and intermediates used in such processes described herein are further exemplified in the following examples, which are offered by way of illustration only and are not limiting.

[0226] In the examples, high-resolution mass spectra were recorded on a Micromass LCT mass spectrometer equipped with an electrospray interface (LC-HRMS).

[0227] 1 H NMR measurements were performed at 300, 400, 500, and 600 MHz, respectively. 1NMR spectroscopy was performed on Bruker Avance III 300, 400, 500, and 600 spectrometers operating at H frequencies. Experiments were typically recorded at 25°C. Chemical shifts are given in ppm using the solvent as the internal standard. Protons on heteroatoms, such as NH and OH protons, are reported only if detected in the NMR and may therefore be missing. The following abbreviations (and their derivatives, e.g., dd: doublet of doublet, etc.) were used: s: singlet; d: doublet; t: triplet; q: quartet; m: multiplet; br, broad; qn, quintet; p: pentet.

[0228] Flash chromatography was performed using either normal phase silica FLASH+® (40M, 25M, or 12M), Biotage® SNAP cartridges KP-Sil (340, 100, 50, or 10), Biotage® SNAP cartridges KP-NH (340, 100, 50, or 10), or Agela® flash column silica-CS cartridges (330, 180, 120, 80), unless otherwise stated.

[0229] Reverse-phase flash chromatography was performed using Agela® C-18 spherical 20-35 μm 100A cartridges unless otherwise stated.

[0230] Purification was carried out by preparative HPLC, preparative SFC or reversed phase flash chromatography on standard equipment using MS or UV-induced fraction collection and conditions as described.

[0231] Generally, all solvents used were of analytical grade and commercially available. Anhydrous solvents were those commonly used for reactions.

[0232] The intermediates and examples named below were named using ChemDraw Professional version 19.0.0.22 from PerkinElmer or Biovia Draw 2020 EE. Please note that different chemical naming software may generate different chemical names for a particular compound. You will understand.

[0233] List of abbreviations AcOH = acetic acid aq. = aqueous d=day DBU = 1,8-diazabicyclo[5.4.0]undec-7-ene DCM = dichloromethane DIPEA = N,N-diisopropylethylamine DMAP = dimethylaminopyridine DMF = dimethylformamide DMSO = dimethyl sulfoxide DMSO-d6 = hexadeuterodimethyl sulfoxide Et2O = diethyl ether EtOAc = ethyl acetate EtOH = ethanol h=time HPLC = High-Performance Liquid Chromatography IPA = 2-propanol IPE = Isopropyl Ether iPrOAc = isopropyl acetate LCMS = Liquid Chromatography Mass Spectrometry MeCN = acetonitrile MeOH = methanol min=minutes MS(ESI) / HRMS(ESI) = Mass spectrometry (electrospray ionization) / High-resolution mass spectrometry MTBE = tert-butyl methyl ether n-BuLi = 1-butyllithium NMP = N-methyl-2-pyrrolidone Pd(dppf)Cl2·CH2Cl2=[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II), complex with dichloromethane rt=room temperature RT=retention time sat.=saturated SFC = Supercritical Fluid Chromatography THF = tetrahydrofuran

[0234] Example 1: Preparation of Compound 1, Form A Step 1: Intermediate 1: 4-chloro-N-[[(4S)-2,2-dimethyl-1,3-dioxolan-4-yl]methyl]phthalazin-1-amine

[0235] [ka]

[0236] To a solution of 1,4-dichlorophthalazine (177 g, 889 mmol, 1.0 equiv.) in anhydrous NMP (450 mL) were added DIPEA (310 mL, 1.78 mmol, 2.0 equiv.) and (S)-(2,2-dimethyl-1,3-dioxolan-4-yl)methanamine (123 g, 938 mmol, 1.05 equiv.) at room temperature, and the mixture was stirred at 110 °C for 5 h. The reaction mixture was cooled to room temperature and poured into HO. The mixture was extracted with 1:1 EtOAc / hexane and washed with HO. The organic layer was evaporated under reduced pressure. The residue was triturated with IPE and filtered to give the title compound (212 g, 81%) as a pale yellow powder. MS (ESI): m / z 294.1 / 296.1 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ1.28(s,3H),1.38(s,3H),3.50-3.77(m,3H),4.00-4.20(m,1H),4.4 0-4.48(m,1H),7.83-7.91(m,1H),7.97-8.04(m,2H),8.05-8.11(m,1H),8.35-8.41(m,1H).

[0237] Step 2: Intermediate 2: 2-[4-[[(4S)-2,2-dimethyl-1,3-dioxolan-4-yl]methylamino]phthalazin-1-yl]-5-(trifluoromethyl)phenol

[0238] [ka]

[0239] To a suspension of Intermediate 1 (203 g, 691 mmol, 1.0 equiv.) and (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (213 g, 1.03 mol, 1.5 equiv.) in 1,4-dioxane (1.7 L) and 2.0 M aqueous NaCO (1.04 L, 2.08 mol, 3.0 equiv.), Pd(dppf)Cl·CHCl (11.3 g, 13.8 mmol, 0.02 equiv.) was added, and the mixture was stirred and refluxed under argon for 6 h. To the reaction mixture were added (2-hydroxy-4-(trifluoromethyl)phenyl)boronic acid (28.5 g, 138 mmol) and Pd(dppf)Cl·CHCl (11.3 g, 13.8 mmol, 0.02 equiv.). After 3 hours, the mixture was cooled to room temperature and poured into H2O and EtOAc. Activated carbon was added to the solvent, and the mixture was stirred and filtered through Celite®. The filtrate was extracted with EtOAc, and the organic layer was evaporated under reduced pressure. The crude mixture was purified by flash chromatography (silica; CHCl3 / MeOH = 100 / 0-19 / 1-8 / 2). The collected fractions were further purified by flash chromatography (NH-silica; CHCl3 / MeOH = 100 / 0-39 / 1-8 / 2) to give the title compound (112 g, 39%) as a brown solid. MS (ESI) m / z 420.2 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ1.29(s,3H),1.40(s,3H),3.60-3.73(m,1H),3.76-3.87(m,2H),4.01-4.10(m,1H) ),4.46-4.55(m,1H),7.25-7.33(m,2H),7.41-7.57(m,2H),7.69-7.91(m,3H)8.32-8.38(m,1H),10.3(br s,1H).

[0240] Step 3: Compound 1: (2S)-3-[[4-[2-hydroxy-4-(trifluoromethyl)phenyl]phthalazin-1-yl]amino]propane-1,2-diol (Form A)

[0241] [ka]

[0242] To a suspension of Intermediate 2 (112 g, 267 mmol) in AcOH (240 mL) was added HO (80 mL), and the mixture was stirred at 80 °C for 5 h. The mixture was cooled to room temperature and evaporated under reduced pressure. The crude mixture was purified by flash chromatography (silica; CHCl / MeOH = 100 / 0 - 90 / 10 - 80 / 20) to give the title compound (65.3 g, 57%) as a colorless solid. The residue (65.3 g + 7.51 g (residue from the previous batch)) was triturated with MeOH and filtered. EtOH was added to the resulting solid, and the solvent was evaporated under reduced pressure to give the title compound (62.7 g, 86%) as a colorless crystalline solid (Form A). MS (ESI): m / z 380.1 [M + H] + . 1 H NMR(400MHz,DMSO-d6)δ3.39-3.49(m,2H),3.51-3.61(m,1H),3.70-3.80(m,1H),3.82-3.91(m,1H),4 .80-4.90(m,1H),5.21-5.29(m,1H),7.26-7.32(m,2H),7.43-7.48(m,1H),7.49-7.54(m,1H),7.71(br s,1H),7.77-7.83(m,1H),7.85-7.92(m,1H),8.32-8.41(m,1H),10.37(br s,1H).

[0243] X-ray diffraction analysis of Form A of Compound 1 X-ray diffraction analysis of Form A of Compound 1 is performed according to standard methods, which can be found, for example, in Kitaigorodsky, AI (1973), "Molecular Crystals and Molecules", Academic Press, New York; Bunn, CW (1948), "Chemical Crystallography", Clarendon Press, London; or Klug, HP & Alexander, LE (1974), "X-ray Diffraction Procedures", John Wiley & Sons, New York.

[0244] X-ray powder diffraction (XRPD) data was measured using corundum as an internal standard. XRPD patterns were determined by mounting the sample on a zero background holder, single crystal silicon, and spreading the sample into a thin layer.

[0245] Powder X-ray diffraction is recorded using a Theta-Theta PANalytical X'Pert PRO (X-ray wavelength 1.5418 Å, nickel-filtered Cu radiation, voltage 45 kV, filament emission 40 mA). Variable divergence and anti-scatter slits and 0.04° incident and diffraction solar slits are used. The sample is rotated during the measurement. A 0.013° step width and 115.770 s count time are used with a PIXcel 1D detector (effective length 3.347° 2-theta) to scan the sample from 2.4 to 50° 2-theta.

[0246] This XRPD pattern is obtained in the Bragg-Brentano configuration.

[0247] The XRPD pattern of Form A of Compound 1 is shown in Figure 1. The most prominent peaks in the XRPD pattern are listed in Table 1. Peak intensities are described herein as vs (very strong), s (strong), m (moderate), and w (weak), corresponding to relative intensity % (based on the most intense peak) of 25-100%, 10-25%, 3-10%, and 1-3%, respectively.

[0248] [Table 1]

[0249] TG / DTA (thermogravimetry / differential thermal analysis) measurements were performed using a TG / DTA 7200 (SII NanoTechnology Inc.). 3.904 mg of Compound 1 Form A was weighed into an aluminum pan. The sample was then heated from 30°C to 300°C at a heating rate of 10°C / min under a nitrogen purge of 200 mL / min. An empty aluminum pan was used as a reference. The output from the TG / DTA analysis is shown in Figure 2. Solid line = DTA analysis, long-dashed line = temperature, short-dashed line = TG analysis. No weight loss was observed from the sample before melting.

[0250] Storage of Compound 1 Form A at 60°C / 75% relative humidity (RH) (open container) for 1 week and at 60°C (closed container, ambient RH) for 1 week showed no change in weight and no change in the XRPD pattern.

[0251] Example 2 - THP-protected Route to Compound 1 Step 1: Intermediates 3, 4 and 5:

[0252] [ka]

[0253] A solution containing 2-[3-(trifluoromethyl)phenoxy]tetrahydro-2H-pyran (534 g, 2.17 mol), N,N,N',N'-tetramethyl-1,2-ethanediamine (266 g, 2.29 mol) in THF (3.2 L) was heated at -10°C with a flow rate of 10 g / min. -1 Separately, n-butyllithium (1.41 L, 1.96 mol, 1.4 M in hexane) was pumped into the reaction tube at 3.9 mL / min at −10° C. -1The product was introduced into the reaction tube and mixed in a helical chamber with a solution of magnesium chloride (251 g, 2.63 mol) in THF (4.78 L) pumped in at room temperature. -1 The mixture was mixed at 12.0 g / ml, and the product was poured into a reaction tube where it was mixed in a helical mixer with a solution of phthalic anhydride (651 g, 4.40 mol) in THF (3.70 L) pumped in. - The product was collected and evaporated to approximately half its weight, then treated with 1 M sodium hydroxide solution (6.4 Kg), stirred at 10 °C for 40 min, the layers were separated, the aqueous layer was extracted with THF (1 L), and the organics were combined to give a THF solution of intermediate 3 (2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid) (13.98 kg), MS (ESI): m / z [M+Na+HO] + 435.2.

[0254] A solution of intermediate 3 in THF was heated to 35°C and treated portionwise with aqueous hydrochloric acid. After stirring for 48 hours, aqueous brine (564 g) was added, the phases were separated, the organic layer was washed with brine (1 kg x 2), the organic layer was separated and partially evaporated to give a solution of intermediate 4 (2-[2-hydroxy-4-(trifluoromethyl)benzoyl]benzoic acid) in THF (1.4 Kg, 24.5 wt% by NMR assay, 50.4% yield).

[0255] 1 H NMR(300MHz,CDCl3)δ8.10-8.15(m,1H),7.56-7.70(m,2H),7.32-7.35(m,1H),7.26(d,1H),7.15-7.20(m,1H),6.93-6.97(m,1H);MS(ESI):m / z[M+Na] + 333.1.

[0256] A solution of intermediate 4 in THF (1.4 kg, 345 g) was diluted with THF (860 mL), then hydrazine hydrate was added (152 g, 3.1 mol) and stirred at 50° C. for 5 hours. The solution was cooled and treated with 3 M hydrochloric acid solution (1021 g), the organics were separated, then saturated brine (556 g) was added, some precipitate formed which was dissolved by diluting with THF (1037 g) and heating to 50° C., the organic layer was separated and washed with saturated brine (800 g×2), the organic layer was partially evaporated, heated to 65° C. and heptane (1757 g) was added slowly. Once the final approximate ratio of THF:heptane (1:1) was reached, the mixture was cooled to 20° C. and the solid was filtered, washed with THF:heptane (2:1, 2×500 mL) and dried to give intermediate 5 (4-[2-hydroxy-4-(trifluoromethyl)phenyl]-1(2H)-phthalazinone) (322.5 g, 93%).

[0257] 1 H NMR(300MHz,CDCl3)δ12.85(s,1H),10.45(s,1H),8.29-8.32(m,1H),7.84-7.87(m,2H),7.53(d,1H),7.20-7.35(m,3H);MS(ESI):m / z[M+H] + 307.0.

[0258] Step 2: Intermediate 6: 2-(4-chloro-1-phthalazinyl)-5-(trifluoromethyl)phenol

[0259] [ka]

[0260] A mixture of intermediate 5 (122 g, 392 mmol) and acetonitrile (1200 mL) was stirred at room temperature, then phosphoryl chloride (72.8 mL, 784 mmol) was added and heated to 50° C. for 22 h. The mixture was cooled to 0-5° C. and water (600 mL) was added while maintaining the temperature below 10° C., followed by a solution of dipotassium phosphate (204.8 g, 1176 mmol in 1200 mL of water), the mixture was warmed to 20° C., and the solid was filtered, washed with water (2×250 mL) and then acetonitrile (4×500 mL), and dried under vacuum at 50° C. to give the title compound (112.9 g, 71%, 80% w / w).

[0261] 1 H NMR(DMSO-d6)δ:10.13-11.11(br s,1H),8.35(ddd,J=8.1,1.0Hz,1H),8.16(ddd,J=8.3,7.1,1.2Hz,1H),8.06(ddd,J=8.3,7.1,1. 2Hz,1H),7.73(ddd,J=8.1,1.0Hz,1H),7.57-7.66(m,1H),7.32-7.40(m,2H);MS(ESI):m / z[M+H] + 325.0 / 327.0.

[0262] Step 3: Compound 1:

[0263] [ka]

[0264] To a stirred mixture of (2S)-3-amino-1,2-propanediol (CAS#61278-21-5) and N-methyl-2-pyrrolidone (160 mL) at 125° C. was added Intermediate 6 (80 g, 197 mmol, 80% w / w) in N-methyl-2-pyrrolidone (400 mL) over 3 hours, followed by stirring at 120° C. for 2 hours. The mixture was cooled to 20° C. and stirred for 16 hours, then water (320 mL) was added over 30 minutes, seed crystals (400 mg, Form A) were added and stirred for 30 minutes, then water (800 mL) was added over 90 minutes, the resulting mixture was stirred for 18 hours, then the solid was filtered, washed with water:isopropyl alcohol (1:1, 400 mL), then water (2×400 mL) and dried at 60° C. to give Compound 1 (62.25 g, 81.7%, 98.0% w / w).

[0265] 1 H NMR(DMSO-d6)δ:10.22-10.46(br s,1H),8.35(d,J=8.1Hz,1H),7.87(ddd,J=8.2,7.1,1.3Hz,1H),7.78(ddd,J=7.6,1.1Hz,1H),7.6 2(t,J=5.6Hz,1H),7.51(d,J=7.6Hz,1H),7.44(dd,J=8.2,0.7Hz,1H),7.25-7.31(m,2H),5.24(br s,1H),4.81(br t,J=5.8Hz,1H),3.81-3.89(m,1H),3.73(m,1H),3.50-3.60(m,1H),3.39-3.45(m,2H);MS(ESI):m / z[M+H] + 380.1.

[0266] Example 3 - Alternative THP-protective Route to Compound 1 Step 1: Intermediate 8: 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one

[0267] [ka]

[0268] 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran (56.3 g, 228.78 mmol) and N1,N1,N2,N2-tetramethylethane-1,2-diamine (37.6 mL, 250.57 mmol) were mixed in THF (200 mL) under N2 at room temperature and cooled to -5 °C. Butyllithium (2.5 M in hexanes, 104 mL, 250.57 mmol) was added over 10 minutes. To this mixture was added a solution of tert-butyl methyl phthalate (40 g, 167.61 mmol) in THF (200 mL) cooled to -78 °C over 10 minutes. The mixture was stirred at -78 °C for an additional 2 hours. The reaction mixture was warmed to -20 °C, and then hydrazine hydrate (16.31 mL, 335.21 mmol) was added. An additional 2 mL of water was added, followed by acetic acid. The reaction mixture was stirred at -20°C overnight. The mixture was partitioned between water (200 mL) and MeTHF (200 mL), the layers were separated, and the aqueous layer was extracted with additional MeTHF (100 mL). The organic layers were combined and concentrated in vacuo. The residue was dissolved in TBME (40 mL), and heptane (160 mL) was added slowly. The precipitate was isolated by filtration to give the crude title compound as a white solid (22.5 g).

[0269] Step 2: Intermediate 9: N-(((S)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)phthalazin-1-amine

[0270] [ka]

[0271] Intermediate 8 (2.00 g, 5.12 mmol) and (S)-(2,2-dimethyl-1,3-dioxolan-4-yl)methanaminium chloride (3.44 g, 20.49 mmol) were mixed in acetonitrile (20 mL). DBU (6.12 mL, 40.99 mmol) and molecular sieves 0.3 nm (water absorption capacity ≥ 20%) (Merck EMD Millipore Corporation) were added. The mixture was stirred at room temperature for approximately 1 hour. ((1H-benzo[d][1,2,3]triazol-1-yl)oxy)tri(pyrrolidin-1-yl)phosphonium hexafluorophosphate (V) (5.33 g, 10.25 mmol) was added, and the reaction mixture was heated to 50 °C and stirred at 50 °C for several hours. The conversion was checked by IPC. The reaction mixture was filtered to remove the molecular sieves. The molecular sieves are rinsed with acetonitrile and the filtrate is evaporated to dryness under vacuum at 40°C. The evaporation residue (18.044 g) containing the product is obtained as an orange oil. It is dissolved in 150 mL of EtOAc and 10% aqueous citric acid solution (100 mL). After phase separation, the organic phase is washed with additional 10% aqueous citric acid solution (2 x 100 mL), then with saturated aqueous NaHCO3 solution (2 x 100 mL) and then dried over MgSO4. The MgSO4 is removed by filtration, washed thoroughly and the filtrate is reduced by evaporation under vacuum at 40°C. The crude product (4.00 g) is obtained as a yellow to pale orange foam. MS (ESI): m / z [M+H] + 504.5.

[0272] Step 3: Compound 1:

[0273] [ka]

[0274] Intermediate 9 is mixed with 5-6 M HCl in IPA (4.66 mL, 25.62 mmol) and iPrOH (20 mL) at 40 °C for about 10 min. An orange solution is present, which is evaporated to dryness. The crude product (4.578 g) is obtained as an orange resin. 200 mg of the crude product is triturated in ethyl acetate / ethanol (1 / 1 vol / vol). The product is isolated by filtration, washed with EtOAc, and dried. The product (105 mg, HCl salt) is obtained as an off-white solid.

[0275] 1 H NMR(500MHz,DMSO)δ10.91(br,1H),10.29(br,1H),9.00(d,1H),8.21-8.02(m,2H),7.70-7.53(m,2H),7.43(s, 1H),7.39-7.33(m,1H),4.00(br,2H,OH),4.00-3.92(m,2H),3.85-3.69(m,2H),3.56-3.46(m,2H),2.50(DMSO). MS(ESI):m / z[M+H] + 380.3.

[0276] Comparative Example 1: Preparation of Compound 2 Step 1: Intermediate 10: 1-Bromo-2-((4-methoxybenzyl)oxy)-4-(trifluoromethyl)benzene

[0277] [ka]

[0278] 2-Bromo-5-(trifluoromethyl)phenol (3.0 g, 12.45 mmol) and 1-(bromomethyl)-4-methoxybenzene (2.53 g, 12.57 mmol) were dissolved in MeCN (30 mL), and potassium carbonate (1.892 g, 13.69 mmol) was added in one portion (no or very weak exotherm). The reaction mixture turned yellow. The reaction mixture was stirred at room temperature overnight. By NMR, the reaction was complete after 16 h. Water and EtOAc were added, and the phases were separated. The aqueous phase was extracted with EtOAc, and the combined organic extracts were washed with brine and evaporated. This gave a pale orange oil that did not crystallize from IPA (approximately 15 mL). Instead, the oil was purified by column chromatography (silica gel, heptane / EtOAc: 20 / 1 as eluent) to give 3.58 g (80%) of the title compound as a colorless oil that crystallized on standing. 1 H NMR (500MHz, DMSO-d6) δ3.76(s,3H),5.23(s,2H),6.95-7.00(m,2H),7.22-7.28(m,1H),7.42(d,2H),7.51(d,1H),7.80-7.86(m,1H).

[0279] Step 2: Intermediate 11: 3-(tert-butyl) 4-methylpyridine-3,4-dicarboxylate

[0280] [ka]

[0281] tert-Butanol (200 mL) was added to 4-(methoxycarbonyl)nicotinic acid (25.0 g, 138 mmol), followed by di-tert-butyl dicarbonate (60.2 g, 276 mmol) and pyridine (25 mL). DMAP (100 mg, catalytic) was added, and the reaction was stirred at 35 °C overnight. Water and iPrOAc were added, and the two phases were separated. The organic extract was washed with two portions of water, evaporated, and the residue was evaporated twice with toluene. The residue was filtered through a column of silica using 40% MTBE in heptane as the mobile phase to give the title compound (27.7 g, 84%) as a pale yellow oil. 1 1H NMR (500MHz, DMSO-d6) δ1.52 (s, 9H), 3.88 (s, 3H), 7.65 (dd, 1H), 8.87 (d, 1H), 8.96 (d, 1H).

[0282] Step 3: Intermediate 12: tert-butyl 4-[2-[(4-methoxyphenyl)methoxy]-4-(trifluoromethyl)benzoyl]pyridine-3-carboxylate

[0283] [ka]

[0284] Intermediate 11 (7.0 g, 29.6 mmol) was dissolved in THF (50 mL) and cooled to -78 °C. In a separate flask, intermediate 10 (10.7 g, 29.6 mmol) was dissolved in THF (50 mL) and n-BuLi (19.4 mL, 31.1 mmol, 1.6 M in hexanes) was added at -78 °C. The pale yellow solution was stirred at -78 °C for 15 seconds and then added dropwise to the first solution via cannula. The reaction mixture was stirred at -78 °C for 10 minutes, then AcOH (1.9 mL in 100 mL of water) was added, followed by EtOAc. The reaction mixture was allowed to reach room temperature and the two phases were separated. The organic extract was washed with water and evaporated to give the title compound (14.4 g, quantitative) as an orange oil. It was used in the next step without further purification. MS (ESI): m / z [M+H] + 488.3.

[0285] Step 4: Intermediate 13: 1-[2-[(4-methoxyphenyl)methoxy]-4-(trifluoromethyl)phenyl]-3H-pyrido[3,4-d]pyridazin-4-one

[0286] [ka]

[0287] Intermediate 12 (41.4 g, 84.9 mmol) was dissolved in THF (300 mL), hydrazine monohydrate (21.1 mL, 340 mmol, 50% in water) was added, and the reaction mixture was stirred at 60° C. for 16 h. Water (100 mL) was added, and the mixture was stirred at room temperature and then poured into water (600 mL). The solid was filtered off and washed with water and MTBE. The product was slurried in refluxing EtOAc (1 L), cooled to room temperature, and filtered to give the title compound (17.3 g, 48%) as an off-white solid. MS (ESI): m / z [M+H] + 428.2. 1 H NMR(500MHz,DMSO-d6)δ3.68(s,3H),5.15(s,2H),6.75(d,2H),7.02(d,2H),7 .27(d,1H),7.51(d,1H),7.65(d,2H),8.93(d,1H),9.48(s,1H),13.23(s,1H).

[0288] Step 5: Intermediate 14: 4-chloro-1-[2-[(4-methoxyphenyl)methoxy]-4-(trifluoromethyl)phenyl]pyrido[3,4-d]pyridazine

[0289] [ka]

[0290] Intermediate 13 (6.0 g, 14.0 mmol) was slurried in 1,4-dioxane (55 mL). Pyridine (9.9 mL, 122 mmol) and phosphoryl trichloride (4.6 mL, 48.9 mmol) were added, and the reaction was stirred at 60° C. for 19 hours. The mixture was cooled to room temperature and then added to trisodium citrate (180 mL, aq., 1 M). The precipitated product was filtered off, washed with water (2×50 mL), and dried under vacuum to give a tan solid. The crude product was slurried in MeCN (80 mL) and heated to 80° C. until dissolved. The mixture was cooled to room temperature, and the formed precipitate was filtered off, washed with MeCN (2×15 mL), and dried to give the title compound (2.57 g, 41%) as a tan solid. MS (ESI): m / z [M+H] + 446.3. 1 1H NMR(500MHz,DMSO-d6)δ3.66(s,3H),5.15(s,2H),6.73(d,2H),6.99(d,2H ),7.58(d,1H),7.65(dd,1H),7.72(d,2H),9.10(d,1H),9.69-9.82(m,1H).

[0291] Step 6: Intermediate 15: (1R,2R)-2-[[1-[2-[(4-methoxyphenyl)methoxy]-4-(trifluoromethyl)phenyl]pyrido[3,4-d]pyridazin-4-yl]amino]cyclohexanol

[0292] [ka]

[0293] Intermediate 14 (2.6 g, 5.7 mmol), (1R,2R)-2-aminocyclohexan-1-ol (1.1 g, 9.2 mmol), and NaHCO (2.4 g, 28.7 mmol) were mixed in IPA (22 mL) and stirred at 80 °C for 3 days. The reaction mixture was poured into water (100 mL) and stirred at room temperature for 2 hours. The solid was filtered off, washed with water, and dried under vacuum at 40 °C to give the title compound (2.9 g, 96%) as a tan solid. MS (ESI): m / z [M+H] + 535.6. 1H NMR(500MHz,DMSO-d6)δ1.30(s,4H),1.69(d,2H),1.97(d,1H),2.12(s,1H),3.61(d,1H),3.66(s,3H),4.19(s,1H),4.83(s ,1H),5.11(s,2H),6.74(d,2H),7.03(d,2H),7.26(d,1H),7.48(d,1H),7.60(s,2H),7.67(d,1H),8.81(d,1H),9.76(s,1H).

[0294] Step 7: Compound 2: 2-(4-(((1R,2R)-2-hydroxycyclohexyl)amino)pyrido[3,4-d]pyridazin-1-yl)-5-(trifluoromethyl)phenol

[0295] [ka]

[0296] Intermediate 15 (2.9 g, 5.5 mmol) was slurried in anhydrous EtOH (99.5%, 7 mL), HCl (4 M in 1,4-dioxane, 20.7 mL, 82.9 mmol) was added, and the reaction was stirred at room temperature for 2 h. The mixture was added dropwise to EtO (150 mL) under stirring to give a precipitate, which was filtered off, washed with EtO, and dried to give a pale yellow solid (HCl salt). The solid was slurried in water (50 mL), made basic (pH = 9) with saturated aqueous NaHCO, and extracted (multiple times) with DCM:MeOH = 9:1. The combined organic extracts were filtered through a phase separator and evaporated to give 1.85 g of an orange semi-solid. The crude material was dissolved in MeCN (20 mL) and IPA (0.5 mL) at 70° C., cooled to room temperature, filtered, washed with MeCN, and dried under vacuum at 40° C. to give the title compound (1.25 g, 56%) as a yellow solid. MS (ESI): m / z [M+H] + 405.3. HRMS(ESI):m / z[M+H] + C 20 H 19 Calculated value for F3N4O2: 405.1538, Found: 405.1538. 1H NMR(500MHz,DMSO-d6)δ1.25-1.41(m,4H),1.72(d,2H),1.99(d,1H),2.13(s,1H),3.56-3.69(m,1H),4 .17-4.28(m,1H),7.24-7.36(m,3H),7.55(d,1H),7.71(d,1H),8.84(d,1H),9.80(s,1H),10.46(s,1H).

[0297] Example 3: Preparation of Compound 2 via THP Protection Route Step 1: Intermediate 17: 1-[2-tetrahydropyran-2-yloxy-4-(trifluoromethyl)phenyl]-3H-pyrido[3,4-d]pyridazin-4-one

[0298] [ka]

[0299] 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran (45.9 g, 186.6 mmol) and N1,N1,N2,N2-tetramethylethane-1,2-diamine (25.9 mL, 173 mmol) were mixed in THF (250 mL) under N2 at room temperature and cooled to 0 °C. Butyllithium (2.4 M in hexanes, 71.9 mL, 173 mmol) was added over 30 min, maintaining the temperature below 5 °C. This mixture was transferred to 3-(tert-butyl)4-methylpyridine-3,4-dicarboxylate (0.5 M in THF, 300 mL, 150 mmol) via Teflon tubing under N2 and cooled to −78 °C over 30 min, maintaining the temperature below −70 °C. The mixture was stirred at −78 °C for an additional 30 min (resulting in the formation of intermediate 16). (Intermediate 16: 1H NMR(500MHz,CDCl3)δ9.17(d,1H),8.81(d,1H),8.19(d,1H),7.54(s,1H),7.40(dd,1H),7.28(CDCl3),7.25(dd,1H),5. 31(t,1H),3.64-3.50(m,2H),1.59-1.45(m,2H),1.45-1.31(m,2H),1.39(s,9H),1.02-0.93(m,1H),0.88-0.76(m,1H). MS(ESI):m / z[M+H] + 452.5.)

[0300] Hydrazine hydrate (14.6 mL, 300 mmol) was added over 4 minutes at −78° C., then the cooling bath was removed and the reaction stirred at room temperature for 1 hour. The reaction mixture was heated to 30° C., and acetic acid (42.9 mL, 750 mmol) in absolute EtOH (43 mL) was added via addition funnel over 15 minutes, and the mixture was diluted with additional EtOH (100 mL) and water (50 mL). The mixture was evaporated to a volume of 200 mL to give a thick suspension, which was cooled to 10° C. and then filtered. The solid was washed with cold EtOH:water 1:1 (110 mL) and cold water (100 mL), air-dried for 30 minutes, then transferred to a flask and dried in vacuo overnight to give the title compound (40.1 g, 68%) as a tan solid. 1 H NMR(500MHz,DMSO)δ0.8-1.62(6H,m),3.41-3.69(2H,m),5.72(1H,d),7.25 -7.39(1H,m),7.49-7.76(3H,m),8.96(1H,dd),9.52(1H,s),13.28(1H,s). MS(ESI):m / z[M+H] + 392.2.

[0301] Step 2: Intermediate 18: (1R,2R)-2-((1-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)pyrido[3,4-d]pyridazin-4-yl)amino)cyclohexan-1-ol

[0302] [ka]

[0303] (1R,2R)-2-Aminocyclohexan-1-ol hydrochloride (291 g, 1916 mmol), ((1H-benzo[d][1,2,3]triazol-1-yl)oxy)tri(pyrrolidin-1-yl)phosphonium hexafluorophosphate (V) (665 g, 1278 mmol), and DBU (382 ml, 2555 mmol) are mixed in a 10 L reactor in acetonitrile (1.25 L). The reaction mass is stirred until a solution is present. A solution of intermediate 17 (250 g, 639 mmol) and DBU (191 ml, 1278 mmol) in THF (1000 mL) [1.4 L, orange solution] is added within 3 hours. IPC after another hour indicates complete conversion. Workup: Add EtOAc (2.5 L) and wash with 10% aqueous citric acid (2 x 2.5 L). The aqueous phase is back-extracted with EtOAc (2.5 L) and the combined organic phases are washed with saturated aqueous NaHCO3 (2 x 2.5 L). The organic phase is reduced by distillation in vacuo at elevated temperature (approximately 70 °C) until a target volume of 1.25 L remains. The concentrated phase of the title compound is cooled to 20 °C. MS (ESI): m / z [M+H] + 489.4.

[0304] Step 3: Compound 2:

[0305] [ka]

[0306] A 5-6N solution of hydrogen chloride in 2-propanol (532 ml, 3194 mmol) is added to the concentrated phase of intermediate 18. A suspension begins to form, and IPC after 0.5 h indicates complete conversion. Acetonitrile (6 L) is added, and an orange suspension is present, which is stirred for 2 h. The product is isolated by filtration, washed with two portions of acetonitrile (1 L), dried over a Nutsche in vacuo, and a stream of N2 is passed through the cake. The crude product (291 g) is obtained as a yellow solid. The crude material is mixed with water (1500 mL), forming a thin suspension of yellow foam. EtOH (anhydrous; 300 mL) is added, and the resulting reaction mass is heated to 60 °C, forming a yellow suspension. Additional EtOH (anhydrous; 50 mL) and water (250 mL) are added. The suspension is cooled from 60 °C to 20 °C within 1 h and stirred at 20 °C for 1 h before being isolated by filtration. The product cake is washed with water (8 L). The product is dried overnight in a vacuum on a nutsche and a stream of N2 (20 L / min) is passed through the cake. The pure product (186 g) is obtained as an off-white, slightly yellow solid. The material is crushed, homogenized in the nutsche during removal, and further dried at elevated temperature (50 °C) in a vacuum oven (<10 mbar). The pure, dried hydrochloride salt of the title compound (153 g, 50%) is obtained as an off-white, slightly yellow crystalline solid.

[0307] Biological and physicochemical data Human NLRP3 speckle formation assay (Test A) To profile compounds for NLRP3 antagonist activity in terms of inhibiting nigericin-induced speck formation, we utilized ASC-GFP reporter monocytes (InvivoGen #thp-ascgfp). This assay is based on NF-kB-dependent expression of an ASC::GFP fusion protein. LPS priming of cells increases ASC::GFP expression, and nigericin recruits ASC::GFP, procaspase-1, and NLRP3 to form ASC-specks, micrometer-sized complexes that are quantified by fluorescence microscopy.

[0308] Preparation of assay reagents: Assay medium: RPMI 1640 (Gibco #72400-021) supplemented with 10% heat-inactivated FBS (Gibco #10270) Cells: THP-ASC-GFP were cultured in RPMI 1640 (Gibco #72400-021) supplemented with 10% heat-inactivated FBS (Gibco #10270) and 100 μg / mL Zeocin (Life Technologies #46-0072) (every other passage) to maintain ASC::GFP expression.

[0309] Step-by-step protocol for performing the assay: Day 1 1. Cells were counted by CEDEX (Innovartis) and diluted to 375,000 cells / mL in assay medium supplemented with 100 nM phorbol 12-myristate 13 acetate (Sigma #P8139). 2. 20 μl of the above cell mixture was dispensed into a black μclear TC-treated (Greiner #781091) 384-well plate using a Multidrop Combi (ThermoFisher). 3. The plates were incubated at 37°C, 5% CO2 for 20 hours.

[0310] Day 2 1.10 μl of LPS (Sigma #L2654) was dispensed at 1 μg / mL using a Multidrop Combi (ThermoFisher). 2. The plate was incubated at 37°C, 5% CO2 for 3 hours. 3. Concentration response curves of 80 nl of test compound in DMSO were prepared and diluted in 20 μl of assay medium supplemented with 68 μM ZVAD-FMK (Promega #7231) in polypropylene 384-well plates (Greiner #781280). 4. 10 μl of the above test compound solution was transferred to the cell plate using a Bravo (Agilent). 5. The plate was incubated at 37°C, 5% CO2 for 30 minutes. 15 μl of nigericin (Sigma #SML1779) at 6.75 μM was dispensed onto the cell plate using a Certus (Gyger) 7. The plate was incubated at 37°C, 5% CO2 for 1 hour. 15 μl of 17.3% formaldehyde (Sigma #F8775) supplemented with Hoechst nucleic acid stain (Life Technologies #H3570) diluted 8.1:5000 was added using a Multidrop (ThermoFisher). 9. The plate was incubated at room temperature for 15 minutes. 10. The plate was washed twice with 40 μl of PBS (Gibco #100100) containing Bluewasher (BlueCatBio). 11. Plates were imaged using ImageXpress (Molecular Devices)

[0311] Image data were processed using Columbus software (Perkin Elmer) using nuclear (nucei) staining to identify cells and spot detection to identify ASC-specks within the cells. Data were further processed using Screener (Genedata AG). Concentration-response data for speck number per cell were fitted using a four-parameter logistic fit to obtain EC 50 The values ​​are reported in Table 2.

[0312] Nigericin-induced (human NLRP3) IL-1β assay (Test B) Compounds were profiled for NLRP3 antagonist activity with respect to the inhibition of nigericin-induced IL-1β release from THP-1 human monocytes. Quantification was performed using a commercially available human IL-1β HTRF detection kit (CisBio, 62HIL1BPEH). The assay uses two anti-IL-1β antibodies in a sandwich assay format. One is labeled with a donor fluorophore (Eu cryptate) and the other with an acceptor (XL). An immune complex containing two antibodies bound to the same IL-1β molecule allows fluorescence resonance energy transfer (FRET) between the donor and acceptor after excitation of the donor with a light source, resulting in fluorescence emission of 665 nM from the acceptor. The fluorescence signal intensity is proportional to the IL-1β concentration in the sample.

[0313] Preparation of assay reagents: Cells: THP human monocytic leukemia cell line. Cells are generally passaged every 2-3 days and maintained at a density of 0.2-0.4. * The cell density was maintained at 10^6 cells / mL.

[0314] Culture and assay medium: RPMI 1640 (Gibco, 72400-021) supplemented with 10% FBS (Sigma, F2442)

[0315] IL-1β Standard: The reconstituted IL-1β standard provided in the CisBio kit was diluted in assay medium to a maximum final concentration of 2 ng / mL in the assay.

[0316] HTRF detection reagents: cAMP-d2 and anti-cAMP cryptate were reconstituted according to the CisBio kit instructions. Immediately before use, the reagents were mixed using the following ratios: 10 / 24 detection buffer (provided with the kit), 14 / 24 PBS (Gibco, 10010), 1 / 120 IL-1β Eu-cryptate antibody, and 1 / 120 IL-1β XL antibody.

[0317] Step-by-step protocol for performing the assay: Day 1 1. 20 nL of test compound dissolved in DMSO was aquostically dispensed (Labcyte Echo) into a white 384-well plate (Greiner; 784075), sealed and stored at room temperature until assayed. 2. 20 nL of 50 μM control compound in DMSO (250 nM final concentration) was added to 100% inhibition control wells using the Echo dispenser, and 20 nL of DMSO was added to 0% control wells. The control compound may be selected from MCC950 (N-[[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)amino]carbonyl]-4-(1-hydroxy-1-methylethyl)-2-furansulfonamide) or any other compound that acts as a full antagonist in the assay. 3. Aliquots of cells were removed from cells grown in continuous culture and counted on a CEDEX (Innovatis). 4. Centrifuge the number of cells required for the experiment at 250 x g for 5 minutes and suspend in assay medium at 37°C for 1.0 min. * Resuspended at 10^6 cells / mL. 5. LPS (Sigma; L2654) was added to a final concentration of 1 μg / mL. 6. Cells were LPS primed en bloc in 50 mL tubes by incubating at 37°C, 5% CO2 and 95% humidity for 3 hours. 7.1.0 * Four microliters of the cell solution at 10^6 cells / mL was dispensed into a white 384-well small-volume plate (Greiner; 784075) using a Multidrop Combi (Thermo Fisher) to obtain 4000 cells / well. 8. Incubate at 37°C, 5% CO2 and 95% humidity for 30 minutes. 9. 4 μL of 40 μM nigericin in assay medium was added with Certus (Gyger) to a final concentration of 20 μM. 10. Incubation for 1 hour at 37°C, 5% CO2 and 95% humidity. 11.4 μL of HTRF detection reagent was added using a Multidrop Combi. 12. Incubate for 3 hours at room temperature protected from light. 13. Homogenous time-resolved fluorescence (HTRF) signals were detected with an Envision (PerkinElmer) or Pherastar (BMG Labtech) reader (λex = 340 nm, λem = 665 and 615 nm).

[0318] Using an IL-1β standard curve, the HTRF data were converted to the amount of IL-1β produced in the samples, which was subsequently used to calculate the concentration response. The concentration response data were analyzed using Screener (Genedata) and fitted using a four-parameter logistic fit. The results of the assay were expressed as IC 50 The concentrations are reported in Table 2 as (μM).

[0319] I C 50 is defined as the concentration at which inhibitory activity reaches 50% of its maximum level. When assays were performed multiple times for the same compound, geometric mean values ​​are reported. To facilitate comparison of efficacy data, efficacy was normalized to the % inhibitory effect of the test compound compared to the inhibition produced by a saturating concentration of control compound (250 nM).

[0320] BzATP-induced (human NLRP3) IL-1β assay (Test C) In a variation of the IL-1β assay, compounds were tested for their ability to inhibit BzATP (2'(3')-O-(4-benzoylbenzoyl)adenosine 5'-triphosphate)-induced IL-1β release from THP-1 human monocytes. As with the nigericin induction assay, quantification was performed using the Human IL-1β HTRF Detection Kit (CisBio, 62HIL1BPEH).

[0321] There were several differences between the nigericin induction assay (Test B) and the BzATP induction assay. Conditions in the BzATP induction assay that had relevant differences compared to the nigericin induction assay included the following: - Cell culture medium: RPMI 1640 (Gibco, 11875-119) supplemented with 10% FBS (Sigma, 171012) and penicillin-streptomycin (Thermo Fisher, 15140-122). Assay medium: RPMI 1640 (Gibco, 22400-105) supplemented with 1% FBS (Sigma, 171012). -Cells were primed with LPS (Sigma, L2630) at a final concentration of 2 μg / mL for 24 hours (instead of 3 hours at 1 μg / mL). -IL-1β production was induced by the addition of BzATP (Sigma, B6396) at a final concentration of 1 mM (instead of nigericin), followed by 30 min of incubation at 37°C, 5% CO2 and 95% humidity.

[0322] The results of the assay are expressed as IC 50 The concentrations are reported in Table 2 as (μM).

[0323] hERG Assay (Test D) Experiments were performed at room temperature on a SyncroPatch 384PE (Nanion Technologies) high-throughput patch clamp platform, using a medium-resistance tip with four patch holes per site. The hERG-expressing Chinese hamster ovary K1 (CHO) cell line was used in an assay-ready format and stored in liquid nitrogen until use. Two vials of cells (10 × 10 per vial) were used. 6The cells were thawed and added to 20 mL of Hepes-buffered saline solution (HBSS). HBSS contained 140 mM NaCl, 4 mM KCl, 10 mM HEPES, and 5 mM glucose (pH 7.4). The internal patch clamp solution was 120 mM KF, 20 mM KCl, 10 mM HEPES, 10 mM EGTA, and 25 μM escin (pH 7.2). After the initial sealing process was completed, a seal enhancer solution containing HBSS supplemented with 10 mM CaCl2 and 1 mM MgCl2 was applied to the cells. The external solution was then replaced (4 times) with an external patch clamp solution containing 80 mM NaCl, 4 mM KCl, 10 mM HEPES, 2 mM CaCl2, 1 mM MgCl2, 5 mM glucose, and 60 mM NMDG (pH 7.4). All solutions were stored at room temperature, except for escin, which was stored at 4°C. All compounds were dispensed into Greiner-bio 384-well plates and tested in a six-point cumulative assay (final DMSO concentration 0.33%). Only wells that passed the pass criteria (seal resistance 30 megaohms, Z prime >0.4, and current size >0.2 nA) were used for this analysis. IC of the hERG assay 50 The results (μM) are reported in Table 2.

[0324] Solubility (Test E) The assay was performed according to the solubility assay described in Wernevik, J. et al., "A Fully Integrated Assay Panel for Early Drug Metabolism and Pharmacokinetics Profiling," Assay and Drug Development Technologies, 2020, 18(4), 157-179, pp. 164-167. Data are reported in Table 2 as solubility (μM). When assays were performed multiple times for the same compound, arithmetic mean values ​​are reported.

[0325] Solubility (Test F) After drying, a 20 mM DMSO solution containing the test compound was diluted 100-fold with disodium hydrogen phosphate-citrate buffer (diluted McIlvaine buffer, pH 6.5). Under these conditions, the theoretical maximum concentration of the test compound was 200 μM. The buffer solution was sonicated, shaken, and stored at 25°C for 24 to 72 hours. The buffer sample was filtered, and the filtrate was diluted with an equal volume of acetonitrile / methanol (1:1, v / v) in a 96-well plate. A 20 mM DMSO solution containing the test compound was diluted 100-fold with acetonitrile / methanol (1:1, v / v) and an equal volume of McIlvaine buffer (pH 6.5) was added to use as a standard solution. The standard and test samples were transferred to a 384-well plate and analyzed by HPLC. The results of the solubility assay are reported in μg / mL in Table 2.

[0326] [Table 2]

[0327] LPS / ATP test Seven-week-old male BALB / cAJcl mice were intraperitoneally administered 0.5 mL of a 4 μg / mL LPS (Sigma-Aldrich Co., LLC, L2630) solution in PBS (Thermo Fisher Scientific Inc., 10010). One hour later, a test article suspension in 0.5% (w / v) aqueous CMC sodium (Nacalai Tesque Inc., 07326-95) was orally administered at a volume of 10 mL / kg. One hour after test article administration, 0.5 mL of a 30 μmol / L ATP (Sigma-Aldrich Co., LLC, A7699) solution in PBS was intraperitoneally administered. Twenty minutes later, the animals were euthanized by cervical dislocation under sevoflurane anesthesia. Immediately after euthanasia, the peritoneal cavity of each animal was washed with 3 mL of ice-cold PBS injected intraperitoneally. The PBS was then collected and the concentration of IL-1β was measured using an ELISA kit (R&D Systems Inc., MLB00C). The test results are shown in Table 3.

[0328] [Table 3]

[0329] Those skilled in the art will appreciate that the above biological assays can be performed using alternative equipment and minor modifications to the protocols without significantly affecting the results.

[0330] Any publications disclosed within this specification are hereby incorporated by reference.

Claims

1. A crystalline form of (2S)-3-[[4-[2-hydroxy-4-(trifluoromethyl)phenyl]phthalazin-1-yl]amino]propane-1,2-diol (Compound 1).

2. 2. The crystalline form of claim 1, which is crystalline form A.

3. An X-ray powder diffraction pattern (CuK) with at least five specific peaks at approximately 2-theta = 13.0, 19.3, 19.8, 22.7, and 24.1° α 2. The crystalline form of claim 1, having a crystalline structure (e.g., a crystalline form of a crystalline substance) that is irradiated with radiation.

4. X-ray powder diffraction pattern (CuK) with at least five specific peaks at 2-theta = 13.0, 19.3, 19.8, 22.7 and 24.1° α 2. The crystalline form of claim 1, having a .alpha.-ray peak (X) and ...

5. The X-ray powder diffraction pattern (CuK) is substantially the same as the X-ray powder diffraction pattern shown in FIG. α 2. The crystalline form of claim 1, having a crystalline structure (e.g., a crystalline form of a crystalline substance) that is irradiated with radiation.

6. 2-(4-(((1R,2R)-2-hydroxycyclohexyl)amino)pyrido[3,4-d]pyridazin-1-yl)-5-(trifluoromethyl)phenol (Compound 2), a crystalline form of the hydrochloride salt.

7. A pharmaceutical composition comprising the crystalline form of any one of claims 1 to 6 and a pharmaceutically acceptable excipient.

8. A crystalline form or pharmaceutical composition according to any one of claims 1 to 7 for use in therapy.

9. 8. The crystalline form or pharmaceutical composition of any one of claims 1 to 7 for use in treating a subject having a disease or condition in which NLRP3 inflammasome activity is implicated.

10. Renal diseases such as acute kidney injury, chronic kidney disease, and diabetic kidney disease; cardiovascular diseases such as coronary atherosclerotic heart disease, cardiomyopathy, myocardial infarction, cardiac hypertrophy, and ischemia-reperfusion injury; liver diseases such as non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, alcoholic steatohepatitis, chronic hepatitis C virus infection, and paracetamol-induced liver injury; autoinflammatory disorders, cryopyrin-associated periodic syndrome, familial cold autoinflammatory syndrome (FCAS), Muckle-Wells syndrome (MWS), chronic infantile neurological, cutaneous, and articular (CINCA) syndrome, and neonatal-onset multisystem inflammatory disease (NOMI).

10. The crystalline form or pharmaceutical composition of any one of claims 1 to 7 for use in treating a subject having a disease or condition selected from the group consisting of: inflammatory diseases such as hidradenitis pneumoniae (HIP), inflammatory skin diseases such as acne vulgaris and hidradenitis suppurativa; inflammatory bowel diseases such as ulcerative colitis (UC) and Crohn's disease; autoimmune diseases such as gout, pseudogout, rheumatoid arthritis (RA), multiple sclerosis (MS), Addison's disease, celiac disease, systemic lupus erythematosus (SLE), and vitiligo; and respiratory diseases such as chronic lung disease, idiopathic pulmonary fibrosis (IPF), chronic obstructive pulmonary disease (COPD), and asthma.

11. 10. A method of inhibiting NLRP3 inflammasome activity in a subject in need thereof, comprising administering to the subject a crystalline form or pharmaceutical composition of any one of claims 1 to 7.

12. A process for the preparation of a compound of formula (Ic) or a salt thereof, comprising: 【Chemistry 1】 (a) reacting 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran with an alkyllithium, optionally n-BuLi, to form (2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)lithium; (b) optionally adding magnesium chloride or magnesium bromide to the reaction product of (a); (c) reacting the product of (a) or (b) with a compound of formula (Ia): 【Chemistry 2】 forming a compound of formula (Ib) or a salt thereof; 【Transformation 3】 (d) reacting the compound of formula (Ib) or a salt thereof with hydrazine, optionally in the presence of an acid such as acetic acid, to form a compound of formula (Ic) or a salt thereof; (e) isolating the compound of formula (Ic) or a salt thereof, The process wherein in formula (Ia), formula (Ib) and formula (Ic), X=CH or N.

13. 1. A process for the preparation of 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof, comprising: (a) reacting 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran with an alkyllithium, optionally n-BuLi, to form (2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)lithium; (b) optionally adding magnesium chloride or magnesium bromide to the reaction product of (a); (c) reacting the product of (a) or (b) with phthalic anhydride to form 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof; (d) reacting 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof with aqueous acid to form 2-[2-hydroxy-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof; (e) reacting 2-[2-hydroxy-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof with hydrazine to form 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof; (f) isolating 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof.

14. 1. A process for the preparation of 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof, comprising: (a) reacting 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran with an alkyllithium, optionally n-BuLi, to form (2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)lithium; (b) optionally adding magnesium chloride or magnesium bromide to the reaction product of (a); (c) reacting the product of (a) or (b) with phthalic anhydride to form 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof; (d) reacting 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof with hydrazine to form 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof; (e) reacting 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof with an acid to form 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof; (f) isolating 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof.

15. 1. A process for the preparation of 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof, comprising: (a) 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof is reacted with POCl ３ to form 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof; (b) isolating 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof.

16. 16. The process of claim 15, wherein the 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof used in step (a) is prepared by the process of claim 13.

17. 16. The process of claim 15, wherein the 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof used in step (a) is prepared by the process of claim 14.

18. 1. A process for the preparation of Compound 1 or a salt thereof, comprising: (a) reacting 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran with an alkyllithium, optionally n-BuLi, to form (2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)lithium; (b) optionally adding magnesium chloride or magnesium bromide to the reaction product of (a); (c) reacting the product of (a) or (b) with phthalic anhydride to form 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof; (d) reacting 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof with aqueous acid to form 2-[2-hydroxy-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof; (e) reacting 2-[2-hydroxy-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof with hydrazine to form 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof; (f) 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof with POCl ３ to form 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof; (g) reacting 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof with (1) (2S)-3-amino-1,2-propanediol or a salt thereof to form Compound 1 or a salt thereof, or (2) diol-protected (2S)-3-amino-1,2-propanediol or a salt thereof, followed by deprotection of the resulting product to form Compound 1 or a salt thereof; (h) isolating Compound 1 or a salt thereof.

19. 1. A process for the preparation of Compound 1 or a salt thereof, comprising: (a) reacting 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran with an alkyllithium, optionally n-BuLi, to form (2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)lithium; (b) optionally adding magnesium chloride or magnesium bromide to the reaction product of (a); (c) reacting the product of (a) or (b) with phthalic anhydride to form 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof; (d) reacting 2-[2-(tetrahydro-2H-pyran-2-yloxy)-4-(trifluoromethyl)benzoyl]benzoic acid or a salt thereof with hydrazine to form 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof; (e) reacting 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof with aqueous acid to form 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof; (f) 4-(2-hydroxy-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof with POCl ３ to form 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof; (g) reacting 2-(4-chlorophthalazin-1-yl)-5-(trifluoromethyl)phenol or a salt thereof with (1) (2S)-3-amino-1,2-propanediol or a salt thereof to form Compound 1 or a salt thereof, or (2) diol-protected (2S)-3-amino-1,2-propanediol or a salt thereof, followed by deprotection of the resulting product to form Compound 1 or a salt thereof; (h) isolating Compound 1 or a salt thereof.

20. 1. A process for the preparation of Compound 1 or a salt thereof, comprising: (a) reacting 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran with an alkyllithium, optionally n-BuLi, to form (2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)lithium; (b) optionally adding magnesium chloride or magnesium bromide to the reaction product of (a); (c) reacting the product of (a) or (b) with tert-butyl methyl phthalate to form tert-butyl 2-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)benzoyl)benzoate; (d) reacting tert-butyl 2-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)benzoyl)benzoate with hydrazine, optionally in the presence of an acid such as acetic acid, to form 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof; (e) reacting 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)phthalazin-1(2H)-one or a salt thereof with diol-protected (2S)-3-amino-1,2-propanediol or a salt thereof in the presence of a coupling reagent, followed by removal of the THP and diol-protecting groups from the resulting product by reaction with an acid to form compound 1 or a salt thereof; (f) isolating Compound 1 or a salt thereof.

21. 1. A process for the preparation of compound 2 or a salt thereof, comprising: (a) reacting 2-(3-(trifluoromethyl)phenoxy)tetrahydro-2H-pyran with an alkyllithium, optionally n-BuLi, to form (2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)lithium; (b) optionally adding magnesium chloride or magnesium bromide to the reaction product of (a); (c) reacting the product of (a) or (b) with 3-(tert-butyl) 4-methylpyridine-3,4-dicarboxylate to form tert-butyl 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)benzoyl)nicotinate or a salt thereof; (d) reacting tert-butyl 4-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)benzoyl)nicotinate or a salt thereof with hydrazine, optionally in the presence of an acid such as acetic acid, to form 1-[2-tetrahydropyran-2-yloxy-4-(trifluoromethyl)phenyl]-3H-pyrido[3,4-d]pyridazin-4-one or a salt thereof; (e) reacting 1-[2-tetrahydropyran-2-yloxy-4-(trifluoromethyl)phenyl]-3H-pyrido[3,4-d]pyridazin-4-one with (1R,2R)-2-aminocyclohexan-1-ol or a salt thereof in the presence of a coupling reagent to form (1R,2R)-2-((1-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)pyrido[3,4-d]pyridazin-4-yl)amino)cyclohexan-1-ol or a salt thereof; (f) removing the THP protecting group from (1R,2R)-2-((1-(2-((tetrahydro-2H-pyran-2-yl)oxy)-4-(trifluoromethyl)phenyl)pyrido[3,4-d]pyridazin-4-yl)amino)cyclohexan-1-ol, or a salt thereof, by reaction with an acid to form compound 2, or a salt thereof; (g) isolating compound 2 or a salt thereof. 【Request Item 22】 【Chemistry 4】 or a salt thereof. 【Request Item 23】 【Chemistry 5】 or a salt thereof. 【Request Item 24】 【Chemistry 6】 or a salt thereof. 【Request Item 25】 【Chemistry 7】 or a salt thereof. 【Request Item 26】 【Chemistry 8】 or a salt thereof, wherein X is CH or N. 【Request Item 27】 【Chemistry 9】 or a salt thereof, wherein X is CH or N. 【Request Item 28】 【Chemistry 10】 or a salt thereof, wherein X is CH or N. 【Request Item 29】 【Chemistry 11】 or a salt thereof. 【Request Item 30】 【Chemistry 12】 or a salt thereof. 【Request Item 31】 【Chemistry 13】 or a salt thereof. 【Request Item 32】 【Chemistry 14】 or a salt thereof.

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