Crystalline form of a pyridazine NLRP3 inhibitor

Abstract

The present disclosure relates to a compound of formula (I) which is in crystalline Form 1, characterized by having a powder X-ray diffractogram displaying peaks expressed as degree 2-Theta angles at about 3.4, 6.8, 10.3, 13.7, and 20.5. The present disclosure also relates to processes for its preparation, as well as a medicament and a pharmaceutical composition comprising it. The present disclosure further concerns the crystalline Form 1 of compound of formula (I) for use as a medicine and more particularly in the prevention and / or in the treatment of Parkinson's disease, frontotemporal dementia, multiple system atrophy, Alzheimer's disease, multiple sclerosis, amyotrophic lateral sclerosis, or brain injury.

Description

[0001] CRYSTALLINE FORM OF A PYRID AZINE NLRP3 INHIBITOR

[0002] FIELD OF THE INVENTION

[0003] Disclosed herein is 2-[6-[(2S)-2-(hydroxymethyl)morpholin-4-yl]pyridazin-3-yl]-3,5- dimethyl-phenol (hereinafter designated as Compound 1) which is in crystalline Form 1. Herein are also provided processes for its preparation, a medicament and a pharmaceutical composition comprising said crystalline Form 1, and this crystalline Form 1 for use as a medicine, and particularly its use as inhibitor of the NOD-like receptor protein 3 (NLRP3) inflammasome pathway, and more particularly its use in the prevention and / or in the treatment of Parkinson’s disease, frontotemporal dementia, multiple system atrophy, Alzheimer’s disease, multiple sclerosis, amyotrophic lateral sclerosis, or brain injury.

[0004] BACKGROUND OF THE INVENTION

[0005] The NOD-like receptor (NLR) family, pyrin domain-containing protein 3 (NLRP3) or NACHT, LRR and PYD domains-containing protein 3 (NALP3), is a cytosolic sensor of diverse pathogen- and host-derived molecules. Upon activation, NLRP3 oligomerizes and recruits an adaptor protein called apoptosis-associated speck like protein (ASC). ASC then polymerizes to form a large aggregate known as ASC speck. In turn, polymerized ASC interacts with the cysteine protease caspase- 1 to form a complex termed the inflammasome. This multicomplex protein forms a platform for the binding, dimerization, and activation of the caspase- 1 protease. Caspase- 1 then cleaves the precursor forms of the pro-inflammatory cytokines ILip and IL 18 (termed pro-ILip and pro-IL18) and thereby activates adapted inflammatory responses. However, this pathway was shown to be associated with various inflammation associated processes and diseases, including:

[0006] - neurodegenerative diseases such as Parkinson’s disease (PD), multiple system atrophy (MSA), Alzheimer’s disease (AD), frontotemporal dementia (FTD), multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS) and brain injury (Guan Y & Han F. Front. Integr., Neurosci. 14 :37, 2020);

[0007] - inflammatory diseases including Muckle-Wells autoinflammatory disorder (Agostini et al., 2004), cryopyrin-associated periodic syndrome (CAPS) (Mortimer et al., Nature Immunol. 2016, 17(10), 1176-1188); sickle cell disease; systemic lupus erythematosus (SLE); liver related diseases, viral hepatitis, non-alcoholic steatohepatitis (NASH), alcoholic steatohepatitis, and alcoholic liver disease (Petrasek et al., J. Clin. Invest. 2012, 122, 3476- 89), and inflammatory arthritis related disorders, such as gout, pseudogout (chondrocalcinosis), osteoarthritis (Ridker et al., N. Engl. J. Med. 2017, 377, 1119-31), and rheumatoid arthritis (Mathews et al., Ann. Rheum. Dis. 2014, 73, 1202-10), acute or chronic arthropathy, and kidney related diseases such as hyperoxaluria (Knaufet et al., Kidney Int. 2013, 84, 895-901), lupus nephritis, hypertensive nephropathy (Krishnan et al., Br. J. Pharmacol. 2016, 173, 752-10 65), hemodialysis related inflammation and diabetic nephropathy (Shahzad et al., Kidney Int. 2015, 87, 74-84);

[0008] - obesity and insulin resistance (Rheinheimer J. et al., Metabolism Clin & Experimental 74: 1-9, 2017), pancreatitis (Fu Q. et al., BioMed Research International Volume 2018, Article ID 12949512018), myocarditis (Toldo S et al, Int J Cardiol 2014);

[0009] - eye diseases, where the NLRP3 inflammasome has been shown to contribute to diabetic retinopathy (Perrone L. et al., J. Cell. Physiol. 221 : 262-272, 2009), acute glaucoma (Chi W. et al. National Academy Science 111 : 11181-11186, 2014), age-related macular degeneration (Tseng W.A. et al., Investigative Ophthal & Visual Science 54: 11-120, 2013), Behcet’s syndrome and dry eye disease (Zheng Q. et al., Experimental Eye Research 134: 133-140, 2015);

[0010] - metabolic, cardiac, skin disorder and cancer, e.g., diabetic cardiomyopathy (Luo B. et al., PLoS ONE 9(8): el04771, 2014), Kawasaki disease (Jia et al. Cell Death and Disease 10:778; 2019; Anzai F. et al., J. Molecular & Cell Cardiology 138: 185-196, 2020), cardiovascular metabolic disorders, atherosclerosis, type I and type II diabetes and related complications, peripheral artery disease (PAD), acute heart failure and hypertension (Ridker et al., N. Engl. J. Med. 2017, 377, 1119-31), wound healing and scar formation, inflammatory skin diseases (Sweeney et al., Br. J. Dermatol. 2015, 173, 1361), asthma, sarcoidosis, age-related macular degeneration, cancer related diseases, e.g., myeloproliferative neoplasms, leukemias, myelodysplastic syndromes (MDS), myelofibrosis, lung cancer, colon cancer (Ridker et al., Lancet 2017, 390, 1833-42); and

[0011] - SARS-Cov-2: NLRP3 inflammasome is a key player in antiviral responses (Zhao C. and Zhao W. 11, 211 : 2020; Freeman & Swartz, Frontiers in Immunol. 11, 1518, 2020).

[0012] Inhibitors of NLRP3 are potential treatments for these conditions with unmet clinical needs. Therefore, there is a need for inhibitors of the NLRP3 inflammasome pathway to provide new or alternative treatments.

[0013] 2-[6-[(2S)-2-(hydroxymethyl)morpholin-4-yl]pyridazin-3-yl]-3,5-dimethyl-phenol

[0014] (Compound 1), of formula (I), depicted below, is an inhibitor of the NLRP3 inflammasome pathway. It may be used in particular as Parkinson’s disease, frontotemporal dementia, multiple system atrophy, Alzheimer’s disease, multiple sclerosis, amyotrophic lateral sclerosis, or brain injury treatment agent.

[0015] Besides its pharmaceutical efficacy, a pharmaceutically active agent has to comply with a variety of additional requirements. For instance, its stability under various environmental conditions, its stability during production of the pharmaceutical formulation or its stability in the final medicament compositions. In addition, when a pharmaceutically active agent is used to prepare a pharmaceutical composition, it should be as pure as possible and its stability in long-term storage must be guaranteed under various environmental conditions. For example, this reduces or avoids the risk that the content of active substance in the medicament be less than that specified.

[0016] Polymorphism occurs where the same chemical entity crystallizes in a different lattice arrangement, resulting in different thermodynamic properties and stabilities specific to the particular polymorphic form. When the chemical entity is a drug, the ability of the chemical entity to exist in more than one crystal form can have a profound effect on the shelf life (stability), solubility, formulation properties, and / or processing properties of the drug. It is thus very important to be able to ensure, from a quality standpoint, that the manufacturing process leads to the specific polymorphic form whose marketing is allowed by regulatory agencies and that formation of other polymorphic forms, with different thermodynamic properties and stabilities, are controlled.

[0017] Further, the availability of a well-defined crystalline form allows the purification of the drug substance by recrystallization. Hence, there is a need to provide the compound of formula (I) under a form which is the most thermodynamically stable form at least under ambient conditions of temperature and pressure and which allows its use and storage at an industrial scale.

[0018] The disclosure relates to a stable crystalline form of the compound of formula (I) which meets the important above-mentioned features.

[0019] ABBREVIATIONS AND DEFINITIONS

[0020] The following abbreviations are used:

[0021] ACN Acetonitrile

[0022] Ar Argon

[0023] DCM Dichloromethane

[0024] DIPEA Diisopropylethylamine

[0025] DMSO-d6Hexadeuterodimethyl sulfoxide

[0026] EtOH Ethanol

[0027] EtOAc Ethylacetate

[0028] H2O Water

[0029] IL-ip Interleukin 1 beta

[0030] LC Liquid chromatography

[0031] LCMS Liquid chromatography / mass spectrometry

[0032] Me-THF 2-methyl tetrahydrofuran

[0033] MeOH Methanol min Minute(s)

[0034] MS Mass Spectrometry

[0035] Na2SO4Sodium sulfate

[0036] NMP N-methyl-2-pyrrolidone

[0037] Pd(PPh3)4Tetrakis(triphenylphosphine)palladium

[0038] Rt Retention time

[0039] THF Tetrahydrofuran

[0040] TNF-a Tumor necrosis factor-a

[0041] °C Degree Celsius mL Milliliter(s) mmol Millimole(s) min Minute(s) h Hour(s)

[0042] A « solvate » refers to a crystalline form with one or more solvent molecules included in the lattice structure.

[0043] « The amorphous phase of a compound » is a solid that lacks the long-range order that is characteristic of a crystal. Consequently, the X-ray diffraction pattern of an amorphous phase does not show diffraction peaks.

[0044] The term « crystalline » refers to any solid substance exhibiting three-dimensional order, which in contrast to an amorphous solid substance, gives a distinctive XRPD pattern with more or less sharp peaks.

[0045] The term « anhydrate » refers to a crystal form of a substance with no water in its structure. By extension, the term « anhydrate » usually refers to a crystal form of a substance with no water and / or solvent in its structure.

[0046] The term « heterosolvate » refers to a crystalline form with more than one type of solvent included in the lattice structure.

[0047] The term « pharmaceutically acceptable » means that which is useful in preparing a pharmaceutical composition that is generally safe, non-toxic, and neither biologically nor otherwise undesirable and includes what is acceptable for veterinary as well as human pharmaceutical use.

[0048] As used herein, « pharmaceutically acceptable excipients » include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like. Except insofar as any conventional excipient is incompatible with the active compounds, its use in a medicament or pharmaceutical composition of the invention is contemplated.

[0049] SUMMARY OF THE INVENTION

[0050] Herein is provided 2-[6-[(2S)-2-(hydroxymethyl)morpholin-4-yl]pyridazin-3-yl]-3,5- dimethyl-phenol (hereinafter designated as Compound 1) in free form in crystalline Form 1, characterized by having a powder-X-ray diffractogram displaying peaks expressed as degree 2-Theta angles at about 3.4, 6.8, 10.3, 13.7, and 20.5 (each time ± 0.2), which optionally further displays the following peaks expressed as degree 2-Theta angles at about: 15.0, 16.1, 17.1, 17.5, 17.9, and 25.3 (each time ± 0.2), optionally further characterized by a powder X- ray diffractogram as substantially illustrated in Figure 1 or Figure 2.

[0051] Herein are further provided processes for the preparation of the crystalline Form 1 of the compound of formula (I).

[0052] Herein are also provided medicaments comprising the crystalline Form 1 of Compound 1, and pharmaceutical compositions comprising the crystalline Form 1 of Compound 1 and at least one pharmaceutically acceptable excipient.

[0053] Herein are further disclosed the crystalline Form 1 of Compound 1 for use as a medicine, for use as an inhibitor of the NOD-like receptor protein 3 (NLRP3) inflammasome pathway, and for use in the prevention and / or in the treatment of Parkinson’s disease, frontotemporal dementia, multiple system atrophy, Alzheimer’s disease, multiple sclerosis, amyotrophic lateral sclerosis, or brain injury.

[0054] Herein is further disclosed the use of the crystalline Form 1 of Compound 1 for the manufacture of a medicament for the prevention and / or the treatment of a disease involving inhibition of the NOD-like receptor protein 3 (NLRP3) inflammasome pathway.

[0055] Herein is further disclosed the use of the crystalline Form 1 of Compound 1 for the manufacture of a medicament for the prevention and / or the treatment of Parkinson’ s disease, frontotemporal dementia, multiple system atrophy, Alzheimer’s disease, multiple sclerosis, amyotrophic lateral sclerosis, or brain injury.

[0056] Herein is further disclosed a method of preventing and / or treating a disease involving inhibition of the NOD-like receptor protein 3 (NLRP3) inflammasome pathway, comprising administering to a subject in need thereof a therapeutically effective amount of the crystalline Form 1 of Compound 1.

[0057] Herein is further disclosed a method of preventing and / or treating Parkinson’s disease, frontotemporal dementia, multiple system atrophy, Alzheimer’s disease, multiple sclerosis, amyotrophic lateral sclerosis, or brain injury, comprising administering to a subject in need thereof a therapeutically effective amount of the crystalline Form 1 of Compound 1. BRIEF DESCRIPTION OF THE DRAWINGS

[0058] Figure 1 is an X-ray powder diffractogram of crystalline Form 1 of 2-[6-[(2S)-2- (hydroxymethyl)morpholin-4-yl]pyridazin-3-yl]-3,5-dimethyl-phenol measured at room temperature (see Example 2).

[0059] Figure 2 is an X-ray powder diffractogram of crystalline Form 1 of 2-[6-[(2S)-2- (hydroxymethyl)morpholin-4-yl]pyridazin-3-yl]-3,5-dimethyl-phenol measured at room temperature and focused on low relative intensities (see Example 2).

[0060] Figure 3 is a Differential Scanning Calorimetry (DSC) thermogram of crystalline Form 1 of 2-[6-[(2S)-2-(hydroxymethyl)morpholin-4-yl]pyridazin-3-yl]-3,5-dimethyl-phenol (see Example 3).

[0061] Figure 4 is a Thermogravimetric Analysis (TGA) thermogram of crystalline Form 1 of 2- [6-[(2S)-2-(hydroxymethyl)morpholin-4-yl]pyridazin-3-yl]-3,5-dimethyl-phenol (see Example 4).

[0062] DETAILED DESCRIPTION

[0063] Crystalline Form of the invention

[0064] As explained above, herein is provided a crystalline form, which is Form 1 of 2-[6-[(2S)-2- (hydroxymethyl)morpholin-4-yl]pyridazin-3-yl]-3,5-dimethyl-phenol (Compound 1) of formula I: characterized by having a powder X-ray diffractogram displaying peaks expressed as degree

[0065] 2-Theta angles at about 3.4, 6.8, 10.3, 13.7, and 20.5 (each time ± 0.2), which optionally further shows the following peaks expressed as degree 2-Theta angles at about 15.0, 16.1, 17.1, 17.5, 17.9, and 25.3 (each time ± 0.2), optionally further characterized by a powder X- ray diffractogram as substantially illustrated in Figure 1 or Figure 2. More particularly, a characteristic X-ray powder diffractogram of the crystalline Form 1 of Compound 1 can be given substantially in Figure 1 or Figure 2, and its characteristic signals are summarized in the following Table I:

[0066] Table

[0067] According to a preferred embodiment, the crystalline Form 1 of Compound 1 presents a powder X-ray diffractogram displaying at least one peak, in particular at least two peaks, preferably at least five peaks, and more preferably at least ten peaks, expressed as degree 2-Theta angle, selected from 3.4, 6.8, 10.3, 13.7, 15.0, 16.1, 17.1, 17.5, 17.9, 20.5, and 25.3 (each time ± 0.2).

[0068] In one embodiment, the crystalline Form 1 of Compound 1 has a differential scanning calorimetry (DSC) thermogram showing a melting endotherm at about 179 °C (± 2 °C) onset temperature and is optionally further characterized by a thermogram as substantially illustrated in Figure 3. As indicated in Figure 3, this melting point temperature is associated with a relatively high enthalpy of fusion AHf(126 J / g).

[0069] Moreover, advantageously, exposure to temperature variations does not alter the crystal structure of crystalline Form 1 of Compound 1 before its melting. In some embodiments, crystalline Form 1 has a TGA pattern with less than 1% weight loss up to 240 °C. In some embodiments, crystalline Form 1 of Compound 1 has a thermogravimetric analysis (TGA) thermogram substantially as shown in Figure 4.

[0070] According to a preferred embodiment, the crystalline Form 1 of Compound 1 is an anhydrate.

[0071] Advantageously, crystalline Form 1 of Compound 1 shows no decomposition up to 240 °C.

[0072] Preparation of the Crystalline Form

[0073] Herein are also provided processes for preparing crystalline Form 1 of 2-[6-[(2S)-2- (hydroxymethyl)morpholin-4-yl]pyridazin-3-yl]-3,5-dimethyl-phenol (Compound 1).

[0074] Crystalline Form 1 of Compound 1 may be obtained by conventional crystallization techniques known to one of skill in the art, such as crystallization by evaporation, crystallization by cooling, or crystallization by adding a non-solvent such as water or heptane.

[0075] In the sense of the present disclosure, a “set temperature” means a temperature which remains the same during the corresponding step.

[0076] In the context of the present disclosure, the expression “almost complete evaporation” of a solvent means that the evaporation is not carried out in full, that is to say that the amount of solvent which is evaporated is decreased but nevertheless still present in a very low content. In other terms, the evaporation must not be carried out dry.

[0077] Crystallization by evaporation

[0078] According to one embodiment, a process for the preparation of crystalline Form 1 of Compound 1 comprises at least the following steps:

[0079] 1) solubilizing Compound 1 in a solvent selected from alcohols, chlorinated hydrocarbons, ketones, acetates, ethers, acetonitrile, and mixtures thereof, optionally in admixture with water, at a set temperature ranging from 18 °C to 80 °C;

[0080] 2) leaving the solution obtained in step 1) at the same temperature as the one set in step 1) for almost complete evaporation;

[0081] 3) isolating the crystalline Form 1 of Compound 1 formed in step 2). In a particular embodiment, the solvent of step 1) is selected from methanol, methanol / water mixture, ethanol, ethanol / water mixture, 2-propanol, 1 -propanol, 1-propanol / water mixture,

[0082] 1 -butanol, 1-butanol / water mixture, di chloromethane, 1,2-di chloropropane, chloroform, 1,1,1 -trichloroethane, tetrachloroethylene, trichloroethylene, carbon tetrachloride, dichloromethane / methanol / acetonitrile mixture, acetone, acetone / water mixture, 2-butanone (also named methyl ethyl ketone or MEK), 2-butanone / water mixture, methyl isobutyl ketone (also named MIBK), methyl isobutyl ketone / water mixture, methyl acetate, methyl acetate / water mixture, ethyl acetate, ethyl acetate / water mixture, isopropyl acetate, isopropyl acetate / water mixture, isobutyl acetate, isobutyl acetate / water mixture, acetonitrile, acetonitrile / water mixture, tetrahydrofuran, and methyl tert-butyl ether (also named MTBE). In a more particular embodiment, the solvent of step 1) is selected from methanol, methanol / water mixture, ethanol, ethanol / water mixture, 2-propanol, 1 -propanol, 1- propanol / water mixture, 1-butanol / water mixture, dichloromethane, dichloromethane / methanol / acetonitrile mixture, acetone, acetone / water mixture,

[0083] 2-butanone, 2-butanone / water mixture, methyl isobutyl ketone / water mixture, methyl acetate, methyl acetate / water mixture, ethyl acetate, ethyl acetate / water mixture, isopropyl acetate / water mixture, isobutyl acetate, isobutyl acetate / water mixture, tetrahydrofuran, and acetonitrile.

[0084] In still another embodiment, the solvent of step 1) is selected from methanol, ethanol, 1- propanol, 2-propanol, 1 -butanol, dichloromethane / methanol / acetonitrile mixture, 2- butanone, methyl isobutyl ketone, methyl acetate, ethyl acetate, isopropyl acetate, isobutyl acetate, acetonitrile, methanol / water mixture, ethanol / water mixture, 1-propanol / water mixture, 1-butanol / water mixture, 2-butanone / water mixture, methyl isobutyl ketone / water mixture, methyl acetate / water mixture, ethyl acetate / water mixture, isopropyl acetate / water mixture, isobutyl acetate / water mixture, tetrahydrofuran, and methyl isobutyl ketone / water mixture.

[0085] When the solvent of step 1) is in admixture with water, the solvent / water volume ratio is typically 99 / 1.

[0086] According to another embodiment, the set temperature of step 1) and step 2) is selected from a range of temperature from 20 °C to 80 °C, or even from 25 °C to 80 °C. Crystallization by cooling

[0087] According to another embodiment, a process for the preparation of crystalline Form 1 of Compound 1 comprises at least the following steps:

[0088] 1) solubilizing or suspending Compound 1 in amorphous form in a solvent selected from alcohols, ketones, acetates, ethers and acetonitrile, at a set temperature which is room temperature;

[0089] 2) optionally purifying the solution or suspension obtained in step 1) by heating at a set temperature ranging from 60 °C to 80 °C, stirring and filtering the solution or suspension;

[0090] 3) heating the solution or suspension obtained in step 1) or in step 2) at a set temperature ranging from 60 °C to 80 °C;

[0091] 4) cooling the solution or suspension obtained in step 2) to a set temperature ranging from -20 °C to 25 °C;

[0092] 5) isolating the crystalline Form 1 of Compound 1 formed in step 4).

[0093] The optional step 2) allows advantageously the elimination of impurities or seeds that may be present in the solution or suspension.

[0094] In a particular embodiment, the solvent of step 1) is selected from ethanol, 2-propanol, 1- propanol, 1 -butanol, acetone, 2-butanone (also named methyl ethyl ketone or MEK), methyl isobutyl ketone (also named MIBK), isopentyl methyl ketone (also named MIAK), methyl acetate, ethyl acetate, isopropyl acetate, isobutyl acetate, acetonitrile, and methyl tert-butyl ether (also named MTBE).

[0095] In a more particular embodiment, the solvent of step 1) is ethanol or 1-butanol.

[0096] In an embodiment, the solvent of step 1) is ethanol.

[0097] According to one variant, the set temperature of step 1) is the same as the one of step 4). According to another variant, the set temperature of step 1) is the same as the one of step 4) and the set temperature of step 2) is the same as the one of step 3).

[0098] Crystallization by adding water as a non-solvent.

[0099] According to another embodiment, a process for the preparation of the crystalline Form 1 of Compound 1 comprises at least the following steps:

[0100] 1) solubilizing or suspending Compound 1 in amorphous form in a solvent selected from alcohols, ketones, acetates, and diethyl-ether, at a set temperature which is room temperature; 2) optionally filtering the solution or suspension obtained in step 1);

[0101] 3) adding water as a non-solvent;

[0102] 4) isolating the crystalline Form 1 of Compound 1 formed in step 3).

[0103] The optional step 2) of filtering allows advantageously to remove impurities or seeds that may be present in the solution or suspension.

[0104] In a particular embodiment, the solvent of step 1) is selected from 2-propanol, 1-propanol, 1 -butanol, acetone, 2-butanone, methyl isobutyl ketone, isopentyl methyl ketone, methyl acetate, ethyl acetate, isopropyl acetate, isobutyl acetate, and diethyl ether.

[0105] Optionally, after step 3) the solvent is left to evaporate.

[0106] Crystallization by adding heptane as a non-solvent.

[0107] According to another embodiment, a process for the preparation of the crystalline Form 1 of Compound 1, comprises at least the following steps:

[0108] 1) solubilizing or suspending Compound 1 in amorphous form in a solvent selected from alcohols, ketones, and acetates, at a set temperature which is room temperature;

[0109] 2) optionally filtering the solution or suspension obtained in step 1);

[0110] 3) adding heptane as a non-solvent;

[0111] 4) isolating the crystalline Form 1 of Compound 1 formed in step 3).

[0112] The optional step 2) of filtering allows advantageously to remove impurities or seeds that may be present in the solution or suspension.

[0113] In a particular embodiment, the solvent of step 1) is selected from ethanol, 2-propanol, 1- propanol, 1 -butanol, acetone, 2-butanone, methyl isobutyl ketone, isopentyl methyl ketone, methyl acetate, ethyl acetate, isopropyl acetate, and isobutyl acetate.

[0114] Pharmaceutical Compositions

[0115] According to another of its aspects, the present invention relates to a pharmaceutical composition comprising at least one crystalline form according to the invention and at least one pharmaceutically acceptable excipient.

[0116] The crystalline form according to the invention may be used for the preparation of medicaments, in particular of medicaments for inhibiting the the NOD-like receptor protein 3 (NLRP3) inflammasome pathway. More particularly, the crystalline form according to the invention may be used for the preparation of medicaments for the prevention and / or treatment of Parkinson’s disease, frontotemporal dementia, multiple system atrophy, Alzheimer’s disease, multiple sclerosis, amyotrophic lateral sclerosis, or brain injury.

[0117] The pharmaceutical compositions may contain more particularly an effective dose of at least one crystalline form according to the invention.

[0118] An “effective dose'' means an amount sufficient to induce a positive modification in the condition to be regulated or treated, but low enough to avoid serious side effects. An effective amount may vary with the pharmaceutical effect to obtain or with the particular condition being treated, the age and physical condition of the end user, the severity of the condition being treated / prevented, the duration of the treatment, the nature of other treatments, the specific compound or composition employed, the route of administration, and like factors.

[0119] The crystalline form according to the invention may be administered in an effective dose by any of the accepted modes of administration in the art.

[0120] In one embodiment, the crystalline form of the invention may be used in a composition intended to be administrated by oral, nasal, sublingual, aural, ophthalmic, topical, rectal, vaginal, urethral, or parenteral injection route.

[0121] The route of administration and the galenic formulation will be adapted by one skilled in the art pursuant to the desired pharmaceutical effect.

[0122] In a preferred embodiment, the crystalline form of the invention may be used in a composition intended to be administrated by oral route.

[0123] One of ordinary skill in the art of therapeutic formulations will be able, without undue experimentation and in reliance upon personal knowledge, to ascertain a therapeutically effective dose of the crystalline form of the invention for a given indication.

[0124] A pharmaceutical composition of the invention may be formulated with any known suitable pharmaceutically acceptable excipients according to the dose, the galenic form, the route of administration and the likes.

[0125] A medicament or pharmaceutical composition of the invention may be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, sprays, ointments, gels, creams, sticks, lotions, pastes, soft and hard gelatine capsules, suppositories, sterile injectable solutions, sterile packages powders and the like. The applications also include a novel kit-of-parts that is suitable for use in the prevention and / or treatment of Parkinson’s disease, frontotemporal dementia, multiple system atrophy, Alzheimer’s disease, multiple sclerosis, amyotrophic lateral sclerosis, or brain injury.

[0126] A kit-of-part according to the invention may comprise (i) the crystalline form according to the invention, and (ii) at least one agent useful for the prevention and / or treatment of Parkinson’s disease, frontotemporal dementia, multiple system atrophy, Alzheimer’s disease, multiple sclerosis, amyotrophic lateral sclerosis, or brain injury, said agent being different from said crystalline form.

[0127] Methods of Treatment

[0128] The crystalline form of the invention may be used in the prevention and / or in the treatment of pathologies involving the NOD-like receptor protein 3 (NLRP3) inflammasome pathway. Thus, is also described a method of treating and / or preventing Parkinson’s disease, frontotemporal dementia, multiple system atrophy, Alzheimer’s disease, multiple sclerosis, amyotrophic lateral sclerosis, or brain injury, including administering to a subject in need thereof a therapeutically effective amount of a crystalline form of the invention.

[0129] According to one embodiment, is also described a method of treating and / or preventing Parkinson’s disease, including administering to a subject in need thereof a therapeutically effective amount of a crystalline form of the invention.

[0130] According to another embodiment, is also described a method of treating and / or preventing frontotemporal dementia, including administering to a subject in need thereof a therapeutically effective amount of a crystalline form of the invention.

[0131] According to another embodiment, is also described a method of treating and / or preventing multiple system atrophy, including administering to a subject in need thereof a therapeutically effective amount of a crystalline form of the invention.

[0132] According to another embodiment, is also described a method of treating and / or preventing Alzheimer’s disease, including administering to a subject in need thereof a therapeutically effective amount of a crystalline form of the invention.

[0133] According to another embodiment, is also described a method of treating and / or preventing multiple sclerosis, including administering to a subject in need thereof a therapeutically effective amount of a crystalline form of the invention. According to another embodiment, is also described a method of treating and / or preventing amyotrophic lateral sclerosis, including administering to a subject in need thereof a therapeutically effective amount of a crystalline form of the invention.

[0134] According to another embodiment, is also described a method of treating and / or preventing brain injury, including administering to a subject in need thereof a therapeutically effective amount of a crystalline form of the invention.

[0135] EXAMPLES

[0136] Example 1: Synthesis of 2-[6-[(2S)-2-(hvdroxymethyl)morpholin-4-yl]pyridazin-3-yl]-

[0137] 3,5-dimethyl-phenol (Compound 1) solid form 1

[0138] Analytical methods used

[0139] NMR:

[0140] The proton magnetic resonance spectra (JH NMR), as described below, are recorded at 400 MHz in DMSO-d6, using the DMSO peak as reference. The chemical shifts (5) are expressed in parts per million (ppm). The signals observed are expressed as follows: s = singlet; d = doublet; t = triplet; m = multiplet or br s = broad singlet; br m = broad multiplet

[0141] LCMS:

[0142] The LCMS characteristics, as described below, indicates the different high-performance liquid chromatography analytical methods used.

[0143] Method A:

[0144] System Waters UPLC-SQD2; ionization: electrospray in positive and / or negative mode (ES+ / -); Column: ACQUITY CSH Cl 8 1.7pm 2.1x50mm; Column temperature: 50 °C Flow: 0.8 mL / min; Solvents: A = H2O (0.1% formic acid); B = ACN (0.1% formic acid) Gradient:

[0145] (min) %A %B

[0146] 0 95 5

[0147] 0.20 95 5

[0148] 3.60 2 98

[0149] 3,70 2 98

[0150] 4,10 95 5

[0151] 5.00 95 5

[0152] Method B :

[0153] System Waters UPLC-SQD2; ionization: electrospray in positive and / or negative mode (ES+ / -); Column: ACQUITY CSH C18+ 1.7pm 2.1x50mm; Column temperature: 60 °C Flow: 1.0 mL / min; Solvents: A = H2O (0.1% formic acid); B = ACN (0.1% formic acid) Gradient:

[0154] (min) %A %B

[0155] 0 97 3

[0156] 2.10 0 100

[0157] 2.45 0 100

[0158] 2.50 97 3

[0159] Mass spectrometry results are reported as the ratio of mass over charge.

[0160] Reactions under microwaves were realized using a Biotage Initiator apparatus.

[0161] Step 1: [(2S)-4-(6-chloropyridazin-3-yl)morpholin-2-yl]methanol

[0162] In a microwave vial, under Ar, to a solution of 3,6-dichloropyridazine (2.6 mmol, 400 mg) in 2.5 mL of NMP was added [(2S)-morpholin-2-yl]methanol; hydrochloride (2.6 mmol, 400 mg) and DIPEA (5.2 mmol, 0.9 mL). The vial was sealed, and the mixture was heated at 180 °C under microwave irradiation for 2.5h. After cooling down the solution, EtOAc was added, and the solution was washed with water (3 times) and brine. The aqueous layer was extracted twice with Me-THF. The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel using DCM / MeOH / ACN (from 100 / 0 / 0 to 96 / 2 / 2) to give the title compound as a yellow solid (214 mg, 36% yield).

[0163] LCMS (method A): Rt = 1.19 min; MS m / z [M+H]+230

[0164] Step 2: 2-[6-[(2S)-2-(hydroxymethyl)morpholin-4-yl]pyridazin-3-yl]-3,5-dimethyl- phenol (Compound 1) solid form 1

[0165] In a microwave vial, under Ar, to a solution of (2 -hydroxy-4, 6-dimethyl-phenyl)boronic acid (0.72 mmol, 120 mg) and [(2S)-4-(6-chloropyridazin-3-yl)morpholin-2-yl]methanol (0.79 mmol, 183 mg) in 4.5 mL of 1,4-di oxane and 1.2 mL of water was added sodium carbonate (1.59 mmol, 132 mg). The solution was purged with Ar for 10 min, then Pd(PPh3)4(0.036 mmol, 42 mg) was added. The vial was sealed, and the mixture was heated at 110 °C under microwave irradiation for 2.5h. After concentration under reduced pressure, EtOAc was added, and the resulting mixture was washed with water (3 times) and brine. The organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel using DCM / MeOH / ACN (from 98 / 1 / 1 to 94 / 2 / 2) to give Compound 1 solid form 1 as a white solid (92 mg, 40% yield).

[0166] LCMS (method B): Rt = 0.92 min; MS m / z [M+H]+316.2

[0167] 'HNMR (400 MHz, DMSO-d6, 30 °C) 5 ppm 2.04 (s, 3 H), 2.23 (s, 3 H), 2.71 (dd, J=12.8, 9.9 Hz, 1 H), 2.97 (td, J=12.3, 3.5 Hz, 1 H), 3.41 - 3.58 (m, 3 H), 3.62 (td, J=11.6, 2.6 Hz, 1 H), 4.00 (dd, J=11.5, 2.3 Hz, 1 H), 4.13 (br d, J=12.9 Hz, 1 H), 4.32 (br d, J=12.6 Hz, 1 H), 4.82 (t, J=5.5 Hz, 1 H), 6.58 (br s, 1 H), 6.59 (br s, 1 H), 7.27 (d, J=9.5 Hz, 1 H), 7.35 (d, J=9.5 Hz, 1 H), 9.39 (s, 1 H)

[0168] Example 2: X-Ray Powder Diffraction (XRPD)

[0169] XRPD analysis was carried out at room temperature on a Bruker D4 ENDEAVOR instrument using the Bragg-Brentano parafocusing geometry. A sealed copper anode X-ray tube was used (X CuKa average = 1.54178 A). A Bruker LYNXEYE linear detector completed the setup. A counting time of few seconds per step in an angular range of from few 2-Theta degrees to several dozen 2-Theta degrees with a 0.017° step size in 29. For each experiment, the powder was deposited on the surface of a sample holder. XRPD analysis of 2-[6-[(2S)-2-(hydroxymethyl)morpholin-4-yl]pyridazin-3-yl]-3,5- dimethyl-phenol solid Form 1 showed a patern consistent with a crystalline material. The XRPD diffractogram of crystalline Form 1 is shown in Figure 1 or Figure 2. Characteristic peaks include one or more of the peaks shown in Table II.

[0170] Table Example 3: Differential Scanning Calorimetry (DSC)

[0171] DSC analysis was carried out on a DSC 2500 calorimeter (TA Instruments). A sample mass of a few mg was deposited in an unsealed aluminum pan and the atmosphere was regulated by a constant nitrogen flow. Analyses have been carried out with a scanning rate of 5 °C / min. An illustrative DSC thermogram generated using crystalline Form 1 of 2-[6-[(2S)-2- (hydroxymethyl)morpholin-4-yl]pyridazin-3-yl]-3,5-dimethyl-phenol is shown in Figure 3. The Form 1 DSC thermogram shows a single endotherm with an onset temperature of Example 4: Thermogravimetric Analysis (TGA)

[0172] Analysis was carried out using a Netzsch TG 209 C instrument. A sample mass of a few mg was deposited in an aluminum crucible. The TGA analysis was conducted under a dry nitrogen stream and the sample was heated from 20 °C to 240 °C at a rate of 5 °C / min.

[0173] An illustrative TGA thermogram generated using crystalline Form 1 of 2-[6-[(2S)-2- (hydroxymethyl)morpholin-4-yl]pyridazin-3-yl]-3,5-dimethyl-phenol is shown in Figure 4.

[0174] The Form 1 TGA thermogram shows less than 1% mass loss up to 240 °C, suggesting an anhydrate solid form.

[0175] Example 5: Pharmacological assessment

[0176] IL-ip secretion assay

[0177] Monocytic THP-1 cells were maintained in RPMI 1640 media (Gibco 21875) + 10% FBS, (Gibco 10500) + 1% penicillin / streptomycin (Gibco 15140-122). Cells were then plated at 40,000 cells per well in 384-well cell culture plates (Coming 3542) and maintained with RPMI1640 (Gibco 11835) + 5% FBS (Gibco 10500). Activation of the NLRP3 inflammasome requires both an NF-kB-dependent priming step and the addition of aNLRP3 activator. The priming step was induced by LPS (lOng / mL, InvivoGen ref tlrl-3pelps) for 3h at 37 °C, then compound, in a 1 :3 serial dilution series in DMSO (final concentration DMSO 0.1%), and the activator nigericin (Sigma Aldrich, ref: SML1779) lOpM (final concentration) were added to the cells and co-incubated for 2 hours. 16 pL supernatant was removed, and IL-ip levels were monitored using an HTRF assay (Human ILlb Kits HTRF Cisbio ref: 62HILBPEH) according to manufacturers’ instructions.

[0178] TNF-a secretion assay

[0179] Monocytic THP-1 cells were maintained in RPMI 1640 media (Gibco 21875) + 10% FBS, (Gibco 10500) + 1% penicillin / streptomycin (Gibco 15140-122). Cells were then plated at 40,000 cells per well in 384-well cell culture plates (Coming 3542) and maintained with RPMI1640 (Gibco 11835) + 5% FBS, (Gibco 10500). TNF-a secretion was triggered by the addition of lOng / mL LPS (InvivoGen, tlrl-3pelps) and cells were incubated for 3 hours. TNF-a levels were monitored directly in the cells plate using an Lumit assay (Lumit TNFa- Human Immunoassay - Promega W6051) according to manufacturers' instructions.

[0180] Data interpretation

[0181] Data were expressed as percentage of inhibition (1%) as compared to a maximum signal control containing no small molecule (1% = 0%) according to the formula: I%=(1 -((Sample- Min)) / ((Max-Min))) x 100, where:

[0182] Sample: Signal obtained in the presence of each compound tested,

[0183] Max: Maximum signal in the absence of compound added,

[0184] Min: Minimum or background signal obtained in the absence of activation signal.

[0185] Relative IC50 (IC50 rel) were obtained from a dose response curve with 10 concentrations. Final IC50 rels are expressed as the geometric mean.

[0186] Relative IC50 values were estimated with Biost@t-SPEED v2.4 internal software based on SAS system using the 4-parameter logistic model according to Ratkovsky and Reedy (I). A. Ratkovsky, T.J. Reedy, Choosing near-linear parameters in the four parameters logistic model radioligands and related assays. Biometrics, 42 (1986), 575-582): Y=A+C / ((l+exp(- B*(log(X)-M)))), where:

[0187] A: Lower asymptote (BOTTOM),

[0188] A+C: Upper asymptote (TOP),

[0189] M: the logarithm of the concentration estimated at the inflexion point (the logarithm of the relative IC50),

[0190] B: Slope at the inflexion point of the curve. The compound of the present invention has significant inhibitory activity on secretion of IL- ip by THP-1 cells with minimal to no interference with the NF-kB pathway as measured by the TNF-a secretion after LPS priming.

[0191] It is therefore apparent that Compound 1 has an inhibitory activity on NOD-like receptor protein 3 (NLRP3) inflammasome.

[0192] Compound 1 may thus be used as inhibitor of the NOD-like receptor protein 3 (NLRP3) inflammasome pathway.

Claims

CLAIMS1. A crystalline form, which is Form 1 of the compound of formula (I):characterized by a powder X-ray diffractogram displaying at least one peak, in particular at least two peaks, preferably at least five peaks, expressed as degree 2-Theta angles, selected from 3.4, 6.8, 10.3, 13.7, 15.0, 16.1, 17.1, 17.5, 17.9, 20.5, and 25.3 (each time ± 0.2).

2. The crystalline form according to claim 1, characterized by a powder X-ray diffractogram displaying peaks expressed as degree 2-Theta angles at 3.4, 6.8, 10.3, 13.7, and 20.5 (each time ± 0.2).

3. The crystalline form according to claim 1 or 2, characterized by a powder X-ray diffractogram further displaying the following peaks expressed as degree 2-Theta angles at 15.0, 16.1, 17.1, 17.5, 17.9, and 25.3 (each time ± 0.2).

4. The crystalline form according to anyone of the preceding claims, further characterized by a powder X-ray diffractogram as substantially illustrated in Figure 1 or Figure 2.

5. The crystalline form according to anyone of the preceding claims, having a single differential scanning calorimetry endotherm with an onset temperature of 179 °C (±2 °C).

6. The crystalline form according to anyone of the preceding claims, having a differential scanning calorimetry thermogram as substantially illustrated in Figure 3.

7. The crystalline form according to anyone of the preceding claims, having a thermogravimetric analysis thermogram showing less than 1% mass loss up to 240 °C.

8. The crystalline form according to anyone of the preceding claims, having a thermogravimetric analysis thermogram as substantially illustrated in Figure 4.

9. The crystalline form according to anyone of the preceding claims, wherein the crystalline form is an anhydrate.

10. A process for the preparation of crystalline Form 1 of the compound of formula (I) as defined in anyone of claims 1 to 9, comprising at least the following steps:1) solubilizing the compound of formula (I) in a solvent selected from alcohols, chlorinated hydrocarbons, ketones, acetates, ethers, acetonitrile, and mixtures thereof, optionally in admixture with water, at a set temperature ranging from 18 °C to 80 °C;2) leaving the solution obtained in step 1) at the same temperature as the one set in step 1) for almost complete evaporation;3) isolating the crystalline Form 1 of the compound of formula (I) formed in step 2).

11. The process of claim 10, wherein the solvent of step 1) is selected from methanol, methanol / water mixture, ethanol, ethanol / water mixture, 2-propanol, 1 -propanol,1-propanol / water mixture, 1-butanol / water mixture, dichloromethane, dichloromethane / methanol / acetonitrile mixture, acetone, acetone / water mixture,2-butanone, 2-butanone / water mixture, methyl isobutyl ketone / water mixture, methyl acetate, methyl acetate / water mixture, ethyl acetate, ethyl acetate / water mixture, isopropyl acetate / water mixture, isobutyl acetate, isobutyl acetate / water mixture, tetrahydrofuran, and acetonitrile.

12. A medicament comprising at least one crystalline form according to anyone of claims 1 to 9.

13. A pharmaceutical composition comprising a crystalline form as defined in anyone of claims 1 to 9, and at least one pharmaceutically acceptable excipient.

14. A crystalline form as defined in anyone of claims 1 to 9, or a composition as defined in claim 13, for use as a medicine.

15. A crystalline form as defined in anyone of claims 1 to 9, or a composition as defined in claim 13, for use in the prevention and / or treatment of Parkinson’s disease, frontotemporal dementia, multiple system atrophy, Alzheimer’s disease, multiple sclerosis, amyotrophic lateral sclerosis, or brain injury.

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