EP4658649A1 - [1,2,4]triazolo[4,3-a]pyridines and [1,2,4]triazolo[4,3-a]pyrazines as nlrp3 inhibitors - Google Patents

Abstract

Provided are compounds for use as inhibitors of the NLRP3 inflammasone pathway, wherein such compounds are as defined by compounds of formula (I) and wherein the radicals R¹, R², R³, R⁴, L and A are defined in the description, and where the compounds may be useful as medicaments, for instance for use in the treatment of a disease or disorder that is associated with NLRP3 inflammasome activity.

Description

[l,2,4]TRIAZOLO[4,3-a]PYRIDINES AND [1,2,4]TRIAZOLO[4,3- aJPYRAZINES AS NLRP3 INHIBITORS

TECHNICAL FIELD

Described herein are [l,2,4]triazolo[4,3-a]pyridines and [l,2,4]triazolo[4,3- a]pyrazines that are useful as inhibitors of the NOD-like receptor protein 3 (NLRP3) inflammasome pathway. Also described herein are processes for the preparation of said compounds, pharmaceutical compositions comprising said compounds, methods of using said compounds in the treatment of various diseases and disorders mediated by the NLRP3 inflammasome pathway.

BACKGROUND

Inflammasomes, considered as central signalling hubs of the innate immune system, are multi-protein complexes that are assembled upon activation of a specific set of intracellular pattern recognition receptors (PRRs) by a wide variety of pathogen- or danger- associated molecular patterns (PAMPs or DAMPs). To date, it was shown that inflammasomes can be formed by nucleotide-binding oligomerization domain (NOD)- like receptors (NLRs) and Pyrin- and HIN200-domain-containing proteins (Van Opdenbosch N and Lamkanfi M. Immunity, 2019 Jun 18;50(6): 1352-1364). The NLRP3 inflammasome is assembled upon detection of environmental crystals, pollutants, host-derived DAMPs and protein aggregates (Tartey S and Kanneganti TD. Immunology, 2019 Apr; 156(4): 329-338). Clinically relevant DAMPs that engage NLRP3 include uric acid and cholesterol crystals that cause gout and atherosclerosis, amyloid-P fibrils that are neurotoxic in Alzheimer’s disease and asbestos particles that cause mesothelioma (Kelley et al., IntJMol Sci, 2019 Jul 6;20(13)). Additionally, NLRP3 is activated by infectious agents such as Vibrio cholerae,' fungal pathogens such as Aspergillus fumigatus and Candida albicans,' adenoviruses, influenza A virus and SARS-CoV-2 (Tartey and Kanneganti, 2019; Fung et al. Emerg Microbes Infect, 2020 Mar 14;9(l):558-570).

Although the precise NLRP3 activation mechanism remains unclear, for human monocytes, it has been suggested that a one-step activation is sufficient while in mice a two-step mechanism is in place. Given the multitude in triggers, the NLRP3 inflammasome requires add-on regulation at both transcriptional and post- transcriptional level (Yang Y et al., Cell Death Dis, 2019 Feb 12; 10(2): 128).

The NLRP3 protein consists of an N-terminal pyrin domain, followed by a nucleotide-binding site domain (NBD) and a leucine-rich repeat (LRR) motif on C- terminal end (Sharif et al., Nature, 2019 Jun; 570(7761):338-343). Upon recognition of PAMP or DAMP, NLRP3 aggregates with the adaptor protein, apoptosis-associated speck-like protein (ASC), and with the protease caspase- 1 to form a functional inflammasome. Upon activation, procaspase-1 undergoes autoproteolysis and consequently cleaves gasdermin D (Gsdmd) to produce the N-terminal Gsdmd molecule that will ultimately lead to pore-formation in the plasma membrane and a lytic form of cell death called pyroptosis. Alternatively, caspase-1 cleaves the pro- inflammatory cytokines pro-IL-ip and pro-IL-18 to allow release of its biological active form by pyroptosis (Kelley et al., 2019).

Dysregulation of the NLRP3 inflammasome or its downstream mediators are associated with numerous pathologies ranging from immune/inflammatory diseases, auto-immune/auto-inflammatory diseases (Cryopyrin-associated Periodic Syndrome (Miyamae T. Paediatr Drugs, 2012 Apr 1; 14(2): 109-17); sickle cell disease; systemic lupus erythematosus (SLE)) to hepatic disorders (e.g. non-alcoholic steatohepatitis (NASH), chronic liver disease, viral hepatitis, alcoholic steatohepatitis, non-alcoholic fatty acid liver disease, and alcoholic liver disease) (Szabo G and Petrasek J. Nat Rev Gastroenterol Hepatol, 2015 Jul;12(7):387-400) and inflammatory bowel diseases (eg. Crohn’s disease, ulcerative colitis) (Zhen Y and Zhang H. Front Immunol, 2019 Feb 28; 10:276). Also, inflammatory joint disorders (e.g. gout, pseudogout (chondrocalcinosis), arthropathy, osteoarthritis, and rheumatoid arthritis (Vande Walle L et al., Nature, 2014 Aug 7;512(7512):69-73) were linked to NLRP3. Additionally, kidney related diseases (hyperoxaluria (Knauf et al., Kidney Int, 2013 Nov;84(5):895-901), lupus nephritis, hypertensive nephropathy (Krishnan et al., Br J Pharmacol, 2016 Feb;173(4):752-65), hemodialysis related inflammation and diabetic nephropathy which is a kidney -related complication of diabetes (Type 1, Type 2 and mellitus diabetes), also called diabetic kidney disease (Shahzad et al., Kidney Int, 2015 Jan;87(l):74-84) are associated to NLRP3 inflammasome activation. Reports link onset and progression of neuroinflammation-related disorders (e.g. brain infection, acute injury, multiple sclerosis, Alzheimer's disease) and neurodegenerative diseases (Parkinsons disease) to NLRP3 inflammasome activation (Sarkar et al., NPJ Parkinsons Dis, 2017 Oct 17;3:30). In addition, cardiovascular or metabolic disorders (e.g. cardiovascular risk reduction (CvRR), atherosclerosis, type I and type II diabetes and related complications (e.g. nephropathy, retinopathy), peripheral artery disease (PAD), acute heart failure and hypertension (Ridker et al., CANTOS Trial Group. N Engl J Med, 2017 Sep 21;377(12): 1119-1131; and Toldo S and Abbate A Nat Rev Cardiol, 2018 Apr; 15 (4): 203 -214) have recently been associated to NLRP3. Also, skin associated diseases were described (e.g. wound healing and scar formation; inflammatory skin diseases, eg. acne, hidradenitis suppurativa (Kelly et al., Br J Dermatol, 2015 Dec;173(6)). In addition, respiratory conditions have been associated with NLRP3 inflammasome activity (e.g. asthma, sarcoidosis, Severe Acute Respiratory Syndrome (SARS) (Nieto-Torres et al., Virology, 2015 Nov;485:330-9)), silicosis, pneumonia, but also age-related macular degeneration (Doyle et al., Nat Med, 2012 May;18(5):791-8). Several cancer related diseases/disorders were described linked to NLRP3 (e.g. myeloproliferative neoplasms, leukemias, myelodysplastic syndromes (MOS), myelofibrosis, lung cancer, colon cancer (Ridker et al., Lancet, 2017 Oct 21;390(10105): 1833-1842; Derangere et al., Cell Death Differ.

2014 Dec;21(12): 1914-24; Basiorka et al., Lancet Haematol, 2018 Sep;5(9): e393- e402, Zhang et al., Hum Immunol, 2018 Jan;79(l):57-62).

Several patent applications describe NLRP3 inhibitors, with recent ones including for instance WO-2020/234715, WO-2021/193897, WO-2022/135567, US- 11,319,319.

There is a need for inhibitors of the NLRP3 inflammasome pathway for example to study neurodegenerative disorders such as Alzheimer’s Disease.

SUMMARY

Described herein are compounds which inhibit the NLRP3 inflammasome pathway.

In some embodiments, provided herein are compounds of Formula (I), or pharmaceutically acceptable salts thereof, wherein

A is CH or N;

L is NR⁵, NR⁵-CH2 or CH2, wherein R⁵ is hydrogen or methyl;

R¹ is

Y is CH2 when L is NR⁵, NR⁵-CH2 or CH2; or Y is NR⁵ or O when L is CH2;

R¹⁰ is hydrogen, C1-2 alkyl, haloCi-2 alkyl, hydroxyCi-2 alkyl, C1-2 alkyloxy, haloCi-2 alkyloxy or -CH2CN;

R¹¹ is hydrogen, methyl, hydroxy or fluoro;

R² is hydrogen, C1-2 alkyl, haloCi-2 alkyl, C1-2 alkyloxy, haloCi-2 alkyloxy, cyclopropyl, halo or cyano; R³ is hydrogen, methyl, ethyl or halo; and

R⁴ is hydrogen, C1-2 alkyl, haloCi-2 alkyl, cyclopropyl, or halo.

In another aspect, there are provided compounds for use as a medicament. In another aspect, there are provided pharmaceutical compositions comprising a therapeutically effective amount of a compound provided herein.

In a further aspect, there are provided compounds and pharmaceutical compositions comprising such compounds for use in the treatment of a disease or disorder mediated by the NLRP3 inflammasome pathway, for example neurodegenerative disorders such as Alzheimer’s Disease.

In another aspect, there is provided the use of the instant compounds in the manufacture of a medicament for the treatment of a disease or disorder mediated by the NLRP3 inflammasome pathway, for example neurodegenerative disorders such as Alzheimer’s Disease.

In another aspect, there is provided a method of treating a disease or disorder mediated by the NLRP3 inflammasome pathway, for example neurodegenerative disorders such as Alzheimer’s Disease. In a further aspect there is provided a method of inhibiting the NLRP3 inflammasome activity in a subject (in need thereof), the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound as provided herein.

DETAILED DESCRIPTION

Described herein are compounds which inhibit the NLRP3 inflammasome pathway.

In some embodiments, provided herein are compounds of Formula (I), or pharmaceutically acceptable salts thereof, wherein

A is CH or N;

L is NR⁵, NR⁵-CH2 or CH2, wherein R⁵ is hydrogen or methyl; Y is CH2 when L is NR⁵, NR⁵-CH2 or CH2; or Y is NR⁵ or O when L is CH2;

R¹⁰ is hydrogen, C1-2 alkyl, haloCi-2 alkyl, hydroxyCi-2 alkyl, C1-2 alkyloxy, haloCi-2 alkyloxy or -CH2CN;

R¹¹ is hydrogen, methyl, hydroxy or fluoro;

R² is hydrogen, C1-2 alkyl, haloCi-2 alkyl, C1-2 alkyloxy, haloCi-2 alkyloxy, cyclopropyl, halo or cyano;

R³ is hydrogen, methyl, ethyl or halo; and

R⁴ is hydrogen, C1-2 alkyl, haloCi-2 alkyl, cyclopropyl, or halo.

In an embodiment,

A is CH or N;

L is NR⁵, or NR⁵-CH2 or CH2, wherein R⁵ is hydrogen or methyl;

R¹ is

Y is CH₂;

R¹⁰ is C1-2 alkyl;

R¹¹ is hydrogen or fluoro;

R² is hydrogen, C1-2 alkyl, haloCi-2 alkyl, C1-2 alkyloxy, haloCi-2 alkyloxy, or halo; in particular hydrogen, methyl, trifluoromethyl; in particular haloCi-2 alkyl, haloCi-2 alkyloxy, or halo; more in particular, methyl or trifluoromethyl;

R³ is hydrogen, methyl, halo, in particular chloro; and

R⁴ is hydrogen, C1-2 alkyl, haloCi-2 alkyl, cyclopropyl, or halo ; in particular, hydrogen, methyl, trifluoromethyl, cyclopropyl or chloro.

In an embodiment

A is CH;

L is NR⁵ or CH2, wherein R⁵ is hydrogen;

R¹ is

Y is CH₂;

R¹⁰ is C1-2 alkyl;

R¹¹ is hydrogen or fluoro; R² is haloCi-2 alkyl, haloCi-2 alkyloxy, or halo;

R³ is hydrogen, methyl or halo; and

R⁴ is hydrogen, C1-2 alkyl, haloCi-2 alkyl, cyclopropyl, or halo.

In an embodiment

L is NH;

R¹⁰ is methyl;

R¹¹ is hydrogen or fluoro;

R² is trifluoromethyl;

R³ is hydrogen, methyl or halo; and

R⁴ is hydrogen, methyl, haloCi-2 trifluoromethyl, cyclopropyl, or chloro.

Compounds of particular interest are (R)-2-(3-((l-methylpiperi din-3 -yl)amino)- [l,2,4]triazolo[4,3-a]pyridin-8-yl)-5-(trifluoromethyl)phenol and rel-(R)-2-(3-((l- methylpiperidin-3-yl)methyl)-[l,2,4]triazolo[4,3-a]pyridin-8-yl)-5- (trifluoromethyl)phenol.

The compounds according to the invention can generally be prepared by a succession of steps, each of which is known to the skilled person. In particular, the compounds can be prepared according to the following synthesis methods.

The compounds of Formula (I) may be synthesized in the form of racemic mixtures of enantiomers which can be separated from one another following art-known resolution procedures. The racemic compounds of Formula (I) may be converted into the corresponding diastereomeric salt forms by reaction with a suitable chiral acid. Said diastereomeric salt forms are subsequently separated, for example, by selective or fractional crystallization and the enantiomers are liberated therefrom by alkali? An alternative manner of separating the enantiomeric forms of the compounds of Formula (I) involves liquid chromatography using a chiral stationary phase or a chiral supercritical fluid chromatography (SFC). Said pure stereochemically isomeric forms may also be derived from the corresponding pure stereochemically isomeric forms of the appropriate starting materials, provided that the reaction occurs stereospecifically.

The absolute configuration of compounds of the invention reported herein was determined by analysis of the racemic mixture by supercritical fluid chromatography (SFC) followed by SFC comparison of the separate enantiomer(s) which were obtained by asymmetric synthesis, followed by vibrational circular dichroism (VCD) analysis of the particular enantiomer(s). Pharmaceutically-acceptable salts include acid addition salts and base addition salts. Such salts may be formed by conventional means, for example by reaction of a free acid or a free base form of a compound as provided herein with one or more equivalents of an appropriate acid or base, optionally in a solvent, or in a medium in which the salt is insoluble, followed by removal of said solvent, or said medium, using standard techniques (e.g. in vacuo, by freeze-drying or by filtration). Salts may also be prepared by exchanging a counter-ion of a compound provided herein in the form of a salt with another counter-ion, for example using a suitable ion exchange resin.

Pharmaceutically acceptable acid addition salts can be formed with inorganic acids and organic acids.

Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like.

Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, sulfosalicylic acid, and the like.

Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases.

Inorganic bases from which salts can be derived include, for example, ammonium salts and metals from columns I to XII of the periodic table. In certain embodiments, the salts are derived from sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc, and copper; particularly suitable salts include ammonium, potassium, sodium, calcium and magnesium salts.

Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like. Certain organic amines include isopropylamine, benzathine, cholinate, diethanolamine, diethylamine, lysine, meglumine, piperazine, and tromethamine.

The instant compounds may contain double bonds and may thus exist as E (entgegeri) and Z (zusammeri) geometric isomers about each individual double bond.

Compounds as provided herein may contain one or more asymmetric carbon atoms and may therefore exhibit enantiomerism or diastereoisomerism. Diastereoisomers may be separated using conventional techniques, e.g. chromatography or fractional crystallisation. The various stereoisomers may be isolated by separation of a racemic or other mixture of the compounds using conventional, e.g. fractional crystallisation or HPLC, techniques. Alternatively the desired isomers may be made by reaction of an appropriate enantiomeric starting material under conditions which will not cause racemisation or epimerisation, or by reaction of an appropriate starting material with a ‘chiral auxiliary’ which can subsequently be removed at a suitable stage, by resolution, including dynamic resolution, for example salt formation with a homochiral acid followed by separation of the diastereomeric salts by conventional means such as crystallization, or by reaction with an appropriate chiral reagent or chiral catalyst.

In the structures shown herein, where the stereochemistry of any particular chiral atom is not specified, then all stereoisomers are contemplated. Where stereochemistry is specified by a solid or dashed wedge representing a particular configuration, then that stereoisomer is so specified and defined.

Absolute configurations are specified according to the Cahn-Ingold-Prelog system. The configuration at an asymmetric atom is specified by either R or S. Resolved compounds whose absolute configuration is not known can be designated by (+) or (-) depending on the direction in which they rotate polarized light.

When a specific stereoisomer is identified, this means that said stereoisomer is substantially free, i.e. associated with less than 50%, preferably less than 20%, more preferably less than 10%, even more preferably less than 5%, in particular less than 2% and most preferably less than 1%, of the other isomers. Thus, when a compound of formula (I) is for instance specified as (R), this means that the compound is substantially free of the (S) isomer.

The compounds may exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like.

Also provided herein are isotopically-labelled compounds wherein one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature (or the most abundant one found in nature). Exemplary isotopes include isotopes of hydrogen, carbon, nitrogen, oxygen, and fluorine, such as ²H, ³H, ⁿC, ¹³C, ¹⁴C , ¹³N, ¹⁵O, ¹⁷O, ¹⁸O, and ¹⁸F. Tritiated (³H) and carbon-14 (¹⁴C) isotopes are useful for their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium may afford therapeutic advantages resulting from greater metabolic stability. Isotopes such as ¹⁵O, ¹³N, ⁿC and ¹⁸F are useful for positron emission tomography (PET) studies to examine substrate receptor occupancy. Isotopically labelled compounds can generally be prepared by following procedures analogous to those disclosed in the Examples hereinafter.

Unless otherwise specified, Ci-_q alkyl groups (where q is the upper limit of the range) defined herein may be straight-chain or be branched-chain.

Cs-q cycloalkyl (where q is the upper limit of the range) refers to an alkyl group that is cyclic, for instance cycloalkyl groups may be monocyclic or, if there are sufficient atoms, bicyclic. In an embodiment, such cycloalkyl groups are monocyclic. Substituents may be attached at any point on the cycloalkyl group.

The term “halo”, when used herein, preferably includes fluoro, chloro, bromo and iodo.

Ci-q alkoxy groups (where q is the upper limit of the range) refers to the radical of formula -OR^a, where R^a is a Ci-_q alkyl group as defined herein.

HaloCi-q alkyl (where q is the upper limit of the range) groups refer to Ci-_q alkyl groups, as defined herein, where such group is substituted by one or more halo. HydroxyCi-q alkyl (where q is the upper limit of the range) refers to Ci-_q alkyl groups, as defined herein, where such group is substituted by one or more (e.g. one) hydroxy (-OH) groups (or one or more, e.g. one, of the hydrogen atoms is replaced with -OH). Similarly, haloCi-_q alkoxy and hydroxyCi-_q alkoxy represent corresponding -OCi-_q alkyl groups that are substituted by one or more halo, or, substituted by one or more (e.g. one) hydroxy, respectively.

The names of the compounds were generated according to the nomenclature rules agreed upon by the Chemical Abstracts Service (CAS) using Advanced Chemical Development, Inc., software (ACD/Name product version 10.01; Build 15494, 1 Dec 2006) or according to the nomenclature rules agreed upon by the International Union of Pure and Applied Chemistry (IUPAC) using Advanced Chemical Development, Inc., software (ACD/Name product version 10.01.0.14105, October 2006). In case of tautomeric forms, the name of the depicted tautomeric form of the structure was generated.

The instant compounds can generally be prepared by a succession of steps, each of which is known to the skilled person. In particular, the compounds can be prepared according to the following synthesis methods.

The compounds of Formula (I) may be synthesized in the form of racemic mixtures of enantiomers which can be separated from one another following art-known resolution procedures. The racemic compounds of Formula (I) may be converted into the corresponding diastereomeric salt forms by reaction with a suitable chiral acid. Said diastereomeric salt forms are subsequently separated, for example, by selective or fractional crystallization and the enantiomers are liberated therefrom by alkalination. An alternative manner of separating the enantiomeric forms of the compounds of Formula (I) involves liquid chromatography using a chiral stationary phase or a chiral supercritical fluid chromatography (SFC). Said pure stereochemically isomeric forms may also be derived from the corresponding pure stereochemically isomeric forms of the appropriate starting materials, provided that the reaction occurs stereospecifically.

The absolute configuration of the compounds reported herein was determined by analysis of the racemic mixture by supercritical fluid chromatography (SFC) followed by SFC comparison of the separate enantiomer(s) which were obtained by asymmetric synthesis, followed by vibrational circular dichroism (VCD) analysis of the particular enantiomer(s).

PREPARATION OF THE COMPOUNDS

Preparation of the compounds

In an aspect of the invention, there is provided a process for the preparation of compounds of the invention, where reference here is made to compounds of Formula (la) and (lb) as defined herein.

Final compounds according to Formula (la), wherein L is NR⁵ can be prepared:

- By deprotecting an Intermediate of Formula (II) in a suitable acidic medium such as, for example, hydrochloric acid solution in 1,4-di oxane, at a suitable temperature such as, for example, room temperature;

Intermediates of Formula (II) can be prepared by reacting an intermediate of Formula (III) with a suitable base such as, for example, sodium hydride, and a suitable electrophile such as, for example, methyl iodide, in a suitable solvent such as, for example, DMF or THF, at a suitable temperature such as, for example, room temperature;

Intermediates of Formula (III) can be prepared by reacting an intermediate of Formula (IV) with a suitable vinylboronic acid or vinyl boronate ester via Suzuki coupling in the presence of a suitable palladium catalyst such as, for example, dichlorobis(triphenylphosphine)palladium(II), in the presence of a suitable base such as, for example, potassium phosphate tribasic, in a suitable solvent such as, for example, a mixture of 1,4-di oxane and water, at a suitable temperature such as, for example, 100 °C;

Alternatively, final compounds according to Formula (la), where L is NR⁵ can be prepared:

- By reacting an Intermediate of Formula (V) with a suitable vinylboronic acid or vinyl boronate ester via Suzuki coupling in the presence of a suitable palladium catalyst such as, for example, dichlorobis(triphenylphosphine)palladium(II), in the presence of a suitable base such as, for example, potassium phosphate tribasic, in a suitable solvent such as, for example, a mixture of 1,4-di oxane and water, at a suitable temperature such as, for example, 100 °C;

Intermediates of Formula (V) can be prepared by reacting an intermediate of Formula (IV) with a suitable base such as, for example, sodium hydride, and a suitable electrophile such as, for example, methyl iodide, in a suitable solvent such as, for example, DMF or THF, at a suitable temperature such as, for example, room temperature;

Final compounds according to Formula (lb), wherein L is CH2 can be prepared:

- By reacting an Intermediate of Formula (VI) with a suitable vinylboronic acid or vinyl boronate ester via Suzuki coupling in the presence of a suitable palladium catalyst such as, for example, dichlorobis(triphenylphosphine)palladium(II), in the presence of a suitable base such as, for example, potassium phosphate tribasic, in a suitable solvent such as, for example, a mixture of 1,4-di oxane and water, at a suitable temperature such as, for example, 100 °C;

Intermediates of Formula (VI) can be prepared by cyclization of an intermediate of Formula (VII) with a suitable dehydrating agent such as, for example, Burgess reagent, in a suitable solvent such as, for example, acetonitrile, at a suitable temperature such as, for example, 80 °C;

Intermediates of Formula (VII) can be prepared by reacting an intermediate of Formula (VIII) with a suitable acid in the presence of a suitable coupling agent such as, for example, DCC, in a suitable solvent such as, for example, DMF, at a suitable temperature such as, for example, room temperature;

Alternatively, intermediates of Formula (VII) can be prepared by reacting an intermediate of Formula (VIII) with a suitable acyl chloride in the presence of a suitable base such as, for example, triethylamine, in a suitable solvent such as, for example, DCM, at a suitable temperature such as, for example, room temperature;

Intermediates according to Formula (IV) can be prepared:

- By reacting an Intermediate of Formula (IX) with iodine in the presence of a suitable base such as, for example, potassium carbonate, in a suitable solvent such as, for example, 1,4-di oxane, at a suitable temperature such as, for example, 80 °C;

Intermediates of Formula (IX) can be prepared by reacting an Intermediate of Formula (VIII) with a suitable thioisocyanate reagent in a suitable solvent such as, for example, dichloromethane, at a suitable temperature such as, for example, room temperature;

Alternatively, intermediates of Formula (IX) can be prepared by reacting an Intermediate of Formula (VIII) with a suitable amine and thiophosgene, with a suitable base such as, for example, triethylamine, in a suitable solvent such as, for example, di chloromethane, at a suitable temperature such as, for example, 0 °C or room temperature;

Intermediates of Formula (VIII) can be prepared by reacting an Intermediate of Formula (X) with hydrazine monohydrate in a suitable solvent such as, for example, ethanol, at a suitable temperature such as, for example, 50 °C. Final compounds according to Formula (Ic) wherein A is N and L is NR⁵ can be prepared:

- By deprotecting an Intermediate of Formula (XI) in a suitable acidic medium such as, for example, hydrochloric acid solution in 1,4-di oxane, at a suitable temperature such as, for example, room temperature;

Intermediates of Formula (XI) can be prepared by reacting an Intermediate of Formula (XII) with iodine in the presence of a suitable base such as, for example, potassium carbonate, in a suitable solvent such as, for example, 1,4- dioxane, at a suitable temperature such as, for example, 80 °C;

Intermediates of Formula (XII) can be prepared by reacting an Intermediate of Formula (XIII) with a suitable thioisocyanate reagent in a suitable solvent such as, for example, dichloromethane, at a suitable temperature such as, for example, room temperature;

Alternatively, intermediates of Formula (XII) can be prepared by reacting an Intermediate of Formula (XIII) with a suitable amine and thiophosgene, with a suitable base such as, for example, triethylamine, in a suitable solvent such as, for example, dichloromethane, at a suitable temperature such as, for example, 0 °C or room temperature;

Intermediates of Formula (XIII) can be prepared by reacting an Intermediate of Formula (XIV) with hydrazine monohydrate in a suitable solvent such as, for example, ethanol, at a suitable temperature such as, for example, 50 °C;

Intermediates of Formula (XIV) can be prepared by reacting an Intermediate of Formula (XV) with a suitable vinylboronic acid or vinyl boronate ester via Suzuki coupling in the presence of a suitable palladium catalyst such as, for example, dichlorobis(triphenylphosphine)palladium(II), in the presence of a suitable base such as, for example, potassium phosphate tribasic, in a suitable solvent such as, for example, a mixture of 1,4-di oxane and water, at a suitable temperature such as, for example, 100 °C.

The skilled chemist will understand that, in the case that R¹ contains a protecting group such as, for example, Boc, deprotection of intermediates of Formula (II) will afford the deprotected compound of Formula (I). Further functionalization of those nor-compounds is possible using an aldehyde coupling partner in the presence of a reductive agent such as, for example, sodium triacetoxyborohydride, in a suitable solvent such as, for example, methanol or dichloromethane, at a suitable temperature such as, for example, 0 °C.

PHARMACOLOGY

The instant compounds are potent inhibitors of the NLRP3 inflammasome pathway. The instant compounds are potent, brain-penetrant, and may be useful in central nervous system diseases such as Parkinson's disease, Alzheimer's disease, dementia, motor neuron disease, Huntington's disease, traumatic brain injury, multiple sclerosis, and amyotrophic lateral sclerosis.

PHARMACEUTICAL COMPOSITIONS AND COMBINATIONS

In an embodiment, further described herein are compositions comprising a pharmaceutically acceptable carrier and, as active ingredient, a therapeutically effective amount of a compound as provided herein. The compounds may be formulated into various pharmaceutical forms for administration purposes. As appropriate compositions there may be cited all compositions usually employed for systemically administering drugs. To prepare the pharmaceutical compositions, an effective amount of the particular compound, optionally in salt form, as the active ingredient is combined in intimate admixture with a pharmaceutically acceptable carrier, which carrier may take a wide variety of forms depending on the form of preparation desired for administration. These pharmaceutical compositions are desirable in unitary dosage form suitable, in particular, for administration orally or by parenteral injection. For example, in preparing the compositions in oral dosage form, any of the usual pharmaceutical media may be employed such as, for example, water, glycols, oils, alcohols and the like in the case of oral liquid preparations such as suspensions, syrups, elixirs, emulsions and solutions; or solid carriers such as starches, sugars, kaolin, diluents, lubricants, binders, disintegrating agents and the like in the case of powders, pills, capsules and tablets. Because of their ease in administration, tablets and capsules represent the most advantageous oral dosage unit forms in which case solid pharmaceutical carriers are obviously employed. For parenteral compositions, the carrier will usually comprise sterile water, at least in large part, though other ingredients, for example, to aid solubility, may be included. Injectable solutions, for example, may be prepared in which the carrier comprises saline solution, glucose solution or a mixture of saline and glucose solution. Injectable suspensions may also be prepared in which case appropriate liquid carriers, suspending agents and the like may be employed. Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations.

The pharmaceutical composition may additionally contain various other ingredients known in the art, for example, a lubricant, stabilising agent, buffering agent, emulsifying agent, viscosity-regulating agent, surfactant, preservative, flavouring or colorant.

It is especially advantageous to formulate the aforementioned pharmaceutical compositions in unit dosage form for ease of administration and uniformity of dosage. Unit dosage form as used herein refers to physically discrete units suitable as unitary dosages, each unit containing a predetermined quantity of active ingredient calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. Examples of such unit dosage forms are tablets (including scored or coated tablets), capsules, pills, powder packets, wafers, suppositories, injectable solutions or suspensions and the like, and segregated multiples thereof.

The daily dosage of the compound will, of course, vary with the compound employed, the mode of administration, the treatment desired and the mycobacterial disease indicated. However, in general, satisfactory results will be obtained when the compound is administered at a daily dosage not exceeding 1 gram, e.g. in the range from 10 to 50 mg/kg body weight.

As used herein, term "pharmaceutical composition" refers to a compound as provided herein or a pharmaceutically acceptable salt thereof, together with at least one pharmaceutically acceptable carrier, in a form suitable for oral or parenteral administration.

As used herein, the term "pharmaceutically acceptable carrier" refers to a substance useful in the preparation or use of a pharmaceutical composition and includes, for example, suitable diluents, solvents, dispersion media, surfactants, antioxidants, preservatives, isotonic agents, buffering agents, emulsifiers, absorption delaying agents, salts, drug stabilizers, binders, excipients, disintegration agents, lubricants, wetting agents, sweetening agents, flavoring agents, dyes, and combinations thereof, as would be known to those skilled in the art (see, for example, Remington The Science and Practice of Pharmacy, 22nd Ed. Pharmaceutical Press, 2013, pp. 1049- 1070). The term "subject" as used herein, refers to an animal, preferably a mammal, most preferably a human, for example who is or has been the object of treatment, observation or experiment.

The term "therapeutically effective amount" as used herein, means that amount of compound that elicits a biological or medicinal response in a subject, for example, reduction or inhibition of an enzyme or a protein activity, or ameliorate symptoms, alleviate conditions, slow or delay disease progression, or prevent a disease, etc. In one non-limiting embodiment, the term "a therapeutically effective amount" refers to the amount of the compound that, when administered to a subject, is effective to (1) at least partially alleviate, inhibit, prevent and/or ameliorate a condition, or a disorder or a disease (i) mediated by NLRP3, or (ii) associated with NLRP3 activity, or (iii) characterised by activity (normal or abnormal) of NLRP3; or (2) reduce or inhibit the activity of NLRP3; or (3) reduce or inhibit the expression of NLRP3. In another nonlimiting embodiment, the term "a therapeutically effective amount" refers to the amount of the compound that, when administered to a cell, or a tissue, or a non-cellular biological material, or a medium, is effective to at least partially reduce or inhibit the activity of NLRP3; or at least partially reduce or inhibit the expression of NLRP3.

As used herein, the term "inhibit", "inhibition" or "inhibiting" refers to the reduction or suppression of a given condition, symptom, or disorder, or disease, or a significant decrease in the baseline activity of a biological activity or process. Specifically, inhibiting NLRP3 or inhibiting NLRP3 inflammasome pathway comprises reducing the ability of NLRP3 or NLRP3 inflammasome pathway to induce the production of IL-1 and/or IL-18. This can be achieved by mechanisms including, but not limited to, inactivating, destabilizing, and/or altering distribution of NLRP3.

As used herein, the term "NLRP3" is meant to include, without limitation, nucleic acids, polynucleotides, oligonucleotides, sense and anti-sense polynucleotide strands, complementary sequences, peptides, polypeptides, proteins, homologous and/or orthologous NLRP molecules, isoforms, precursors, mutants, variants, derivatives, splice variants, alleles, different species, and active fragments thereof.

As used herein, the term "treat", "treating" or "treatment" of any disease or disorder refers to alleviating or ameliorating the disease or disorder (i.e., slowing or arresting the development of the disease or at least one of the clinical symptoms thereof); or alleviating or ameliorating at least one physical parameter or biomarker associated with the disease or disorder, including those which may not be discernible to the patient.

As used herein, the term "prevent", "preventing" or "prevention" of any disease or disorder refers to the prophylactic treatment of the disease or disorder; or delaying the onset or progression of the disease or disorder. As used herein, a subject is "in need of’ a treatment if such subject would benefit biologically, medically or in quality of life from such treatment.

In an embodiment, there is provided a compound, according to any one of the embodiments described herein, for use as a medicament.

In an embodiment, there is provided a compound, according to any one of the embodiments described herein (and/or pharmaceutical compositions comprising such compound, according to any one of the embodiment described herein) for use in the treatment of a disease or disorder associated with NLRP3 activity (including inflammasome activity); in the treatment of a disease or disorder in which the NLRP3 signalling contributes to the pathology, and/or symptoms, and/or progression, of said disease/disorder; in inhibiting NLRP3 inflammasome activity (including in a subject in need thereof); and/or as an NLRP3 inhibitor.

In an embodiment, there is provided the use of compounds as provided herein, according to any one of the embodiments described herein (and/or pharmaceutical compositions comprising such compound, according to any one of the embodiment described herein): in the treatment of a disease or disorder associated with NLRP3 activity (including inflammasome activity); in the treatment of a disease or disorder in which the NLRP3 signalling contributes to the pathology, and/or symptoms, and/or progression, of said disease/disorder; in inhibiting NLRP3 inflammasome activity (including in a subject in need thereof); and/or as an NLRP3 inhibitor.

In an embodiment, there is provided the use of compounds as provided herein, according to any one of the embodiments described herein (and/or pharmaceutical compositions comprising such compounds, according to any one of the embodiment described herein), in the manufacture of a medicament for: the treatment of a disease or disorder associated with NLRP3 activity (including inflammasome activity); the treatment of a disease or disorder in which the NLRP3 signalling contributes to the pathology, and/or symptoms, and/or progression, of said disease/disorder; and/or inhibiting NLRP3 inflammasome activity (including in a subject in need thereof).

In an embodiment, there is provided a method of treating a disease or disorder in which the NLRP3 signalling contributes to the pathology, and/or symptoms, and/or progression, of said disease/disorder, comprising administering a therapeutically effective amount of a compound as provided herein, according to any one of the embodiments described herein (and/or pharmaceutical compositions comprising such compound, according to any one of the embodiment described herein), for instance to a subject (in need thereof). In a further embodiment, there is provided a method of inhibiting the NLRP3 inflammasome activity in a subject (in need thereof), the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound as provided herein, according to any one of the embodiments described herein (and/or pharmaceutical compositions comprising such compound, according to any one of the embodiment described herein).

The instant compounds may have the advantage that they may be more efficacious than, be less toxic than, be longer acting than, be more potent than, produce fewer side effects than, be more easily absorbed than, and/or have a better pharmacokinetic profile (e.g. higher oral bioavailability and/or lower clearance) than, and/or have other useful pharmacological, physical, or chemical properties over, compounds known in the prior art, whether for use in the above-stated indications or otherwise.

For instance, the instant compounds may have the advantage that they have a good or an improved thermodynamic solubility (e.g. compared to compounds known in the prior art; and for instance as determined by a known method and/or a method described herein). The instant compounds may have the advantage that they will block pyroptosis, as well as the release of pro-inflammatory cytokines (e.g. IL-10) from the cell. The instant compounds may also have the advantage that they avoid side-effects, for instance as compared to compounds of the prior art, which may be due to selectivity of NLRP3 inhibition.

Compounds as provided herein may also have the advantage that they have good or improved in vivo pharmacokinetics and oral bioavailability. They may also have the advantage that they have good or improved in vivo efficacy. Specifically, the instant compounds may also have advantages over prior art compounds when compared in the tests outlined hereinafter.

EXPERIMENTAL PART

EXAMPLES

Several methods for preparing the Compounds of this disclosure are illustrated in the following examples. Unless otherwise noted, all starting materials were obtained from commercial suppliers and used without further purification, or alternatively can be synthesized by a skilled person by using published methods.

Table 1 : Abbreviations

Preparation of intermediates

For intermediates that were used in a next reaction step as a crude or as a partially purified intermediate, in some cases no mol amounts are mentioned for such intermediate in the next reaction step or alternatively estimated mol amounts or theoretical mol amounts for such intermediate in the next reaction step are indicated in the reaction protocols described below.

2-Iodo-5-(trifluoromethyl)phenol [102771-00-6] Il

Sodium hydride [CAS 7646-69-7] (2.47 g, 61.69 mmol) was suspended in anhydrous toluene (92 mL) at 0°C under nitrogen atmosphere. Then 3- trifluoromethylphenol [CAS 98-17-9] (3.8 mL, 30.84 mmol) was added dropwise. The mixture was stirred at 0 °C for 30 min. Then iodine [7553-56-2] (7.83 g, 30.84 mmol) was added portionwise and the mixture was stirred from 0°C to rt for 3 h. The mixture was acidified at 0 °C with 37% HC1 sol. in water until pH reached 4-5, then was extracted with EtOAc and washed twice with brine. The organic layer was separated, dried (MgSO4), filtered and the solvents evaporated in vacuo. The crude product was purified by flash column chromatography (silica 80 g; EtOAc in heptane 0/100 to 10/90). The desired fractions were collected and concentrated in vacuo to yield Intermediate 1 (6.5 g, yield 72 %) as a colourless oil. l-Iodo-2-(methoxymethoxy)-4-(trifluoromethyl)benzene 12

Intermediate 1 (6.5 g, 22.6 mmol) and K2CO3 [CAS 584-08-7] (5.3 g, 38.4 mmol) were dissolved in anhydrous DMF (95 mL). The reaction mixture was stirred at rt for 30 minutes. Then chloromethyl methyl ether [CAS 107-30-2] (2.4 mL, 29.3 mmol) was added dropwise and the reaction mixture was stirred at rt for 16h. The reaction mixture was diluted with water and extracted with EtOAc. The organic layers were combined, dried (MgSO4), filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (silica 80 g; heptane 100%). The desired fractions were collected and concentrated in vacuo to yield the Intermediate 2 (5.5 g, yield 72%) as a colorless oil.

2-(2-(Methoxymethoxy)-4-(trifluoromethyl)phenyl)-4,4,5,5-tetramethyl-l,3,2- dioxaborolane 13

Isopropyl magnesium chloride (2 M in THF) [CAS 1068-55-9] (7.2 mL, 14.46 mmol) was added dropwise to a stirred solution of Intermediate 2 (4 g, 12.05 mmol) in anhydrous THF (96 mL) at 0 °C under nitrogen atmosphere. The reaction mixture was stirred at 0 °C for 2 h. Afterwards, 2-isopropoxy -4,4,5, 5-tetramethyl-l, 3,2- dioxaborolane [CAS 61676-62-8] (3.7 mL, 18.07 mmol) was added dropwise to the mixture. The reaction mixture was slowly warmed up to rt and stirred for 18 h. The reaction was quenched with NH4C1 (saturated in water) and extracted with EtOAc. The organic layer was separated, dried (MgSO4), filtered and the solvents evaporated in vacuo. The crude product was purified by flash column chromatography (silica 80 g; EtOAc in heptane 0/100 to 20/80). The desired fractions were collected and concentrated in vacuo to yield Intermediate 3 (2.6 g, yield 63 %) as a colorless oil. 3-Methyl-5-(tnfluoromethyl)phenol [934180-46-8] 14

Lithium hydroxide hydrate (2.8 g, 65.24 mmol) was added to a stirred solution of Tris(dibenzylideneacetone)dipalladium(0) [CAS 51364-51-3] (616 mg, 0.652 mmol) and BippyPhos [CAS 894086-00-1] (681 mg, 1.31 mmol) in 50 mL of 1,4-dioxane and 5 ml of distilled water (previously bubbled with nitrogen for 5 min). The mixture was stirred at rt for 5 min, then 3-bromo-5-methylbenzotrifluoride [CAS 86845-28-5] (5.25 g, 21.74 mmol) was added. The reaction mixture was stirred at 100 °C for 16 h. The mixture was filtered over a pad of celite® and was washed with AcOEt. The filtrate was washed with HC1 (2 M in water). The organic layer was separated, dried (MgSO4), filtered and the solvents evaporated in vacuo. The crude product was purified by flash column chromatography (silica 120g; AcOEt in heptane 0/100 to 2/98). The desired fractions were collected and concentrated in vacuo to yield Intermediate 4 (3.4 g, yield 80%) as a yellow oil.

2-Iodo-3-methyl-5-(trifluoromethyl)phenol 15

Sodium hydride (60% dispersion in mineral oil, 1.59 g, 39.7 mmol) was added to a stirred solution of Intermediate 4 (3.4 g, 19.9 mmol) in 60 mL of anhydrous toluene at 0 °C under nitrogen atmosphere. The mixture was stirred at 0 °C for 30 min. Then iodine (5.05 g, 19.87 mmol) was added portionwise and the mixture was stirred from 0 °C for 3 h. The mixture was acidified at 0 °C with cone. HC1 until pH 4-5, then was extracted with AcOEt and washed twice with brine. The organic layer was separated, dried (MgSO4), filtered and the solvents evaporated in vacuo to yield Intermediate 5 (6.5 g, yield 74%) as a brown oil which was used as such without further purification. 2-Iodo-l-(methoxymethoxy)-3-methyl-5-(trifluoromethyl)benzene 16

Intermediate 5 (9.05 g, 29.96 mmol) was dissolved in DCM [75-09-2] (300 mL) and cooled to 0°C. To this solution was added N,N-Diisopropylethylamine [7087-68-5] (6.34 mL, 35.96 mmol) followed by dropwise addition of chloromethyl methyl ether [107-30-2] (2.8 mL, 35.96 mmol). This was allowed to gradually warm to rt and react for 16 h. The mixture was concentrated and the residue was purified by flash column chromatography (Load in silica 120g; DCM in heptane from 0/100 to 3/97). The desired fractions were collected and concentrated in vacuo to yield Intermediate 6 (7.39 g, yield: 71%) as a yellowish oil.

2-(2-(Methoxymethoxy)-6-methyl-4-(trifluoromethyl)phenyl)-4,4,5,5-tetramethyl-

1,3,2-dioxaborolane 17

Intermediate 6 (7.4 g, 21.34 mmol) was added dropwise to a stirred solution of Palladium(II) acetate [3375-31-3] (489 mg, 2.13 mmol), CyJohnPhos [247940-06-3] (787 mg, 0.57 mmol) and triethylamine [121-44-8] (15 mL, 106.72 mmol) and anhydrous 1,4-di oxane (92 mL) (previously bubbled with nitrogen for 5 min) in a sealed tube. The mixture was stirred at rt for 5 min, then 4, 4,5,5- Tetramethyl[l,3,2]dioxaborolane [25015-63-8] (16 mL, 106.72 mmol) was added. The reaction mixture was stirred at 100°C for 16h. The mixture was filtered over a pad of celite® and was washed with EtOAc. The filtrate was washed with sat. aqueous NH4C1. The organic layer was separated, dried (MgSO4), filtered and the solvents evaporated in vacuo. The crude product was purified by flash column chromatography (silica 120g; EtOAc in heptane 0/100 to 1/99). The desired fractions were collected and concentrated in vacuo to yield Intermediate 7 (6.87 g, yield 90%) as a orange solid. 2-Hydrazineyl-3-iodopyr idine 18- 1

Hydrazine monohydrate (1.8 mL, 1 g/mL, 35.9 mmol) was added to stirred solution of 2-fluoro-3 -iodopyridine (2 g, 9 mmol) in 14 mL of EtOH and the reaction mixture was stirred at 50 °C for 16 h. The solvent was evaporated in vacuo. Then was washed with water and the white solid that formed was filtered, washed with water. The product was dissolved in DCM, dried (MgSCU), filtered and concentrated in vacuo to yield Intermediate 8 (1.2 g, yield 57%) as a white solid.

The following intermediates were made accordingly from the corresponding commercially available fluoro analogues. The skilled chemist will understand that depending on the electronic of the system, reactivity may be different resulting in a different temperature for the reaction. tert- Butyl 3-isothiocyanatopiperidine-l-carboxylate 19

A tube was charged with l-Boc-3 -aminopiperidine (1 g, 5 mmol) in 20 mL of anhydrous DCM. The mixture was cooled to -70 °C and l,l'-thiocarbonyldi -2(177)- pyridone (1.4 g, 6 mmol) was added in one portion. The mixture was allowed to rise to rt and was further stirred for 16 h. The reaction mixture was washed with a 10 % aqueous solution of K2CO3. The organic layer was dried over MgSC , filtered off and the solvent removed by evaporation. A purification was performed by column chromatography on a 25g SFAR cartridge in a biotage system using 100% DCM (12cv). The product fractions were collected and concentrated in vacuo to yield Intermediate 9 (1.2 g, assumed quant, yield) as a clear oil.

Alternatively, a flask was charged with l-Boc-3 -aminopiperidine (5 g, 25 mmol) and Et3N (10.8 mL, 0.7 g/mL, 74.9 mmol) in anhydrous DCM (100 mL). The mixture was cooled to -70 °C and thiophosgene (2.3 mL, 1.5 g/mL, 30 mmol) was added dropwise. The mixture was allowed to rise to room temperature and was further stirred at room temperature for 2h. The reaction mixture was washed with water. The organic layer was dried over MgSO4, filtrated and the solvent removed by evaporation. A purification was performed by column chromatography on a 10g SFAR cartridge in a biotage system using a gradient from 0% till 50% EtOAc in heptane (12cv). The product fractions were collected and concentrated in vacuo to yield Intermediate 9 (5.6 g, yield 93%) as an oil. tert- Butyl 3-(2-(3-iodopyridin-2-yl)hydrazine-l-carbothioamido)piperidine-l- carboxylate II 0-1

A VLT tube was charged with Intermediate 9 (468 mg, 1.9 mmol) in 10 mL of anhydrous DCM. The mixture was cooled on an ice bath to 0 - 5 °C. A solution of Intermediate 8-1 (412.6 mg, 1.8 mmol) in 5 mL DCM was added dropwise to the cooled mixture and the mixture was further stirred for 2 h at rt. The precipitated product was filtered off and dried to yield Intermediate 10-1 (817 mg, yield 98%) as a white solid.

The following intermediates were made accordingly: tert-butyl 3-(2-(2-(3-iodopyridin-2-yl)hydrazineyl)-2-oxoethyl)piperidine-l- carboxylate Ill DCC [538-75-0] (300 mg, 1.454 mmol) was added to a stirred solution of N- Boc-3 -piperidineacetic acid [183483-09-2] (328 mg, 1.348 mmol) and Intermediate 8-1 (330 mg, 1.404 mmol) and the resulting mixture was stirred at room temperature for 1 h. Volatiles were removed in vacuo and ACN was added. The precipitate was filtered off and washed with ACN. The filtrate was concentrated under reduced pressure to afford Intermediate 11 (620 mg, assumed quant, yield) as a yellow froth which was used without further purification for the next step. tert-butyl 3-((8-iodo-[l,2,4]triazolo[4,3-a]pyi'idin-3-yl)methyl)piperidine-l- carboxylate 112

Burgess reagent [29684-56-8] (700 mg, 2.937 mmol) was added to a stirred solution of Intermediate 11 (610 mg, 1.325 mmol) in 40 mL of anhydrous acetonitrile and the resulting mixture was heated at 80 °C for 5 h. Upon cooling to room temperature, the mixture was evaporated under reduced pressure and directly subjected to column chromatography on silica gel with Hept/EtOAc (1 :0 to 0: 1) to afford Intermediate 12 (503 mg, yield 86%) as an off-white solid. tert-butyl (31?,51?)-3-fluoro-5-(2-(3-iodopyridin-2-yl)hydrazine-l carbothioamido)piperidine-l-carboxylate 113 tert-butyl (3R, 5A)-3-amino-5-fluoropiperidine-l-carboxylate (510.8 mg, 2.3 mmol) and Et3N (0.9 mL, 0.7 g/mL, 6.4 mmol) was dissolved in dry DCM (20 mL). The mixture was cooled on an ice bath to 0-5°C. Thiophosgene (163.1 pL, 1.5 g/mL, 2.1 mmol) was added dropwise to the mixture at 0-5°C. The mixture was allowed to rise to rt and further stirred for 2h. The mixture was cooled to 0-5°C and Intermediate 8-1 (300 mg, 1.3 mmol) dissolved in 5 mL DCM was added dropwise to the cooled mixture. The mixture was further stirred at r.t. for 16 h. The mixture was washed with water and the organic layer was separated, dried (MgSO4), filtered and concentrated in vacuo. A purification was performed by column chromatography on a 25g SFAR cartridge in a Biotage system using a gradient from 0% till 50% DCM/MeOH (90/10) in DCM (12cv). The product fractions were collected and concentrated in vacuo to yield Intermediate 13 (598 mg, yield 57%) as a yellow foam. tert-butyl 3-((8-iodo-[l,2,4]triazolo[4,3-a]pyi'idin-3-yl)amino)piperidine-l- carboxylate 114-1

Potassium carbonate (224 mg, 1.62 mmol) and iodine (229 mg, 0.90 mmol) were added to a stirred solution of Intermediate 10-1 (430 mg, 0.90 mmol) in 8 mL of 1,4-di oxane. The reaction mixture was stirred at 80 °C for 4 h. The solvent was evaporated in vacuo. The crude product was purified by flash column chromatography (silica 25g; MeOH in DCM from 0/100 to 5/95). The desired fractions were collected and concentrated in vacuo to yield Intermediate 14-1 (330 mg, yield 79%) as a brown sticky oil. The following intermediates were made accordingly:

tert-butyl 3-((8-(2-hydroxy-4-(trifluoromethyl)phenyl)-[l,2,4]triazolo[4,3- a]pyridin-3-yl)amino)piperidine-l-carboxylate 115-1 Tetrakis(triphenylphosphine)palladium (0) [14221-01-3] (0.087 g, 0.074 mmol) was added to a stirred suspension of Intermediate 14-1 (0.33 g, 0.74 mmol), 2 -hydroxy - 4-trifluoromethylphenylboronic acid [1072951-50-8] (0.26 g, 1.01 mmol) and Na2CO3 (0.24 g, 2.23 mmol) in 6 mL of 1,4-di oxane and 1.5 mL of water (previously bubbled with nitrogen for 5 min) in a sealed tube. The reaction mixture was stirred at 100 °C for 16 h. Then, the mixture was filtered over celite® pad and solvents were concentrated in vacuo. The crude was purified by flash column chromatography (SiO2 25 g; EtOAc in Heptane from 0/100 to 50/50). The desired fractions were collected and concentrated in vacuo to yield Intermediate 15-1 (350 mg, yield 97%) as a brown foam solid. The following intermediates were made accordingly: tc/7-but l (l?)-3-((8-(2-(methoxymethoxy)-4-(trifluoromethyl)phenyl)- [l,2,4]triazolo[4,3-«]pyi'idin-3-yl)(methyl)amino)piperidine-l-carboxylate 116

NaH (60% dispersion in mineral oil) [7646-69-7] (38.1 mg, 0.95 mmol, 2 equiv) was added portionwise into a mixture of Intermediate 15-14 (248.5 mg, 0.48 mmol, 1 equiv) in dry DMF (10 mL) at 0 °C and flushed with nitrogen. The reaction was stirred at rt for 15 min. After cooling again to 0 °C, iodomethane [74-88-4] (89 pL, 2.28 g/mL, 1.43 mmol, 3 equiv) was added and then the reaction mixture was allowed to stir at rt for 2.5 h. The reaction mixture was quenched with sat. aq. NaHCO3 solution and extracted with EtOAc three times. The combined organic layers were dried over MgSO4, filtered off and concentrated in vacuo. The crude product was purified by flash column chromatography (Combiflash, 0 to 100% EtOAc:Hept, over 15 min, silica 24 g, 35 mL/min, liquid deposit using DCM, product comes with ca. 70% EtOAc) yielding Intermediate 16 (210 mg, yield 68%) as a yellow oil. tc/7-but l (l?)-3-((8-(2-hydroxy-4-(trifluoromethyl)phenyl)-6-methyl- [l,2,4]triazolo[4,3-«]pyi'idin-3-yl)amino)piperidine-l-carboxylate 117

A vial was charged with Intermediate 15-6 (180 mg, 0.352 mmol), Trimethylboroxine 50 % w/w in THF (0.128 mL, 0.898 g/mL, 0.457 mmol) and sodium tert-butoxide (0.527 mL, 2 M, 1.055 mmol) in dry 1,4-dioxane (5 mL, 1.03 g/mL, 58.452 mmol). The solution was degazed under N2 atmosphere for 5 minutes. Xphos Pd G4 (8.299 mg, 9.645 pmol) was added. The tube was sealed and the mixture reaction was stirred at 100 °C for 5h (After 4h, RM is purged again with N2 for 5 min and some more Xphos Pd G4 was added). Water (20 mL) and EtOAc (25 mL) were added. Aqueous layer was extracted with EtOAc (3 x 30 mL). Organic layers were collected, dried over MgSO4, filtered and concentrated under vacuum. Then, a purification was performed by column chromatography on a 10g SFAR cartridge in a biotage system using a gradient from 0% till 100% EtOAc in heptane (12cv). The product fractions were collected and concentrated in vacuo to become Intermediate 17 (100 mg, yield 58%). tc/7-butyl (l?)-3-((8-(2-hydroxy-4-(trifluoromethyl)phenyl)-6-cyclopropyl- [l,2,4]triazolo[4,3-«]pyi'idin-3-yl)amino)piperidine-l-carboxylate 118

Intermediate 18 was made by analogy with Intermediate 17, using potassium cyclopropyltrifluoroborate as coupling partner.

2-fluoro-3-(2-(methoxymethoxy)-4-(trifluoromethyl)phenyl)pyrazine 119

A 10-20 mL MW vial was charged with 2-fluoro-3 -iodopyrazine [206278-26-4] (741.2 mg, 3.31 mmol, 1 equiv), Intermediate 3 (2.2 g, 6.62 mmol, 2 equiv), Pd(PPhs)4 [14221-01-3] (382.4 mg, 0.33 mmol, 10 mol%) and K2CO3 [584-08-7] (686.0 mg, 4.96 mmol, 1.5 equiv) in a mixture of DME [110-71-4] (8.6 mL) and DI water (2.5 mL). The reaction mixture was heated at 90 °C overnight. The reaction mixture was filtered off and poured into water and EtOAc. The organic layer was separated, and the aqueous layer was extracted again with EtOAc twice. The combined organic layers were dried over MgSO4, filtered off and concentrated in vacuo. The crude product was purified by flash column chromatography (Combiflash, 0 to 40% EtOAc:Hept, over 15 min, silica 40 g, 40 mL/min, liquid deposit using DCM, product comes with ca. 20- 30% EtOAc, tubes 9 to 19) yielding Intermediate 19 (1.27 g, 89% purity, quant.) as a yellow solid. 2-hydrazineyl-3-(2-(methoxymethoxy)-4-(trifluoromethyl)phenyl)pyrazine 120

Hydrazine monohydrate [10217-52-4] (0.94 mL, 1.03 g/mL, 19.44 mmol, 5.5 equiv) was added to stirred solution of Intermediate 19 (1.2 g, 3.53 mmol, 1 equiv) in EtOH (5.5 mL) and the reaction mixture was stirred at 50 °C for 16 h. The reaction mixture was cooled at 0 °C and the resulting precipitate was filtered. The filtrate was concentrated in vacuo. The resulting residue was further triturated in ACN at 0 °C and filtered yielding Intermediate 20 (853 mg, yield 77%) as an off-white solid. tert-butyl (R)-3-(2-(3-(2-(methoxymethoxy)-4-(trifluoromethyl)phenyl)pyrazin-2- yl)hydrazine-l-carbothioamido)piperidine-l-carboxylate 121

A flask was charged with Intermediate 9 (536 mg, 2.21 mmol, 1.1 equiv) in dry DCM (7 mL). The reaction mixture was cooled to 0 °C in an ice bath. A solution of Intermediate 20 (631.89 mg, 2.01 mmol, 1 equiv) in dry DCM (7.5 mL) was added dropwise and the reaction mixture was stirred at rt for 5 h. The reaction mixture was concentrated in vacuo. The crude product was purified by flash column chromatography (Combiflash, 0 to 3% MeOH:DCM, over 15 min, silica 24 g, 35 mL/min, liquid deposit using DCM, product comes with ca. 1-2% MeOH, tubes 13 to 17) yielding Intermediate 21 (763.7 mg, yield 53%, 78% purity) as a bright yellow foam. tert-butyl (l?)-3-((8-(2-(methoxymethoxy)-4-(trifluoromethyl)phenyl)- [l,2,4]triazolo[4,3-«]pyi’azin-3-yl)amino)piperidine-l-carboxylate 122

To a solution of Intermediate 21 (735 mg, 1.32 mmol, 1 equiv) in dry 1,4- dioxane (9 mL) was added K2CO3 [584-08-7] (365 mg, 2.64 mmol, 2 equiv) and iodine [7553-56-2] (368.7 mg, 1.45 mmol, 1.1 equiv). The mixture was heated at 80 °C for 4.5 h. The reaction mixture was concentrated in vacuo. The residue was pardoned between water and DCM. The aqueous layer was extracted with DCM twice. The combined organic layer were dried over MgSO4, filtered off and concentrated in vacuo. The crude product was purified by flash column chromatography (Combiflash, 0 to 5% MeOH:DCM, over 60 min, silica 40 g, 40 mL/min, liquid deposit using DCM, product comes with ca. 2-3% MeOH, tubes 10 to 24) yielding Intermediate 22 (77.6 mg, yield 9%, 81% purity) as a brown solid.

2-(3-(piperidin-3-ylamino)-[l,2,4]triazolo[4,3-«]pyi'idin-8-yl)-5-

(trifluoromethyl)phenol hydrochloride 123-1

HC1 4N in dioxane [7647-01-0] (2.75 mL, 11.00 mmol) was added to a stirred solution of Intermediate 15-1 (350 mg, 0.73 mmol) in Dioxane (0.4 mL) at 0°C. The mixture was stirred at rt for Ih. The solvent was concentrated in vacuo to yield Intermediate 23-1 (330 mg, estim. quant, yield) as a brown foam solid.

Alternatively, HC1 (6M in iPrOH) (2.7 mL, 6 M, 16.4 mmol) was added to a soluton of Intermediate 15-1 (783 mg, 1.6 mmol) in iPrOH (10 mL) and the mixture was heated at 80 °C for 30'. The mixture was cooled and concentrated in vacuo. The product was triturated in dipe, filtered off and dried under vacuum to become Intermediate 23-1 (578 mg, yield 93%). The following intermediates were made accordingly:

Preparation of final compounds

The preparation of final compounds can be done using racemic mixture followed by SFC or directly the enantiopure material. In case of racemic mixtures, absolute configurations were attributed by synthesizing the enantiopure materials using the same sequence.

(5)-2-(3-((l-methylpiperidin-3-yl)amino)-[l,2,4]triazolo[4,3-«]pyi'idin-8-yl)-5-

(trifluoromethyl)phenol 1 and (l?)-2-(3-((l-methylpiperidin-3-yl)amino)- [l,2,4]triazolo[4,3-«]pyi'idin-8-yl)-5-(trifluoromethyl)phenol 2

Sodium triacetoxyborohydride [56553-60-7] (240 mg, 1.10 mmol) was added to a stirred solution of Intermediate 23-1 (330 mg, 0.73 mmol), triethylamine [121-44-8] (513 pL, 3.66 mmol) and formaldehyde (37% aqueous solution) [50-00-0] (109 pL, 1.47 mmol) in methanol (10.2 mL) at 0 °C under nitrogen atmosphere. The mixture was stirred at rt for 16h. Solvent was evaporated in vacuo. The crude was purified by reverse phase using as a column: Phenomenex Gemini C18 30x100mm 5pm Column; from 81% [65mM NH4OAc + ACN (90: 10)] - 19% [ACN:MeOH (1 : 1)] to 45% [65mM NH4OAc + ACN (90: 10)] - 55% [ACN:MeOH (1 : 1)]; Method: MMP4AC. Fractions of Pl were collected and concentrated in vacuo. The impure product was repurified by reverse phase using as a column: Phenomenex Gemini C18 30x100mm 5pm Column; from 95% [25mM NH4HCO3] - 5% [ACN:MeOH (1 : 1)] to 0% [25mM NH4HCO3] - 100% [ACNMeOH (1 : 1)]; Method: M5BIC. Fractions of Pl were collected and extracted with DCM. The combined organic layers were dried over anh. MgSO4, filtered and concentrated vacuo. SFC purification was made using as a column: i-amylose-1 column with ISOC 40% 2 Propanol + 0.1% DEA to collect and concentrate to yield Compound 1 (54.3 mg, yield 43%) as a yellowish foam solid and Compound 2 (49.6 mg, yield 39%) as a yellowish foam solid.

The following compounds were made accordingly:

Analytical data

Example A - LCMS

The High-Performance Liquid Chromatography (HPLC) measurement was performed using a LC pump, a diode-array (DAD) or a UV detector and a column as specified in the respective methods. If necessary, additional detectors were included (see table of methods below).

Flow from the column was brought to the Mass Spectrometer (MS) which was configured with an atmospheric pressure ion source. It is within the knowledge of the skilled person to set the tune parameters (e.g. scanning range, dwell time. . .) in order to obtain ions allowing the identification of the compound’s nominal monoisotopic molecular weight (MW). Data acquisition was performed with appropriate software.

Compounds are described by their experimental retention times (Rt) and ions. If not specified differently in the table of data, the reported molecular ion corresponds to the [M+H]+ (protonated molecule) and/or [M-H]- (deprotonated molecule). In case the compound was not directly ionizable the type of adduct is specified (i.e. [M+NH4]+, [M+HCOO]-, etc. . .). For molecules with multiple isotopic patterns (Br, Cl), the reported value is the one obtained for the lowest isotope mass. All results were obtained with experimental uncertainties that are commonly associated with the method used.

Hereinafter, “SQD” means Single Quadrupole Detector, “MSD” Mass Selective Detector, “RT” room temperature, “BEH” bridged ethylsiloxane/silica hybrid, “DAD” Diode Array Detector, ”HSS” High Strength silica.

LCMS data for all final compounds are depicted below:

Example B - NMR

1H NMR spectra were recorded on 400 MHz Bruker Avance III and Avance NEO spectrometers. DMSO-de was used as solvent, unless otherwise mentioned. The chemical shifts are expressed in ppm relative to tetramethylsilane.

Example C - Pharmaceutical Compositions

A compound of the invention (for instance, a compound of the examples) is brought into association with a pharmaceutically acceptable carrier, thereby providing a pharmaceutical composition comprising such active compound. A therapeutically effective amount of a compound of the invention (e.g. a compound of the examples) is intimately mixed with a pharmaceutically acceptable carrier, in a process for preparing a pharmaceutical composition. Example D - Biological Examples

The activity of a compound according to the present invention can be assessed by in vitro methods. A compound the invention exhibits valuable pharmacological properties, e.g. properties susceptible to inhibit NLRP3 activity, for instance as indicated the following test, and are therefore indicated for therapy related to NLRP3 inflammasome activity. PBMC assay

Peripheral venous blood was collected from healthy individuals and human peripheral blood mononuclear cells (PBMCs) were isolated from blood by Ficoll-Histopaque (Sigma-Aldrich, A0561) density gradient centrifugation. After isolation, PBMCs were stored in liquid nitrogen for later use. Upon thawing, PBMC cell viability was determined in growth medium (RPMI media supplemented with 10% fetal bovine serum, 1% Pen-Strep and 1% L-glutamine). Compounds were spotted in a 1 :3 serial dilution in DMSO and diluted to the final concentration in 30 pl medium in 96 well plates (Falcon, 353072). PBMCs were added at a density of 7.5 x 104 cells per well and incubated for 30 min in a 5% CO2 incubator at 37 °C. LPS stimulation was performed by addition of 100 ng/ml LPS (final concentration, Invivogen, tlrl-smlps) for 6 h followed by collection of cellular supernatants and the analysis of IL-ip (pM) and TNFa cytokines levels (pM) via MSD technology according to manufacturers’ guidelines (MSD, K151A0H).

The IC50 values (for IL-ip) and EC50 values (TNFa) were obtained on compounds of the invention/examples, and are depicted in the following table:

Example F - Further Testing

One or more compound(s) may be tested in a number of other assays to evaluate, amongst other properties, permeability, stability (including metabolic stability and blood stability) and solubility. Metabolic stability test in liver microsomes and hepatocytes

The metabolic stability of a test compound is tested by using liver microsomes (0.5 mg/ml protein) from human and preclinical species incubated up to 60 minutes at 37°C with 1 pM test compound.

The in vitro metabolic half-life (ti/2 ) is calculated using the slope of the log- linear regression from the percentage parent compound remaining versus time relationship (K), ti/2 = - ln(2)/ K.

The in vitro intrinsic clearance (Clint) (ml/min/mg microsomal protein) is calculated using the following formula:

0.693 V_inc

Where: Vine = incubation volume,

Wmic prot,inc weight of microsomal protein in the incubation.

The metabolic stability of a test compound is tested using liver hepatocytes (1 milj cells) from human and preclinical species incubated up to 120 minutes at 37°C with 1 pM test compound.

The in vitro metabolic half-life (ti/2) is calculated using the slope of the log-linear regression from the percentage parent compound remaining versus time relationship (K), ti/2 = - ln(2)/ K.

The in vitro intrinsic clearance (Clint) (pl/min/million cells) is calculated using the following formula:

0.693 V_inc

Clint ~ - X - x lOOO ti/2 # cells_inc

Where: Vine = incubation volume,

# cellS/n_C = number of cells (xlO⁶) in the incubation

Pharmacokinetics

The compound of interest is spiked at a certain concentration in plasma or blood from the agreed preclinical species; then after incubating to predetermined times and conditions (37°C, 0°C (ice) or room temperature) the concentration of the test compound in the blood or plasma matrix can then be determined with LCMS/MS.

Claims

Claims

1. A compound of formula (I), or pharmaceutically acceptable salts thereof, wherein

Y is CH2 when L is NR⁵, NR⁵-CH2 or CH2; or Y is NR⁵ or O when L is CH2;

R¹⁰ is hydrogen, C1-2 alkyl, haloCi-2 alkyl, hydroxyCi-2 alkyl, C1-2 alkyloxy, haloCi-2 alkyloxy or -CH2CN;

R¹¹ is hydrogen, methyl, hydroxy or fluoro;

R² is hydrogen, C1-2 alkyl, haloCi-2 alkyl, C1-2 alkyloxy, haloCi-2 alkyloxy, cyclopropyl, halo or cyano;

R³ is hydrogen, methyl, ethyl or halo; and

R⁴ is hydrogen, C1-2 alkyl, haloCi-2 alkyl, cyclopropyl, or halo.

2. The compound of claim 1, wherein A is CH; wherein R⁵ is hydrogen;

Y is CH₂;

R¹⁰ is C1-2 alkyl;

R¹¹ is hydrogen or fluoro;

R² is haloCi-2 alkyl, haloCi-2 alkyloxy, or halo;

R³ is hydrogen, methyl or halo; and

R⁴ is hydrogen, C1-2 alkyl, haloCi-2 alkyl, cyclopropyl, or halo.

3. The compound of claim 1 wherein L is NH;

R¹⁰ is methyl;

R¹¹ is hydrogen or fluoro;

R² is trifluoromethyl;

R³ is hydrogen, methyl or halo; and

R⁴ is hydrogen, methyl, haloCi-2 trifluoromethyl, cyclopropyl, or chloro.

4. The compound of claim 1 wherein the compound is (R)-2-(3-((l-methylpiperi din-3 - yl)amino)-[l,2,4]triazolo[4,3-a]pyridin-8-yl)-5-(trifluoromethyl)phenol.

5. The compound of claim 1 wherein the compound is rel-(R)-2-(3-((l-methylpiperidin- 3-yl)methyl)-[l,2,4]triazolo[4,3-a]pyridin-8-yl)-5-(trifluoromethyl)phenol.

6. A pharmaceutical composition comprising a therapeutically effective amount of a compound as defined in any one of claims 1 to 5 and a pharmaceutically acceptable carrier.

7. A process for preparing a pharmaceutical composition as defined in claim 6, characterized in that a pharmaceutically acceptable carrier is intimately mixed with a therapeutically effective amount of a compound as defined in any one of claims