EP4615841A1 - Triazinone derivatives as nlrp3 inhibitors - Google Patents

Abstract

The invention relates to novel compounds having the general formula (I) wherein R¹, R², R³, R⁴ and R⁵ are as described herein, composition including the compounds and methods of using the compounds.

Description

Novel Compounds

Field of the Invention

The present invention relates to organic compounds useful for therapy and/or prophylaxis in a mammal, and in particular to compounds that modulate NLRP3 inhibition.

The present invention provides novel compounds of formula I wherein,

R¹ is H, alkyl, alkoxy, -CN, halo, haloalkyl, haloalkoxy, acetyl or SFe;

R⁵ is H; or R¹ and R⁵, and the atoms to which they are bonded, form either an 4-6 membered heterocycle ring comprising a single O heteroatom optionally substituted with one or two substituents independently selected from halo or alkyl, or R¹ and R⁵, and the atoms to which they are bonded, form a 4-6 membered cycloalkyl ring optionally substituted with 1 to 2 substituents independently selected from halo or alkyl;

R² is H, halo or alkyl and R³ is H or alkyl, wherein only one of R² and R³ can be H;

R⁴ is oxetane, alkyl, or -(CH2)_n-R⁶ wherein R⁶ is hydroxy or methoxy and n is greater than 1; and pharmaceutically acceptable salts thereof. Furthermore, the invention includes all racemic mixtures, all their corresponding enantiomers and/or optical isomers.

Background of the Invention

The NOD-like receptor (NLR) family, pyrin domain-containing protein 3 (NLRP3) inflammasome is a component of the inflammatory process, and its aberrant activity is pathogenic in inherited disorders such as cryopyrin-associated periodic syndromes (CAPS) and complex diseases such as multiple sclerosis, type 2 diabetes, Alzheimer’s disease and atherosclerosis.

NLRP3 is an intracellular signaling molecule that senses many pathogen-derived, environmental and host-derived factors. Upon activation, NLRP3 binds to apoptosis-associated speck-like protein containing a caspase activation and recruitment domain (ASC). ASC then polymerises to form a large aggregate known as an ASC speck. Polymerised ASC in turn interacts with the cysteine protease caspase-1 to form a complex termed the inflammasome. This results in the activation of caspase- 1, which cleaves the precursor forms of the proinflammatory cytokines IL-ip and IL- 18 (termed pro-IL-ip and pro-IL-18 respectively) to thereby activate these cytokines. Caspase-1 also mediates a type of inflammatory cell death known as pyroptosis. The ASC speck can also recruit and activate caspase-8, which can process pro-IL-ip and pro-IL- 18 and trigger apoptotic cell death.

Caspase- 1 cleaves pro-IL-ip and pro-IL-18 to their active forms, which are secreted from the cell. Active caspase- 1 also cleaves gasdermin-D to trigger pyroptosis. Through its control of the pyroptotic cell death pathway, caspase- 1 also mediates the release of alarmin molecules such as IL-33 and high mobility group box 1 protein (HMGB1). Caspase-1 also cleaves intracellular IL-1R2 resulting in its degradation and allowing the release of IL-la. In human cells caspase-1 may also control the processing and secretion of IL-37. A number of other caspase-1 substrates such as components of the cytoskeleton and glycolysis pathway may contribute to caspase-1- dependent inflammation.

NLRP3 -dependent ASC specks are released into the extracellular environment where they can activate caspase-1, induce processing of caspase-1 substrates and propagate inflammation. Active cytokines derived from NLRP3 inflammasome activation are important drivers of inflammation and interact with other cytokine pathways to shape the immune response to infection and injury. For example, IL-ip signalling induces the secretion of the pro-inflammatory cytokines IL-6 and TNF. IL-ip and IL- 18 synergise with IL-23 to induce IL- 17 production by memory CD4 Th 17 cells and by y6 T cells in the absence of T cell receptor engagement. IL- 18 and IL-12 also synergise to induce IFN-y production from memory T cells and NK cells driving a Thl response.

The inherited CAPS diseases Muckle-Wells syndrome (MWS), familial cold autoinflammatory syndrome (FCAS) and neonatal -onset multisystem inflammatory disease (NOMID) are caused by gain-of-function mutations in NLRP3, thus defining NLRP3 as a critical component of the inflammatory process. NLRP3 has also been implicated in the pathogenesis of a number of complex diseases, notably including metabolic disorders such as type 2 diabetes, atherosclerosis, obesity and gout.

A role for NLRP3 in diseases of the central nervous system is emerging, and lung diseases have also been shown to be influenced by NLRP3. NLRP3 has also been suggested to have a role in a number of central nervous system conditions, including Parkinson's disease (PD), Alzheimer's disease (AD), dementia, Huntington's disease, cerebral malaria, brain injury from pneumococcal meningitis (Walsh et al., Nature Reviews, 15: 84-97, 2014, and Dempsey et al. Brain. Behav. Immun. 201761 : 306-316). NLRP3 has also been shown to play a role in a number of lung diseases including chronic obstructive pulmonary disorder (COPD), asthma (including steroid-resistant asthma), asbestosis, and silicosis (De Nardo et al., Am. J. Pathol., 184: 42-54, 2014 and Kim et al. Am J Respir Crit Care Med. 2017 196(3): 283-97). Furthermore, NLRP3 has a role in the development of liver disease, kidney disease and aging. Many of these associations were defined using Nlrpi-^ mice, but there have also been insights into the specific activation of NLRP3 in these diseases. In type 2 diabetes mellitus (T2D), the deposition of islet amyloid polypeptide in the pancreas activates NLRP3 and IL-ip signalling, resulting in cell death and inflammation.

Several small molecules have been shown to inhibit the NLRP3 inflammasome. Glyburide inhibits IL-ip production at micromolar concentrations in response to the activation of NLRP3 but not NLRC4 or NLRP1. Other previously characterised weak NLRP3 inhibitors include parthenolide, 3,4-methylenedioxy-P-nitrostyrene and dimethyl sulfoxide (DMSO), although these agents have limited potency and are nonspecific.

Current treatments for NLRP3-related diseases include biologic agents that target IL-1. These are the recombinant IL-1 receptor antagonist anakinra, the neutralizing IL-ip antibody canakinumab and the soluble decoy IL-1 receptor rilonacept. These approaches have proven successful in the treatment of CAPS, and these biologic agents have been used in clinical trials for other IL-ip-associated diseases.

There is a need to provide compounds with improved pharmacological and/or physiological and/or physicochemical properties and/or those that provide a useful alternative to known compounds.

Summary of the Invention

The present invention provides novel compounds of formula I wherein,

R¹ is H, alkyl, alkoxy, -CN, halo, haloalkyl, haloalkoxy, acetyl or SFe;

R⁵ is H; or R¹ and R⁵, and the atoms to which they are bonded, form either an 4-6 membered heterocycle ring comprising a single O heteroatom optionally substituted with one or two substituents independently selected from halo or alkyl, or R¹ and R⁵, and the atoms to which they are bonded, form a 4-6 membered cycloalkyl ring optionally substituted with 1 to 2 substituents independently selected from halo or alkyl;

R² is H, halo or alkyl and R³ is H or alkyl, wherein only one of R² and R³ can be H; R⁴ is oxetane, alkyl, or -(CH2)_n-R⁶ wherein R⁶ is hydroxy or methoxy and n is greater than 1; and pharmaceutically acceptable salts thereof.

The term “alkyl” denotes a monovalent linear or branched saturated hydrocarbon group of 1 to 6 carbon atoms. In some embodiments, if not otherwise described, alkyl comprises 1 to 6 carbon atoms (Ci-6-alkyl), or 1 to 4 carbon atoms (Ci-4-alkyl). Examples of Ci-6-alkyl include methyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, tert-butyl and pentyl. Particular alkyl group is methyl.

The term “alkoxy” denotes a group of the formula -O-R’, wherein R’ is a Ci-6-alkyl group. Examples of Ci-6-alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy and tert-butoxy.

The term “cycloalkyl” denotes monocyclic or polycyclic saturated or partially unsaturated, non-aromatic hydrocarbon. In some embodiments, unless otherwise described, cycloalkyl comprises 3 to 8 carbon atoms, 3 to 6 carbon atoms, or 3 to 5 carbon atoms. In some embodiments, cycloalkyl is a saturated monocyclic or polycyclic hydrocarbon. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and octahydropentalenyl, and the like. Particular example is cyclobutyl. Another particular example is cyclopentyl.

The term “halogen”, “halide” and “halo” are used interchangeably herein and denote fluoro, chloro, bromo or iodo. Particular halogens are fluoro and chloro. Preferred halogen is fluoro.

The term “haloalkyl” denotes a Ci-6-alkyl group wherein at least one of the hydrogen atoms of the Ci-6-alkyl group has been replaced by the same or different halogen atoms. Example of haloalkyl include fluoromethyl, difluoromethyl and trifluoromethyl. Particular example is tri fluoromethyl.

The term “haloalkoxy” denotes a Ci-6-alkoxy group wherein at least one of the hydrogen atoms of the Ci-6-alkoxy group has been replaced by the same or different halogen atoms. Examples of haloalkoxy are difluoromethoxy, trifluoromethoxy, difluoroethoxy and trifluoroethoxy. Particular examples are difluoromethoxy and trifluromethoxy. The term “heterocycle ring” denotes a monovalent saturated or partly unsaturated mono- or bicyclic ring system of 4 to 9 ring atoms, comprising 1, 2, or 3 ring heteroatoms selected from N, O and S, the remaining ring atoms being carbon. Examples for monocyclic saturated heterocycle rings are azetidinyl, diazepanyl, pyrrolidinyl, tetrahydrofuranyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, and piperazinyl. Examples of polycyclic saturated heterocycle rings are azaspiroheptanyl, diazaspiroheptanyl. azaspirooctanyl, diazospirooctanyl, diazaspirononanyl, oxaazaspirooctanyl, and oxadiazaspirononanyl. Particular example of a heterocycle ring is a saturated furanyl ring.

The term “hydroxy” denotes a -OH group.

The term “nitrile” denotes a -C=N group.

The term “pharmaceutically acceptable salts" refers to those salts which retain the biological effectiveness and properties of the free bases or free acids, which are not biologically or otherwise undesirable. The salts are formed with inorganic acids such as trifluoroacetic acid, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, particularly hydrochloric acid, and organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, N-acetylcystein. In addition these salts may be prepared from addition of an inorganic base or an organic base to the free acid. Salts derived from an inorganic base include, but are not limited to, the sodium, potassium, lithium, ammonium, calcium, magnesium salts. Salts derived from organic bases include, but are not limited to salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, lysine, arginine, N-ethylpiperidine, piperidine, polyamine resins. The compound of formula I can also be present in the form of zwitterions. Particularly preferred pharmaceutically acceptable salts of compounds of formula I are the salts formed with formic acid and the salts formed with hydrochloric acid yielding a hydrochloride, dihydrochloride or trihydrochloride salt.

The abbreviation uM means microMolar and is equivalent to the symbol pM. The abbreviation uL means microliter and is equivalent to the symbol pL.

The abbreviation ug means microgram and is equivalent to the symbol pg.

The compounds of formula I can contain several asymmetric centers and can be present in the form of optically pure enantiomers, mixtures of enantiomers such as, for example, racemates, optically pure diastereoisomers, mixtures of diastereoisomers, diastereoisomeric racemates or mixtures of diastereoisomeric racemates.

According to the Cahn-Ingold-Prelog Convention the asymmetric carbon atom can be of the "R" or "S" configuration.

Also an embodiment of the present invention provides compounds according to formula I as described herein and pharmaceutically acceptable salts or esters thereof, in particular compounds according to formula I as described herein and pharmaceutically acceptable salts thereof, more particularly compounds according to formula I as described herein.

An embodiment of the present invention provides compounds according to formula I as described herein, where R¹ is H, alkyl, alkoxy, halo, haloalkyl, or haloalkoxy;

An embodiment of the present invention provides compounds according to formula I as described herein, wherein R¹ is haloalkyl or haloalkoxy.

An embodiment of the present invention provides compounds according to formula I as described herein, wherein R² is H and R³ is alkyl.

An embodiment of the present invention provides compounds according to formula I as described herein, wherein R⁴ is alkyl or -(CH2)_n-R⁶ wherein R⁶ is hydroxy and n is 2.

An embodiment of the present invention provides compounds according to formula I as described herein, wherein R⁴ is alkyl.

An embodiment of the present invention provides compounds according to formula I as described herein, wherein R¹ and R⁵, with the atoms to which they are attached, form a 4-to-5 membered cycloalkyl or a 5-membered heterocycle comprising a single O heteroatom. An embodiment of the present invention provides compounds according to formula I as described herein, wherein

R¹ is haloalkyl or haloalkoxy;

R⁵ is H; or R¹ and R⁵, with the atoms to which they are attached, form a 4-to-5 membered cycloalkyl or a 5-membered heterocycle comprising a single O heteroatom;

R² is H;

R³ is alkyl;

R⁴ is alkyl or -(CH2)_n-R⁶ wherein R⁶ is hydroxy and n is 2. and pharmaceutically acceptable salts thereof.

An embodiment of the present invention provides compounds according to formula I as described herein, wherein

R¹ is haloalkyl or haloalkoxy;

R⁵ is H; or R¹ and R⁵, with the atoms to which they are attached, form a 4-to-5 membered cycloalkyl or a 5-membered heterocycle comprising a single O heteroatom;

R² is H;

R³ is alkyl;

R⁴ is alkyl; and pharmaceutically acceptable salts thereof.

Particular examples of compounds of formula I as described herein are selected from

3-[2-Hydroxy-4-(trifluoromethoxy)phenyl]-4-methyl-6-(6-methyl-3,3a,4,5,7,7a- hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl)-l,2,4-triazin-5-one; formic acid;

6-[(3aR,7aS)-6-methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-

3-[2-hydroxy-4-(tri fluoromethoxy )phenyl]-4-m ethyl- 1, 2, 4-tri azin-5 -one; 6-[(3aS,7aR)-6-methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-

3-[2-hydroxy-4-(trifluoromethoxy)phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aR,7aS)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-

3-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]- 3-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-4-methyl-l,2,4-triazin-5-one;

3-[4-(Difluoromethoxy)-2-hydroxy-phenyl]-4-methyl-6-(6-methyl-3,3a,4,5,7,7a- hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl)-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]- 3-[4-(difluoromethoxy)-2-hydroxy-phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aR,7aS)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-

3-[4-(difluoromethoxy)-2-hydroxy-phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aR,7aS)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-

3-[2-hydroxy-4-(trifluoromethyl)phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-

3-[2-hydroxy-4-(trifluoromethyl)phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aR,7aS)-6-methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]- 3-(2-hydroxy-3-bicyclo[4.2.0]octa-l,3,5-trienyl)-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-

3-(2-hydroxy-3-bicyclo[4.2.0]octa-l,3,5-trienyl)-4-methyl-l,2,4-triazin-5-one; and pharmaceutically acceptable salts thereof.

Other particular examples of compounds of formula I as described herein are selected from

3-(4-Hydroxyindan-5-yl)-4-methyl-6-(6-methyl-3,3a,4,5,7,7a-hexahydro-2H- pyrrolo[2,3 -c]pyridin- 1 -yl)- 1 ,2,4-triazin-5-one; 6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]- 3-(4-hydroxyindan-5-yl)-4-methyl-l,2,4-triazin-5-one;

6-[(3aR,7aS)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]- 3-(4-hydroxyindan-5-yl)-4-methyl-l,2,4-triazin-5-one;

6-[6-(2-Hydroxyethyl)-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3- [2-hydroxy-4-(trifluoromethoxy)phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-(2-Hydroxyethyl)-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3- c]pyridin-l-yl]-3-[2-hydroxy-4-(trifluoromethoxy)phenyl]-4-methyl-l,2,4-triazin-

5-one;

6-[(3aR,7aS)-6-(2-Hydroxyethyl)-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3- c]pyridin-l-yl]-3-[2-hydroxy-4-(trifluoromethoxy)phenyl]-4-methyl-l,2,4-triazin-

5-one; and pharmaceutically acceptable salts thereof.

Preferred examples of compounds of formula I as described herein are selected from

6-[(3aR,7aS)-6-methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]- 3-[2-hydroxy-4-(tri fluoromethoxy )phenyl]-4-m ethyl- 1, 2, 4-tri azin-5 -one;

6-[(3aR,7aS)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]- 3-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-4-methyl-l,2,4-triazin-5-one;

6-[(3aR,7aS)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]- 3-[4-(difluoromethoxy)-2-hydroxy-phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]- 3-[2-hydroxy-4-(trifluoromethyl)phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]- 3-(2-hydroxy-3-bicyclo[4.2.0]octa-l,3,5-trienyl)-4-methyl-l,2,4-triazin-5-one; and pharmaceutically acceptable salts thereof. Other preferred examples of compounds of formula I as described herein are selected from

6-[(3aS,7aR)-6-methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]- 3-[2-hydroxy-4-(tri fluoromethoxy )phenyl]-4-m ethyl- 1, 2, 4-tri azin-5 -one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]- 3-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]- 3-[4-(difluoromethoxy)-2-hydroxy-phenyl]-4-methyl-l,2,4-triazin-5-one; and pharmaceutically acceptable salts thereof.

Another preferred examples of compounds of formula I as described herein is 6- [(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-(4- hydroxyindan-5-yl)-4-methyl-l,2,4-triazin-5-one, or a pharmaceutically acceptable salt thereof.

Most preferred examples of compounds of formula I as described herein are selected from

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]- 3-[2-hydroxy-4-(trifluoromethyl)phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]- 3-(2-hydroxy-3-bicyclo[4.2.0]octa-l,3,5-trienyl)-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]- 3-[2-hydroxy-4-(tri fluoromethoxy )phenyl]-4-m ethyl- 1,2, 4-tri azin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]- 3-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]- 3-[4-(difluoromethoxy)-2-hydroxy-phenyl]-4-methyl-l,2,4-triazin-5-one; 6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]- 3-(4-hydroxyindan-5-yl)-4-methyl-l,2,4-triazin-5-one; and pharmaceutically acceptable salts thereof.

Another embodiment of the invention provides a pharmaceutical composition or medicament containing a compound of the invention and a therapeutically inert carrier, diluent or excipient, as well as a method of using the compounds of the invention to prepare such composition and medicament. In one example, the compound of formula I may be formulated by mixing at ambient temperature at the appropriate pH, and at the desired degree of purity, with physiologically acceptable carriers, i.e., carriers that are non-toxic to recipients at the dosages and concentrations employed into a galenical administration form. The pH of the formulation depends mainly on the particular use and the concentration of compound, but preferably ranges anywhere from about 3 to about 8. In one example, a compound of formula I is formulated in an acetate buffer, at pH 5. In another embodiment, the compound of formula I is sterile. The compound may be stored, for example, as a solid or amorphous composition, as a lyophilized formulation or as an aqueous solution.

Compositions are formulated, dosed, and administered in a fashion consistent with good medical practice. Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners.

The compounds of the invention may be administered by any suitable means, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal and epidural and intranasal, and, if desired for local treatment, intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.

The compounds of the present invention may be administered in any convenient administrative form, e.g., tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions may contain components conventional in pharmaceutical preparations, e.g., diluents, carriers, pH modifiers, sweeteners, bulking agents, and further active agents. A typical formulation is prepared by mixing a compound of the present invention and a carrier or excipient. Suitable carriers and excipients are well known to those skilled in the art and are described in detail in, e.g., Ansel, Howard C., et al., Ansel’s Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, Alfonso R., et al. Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C. Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005. The formulations may also include one or more buffers, stabilizing agents, surfactants, wetting agents, lubricating agents, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, glidants, processing aids, colorants, sweeteners, perfuming agents, flavoring agents, diluents and other known additives to provide an elegant presentation of the drug (i.e., a compound of the present invention or pharmaceutical composition thereof) or aid in the manufacturing of the pharmaceutical product (i.e., medicament).

The compounds of formula I and their pharmaceutically acceptable salts can be processed with pharmaceutically inert, inorganic or organic adjuvants for the production of tablets, coated tablets, dragees, hard gelatin capsules, injection solutions or topical formulations Lactose, com starch or derivatives thereof, talc, stearic acid or its salts etc. can be used, for example, as such adjuvants for tablets, dragees and hard gelatin capsules.

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

Suitable adjuvants for the production of solutions and syrups are, for example, water, polyols, saccharose, invert sugar, glucose, etc.

Suitable adjuvants for injection solutions are, for example, water, alcohols, polyols, glycerol, vegetable oils, etc.

Suitable adjuvants for suppositories are, for example, natural or hardened oils, waxes, fats, semi-solid or liquid polyols, etc.

Suitable adjuvants for topical ocular formulations are, for example, cyclodextrins, mannitol or many other carriers and excipients known in the art. Moreover, the pharmaceutical preparations can contain preservatives, solubilizers, viscosity-increasing substances, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavorants, salts for varying the osmotic pressure, buffers, masking agents or antioxidants. They can also contain still other therapeutically valuable substances.

The dosage can vary in wide limits and will, of course, be fitted to the individual requirements in each particular case. In general, in the case of oral administration a daily dosage of about 0.1 mg to 20 mg per kg body weight, preferably about 0.5 mg to 4 mg per kg body weight (e.g. about 300 mg per person), divided into preferably 1-3 individual doses, which can consist, for example, of the same amounts, should it be appropriate. In the case of topical administration, the formulation can contain 0.001% to 15% by weight of medicament and the required dose, which can be between 0.1 and 25 mg in can be administered either by single dose per day or per week, or by multiple doses (2 to 4) per day, or by multiple doses per week It will, however, be clear that the upper or lower limit given herein can be exceeded when this is shown to be indicated.

An embodiment of the present invention is a compound according to formula I as described herein for use as a therapeutically active substance.

An embodiment of the present invention is a compound according to formula I as described herein for use in the treatment or prevention of a disease, disorder or condition, wherein the disease, disorder or condition is responsive to NLRP3 inhibition.

An embodiment of the present invention is a compound according to formula I as described herein for the treatment or prophylaxis of a disease, disorder or condition, wherein the disorder or condition is responsive to NLRP3 inhibition.

As used herein, the term “NLRP3 inhibition” refers to the complete or partial reduction in the level of activity of NLRP3 and includes, for example, the inhibition of active NLRP3 and/or the inhibition of activation of NLRP3.

There is evidence for a role of NLRP3 -induced IL-1 and IL- 18 in the inflammatory responses occurring in connection with, or as a result of, a multitude of different disorders (Menu et al., Clinical and Experimental Immunology, 166: 1-15, 2011; Strowig et al., Nature, 481 : 278- 286, 2012). In one embodiment, the disease, disorder or condition is selected from:

(i) inflammation;

(ii) an auto-immune disease;

(iii) cancer;

(iv) an infection;

(v) a central nervous system disease;

(vi) a metabolic disease;

(vii) a cardiovascular disease;

(viii) a respiratory disease;

(ix) a liver disease;

(x) a renal disease;

(xi) an ocular disease;

(xii) a skin disease;

(xiii) a lymphatic condition;

(xiv) a psychological disorder;

(xv) graft versus host disease;

(xvi) allodynia;

(xvii) a condition associated with diabetes; and

(xviii) any disease where an individual has been determined to carry a germline or somatic non-silent mutation in NLRP3

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

(i) cancer;

(ii) an infection;

(iii) a central nervous system disease;

(iv) a cardiovascular disease;

(v) a liver disease;

(vi) an ocular disease; or

(vii) a skin disease.

In a further typical embodiment of the invention, the disease, disorder or condition is inflammation. Examples of inflammation that may be treated or prevented include inflammatory responses occurring in connection with, or as a result of: (i) a skin condition such as contact hypersensitivity, bullous pemphigoid, sunburn, psoriasis, atopical dermatitis, contact dermatitis, allergic contact dermatitis, seborrhoetic dermatitis, lichen planus, scleroderma, pemphigus, epidermolysis bullosa, urticaria, erythemas, or alopecia;

(ii) a joint condition such as osteoarthritis, systemic juvenile idiopathic arthritis, adult-onset Still’s disease, relapsing polychondritis, rheumatoid arthritisjuvenile chronic arthritis, gout, or a seronegative spondyloarthropathy (e.g. ankylosing spondylitis, psoriatic arthritis or Reiter’s disease);

(iii) a muscular condition such as polymyositis or myasthenia gravis;

(iv) a gastrointestinal tract condition such as inflammatory bowel disease (including Crohn’s disease and ulcerative colitis), colitis, gastric ulcer, Coeliac disease, proctitis, pancreatitis, eosinopilic gastro-enteritis, mastocytosis, antiphospholipid syndrome, or a food-related allergy which may have effects remote from the gut (e.g., migraine, rhinitis or eczema);

(v) a respiratory system condition such as chronic obstructive pulmonary disease (COPD), asthma (including eosinophilic, bronchial, allergic, intrinsic, extrinsic or dust asthma, and particularly chronic or inveterate asthma, such as late asthma and airways hyper-responsiveness), bronchitis, rhinitis (including acute rhinitis, allergic rhinitis, atrophic rhinitis, chronic rhinitis, rhinitis caseosa, hypertrophic rhinitis, rhinitis pumlenta, rhinitis sicca, rhinitis medicamentosa, membranous rhinitis, seasonal rhinitis e.g. hay fever, and vasomotor rhinitis), sinusitis, idiopathic pulmonary fibrosis (IPF), sarcoidosis, farmer’s lung, silicosis, asbestosis, volcanic ash induced inflammation, adult respiratory distress syndrome, hypersensitivity pneumonitis, or idiopathic interstitial pneumonia;

(vi) a vascular condition such as atherosclerosis, Behcet’s disease, vasculitides, or Wegener’s granulomatosis;

(vii) an autoimmune condition such as systemic lupus erythematosus, Sjogren’s syndrome, systemic sclerosis, Hashimoto’s thyroiditis, type I diabetes, idiopathic thrombocytopenia purpura, or Graves disease; (viii) an ocular condition such as uveitis, allergic conjunctivitis, or vernal conjunctivitis;

(ix) a nervous condition such as multiple sclerosis or encephalomyelitis;

(x) an infection or infection-related condition, such as Acquired Immunodeficiency Syndrome (AIDS), acute or chronic bacterial infection, acute or chronic parasitic infection, acute or chronic viral infection, acute or chronic fungal infection, meningitis, hepatitis (A, B or C, or other viral hepatitis), peritonitis, pneumonia, epiglottitis, malaria, dengue hemorrhagic fever, leishmaniasis, streptococcal myositis, mycobacterium tuberculosis (including mycobacterium tuberculosis and HIV co-infection), mycobacterium avium intracellulare, pneumocystis carinii pneumonia, orchitis/epidydimitis, legionella, Lyme disease, influenza A, Epstein-Barr virus infection, viral encephalitis/aseptic meningitis, or pelvic inflammatory disease;

(xi) a renal condition such as mesangial proliferative glomerulonephritis, nephrotic syndrome, nephritis, glomerular nephritis, obesity related glomerulopathy, acute renal failure, acute kidney injury, uremia, nephritic syndrome, kidney fibrosis including chronic crystal nephropathy, or renal hypertension;

(xii) a lymphatic condition such as Castleman’s disease;

(xiii) a condition of, or involving, the immune system, such as hyper IgE syndrome, lepromatous leprosy, familial hemophagocytic lymphohistiocytosis, or graft versus host disease;

(xiv) a hepatic condition such as chronic active hepatitis, non-alcoholic steatohepatitis (NASH), alcohol-induced hepatitis, non-alcoholic fatty liver disease (NAFLD), alcoholic fatty liver disease (AFLD), alcoholic steatohepatitis (ASH), primary biliary cirrhosis, fulminant hepatitis, liver fibrosis, or liver failure;

(xv) a cancer, including those cancers listed above;

(xvi) a bum, wound, trauma, haemorrhage or stroke;

(xvii) radiation exposure; (xviii) a metabolic disease such as type 2 diabetes (T2D), atherosclerosis, obesity, gout or pseudo-gout; and/or

(xix) pain such as inflammatory hyperalgesia, pelvic pain, allodynia, neuropathic pain, or cancer-induced bone pain.

An embodiment of the present invention is a compound according to formula I as described herein for the treatment or prophylaxis of a disease, disorder or condition selected from:

(i) inflammation;

(ii) an auto-immune disease;

(iii) cancer;

(iv) an infection;

(v) a central nervous system disease;

(vi) a metabolic disease;

(vii) a cardiovascular disease;

(viii) a respiratory disease;

(ix) a liver disease;

(x) a renal disease;

(xi) an ocular disease;

(xii) a skin disease;

(xiii) a lymphatic condition;

(xiv) a psychological disorder;

(xv) graft versus host disease;

(xvi) allodynia;

(xvii) a condition associated with diabetes; and

(xviii) any disease where an individual has been determined to carry a germline or somatic non-silent mutation in NLRP3.

An embodiment of the present invention is the use of a compound according to formula I as described herein in the treatment or prophylaxis of a disease, disorder or condition, wherein the disease, disorder or condition is responsive to NLRP3 inhibition. An embodiment of the present invention is the use of a compound according to formula I as described herein in the treatment or prophylaxis of a disease, disorder or condition selected from Alzheimer’s disease and Parkinson’s disease.

An embodiment of the present invention is the use a compound according to formula I as described herein for use in the treatment or prophylaxis of a disease, disorder or condition selected from Asthma or COPD.

An embodiment of the present invention is a compound according to formula I as described herein for the treatment or prophylaxis of a disease, disorder or condition selected from Alzheimer’s disease and Parkinson’s disease.

An embodiment of the present invention is a compound according to formula I as described herein for the treatment or prophylaxis of a disease, disorder or condition selected from Asthma or COPD.

An embodiment of the present invention is the use of a compound according to formula I as described herein for preparation of a medicament for the treatment or prophylaxis of a disease, disorder or condition selected from Alzheimer’s disease and Parkinson’s disease.

An embodiment of the present invention is the use of a compound according to formula I as described herein for the preparation of a medicament for the treatment or prophylaxis of a disease, disorder or condition selected from Asthma or COPD.

An embodiment of the present invention is a method of treatment or prophylaxis of a disease, disorder or condition selected from Alzheimer’s disease and Parkinson’s disease, which method comprises administering an effective amount of a compound according to formula I as described herein.

An embodiment of the present invention is a method of treatment or prophylaxis of a disease, disorder or condition selected from Asthma or COPD, which method comprises administering an effective amount of a compound according to formula I as described herein.

An embodiment of the present invention relates to a method of inhibiting NLRP3, which method comprises administering an effective amount of a compound according to formula I as described herein. Also an embodiment of the present invention are compounds of formula I as described herein, when manufactured according to any one of the described processes.

An embodiment of the present invention is a pharmaceutical composition comprising a compound according to formula I as described herein and a therapeutically inert carrier.

Assay Procedures

NLRP3 and Pyroptosis

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

THP-1 Cells: Culture and Preparation

THP-1 cells (ATCC # TIB-202) were grown in RPMI containing L-glutamine (Gibco #11835) supplemented with ImM sodium pyruvate (Sigma # S8636) and penicillin (lOOunits/ml) / streptomycin (O.lmg/ml) (Sigma # P4333) in 10% Fetal Bovine Serum (FBS) (Sigma # F0804). The cells were routinely passaged and grown to confluency (~10⁶cells/ml). On the day of the experiment, THP-1 cells were harvested and resuspended into RPMI medium (without FBS). The cells were then counted and viability (>90%) checked by Trypan blue (Sigma # T8154). Appropriate dilutions were made to give a concentration of 625,000cells/ml. To this diluted cell solution was added LPS (Sigma # L4524) to give a Ipg/ml Final Assay Concentration (FAC). 40pl of the final preparation was aliquoted into each well of a 96-well plate. The plate thus prepared was used for compound screening. THP-1 Cells Pyroptosis Assay

The following method step-by-step assay was followed for compound screening.

1. Seed THP-1 cells (25,000cells/well) containing l.Opg/ml LPS in 40pl of RPMI medium (without FBS) in 96-well, black walled, clear bottom cell culture plates coated with poly-D- lysine (VWR # 734-0317)

2. Add 5 pl compound (8 points half-log dilution, with lOpM top dose) or vehicle (DMSO 0.1% FAC) to the appropriate wells

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

4. Add 5 l nigericin (Sigma # N7143) (FAC 5pM) to all wells

5. Incubate for Ihr at 37°C, 5% CO2

6. At the end of the incubation period, spin plates at 300xg for 3mins and remove supernatant

7. Then add 50 pl of resazurin (Sigma # R7017) (FAC 100 pM resazurin in RPMI medium without FBS) and incubate plates for a further 1-2 hours at 37 °C and 5% CO2

8. Plates were read in an Envision reader at Ex 560nm and Em 590nm

9. IC50 data is fitted to a non-linear regression equation (log inhibitor vs response-variable slope 4-parameters)

The results of the pyroptosis assay are summarised in Table 1 below as THP IC50.

Human Whole Blood IL- l b Release Assay

For systemic delivery, the ability to inhibit NLRP3 when the compounds are present within the bloodstream is of great importance. For this reason, the NLRP3 inhibitory activity of a number of compounds in human whole blood was investigated in accordance with the following protocol. Human whole blood in Li-heparin tubes was obtained from healthy donors from a volunteer donor panel.

1. Plate out 80pl of whole blood containing Ipg/ml of LPS in 96-well, clear bottom cell culture plate (Coming # 3585)

2. Add 1 Opl compound (8 points half-log dilution with lOpM top dose) or vehicle (DMSO 0.1% FAC) to the appropriate wells

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

4. Add 1 Opl nigericin (Sigma # N7143) (lOpM FAC) to all wells

5. Incubate for Ihr at 37°C, 5% CO2

6. At the end of the incubation period, spin plates at 300xg for 5mins to pellet cells and remove 20pl of supernatant and add to 96-well v-bottom plates for IL-ip analysis (note: these plates containing the supernatants can be stored at -80°C to be analysed at a later date)

7. IL-ip was measured according to the manufacturer protocol (Perkin Elmer- AlphaLisa IL-1 Kit AL220F-5000)

8. IC50 data is fitted to a non-linear regression equation (log inhibitor vs response-variable slope 4-parameters)

The results of the human whole blood assay are summarised in Table 1 below as HWB IC50. hERG screening assay

In the drug development process of small molecules, one of the most frequent adverse side effects, leading to the failure of drugs, is the cardiac arrhythmias. Such failure is often related to the capacity of the drug to inhibit the human ether-a-go-go-related gene (hERG) cardiac potassium channel. Having no or low inhibition of the hERG cardiac potassium channel is therefore considered as beneficial. Cells

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

Experimental solutions

The extracellular solution contains (in mM): NaCl 150; KC1 4; CaCh 1; MgCh 1; HEPES 10; pH 7.2-7.4 with NaOH, osmolarity 290-330 mOsm. The internal solution contains (in mM): KC1, 10; KF, 100; NaCl, 10; HEPES, 10; EGTA, 20; pH = 7.0-7.4 with KOH, osmolarity 260- 300 mOsm.

Electrophysiology

The effects of a compound on hERG K+-currents parameters will be evaluated at 2 concentrations in at least 4 cells.

The hERG test is performed using automated patch clamp system SynchroPatch® 384 (Nanion Technologies GmbH, Germany). K+ currents are measured with the patch-voltage- clamp technique in the whole-cell configuration at 35-37°C.

Cells were held at a resting voltage of -80 mV and they were stimulated by a voltage pattern shown in Figure 1 (pulse pattern used to elicit outward K⁺ current at 35-37°C) to activate hERG channels and conduct outward IKhERG current, at a stimulation frequency of 0.1 Hz (6 bpm)

Data analysis The amplitudes of IKhERG were recorded in each concentration of drug and they were compared to the vehicle control values (taken as 100%) to define fractional blocks. The concentration-response data were fitted with the following relationship:

Concentration-response curves were fitted by non-linear regression analysis using

EworkBook suite (ID Business Solutions Ltd, UK). Data fit was done with the 4 Parameter

Logistic Model (fit = (A+(B/(l+((x/C)^AD)))), where A=0 and B=100).

The results of the hERG assay are summarised in Table 2 below as hERG IC20.

Transcellular P-gp Assay:

The general assay uses transfected LLC-PK1 cells (porcine kidney epithelial cells) overexpressing human or mouse P-gp, cultured on 96 well semi-permeable filter membrane plates, where they form a polarized monolayer with tight junctions, and act as a barrier between the apical and basolateral compartment.

P-gp is expressed in the apical-facing membrane of the monolayer.

The tightness of the cell monolayer and functional activity of P-gp are confirmed by addition of a cell-impermeable marker, Lucifer yellow, and a reference P-gp substrate, edoxaban, respectively.

PAMPA:

PAMPA (Parallel Artificial Membrane Permeability Assay) is a first line permeability screen for drug candidates. The PAMPA assay mimics the transcellular absorption conditions using an artificial phospholipid membrane. This assay determines a permeability value that can be used for compound optimization and ranking purposes as well as input parameters for in silico models to predict intestinal absorption. The donor concentration is measured at t-start (reference) and compared with the donor and acceptor concentration after a certain time (t-end) to calculate the extent of passage of the compound through the membrane.

Microsomal Stability:

Incubations of test compounds at 1 pM in microsomes (0.5 mg/mL) plus cofactor NADPH are performed in 96 well plates at 37°C on a TEC AN (Tecan Group Ltd, Switzerland) automated liquid handling system. After a 10 minutes pre-incubation step of the test compound with the microsomes, the enzymatic reaction is started by the addition of cofactors. At 1, 3, 6, 9, 15, 25, 35 and 45 minutes, aliquots of the incubations are removed and quenched with 1 :3 (v/v) acetonitrile containing internal standard. Samples are then cooled and centrifuged before analysis of the supernatant by LC-MS/MS 2.

Metabolic Stability in Hepatocytes:

Assay descriptions:

Biological materials. Cryopreserved hepatocytes [mouse, rat, rabbit, monkey and human (male and female; mixed)] are obtained. Viability of hepatocytes after reconstitution is at least 80% throughout the study. Ready-to-use rat/human HepatoPac® cultures [long-term hepatocyte cocultures; pooled (n=5 for male and n=5 for female for human)] with stromal mouse fibroblasts (negative control; pooled) with the plates for incubations, application medium and maintenance medium are acquired.

Metabolism by suspended hepatocytes. Primary pooled cryopreserved hepatocytes are reconstituted in pre-warmed William’s E media containing 10% FCS, 0.05 mg/mL streptomycin and 50 U/mL penicillin and 0.4 mM L-glutamine; and 0.01 mg/mL gentamicin, 0.048 mg/mL hydrocortisone and 0.004 mg/mL insulin, to a final suspension density of 1 x 106 cells/mL. The incubation was performed fully automatically with Liquid Handling System (Tecan) equipped with a CO2 incubator with an orbital shaker. After the addition of a test compound at e.g. 1 pM to the wells (1 x 105 cells/well), the 96-well hepatocyte suspension culture plates are incubated in a 5% CO2 at 37°C. Samples are quenched by addition of acetonitrile (including an internal standard) to the incubation well at the designated time points up to 2 h.

Metabolism by HepatoPac®. Incubations for a test article (at e.g. 1 pM, 0.1% v/v DMSO) as conducted in suspension assays are performed in 96-well plates containing either a co-culture of adherent hepatocytes with mouse fibroblast control cells or control cells alone (5% CO2 atmosphere and 37°C). The incubation media in human HepatoPac® is identical with that in suspended hepatocytes. At defined time points (2, 18, 26, 48, 72 and 96 h), whole wells are quenched with ice-cold acetonitrile containing an internal standard.

Samples are then centrifuged appropriately and the supernatant analyzed by LC-MS/MS. The incubation is conducted in n=l or 2.

Table 1: NLRP3 inhibitory activity Table 2: hERG inhibition assay

The invention will now be illustrated by the following examples which have no limiting character.

In case the preparative examples are obtained as a mixture of enantiomers or diastereoisomers, the pure enantiomers or diastereomers can be obtained by methods described herein or by methods known to those skilled in the art, such as e.g. chiral chromatography or crystallization.

Experimental Methods

Abbreviations: Examples

All examples and intermediates were prepared under nitrogen atmosphere if not specified otherwise.

Synthesis of intermediates:

Intermediate 1 :

3-Chloro-4-methyl-6-(6-methyl-3,3a,4,5,7,7a-hexahydro-2Z/-pyrrolo[2,3-c]pyridin-l-yl)- l,2,4-triazin-5-one

Step A: 2-[(4-Methoxyphenyl)methyl]-4-methyl-6-(6-methyl-3, 3 a, 4,5,7, 7a-hexahydro-2//- pyrrolo[2,3 -c]pyridin- 1 -yl)- 1 ,2,4-triazine-3 ,5-dione

A solution of 6-bromo-2-[(4-methoxyphenyl)methyl]-4-methyl-l, 2, 4-triazine-3, 5-dione (1.66 g, 4.84 mmol, 1.1 eq), commercially available 6-methyl-l,2,3,3a,4,5,7,7a-octahydropyrrolo[2,3- c]pyridine;dihydrochloride (CAS # 2126160-15-2, 1.10 g, 4.39 mmol, 1.0 eq), cesium carbonate (7.14 g, 21.9 mmol, 5.0 eq) in MeCN (20 mL) and water (80.0 pL, 4.44 mmol, 1.01 eq) was sparged (bubbling N2, while sonicating for 5 min), palladium (II) acetate (40.0 mg, 0.18 mmol, 0.04 eq) and Xantphos (103 mg, 0.18 mmol, 0.04 eq) in MeCN (20 mL) then added and the reaction mixture was heated to 50 °C for 1 hour. After this time the reaction temperature was increased to 85 °C and stirred at this temperature for 18 h. The reaction mixture was cooled, filtered through celite, rinsing with EtOAc and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (40 g cartridge, 0-10% MeOH [0.7 M NH3]: EtOAc) to afford the title compound (702 mg, 38%) as an orange solid. LCMS: m/z 386.3 [M+H]⁺, ESI pos.

Step B: 4-Methyl-6-(6-methyl-3, 3a, 4,5,7, 7a-hexahydro-2J/-pyrrolo[2,3-c]pyri din-l-yl)-2J/-l, 2,4- triazine-3, 5-dione To a solution of 2-[(4-methoxyphenyl)methyl]-4-methyl-6-(6-methyl-3, 3a, 4,5,7, 7a-hexahydro- 2J/-pyrrolo[2,3-c]pyridin-l-yl)-l, 2, 4-triazine-3, 5-dione (step A) (603 mg, 1.56 mmol, 1.0 eq) in DCM (3 mL) and MeCN (1.5 mL) was added trifluoromethanesulfonic acid (360 pL, 4.07 mmol, 2.6 eq). The reaction mixture was heated to 35 °C and stirred for 18 hours. The reaction mixture was loaded onto silica and purified by silica gel chromatography (40 g cartridge, 0-15% MeOH [0.7 M NH3]: Ethyl acetate) to afford the title compound (407 mg, 98% yield) as an orange solid. LCMS: m/z 266.2 [M+H]⁺, ESI pos.

Step C: 3-Chloro-4-methyl -6-(6-methyl-3, 3a, 4,5,7, 7a-hexahydro-2J/-pyrrolo[2,3-c]pyri din-1 -yl)- l,2,4-triazin-5-one

Aforementioned 4-methyl-6-(6-methyl-3, 3a, 4,5,7, 7a-hexahydro-2J/-pyrrolo[2,3-c]pyri din-l-yl)- 2J/-1, 2, 4-triazine-3, 5-dione (step B) (503 mg, 1.90 mmol, 1.0 eq) was added to a stirred solution of phosphorus oxychloride (5.0 mL, 53.6 mmol, 28.3 eq) and the reaction was heated to 100 °C for 16 h. The reaction was diluted with further phosphorus oxychloride (5.0 mL, 53.6 mmol, 28.3 eq) and heated to 100 °C for an additional 24 h. The reaction mixture was cooled and concentrated in vacuo, then the resulting brown oil was taken up in MeCN (~30 mL) and added portionwise to a vigorously stirred solution of EtOAc (50 mL) and K3PO4 (50% aq, 100 mL). The pH of the aqueous solution was then adjusted to -pH 12 by portionwise addition of solid K3PO4, and the organic layer was separated. The aqueous phase was extracted with EtOAc (2 x 50 mL) and the combined organic layer were dried (MgSO4), filtered, and concentrated in vacuo to afford the title compound (414 mg, 73%) light yellow solid. LCMS: m/z 284.2/286.2 [M+H]⁺, ESI pos.

Intermediate 2:

3-(4,4,5,5-Tetramethyl-l,3,2-dioxaborolan-2-yl)bicyclo[4.2.0]octa-l(6),2,4-trien-2-ol

A mixture of 3-bromobicyclo[4.2.0]octa-l(6),2,4-trien-2-ol (317 mg, 1.59 mmol, 1.0 eq), bis(pinacolato)diboron (2028.8 mg, 7.99 mmol, 5.02 eq), Pd(dppf)C12.DCM Complex (158.5 mg, 0.19 mmol, 0.12 eq), and potassium acetate (475.5 mg, 4.85 mmol, 3.04 eq) in 1,4-Dioxane (15 mL) was degassed with N2 for 5 mins, then was heated to 90 °C and stirred for 2 h. The reaction was allowed to cool to rt, then was concentrated in vacuo and purified by column chromatography on silica gel (40 g cartridge, 0-100% EtOAc/isohexane) to give the title compound (144.2 mg, 14% yield) as a white solid. A further batch of the title compound (366 mg, 25% yield) was also isolated as a white solid. 'H NMR (500 MHz, DMSO) 8 8.55 (s, 1H), 7.36 (d, 1H), 6.58 (d, 1H), 3.12 - 2.97 (m, 4H), 1.28 (s, 12H). LCMS m/z 247.7 [M+H]⁺ ESI pos.

Intermediate 3:

2-[4-(Difluoromethoxy)-2-methoxy-phenyl]-4,4,5,5-tetramethyl-l,3,2-dioxaborolane

Step A: 1 -Bromo-4-(difluoromethoxy)-2-methoxy-benzene

A solution of commercially available 4-bromo-3 -methoxyphenol (CAS # 102127-34-4, 2.0 g, 9.85 mmol, 1.0 eq), cesium carbonate (16.05 g, 49.3 mmol, 5.0 eq) and sodium chlorodifluoroacetate (4.51 g, 29.6 mmol, 3.0 eq) in DMF (50 mL) and Water (5 mL) was placed under a nitrogen atmosphere and stirred at 90 °C for 16 h. sodium chlorodifluoroacetate (4.51 g, 29.6 mmol, 3.0 eq) was added to the reaction mixture which was heated at 90 °C for 3 h. The reaction mixture was allowed to cool to RT, diluted with DCM (20 mL) and 10% aq. LiCl (200 ml) and the layers separated. The aqueous layer was extracted with DCM (2 x 20 mL) after which the combined organics were concentrated in vacuo. The crude product was purified by silica gel chromatography (40 g cartridge, 0-50% (EtOAc/isohexane)) to afford the title compound (847 mg, 30% yield) as a clear colourless oil. ^XH NMR (500 MHz, CDCh) 6 7.50 (d, 1H), 6.68 (d, 1H), 6.65 - 6.33 (m, 2H), 3.89 (s, 3H).

Step B: 2-[4-(Difluoromethoxy)-2-methoxy-phenyl]-4,4,5,5-tetramethyl-l,3,2-dioxaborolane

A solution of l-bromo-4-(difluoromethoxy)-2-methoxy-benzene (step A) (2.05 g, 8.10 mmol, 1.0 eq), bis(pinacolato)diboron (2.67 g, 10.5 mmol, 1.3 eq), Potassium acetate (2.67 g, 27.2 mmol, 3.35 eq), Xphos (82.0 mg, 0.17 mmol, 0.02 eq) and Xphos Pd G3 (410.0 mg, 0.48 mmol, 0.06 eq) in isopropyl acetate (50 mL) was heated to 90 °C under a nitrogen atmosphere and stirred at this temperature for 16. The reaction mixture was allowed to cool to RT, diluted with water (50 mL) and DCM (100 mL) after which the layers were separated. The aqueous layer was extracted with DCM (2 x 100 mL) after which the combined organics were concentrated in vacuo. The crude product was purified by silica gel chromatography (24 g column, 0-50% (EtOAc/isohexane)) to afford the title compound (1.65 g, 58% yield) as a brown oil. NMR (500 MHz, DMSO) 6 7.57 (d, 1H), 7.32 (t, 1H), 6.76 - 6.68 (m, 2H), 3.75 (s, 3H), 1.26 (s, 12H).

Intermediate 4:

5-(4,4,5,5-Tetramethyl-l,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzofuran-4-ol

Step A: 5-Bromo-2,3 -dihydrob enzofuran-4-ol

To a solution of 2, 3 -dihydrob enzofuran-4-ol (CAS # 144822-82-2, 2.00 g, 14.7 mmol, 1 eq) in methanol (40 mL) was added pyridine tribromide (4.70 g, 14.7 mmol, 1 eq) at -40 °C. The resulting mixture was stirred at -40 °C for 0.5 hour, then warmed to 20 °C and stirred for 16 hours. After reaction completion, the reaction mixture was dissolved in EtOAc (100 mL). The organic layer was washed with 1 N hydrochloric acid (100 mL x 2), followed by brine (100 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (SiCh, PE: EtOAc = 15: 1 to 10: 1) to afford the title compound (1.90 g, 60% yield) as a yellow solid. LCMS: m/z 212.8 [M-H]', ESI neg.

Step B: 5-(4,4,5,5-Tetramethyl-l,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzofuran-4-ol

To a solution of aforementioned 5-bromo-2,3-dihydrobenzofuran-4-ol (58.4 g, 271 mmol, 1.00 eq) in dioxane (600 mL) was added B2Pin2 (69.0 g, 271 mmol, 1.00 eq), KO Ac (58.6 g, 597 mmol, 2.20 eq) and Pd(PPh3)2Ch (9.94 g, 13.6 mmol, 0.05 eq). Then the mixture was stirred at 90 °C for 2 hrs. After reaction completion, the reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, Petroleum ether/Ethyl acetate=10/l to 5/1) to give a yellow solid. Target (9.72 g, 13% yield, 97% purity) was obtained as yellow solid. ¹H NMR (400 MHz, CDCh): 6 = 8.01 (s, 1H), 7.44 (d, 1H), 6.41 (d, 1H), 4.62 (t, 2H), 3.17 (t, 2H), 1.36 (s, 12H).

Syntheses of Examples

Example 1:

3-[2-Hydroxy-4-(trifluoromethoxy)phenyl]-4-methyl-6-(6-methyl-3,3a,4,5,7,7a-hexahydro-

2//-pyrrolo|2.3-c|pyridin-l-yl)-l .2.4-triazin-5-one

To a stirred solution of aforementioned 3-chloro-4-methyl-6-(6-methyl-3, 3a, 4,5,7, 7a-hexahydro- 2J/-pyrrolo[2,3-c]pyridin-l-yl)-l,2,4-triazin-5-one Intermediate 1 (206 mg, 0.73 mmol, 1.0 eq), commercially available [2-hydroxy-4-(trifluoromethoxy)phenyl]boronic acid (CAS # 1309768- 22-6, 178 mg, 0.80 mmol, 1.10 eq) in 1,4-Dioxane (3 mL) and saturated aqueous sodium carbonate (750 pL, which had been sparged for 2 min (N2 bubbling with sonication)), xphos Pd g3 (32.0 mg, 0.04 mmol, 0.05 eq) was added and the reaction was heated at 80 °C and stirred for 6 h. The reaction was cooled degassed and additional saturated aqueous sodium carbonate (250 pL, 2.67 mmol, 3.67 eq) and xphos Pd g3 (35.0 mg, 0.04 mmol, 0.06 eq) was added and the reaction was heated at 80 °C and stirred for a further 18 h. The reaction mixture was concentrated and loaded onto some celite then added to a plug of celite and rinsed with EtOAc (100 mL). The solvent was removed, and the crude sample was dissolved in DMSO, filtered and purified by reversed phase preparative HPLC (Waters 2767 Sample Manager, Waters 2545 Binary Gradient Module, Waters Systems Fluidics Organiser, Waters 515 ACD pump, Waters 515 Makeup pump, Waters 2998 Photodiode Array Detector, Waters QDa) on a Waters XBridge BEH Cis ODB prep column, 130 A, 5 pm, 30 mm X 100 mm, flow rate 40 mL min'¹ eluting with a 0.3% Ammonia in water-MeCN gradient over 12.5 mins using UV across all wavelengths with PDA as well as a QDA and ELS detector. At-column dilution pump gives 2 mL min'¹ Methanol over the entire method, which is included in the following MeCN percentages. Gradient information: 0.0-0.5 min, 5% MeCN; 0.5- 10.5 min, ramped from 5% MeCN to 30% MeCN; 10.5-10.6 min, ramped from 30% MeCN to 100% MeCN; 10.6-12.5 min, held at 100% MeCN. The clean fractions were combined and evaporated to afford the title compound (86 mg, 29%) as a light-yellow solid. LCMS m/z 426.2 [M+H]⁺, ESI pos.

Chiral separation of Example 1:

Example 1 (80.6 mg) was dissolved in MeOH (3 mL), filtered and was then separated by chiral SFC on a Waters prep 100 with a PDA and a QDA detectors, 40 °C, 120 bar. The column was a Lux® 5 pM Amylose- 1, LC Column 250 x 21.2 mm, AXIA™ Packed (Phenomenex®); flow rate 65 mL/ min of 30 % ethanol (0.4 % triethylamine), 70 % CO2. The clean fractions were pooled, rinsed with methanol and concentrated to dryness using a rotary evaporator. The residues were redissolved in methanol, transferred into final vials and evaporated on a Biotage VI 0. The samples were then further dried in a vacuum oven at 30 °C/5 mbar over night to afford Enantiomer 1A (30.0 mg, 9.23% yield) and IB (29.8 mg, 9.17% yield) both as colourless glasses for which the stereochemistry was arbitrarily assigned.

Enantiomer 1 A : 6- 1 (3a.S.7a/?)-6-M et hy 1-3.3a.4.5.7.7a-hexa hyd ro-2//-pyrrolo [2 ,3-c] pyridin- l-yl]-3-[2-hydroxy-4-(trifluoromethoxy)phenyl]-4-methyl-l,2,4-triazin-5-one

'H NMR (500 MHz, DMSO, 353.15 Kelvin) 8 7.40 (d, 1H), 6.88 - 6.83 (m, 2H), 4.72 - 4.61 (m, 1H), 3.86 - 3.78 (m, 1H), 3.75 - 3.65 (m, 1H), 3.17 (s, 3H), 2.94 (ddd, 1H), 2.48 - 2.44 (m, 1H), 2.35 - 2.28 (m, 1H), 2.17 (s, 3H), 2.07 (td, 1H), 2.03 - 1.77 (m, 4H), 1.67 (dq, 1H). Note: Phenol is exchanging; LCMS m/z 426.4 [M+H]⁺ ESI pos.

Enantiomer IB: 6-|(3:i/?.7a.S)-6-Methyl-3.3a.4.5.7.7a-hexahydro-2//-pyrrolo|2.3-c|pyridin- l-yl]-3-[2-hydroxy-4-(trifluoromethoxy)phenyl]-4-methyl-l,2,4-triazin-5-one

'H NMR (500 MHz, DMSO, 353.15 Kelvin)) 8 7.40 (d, 1H), 6.88 - 6.83 (m, 2H), 4.72 - 4.61 (m, 1H), 3.86 - 3.78 (m, 1H), 3.75 - 3.65 (m, 1H), 3.17 (s, 3H), 2.94 (ddd, 1H), 2.48 - 2.44 (m, 1H), 2.35 - 2.28 (m, 1H), 2.17 (s, 3H), 2.07 (td, 1H), 2.03 - 1.77 (m, 4H), 1.67 (dq, 1H). Note: Phenol is exchanging; LCMS m/z 426.4 [M+H]⁺ ESI pos.

Examples 2A and 2B:

Enantiomer 2A and Enantiomer 2B of 3-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-4- methyl-6-(6-methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl)-l,2,4-triazin-5- one

To a stirred solution of aforementioned 3-chloro-4-methyl-6-(6-methyl-3, 3a, 4,5,7, 7a-hexahydro- 2J/-pyrrolo[2,3-c]pyridin-l-yl)-l,2,4-triazin-5-one Intermediate 1 (206 mg, 0.73 mmol, 1.0 eq), 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-2,3-dihydrobenzofuran-4-ol Intermediate 4 (209.3 mg, 0.8 mmol, 1.1 eq) in 1,4-Dioxane (3 mL) and saturated aqueous sodium carbonate (750.0 uL) which had been sparged for 2 min (N2 bubbling with sonication) was added Xphos Pd g3 (32.0 mg, 0.04 mmol, 0.05 eq) and the reaction was stirred at 80 °C for 20 h. The reaction mixture was concentrated and loaded onto some celite then added to a plug of celite and rinsed with EtOAc (100 mL). The solvent was removed, and the crude sample was dissolved in DMSO (4.6 mL), filtered and purified by reversed phase preparative HPLC (Waters 2767 Sample Manager, Waters 2545 Binary Gradient Module, Waters Systems Fluidics Organiser, Waters 515 ACD pump, Waters 515 Makeup pump, Waters 2998 Photodiode Array Detector, Waters QDa) on a Waters XBridge BEH Cis ODB prep column, 130 A, 5 pm, 30 mm X 100 mm, flow rate 40 mL min-1 eluting with a 0.3% Ammonia in water-MeCN gradient over 12.5 mins using UV across all wavelengths with PDA as well as a QDA and ELS detector. At-column dilution pump gives 2 mL min'¹ Methanol over the entire method, which is included in the following MeCN percentages. Gradient information: 0.0-0.5 min, 5% MeCN; 0.5-10.5 min, ramped from 5% MeCN to 30% MeCN; 10.5-10.6 min, ramped from 30% MeCN to 100% MeCN; 10.6-12.5 min, held at 100% MeCN. The clean fractions were evaporated in a Genevac and then dissolved in MeOH (5 mL) with sonication, filtered and was then separated by chiral SFC on a Waters prep 15 with UV detection by DAD at 210 - 400 nm, 40 °C, 120 bar. The column was a Phenomenex Lux Al 10X250 mm, 5pm, flow rate 15 mL/min at 60% MeOH (0.2% DEA), 40% CO2. The clean fractions were pooled, rinsed with methanol and concentrated to dryness using a rotary evaporator. The residues were re-dissolved in methanol, transferred into final vials and evaporated on a Biotage V10. The samples were then further dried in a vacuum oven at 30 °C/5 mbar over the weekend to afford Enantiomer 2A (13.9 mg, 4.74%) and 2B (14.2 mg, 5%) both as a light brown freeze-dried solid for which the stereochemistry was arbitrarily assigned.

Enantiomer 2A: 6-|(3:iS.7:iR)-6-Metliyl-3.3a.4.5.7.7a-liexaliydro-2//-pyrrolo|2.3-c|pyridin- l-yl]-3-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-4-methyl-l,2,4-triazin-5-one 'H NMR (500 MHz, DMSO) 8 7.02 (d, 1H), 6.35 (d, 1H), 4.70 - 4.63 (m, 1H), 4.58 (t, 2H), 3.85

- 3.77 (m, 1H), 3.74 - 3.63 (m, 1H), 3.18 (s, 3H), 3.15 (t, 2H), 2.96 - 2.91 (m, 1H), 2.48 - 2.44 (m, 1H), 2.34 - 2.27 (m, lH), 2.17 (s, 3H), 2.07 (td, 1H), 2.02 - 1.78 (m, 4H), 1.70 - 1.63 (m, 1H). Note: Phenol is exchanging. LCMS m/z 384.2 [M+H]⁺ ESI pos.

Enantiomer 2B : 6- [(3a/?,7a5)-6-Methyl-3,3a,4,5,7,7 a-hexa hy d ro-2//-py r rolo [2 ,3-c] pyridin- l-yl]-3-(4-hydroxy-2,3-dihydrobenzofuran-5-yl)-4-methyl-l,2,4-triazin-5-one

'H NMR (500 MHz, DMSO) 8 6.99 (d, 1H), 6.29 (d, 1H), 4.69 - 4.62 (m, 1H), 4.55 (t, 2H), 3.84

- 3.77 (m, 1H), 3.72 - 3.64 (m, 1H), 3.19 (s, 3H), 3.12 (t, 2H), 2.96 - 2.91 (m, 1H), 2.49 - 2.43 (m, 1H), 2.35 - 2.27 (m, lH), 2.16 (s, 3H), 2.07 (td, 1H), 2.02 - 1.76 (m, 4H), 1.69 - 1.64 (m, 1H). Note: Phenol is exchanging. LCMS m/z 384.2 [M+H]⁺ ESI pos.

Example 3:

3-[4-(Difluoromethoxy)-2-hydroxy-phenyl]-4-methyl-6-(6-methyl-3,3a,4,5,7,7a-hexahydro-

2//-pyrrolo|2.3-c|pyridin-l-yl)-l .2.4-triazin-5-one

Step A: 3-[4-(Difluoromethoxy)-2-methoxy-phenyl]-4-methyl-6-(6-methyl-3,3a,4,5,7,7a- hexahydro-2Z7-pyrrolo[2,3-c]pyridin-l-yl)-l,2,4-triazin-5-one

To a stirred solution of aforementioned 3-chloro-4-methyl-6-(6-methyl-3, 3a, 4,5,7, 7a-hexahydro- 2Z7-pyrrolo[2,3-c]pyridin-l-yl)-l,2,4-triazin-5-one Intermediate 1 (294 mg, 1.04 mmol, 1.0 eq), 2-[4-(difluoromethoxy)-2-methoxy-phenyl]-4,4,5,5-tetramethyl-l,3,2-dioxaborolane

Intermediate 3 (377 mg, 1.26 mmol, 1.21 eq) in 1,4-Dioxane (5 mL) and saturated aqueous sodium carbonate (1.0 mL, 10.7 mmol, 10.3 eq) which had been sparged for 2 min (N2 bubbling with sonication) was added xphos Pd g3 (45.0 mg, 0.05 mmol, 0.05 eq) and the reaction was heated to 80 °C and left to stir for 2.5 h. The reaction mixture was concentrated and loaded onto Celite and purified by column reverse-phase chromatography (Cis, 40 g, 20-100% MeCN: 10 mM aq. ammonium bicarbonate) to afford the title compound (276 mg, 62% yield) as a light-yellow solid. LCMS m/z 421.8 [M+H]⁺, ESI pos.

Step B: 3-[4-(Difluoromethoxy)-2-hydroxy-phenyl]-4-methyl-6-(6-methyl-3,3a,4,5,7,7a- hexahydro-2J/-pyrrolo[2,3-c]pyridin-l-yl)-l,2,4-triazin-5-one

3 - [4-(difluoromethoxy)-2-methoxy-phenyl] -4-methyl-6-(6-methyl-3 , 3 a, 4, 5 ,7,7a-hexahydro-27/- pyrrolo[2,3-c]pyridin-l-yl)-l,2,4-triazin-5-one (step A) (296 mg, 0.7 mmol, 1.0 eq), potassium carbonate (292 mg, 2.11 mmol, 3.01 eq) and NMP (3 m ) were placed in a microwave vial, sonicated and sealed, benzenethiol (76.0 pL, 0.74 mmol, 1.06 eq) was added and the reaction mixture irradiated in a biotage microwave for 65 mins at 120 °C. The reaction mixture was filtered and then added directly onto a basic RP column (Cis, 43 g cartridge, 10-100%, eluting at 35% to give, after concentration to dryness, to afford the title compound in 2 fractions: the first (114 mg, 39% yield) as a light brown solid. A second fraction was also obtained (100 mg, 34% yield) as a white solid.

Chiral separation of Example 3:

Example 3 (190.1 mg) was dissolved to 19 mg/mL in DCM/DMSO/MeOH with sonication, filtered and was then separated by chiral SFC on a Sepiatec with UV detection by DAD at 220 nm, 40 °C, 120 bar. The column was Chiralpak IG 10X250mm, 5um, flow rate 20mL/ min at 30% MeOH (0.5% DEA), 70% CO2. The clean fractions were pooled, rinsed with methanol and concentrated to dryness using a rocket evaporator at 40 °C. The residues were re-dissolved in methanol transferred into final vials and evaporated on a Biotage VI 0. The samples were then further dried in a vacuum oven at 30 °C/ 5 mbar over night to afford Enantiomer 3A (64.6 mg, 22% yield) and Enantiomer 3B (66.2 mg, 23% yield) both as light brown solids for which the stereochemistry was arbitrarily assigned.

Enantiomer 3A: 6-|(3:iS.7:iR)-6-Metliyl-3.3a.4.5.7.7a-liexaliydro-2//-pyrrolo|2.3-c|pyridin- l-yl]-3-[4-(difluoromethoxy)-2-hydroxy-phenyl]-4-methyl-l,2,4-triazin-5-one

'H NMR (500 MHz, DMSO, 353.15 Kelvin) 8 10.31 (s, 1H), 7.32 (d, 1H), 7.18 (t, 1H), 6.75 - 6.68 (m, 2H), 4.70 - 4.61 (m, 1H), 3.86 - 3.78 (m, 1H), 3.73 - 3.64 (m, 1H), 3.17 (s, 3H), 2.97 - 2.89 (m, 1H), 2.48 - 2.43 (m, 1H), 2.35 - 2.26 (m, 1H), 2.17 (s, 3H), 2.11 - 1.77 (m, 5H), 1.70 - 1.63 (m, 1H); LCMS m/z 408.3 [M+H]⁺ ESI pos.

Enantiomer 3B : 6- [(3aR,7aS)-6-Methyl-3,3a,4,5,7,7 a-hexa hy d ro-2//-py r rolo [2 ,3-c] pyridin- l-yl]-3-[4-(difluoromethoxy)-2-hydroxy-phenyl]-4-methyl-l,2,4-triazin-5-one

'H NMR (500 MHz, DMSO, 353.15 Kelvin) 8 7.29 (d, 1H), 7.16 (t, 1H), 6.70 - 6.61 (m, 2H), 4.71 - 4.63 (m, 1H), 3.85 - 3.78 (m, 1H), 3.72 - 3.65 (m, 1H), 3.18 (s, 3H), 2.98 - 2.90 (m, 1H), 2.48 - 2.44 (m, 1H), 2.34 - 2.27 (m, 1H), 2.16 (s, 3H), 2.10 - 1.77 (m, 5H), 1.70 - 1.64 (m, 1H). Note: Phenol proton is exchanging); LCMS m/z 408.2 [M+H]⁺ ESI pos.

Example 4A and 4B:

Enantiomer 4A and Enantiomer 4B of 3-[2-hydroxy-4-(trifluoromethyl)phenyl]-4-methyl-6- (6-inetliyl-3.3:i.4.5.7.7:i-liexahydro-2//-pyrrolo|2.3-c|pyridiⁿ-l-yl)-l .2.4-tri:izin-5-one

To a stirred solution of aforementioned 3-chloro-4-methyl-6-(6-methyl-3, 3a, 4,5,7, 7a-hexahydro- 2J/-pyrrolo[2,3-c]pyridin-l-yl)-l,2,4-triazin-5-one Intermediate 1 (140.0 mg, 0.49 mmol, 1.0 eq), [2-hydroxy-4-(trifluoromethyl)phenyl]boronic acid (112 mg, 0.54 mmol, 1.1 eq) in 1,4-dioxane (2.5 mL) and saturated aqueous sodium carbonate (500 pL) which had been sparged for 2 min (N2 bubbling with sonication) was added xphos Pd g3 (21.0 mg, 0.02 mmol, 0.05 eq) and the reaction was heated to 80 °C and left to stir for 2 h. The reaction mixture was cooled to r.t., sparged and additional xphos Pd g3 (11.0 mg, 0.01 mmol, 0.03 eq) was added and heated at 80 °C for an additional 2 h. The sample was cooled to r.t., sparged, additional saturated aqueous sodium carbonate (250 pL,), [2-hydroxy-4-(trifluoromethyl)phenyl]boronic acid (51.0 mg, 0.25 mmol, 0.5 eq) and xphos Pd g3 (11.0 mg, 0.01 mmol, 0.03 eq) were added and the reaction was heated at 80 °C for a further 2 h. The reaction mixture was concentrated and loaded onto Celite then added to a plug of Celite and rinsed with EtOAc (100 mL). The solvent was removed, and the crude sample (345 mg) was dissolved in DMSO (1.3 mL), filtered and purified by reversed phase preparative HPLC (Waters 2767 Sample Manager, Waters 2545 Binary Gradient Module, Waters Systems Fluidics Organiser, Waters 515 ACD pump, Waters 515 Makeup pump, Waters 2998 Photodiode Array Detector, Waters QDa) on a Waters X-Select CSH Cis ODB prep column, 130 A, 5 pm, 30 mm X 100 mm, flow rate 40 mL min-1 eluting with a 0.1% Formic acid in water-MeCN gradient over 8.5 mins using UV across all wavelengths with PDA as well as a QDA and ELS detector. At- column dilution pump gives 2 mL min-1 Methanol over the entire method, which is included in the following MeCN percentages. Gradient information: 0.0-0.5 min, 10% MeCN; 0.5-5.5 min, ramped from 10% MeCN to 40% MeCN; 5.5 -5.6 min, ramped from 40% MeCN to 100% MeCN; 5.6-8.5 min, held at 100% MeCN. The clean fractions were evaporated in a Genevac and then the residue was dissolved in MeOH (2 mL) with sonication, filtered and was then separated by chiral SFC on a Waters prep 15 with UV detection by DAD at 210 - 400 nm, 40 °C, 120 bar. The column was IG 10X250mm, 5pm, flow rate 15mL/ min at 20% EtOH (0.5% DEA), 80% CO2. The clean fractions were pooled, rinsed with methanol, and concentrated to dryness using a rotary evaporator. The residues were re-dissolved in methanol transferred into final vials and evaporated on a Biotage V10. The samples were then further dried in a vacuum oven at 30 °C/5 mbar over the weekend to afford Enantiomer 4A (15.9 mg, 8% yield) and 4B (17.1 mg, 8% yield) both as a light brown freeze-dried solid for which the stereochemistry was arbitrarily assigned.

Enantiomer 4 A : 6- [(3aR,7aS)-6-Methyl-3,3a,4,5,7,7 a-hexa hy d ro-2//-py r rolo [2 ,3-c] pyridin- l-yl]-3-[2-hydroxy-4-(trifluoromethyl)phenyl]-4-methyl-l,2,4-triazin-5-one

'H NMR (500 MHz, DMSO, 353.15 Kelvin) 8 7.52 (d, 1H), 7.31 - 7.18 (m, 2H), 4.71 - 4.64 (m, 1H), 3.86 - 3.80 (m, 1H), 3.75 - 3.66 (m, 1H), 3.17 (s, 3H), 2.99 - 2.91 (m, 1H), 2.48 - 2.44 (m, 1H), 2.37 - 2.27 (m, 1H), 2.17 (s, 3H), 2.12 - 2.02 (m, 1H), 2.02 - 1.77 (m, 4H), 1.71 - 1.63 (m, 1H). Note: The phenol proton is exchanging. LCMS m/z 410.2 [M+H]⁺ ESI pos.

Enantiomer 4B: 6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin- l-yl]-3-[2-hydroxy-4-(trifluoromethyl)phenyl]-4-methyl-l,2,4-triazin-5-one

'H NMR (400 MHz, DMSO, 363.15 Kelvin) 8 10.34 (s, 1H), 7.52 (d, 1H), 7.24 - 7.19 (m, 2H), 4.72 - 4.62 (m, 1H), 3.89 - 3.80 (m, 1H), 3.77 - 3.65 (m, 1H), 3.18 (s, 3H), 2.98 - 2.91 (m, 1H), 2.48 - 2.44 (m, 1H), 2.38 - 2.27 (m, 1H), 2.18 (s, 3H), 2.13 - 1.78 (m, 5H), 1.72 - 1.64 (m, 1H). Note: The phenol proton is exchanging; one proton is partially obscured by the water peak. LCMS m/z 410.2 [M+H]⁺ ESI pos. Example 5A and 5B:

Enantiomer 5A and Enantiomer 5B of 3-(2-hydroxy-3-bicyclo[4.2.0]octa-l,3,5-trienyl)-4- methyl-6-(6-methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyi'idin-l-yl)-l,2,4-triazin-5- one

A mixture of 3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)bicyclo[4.2.0]octa-l(6),2,4-trien-2- ol Intermediate 2 (0.15 g, 0.61 mmol, 1.1 eq), 3-chloro-4-methyl-6-(6-methyl-3,3a,4,5,7,7a- hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl)-l,2,4-triazin-5-one Intermediate 1 (157.0 mg, 0.55 mmol, 1.0 eq), meCgPPh Pd G3 (CAS # 2230788-58-4, 18.3 mg, 0.03 mmol, 0.05 eq) and potassium carbonate (0.23 g, 1.66 mmol, 3.0 eq) in 1,4-Dioxane (4 mL) and Water (1 mL) was degassed for 5 mins with nitrogen, then the reaction was heated to 90 °C for 1 h. The reaction mixture was allowed to cool to rt and was concentrated in vacuo. The crude material was purified by column chromatography on silica gel (24 g cartridge, 0-10% (0.7 M NH3)MeOH/DCM), then by RP chromatography on Cis (4 g cartridge, 0-20% MeCN/lhO (0.1% Formic acid)) to afford an off white solid (17.7 mg). The white solid was dissolved in MeOH (1 mL) with sonication, filtered and was then separated by chiral SFC on a Waters prep 15 with UV detection by DAD at 210 - 400 nm, 40 °C, 120 bar. The column was Al 10X250 mm, 5 pm, flow rate 15mL/ min at 40% MeOH (neutral) 60% CO2. The clean fractions were pooled, rinsed with methanol, and concentrated to dryness using a rotary evaporator. The residues were re-dissolved in methanol transferred into final vials and evaporated on a Biotage V10. The samples were then further dried in a vacuum oven at 30 °C/5 mbar over the weekend to afford Enantiomer 5A (5.8 mg, 3%) and 5B (5.3 mg, 3%) both as off-white solids for which the stereochemistry was arbitrarily assigned.

Enantiomer 5 A : 6- [ ( 3 a.S', 7 al?)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo [2 ,3- c] pyridin- l-yl]-3-(2-hydroxy-3-bicyclo[4.2.0]octa-l,3,5-trienyl)-4-methyl-l,2,4-triazin-5-one 'l l NMR

(500 MHz, DMSO) 8 7.08 (d, 1H), 6.65 (d, 1H), 4.76 - 4.56 (m, 1H), 3.85 - 3.70 (m, 1H), 3.68 - 3.58 (m, 1H), 3.14 (s, 3H), 3.10 - 3.03 (m, 4H), 2.90 (dd, 1H), 2.48 - 2.43 (m, 1H), 2.34 - 2.23 (m, 1H), 2.14 (s, 3H), 2.06 - 1.74 (m, 5H), 1.69 - 1.62 (m, 1H). One exchangeable proton not observed. LCMS m/z 368.2 [M+H]⁺ ESI pos. Enantiomer 5B : 6- [(3al?, 7 aA)-6-Methyl-3,3a,4,5,7,7 a-hexa hy d ro-2//-py r rolo [2 ,3- c] pyridin- l-yl]-3-(2-hydroxy-3-bicyclo[4.2.0]octa-l,3,5-trienyl)-4-methyl-l,2,4-triazin-5-one 'l l NMR (500 MHz, DMSO) 8 7.07 (d, 1H), 6.63 (d, 1H), 4.80 - 4.56 (m, 1H), 3.85 - 3.68 (m, 1H), 3.68 - 3.57 (m, 1H), 3.14 (s, 3H), 3.11 - 3.01 (m, 4H), 2.90 (dd, 1H), 2.48 - 2.42 (m, 1H), 2.34 - 2.25 (m, 1H), 2.14 (s, 3H), 2.07 - 1.74 (m, 5H), 1.69 - 1.59 (m, 1H). One exchangeable proton not observed. LCMS m/z 368.2 [M+H]⁺ ESI pos.

Example 6: 3-(4-Hydroxyindan-5-yl)-4-methyl-6-(6-methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3- c]pyridin-l-yl)-l,2,4-triazin-5-one

A mixture of 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)indan-4-ol (CAS # 2795101-92-5, 287.1 mg, 1.1 mmol), 3-chloro-4-methyl-6-(6-methyl-3,3a,4,5,7,7a-hexahydro-2J/-pyrrolo[2,3- c]pyridin-l-yl)-l,2,4-triazin-5-one Intermediate 1 (232.0 mg, 0.74 mmol, 1.0 eq), XPhos Pd G3 (62.4 mg, 0.07 mmol, 0.1 eq) and potassium carbonate (305.1 mg, 2.21 mmol, 3.0 eq) in MeCN (4 mL) and water (1 mL) was degassed for 5 mins with nitrogen, then the reaction was heated to 80 °C for 1 h. The reaction was allowed to cool to rt and was concentrated in vacuo. The crude material was purified by column chromatography on silica gel (40 g cartridge, 0-10% (0.7M NH3) MeOH/DCM) to give the title compound (93.3 mg, 30% yield) as an off white solid. LCMS m/z 382.3 [M+H]⁺ ESI pos.

Chiral separation of Example 6:

Example 6 (71.5 mg) was dissolved in MeOH, filtered and was then separated by chiral SFC on a Waters prep 100 with a PDA and a QDa detectors, 40 °C, 120 bar. The column was a Lux Amylose-1, 5 pM, 21 mm X 250 mm; flow rate 65 mL/min of 65 % MeOH (0.5% DEA), 35% CO2. The clean fractions were pooled, rinsed with methanol and concentrated to dryness using a rotary evaporator. The residues were re-dissolved in methanol, transferred into final vials and evaporated on a Biotage V10. The samples were then further dried in a vacuum oven at 30 °C/ 5 mbar overnight to afford Enantiomer 6A and Enantiomer 6B as yellowish glass. The samples were then co-evaporated with MeCN, followed by MeOH, to remove trace DEA and dried in a desiccator to give Enantiomer 6A (31.6 mg, 11% yield) and Enantiomer 6B (23.7 mg, 8% yield) both as off-white solids for which the stereochemistry was arbitrarily assigned.

Enantiomer 6A: 6-[(3aS, 7al?)-6-Methyl-3, 3a, 4,5,7, 7a-hexahydro-2//-pyrrolo|2.3-c|pyridin- l-yl]-3-(4-hydroxyindan-5-yl)-4-methyl-l,2,4-triazin-5-one

^XH NMR (500 MHz, DMSO) 8 9.31 (s, 1H), 7.03 (d, 1H), 6.80 (d, 1H), 4.77 - 4.54 (m, 1H), 3.87

- 3.70 (m, 1H), 3.70 - 3.57 (m, 1H), 3.12 (s, 3H), 2.95 - 2.86 (m, 3H), 2.83 (t, 2H), 2.49 - 2.42 (m, 1H), 2.34 - 2.25 (m, 1H), 2.14 (s, 3H), 2.07 - 1.76 (m, 7H), 1.70 - 1.63 (m, 1H). LCMS m/z

382.2 [M+H]⁺ ESI pos.

Enantiomer 6B: 6-[(3aR, 7a5)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin- l-yl]-3-(4-hydroxyindan-5-yl)-4-methyl-l,2,4-triazin-5-one

^XH NMR (500 MHz, DMSO) 8 9.28 (s, 1H), 7.03 (d, 1H), 6.81 (d, 1H), 4.82 - 4.49 (m, 1H), 3.88

- 3.69 (m, 1H), 3.69 - 3.51 (m, 1H), 3.12 (s, 3H), 2.96 - 2.86 (m, 3H), 2.83 (t, 2H), 2.49 - 2.42 (m, 1H), 2.34 - 2.25 (m, 1H), 2.14 (s, 3H), 2.07 - 1.76 (m, 7H), 1.69 - 1.62 (m, 1H). LCMS m/z

382.3 [M+H]⁺ ESI pos.

Example 7:

6-[6-(2-Hydroxyethyl)-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-[2- hydroxy-4-(trifluoromethoxy)phenyl] -4-methyl- 1 ,2,4-triazin-5-one

Step A: /c/7-Butyl pyrrolo[2,3-c]pyridine-l -carboxylate

A solution of 6-azaindole (10.0 g, 84.7 mmol, 1.0 eq) and di -t-butyl di carb onate (27.8 g, 127.24 mmol, 1.5 eq) in THF (240 mL) and triethylamine (20.0 mL, 143.49 mmol, 1.7 eq) was stirred at rt for 1.5 h. The reaction mixture was concentrated in vacuo to give the title compound (34.1 g, quant.) as a light orange oil. LCMS: m/z 162.5 [M-tBu+H]⁺, ESI pos.

Step B: /crt-Butyl 2,3,3a,4,5,6,7,7a-octahydropyrrolo[2,3-c]pyridine-l-carboxylate

A solution of tert-butyl pyrrolo[2,3-c]pyridine-l -carboxylate (34.1 g, 84.37 mmol, 1.0 eq) and Rh/C Type 20D (5.2 g, 2.53 mmol, 0.03 eq) in ethanol (120 mL) and acetic acid (30 mL) was placed under a hydrogen atmosphere and stirred under hydrogen at 50 °C and 5 bar pressure for 2 days. The reaction mixture was filtered through a plug of celite and the plug rinsed with ethanol (2 x 50 mL). The filtrate was diluted with toluene (100 mL) and concentrated to provide an oil which was further azeotroped with toluene (2 x 100 mL). The reaction mixture was diluted with a mixture of chloroform: iPrOH (9:1, 100 mL) and 2M NaOH added until a pH of 8-9 was achieved (-100 mL). The separated aqueous layer was further extracted with the solvent mix (2 x 100 mL). The combined extracts were dried (TsfeSCU), filtered and concentrated to provide the title compound (20.6 g, 91% yield) as a viscous yellow oil. LCMS: m/z 171.5 [M-tBu+H]⁺, ESI pos.

Step C: tert- Butyl 6-benzyl-3,3a,4,5,7,7a-hexahydro-2J/-pyrrolo[2,3-c]pyridine-l-carboxylate To a stirred suspension of tert-butyl 2,3,3a,4,5,6,7,7a-octahydropyrrolo[2,3-c]pyridine-l- carboxylate (9.2 g, 34.55 mmol, 1.0 eq) and potassium carbonate (10.6 g, 76.6 mmol, 2.22 eq) in MeCN (150 mL) was added benzyl bromide (4.3 mL, 36.2 mmol, 1.05 eq) by dropwise addition and the reaction was left to stir at rt for 2.5 h. Benzyl bromide (1.2 mL, 10.09 mmol, 0.29 eq) was added and the reaction left to stir for -16 h. The reaction mixture was diluted with EtOAc (250 mL) and water (250 mL) and the phases were separated. The aqueous layer was extracted again with EtOAc (3 x 100 mL) and the combined organic layers were washed with NaHCOs (2 x 100 mL), dried with MgSO4 and concentrated in vacuo to provide the title compound (13.5 g, quant). -1 g was purified by column chromatography on silica gel (40 g cartridge, 0-10% MeOH (0.7M NH3)/DCM) to give the title compound (827 mg) as an off-white solid. LCMS m/z 317.3 [M+H]⁺ ESI pos.

Step D: 6-Benzyl-l,2,3,3a,4,5,7,7a-octahydropyrrolo[2,3-c]pyridine; dihydrochloride salt tert-butyl 6-benzyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridine-l-carboxylate (827.0 mg, 2.61 mmol, 1.0 eq) was dissolved in DCM (10 mL) and 4M hydrochloric acid in dioxane (2.6 mL, 10.4 mmol, 3.98 eq) was added drop wise. The reaction mixture was stirred at rt for -16 h. The reaction mixture was concentrated in vacuo then dried in a vacuum oven for 1 day at 50 °C to provide the title compound (870.0 mg, 86% yield) as a light yellow waxy solid. LCMS m/z 217.2 [M+H]⁺ ESI pos.

Step E: 6-(6-Benzyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl)-2-[(4- methoxyphenyl)methyl]-4-methyl-l, 2, 4-triazine-3, 5-dione

A mixture of 6-benzyl-l,2,3,3a,4,5,7,7a-octahydropyrrolo[2,3-c]pyridine dihydrochloride salt (870.0 mg, 3.01 mmol, 1.0 eq), 6-bromo-2-[(4-methoxyphenyl)methyl]-4-m ethyl- 1,2, 4-triazine- 3, 5-dione (1.19 g, 3.65 mmol, 1.21 eq), CS2CO3 (7.82 g, 24.0 mmol, 7.98 eq) in MeCN (14 mL) was sparged (bubbling N2 and sonication for 2 min). Pd(OAc)2 (54.0 mg, 0.24 mmol, 0.08 eq) and Xanthos (140.0 mg, 0.24 mmol, 0.08 eq) were added and the mixture was heated to 50 °C and stirred for 1 h. The reaction was then heated to 80 °C and stirred for -18 h. The reaction mixture allowed to cool to rt, then was filtered and the precipitate washed with MeCN (2 x 50 mL). The filtrate was concentrated in vacuo and the resulting residue was taken up in EtOAc (100 mL) and HC1 (100 mL, IM Aq). The aqueous layer was separated and the organic layer was washed with HC1 (2 x 100 mL, IM Aq). The aqueous phases were combined and the pH was adjusted to -9-10 by addition of solid NaOH. The aqueous material was extracted with EtOAc (3 x 100 mL) and the combined organic layers were dried with MgSO4 and concentrated in vacuo. The crude product was purified by column chromatography on silica gel (40 g cartridge, 0-10% MeOH (0.7M NHsj/DCM) to give the title compound (697.0 mg, 48% yield) as a viscous dark red oil. LCMS m/z 462.3 [M+H]⁺ ESI pos.

Step F: 6-(6-Benzyl -3, 3a, 4,5,7, 7a-hexahydro-2J7-pyrrolo[2,3-c]pyridin-l-yl)-4-methyl -277-1,2, 4- triazine-3, 5-dione

Trifluorom ethanesulfonic acid (320.0 uL, 3.62 mmol, 2.52 eq) was added dropwise to a stirred solution of 6-(6-benzyl-3,3a,4,5,7,7a-hexahydro-277-pyrrolo[2,3-c]pyridin-l-yl)-2-[(4- methoxyphenyl)methyl]-4-methyl- 1, 2, 4-triazine-3, 5-dione (697.0 mg, 1.43 mmol, 1.0 eq) in DCM (8 mL) and MeCN (4 mL) at rt. The reaction was heated to 35 °C and stirred for 22 h. The reaction was diluted with H2O (100 mL) and DCM (100 mL) and transferred to a separating funnel. The separated organic layer was further extracted with IM Aq HC1 (100 mL). The aqueous layers were combined in a vigorously stirring conical flask, cooled to 0 °C and basified with sodium phosphate tribasic until a pH of -8-9 was achieved. The mixture was again transferred to a separating funnel and diluted with DCM (200 mL). The separated aqueous layer was further extracted with DCM (2 x 100 mL) and the combined organic layers were dried with MgSCU and concentrated in vacuo to provide the title compound (420.0 mg, 84% yield) as an off-white solid. LCMS m/z 341.8 [M+H]⁺ ESI pos.

Step G: 6-(6-Benzyl-3, 3a, 4,5,7, 7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl)-3 -chi oro-4-methyl- l,2,4-triazin-5-one

Phosphorus oxychloride (5.0 mL, 53.64 mmol, 43.6 eq) was added to 6-(6-benzyl-3,3a,4,5,7,7a- hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl)-4-methyl-2J/-l, 2, 4-triazine-3, 5-dione (420.0 mg, 1.23 mmol, 1.0 eq) and the solution was stirred at 105 °C. The resulting brown opaque solution was vigorous stirred for 3 days. The reaction mixture was concentrated in vacuo, then the resulting brown oil was taken up in MeCN (~10 mL) and added portion-wise to a vigorously stirred solution of EtOAc (50 mL) and K3PO4 (25 wt% Aq, 50 mL). The pH was monitored during addition to ensure the aqueous material remained basic (~pH 8). The pH of the aqueous solution was then adjusted to -pH 12 by portion-wise addition of solid K3PO4, and the organic material was separated. The aqueous material was extracted again with EtOAc (2 x 100 mL) and the combined organic layers were dried (MgSO4), filtered, and concentrated in vacuo. The crude product was purified by column chromatography on silica gel (24 g cartridge, 0-10% MeOH(0.7MNH3)/DCM) to provide the title compound (413.0 mg, 90% yield) as a light brown solid. LCMS m/z 360.2/362.1 [M+H]⁺ ESI pos.

Step H: 6-(6-Benzyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl)-3-[2-benzyloxy-4- (trifluoromethoxy)phenyl]-4-methyl-l,2,4-triazin-5-one

A mixture of [2-benzyloxy-4-(trifluoromethoxy)phenyl]boronic acid (314.0 mg, 1.01 mmol, 1.21 eq), 6-(6-benzyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl)-3-chloro-4-methyl- l,2,4-triazin-5-one (299.0 mg, 0.83 mmol, 1.0 eq) and sodium hydrogen carbonate (222.0 mg, 2.64 mmol, 3.18 eq) in water (4 mL) and MeCN (4 mL) was degassed for 5 mins. XPhos Pd G3 (75.0 mg, 0.09 mmol, 0.11 eq) was added and the reaction mixture was heated at 85 °C for 3 h. The reaction was cooled to rt, filtered through celite and concentrated in vacuo. The reaction was diluted with EtOAc (100 mL) and IM aq HC1 (100 mL) and transferred to a separating funnel. The separated organic layer was further extracted with IM aq HC1 (100 mL). The aqueous layers were combined in a vigorously stirring conical flask, cooled to 0 °C and basified with NaOH until a pH of ~8-9 was achieved. The mixture was again transferred to a separating funnel and diluted with DCM (200 mL). The separated aqueous layer was further extracted with DCM (2 x 100 mL) and the combined organic layers were dried with Na2SO4, filtered, and concentrated in vacuo to provide the title compound (423.0 mg, 75% yield) as a light-yellow solid. LCMS m/z 592.2 [M+H]⁺ ESI pos.

Step I: 6-(2,3,3a,4,5,6,7,7a-Octahydropyrrolo[2,3-c]pyridin-l-yl)-3-[2-hydroxy-4-

(trifluoromethoxy)phenyl]-4-methyl-l,2,4-triazin-5-one

Pd/C (Type 39) (190.0 mg, 0.18 mmol, 0.25 eq) and Pd/C (Type 87) (380.0 mg, 0.18 mmol, 0.25 eq) were added to a stirred solution of 6-(6-benzyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3- c]pyridin-l-yl)-3-[2-benzyloxy-4-(trifluoromethoxy)phenyl]-4-methyl-l,2,4-triazin-5-one (423.0 mg, 0.71 mmol, 1.0 eq) in 1,4-Dioxane (7 mL). The hydrogenation vessel was placed under an atmosphere of hydrogen gas (2 bar) at 50 °C and vigorously stirred for 4 h. The reaction was filtered through a plug of celite, rinsing with dioxane then MeOH, and concentrated to dryness to give the title compound (246.0 mg, 80% yield) as a light-yellow solid. LCMS m/z 412.2 [M+H]⁺ ESI pos.

Step J : 6-[6-(2-Hydroxyethyl)-3,3a,4,5,7,7a-hexahydro-2J/-pyrrolo[2,3-c]pyridin-l-yl]-3-[2- hydroxy-4-(trifluoromethoxy)phenyl]-4-methyl-l,2,4-triazin-5-one

2-Iodoethanol (19.8 mg, 0.12 mmol, 0.95 eq) in DMF (215 uL) was added dropwise to a stirred solution of 6-(2,3,3a,4,5,6,7,7a-octahydropyrrolo[2,3-c]pyridin-l-yl)-3-[2-hydroxy-4- (trifluoromethoxy)phenyl]-4-methyl-l,2,4-triazin-5-one (50.0 mg, 0.12 mmol, 1.0 eq) and DIPEA (32.0 uL, 0.18 mmol, 1.51 eq) in DMF (230 uL) at rt, and the reaction mixture was stirred for 48 h. The reaction mixture was transferred to a separating funnel and diluted with EtOAc (25 mL) and IM aq HC1 (25 mL). The separated organic layer was further extracted with IM aq HC1 (25 mL). The combined aqueous layers were washed with EtOAc (25 mL). The aqueous layer was basified with sat aq NaHCOs until a pH of ~8 was achieved and then extracted with DCM (3 x 25 mL). The combined organic layers were dried (Na2SO4), filtered and concentrated to provide the crude product. The crude product was purified by column chromatography on silica gel (4 g cartridge, 0-10% MeOH (0.7M NHs)/DCM) to give the title compound (25.7 mg, 46% yield) as a white solid. LCMS m/z 456.3 [M+H]⁺ ESI pos.

Chiral separation of Example 7:

Example 7 (24 mg) was dissolved to 8 mg/mL in 2 mL MeOH and 1 mL DCM with sonication and heating, filtered and was then separated by chiral SFC on a Waters Prep 100 with a PDA and a QDA detector, 40 °C, 120 bar. The column was a ChiralpaK IC, 21 x 250 mm, 5 pm, flow rate 65mL/ min at 30% MeOH (0.3% DEA), 70% CO2. The clean fractions were pooled, rinsed with methanol, and concentrated to dryness using a rotary evaporator. The residues were re-dissolved in methanol, transferred into final vials and evaporated on a Biotage V10 to afford Example 7A (1.3 mg, 2 % yield) and Example 7B (5.2 mg, 9% yield) both as off white solids for which the stereochemistry was arbitrarily assigned.

Example 7A: 6-[(3aS, 7i//?)-6-(2-Hydroxyethyl)-3.3a.4.5.7.7a-hexahydro-2//-pyrrolo|2.3- c]pyridin-l-yl]-3-[2-hydroxy-4-(trifluoromethoxy)phenyl]-4-methyl-l,2,4-triazin-5-one

'H NMR (500 MHz, DMSO) 8 7.40 (d, 1H), 6.95 - 6.77 (m, 2H), 4.79 - 4.52 (m, 1H), 4.41 - 4.28 (m, 1H), 3.90 - 3.56 (m, 2H), 3.47 (t, 2H), 3.14 (s, 3H), 3.07 - 2.99 (m, 1H), 2.66 - 2.56 (m, 1H), 2.38 (t, 2H), 2.34 - 2.24 (m, 1H), 2.20 - 2.08 (m, 1H), 2.05 - 1.75 (m, 4H), 1.69 - 1.60 (m, 1H). One exchangeable proton not observed. LCMS m/z 456.3 [M+H]⁺ ESI pos.

Example 7B: 6-[(3aR, 7i/.S)-6-(2-Hydroxyethyl)-3.3a.4.5.7.7a-hexahydro-2//-pyrrolo|2.3- c]pyridin-l-yl]-3-[2-hydroxy-4-(trifluoromethoxy)phenyl]-4-methyl-l,2,4-triazin-5-one

'H NMR (500 MHz, DMSO) 8 7.27 (d, 1H), 6.73 - 6.48 (m, 2H), 4.79 - 4.55 (m, 1H), 4.48 - 4.19 (m, 1H), 3.81 - 3.69 (m, 1H), 3.67 - 3.58 (m, 1H), 3.46 (t, 2H), 3.18 (s, 3H), 3.05 - 2.99 (m, 1H), 2.62 - 2.56 (m, 1H), 2.37 (t, 2H), 2.34 - 2.24 (m, 1H), 2.19 - 2.09 (m, 1H), 2.04 - 1.73 (m, 4H), 1.69 - 1.60 (m, 1H). One exchangeable proton not observed. LCMS m/z 456.3 [M+H]⁺ ESI pos.

Example A

A compound of formula I can be used in a manner known per se as the active ingredient for the production of tablets of the following composition:

Per tablet

Active ingredient 200 mg

Microcrystalline cellulose 155 mg

Com starch 25 mg

Talc 25 mg

Hydroxypropylmethylcellulose 20 mg

425 mg Example B

A compound of formula I can be used in a manner known per se as the active ingredient for the production of capsules of the following composition:

Per capsule

Active ingredient 100.0 mg

Com starch 20.0 mg

Lactose 95.0 mg

Talc 4.5 mg

Magnesium stearate 0.5 mg

220.0 mg

Claims

Claims ompound of formula I wherein,

R¹ is H, alkyl, alkoxy, -CN, halo, haloalkyl, haloalkoxy, acetyl or SFe;

R⁵ is H; or R¹ and R⁵, and the atoms to which they are bonded, form either an 4-6 membered heterocycle ring comprising a single O heteroatom optionally substituted with one or two substituents independently selected from halo or alkyl, or R¹ and R⁵, and the atoms to which they are bonded, form a 4-6 membered cycloalkyl ring optionally substituted with 1 to 2 substituents independently selected from halo or alkyl;

R² is H, halo or alkyl and R³ is H or alkyl, wherein only one of R² and R³ can be H;

R⁴ is oxetane, alkyl, or -(CH2)_n-R⁶ wherein R⁶ is hydroxy or methoxy and n is greater than 1; and pharmaceutically acceptable salts thereof. ompound according to claim 1, wherein R¹ is haloalkyl or haloalkoxy. ompound according to claim 1 or 2, wherein R² is H and R³ is alkyl. ompound according to any of claims 1 to 3, wherein R⁴ is alkyl or -(CH2)_n-R⁶ wherein R⁶ydroxy and n is 2. ompound according to any of claims 1 to 4, wherein R⁴ is alkyl. ompound according to claim 1, wherein

R¹ is haloalkyl or haloalkoxy;

R⁵ is H; or R¹ and R⁵, with the atoms to which they are attached, form a 4-to-5 membered cycloalkyl or a 5-membered heterocycle comprising a single O heteroatom;

R² is H;

R³ is alkyl;

R⁴ is alkyl or or -(CH2)_n-R⁶ wherein R⁶ is hydroxy and n is 2. and pharmaceutically acceptable salts thereof. ompound according to claim 1, wherein

R¹ is haloalkyl or haloalkoxy;

R⁵ is H; or R¹ and R⁵, with the atoms to which they are attached, form a 4-to-5 membered cycloalkyl or a 5-membered heterocycle comprising a single O heteroatom;

R² is H;

R³ is alkyl;

R⁴ is alkyl; and pharmaceutically acceptable salts thereof. ompound according to any of claims 1 to 7, wherein the compound is selected from

3-[2-Hydroxy-4-(trifluoromethoxy)phenyl]-4-methyl-6-(6-methyl-3,3a,4,5,7,7a- hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl)-l,2,4-triazin-5-one; formic acid; 6-[(3aR,7aS)-6-methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-[2- hydroxy-4-(trifluoromethoxy)phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-[2- hydroxy-4-(trifluoromethoxy)phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aR,7aS)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-(4- hydroxy-2, 3-dihydrobenzofuran-5-yl)-4-methyl-l, 2, 4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-(4- hydroxy-2,3-dihydrobenzofuran-5-yl)-4-methyl-l,2,4-triazin-5-one;

3-[4-(Difluoromethoxy)-2-hydroxy-phenyl]-4-methyl-6-(6-methyl-3,3a,4,5,7,7a- hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl)-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-[4- (difluoromethoxy)-2-hydroxy-phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aR,7aS)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-[4- (difluoromethoxy)-2-hydroxy-phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aR,7aS)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-[2- hydroxy-4-(trifluoromethyl)phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-[2- hydroxy-4-(trifluoromethyl)phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aR,7aS)-6-methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-(2- hydroxy-3-bicyclo[4.2.0]octa-l,3,5-trienyl)-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-(2- hydroxy-3-bicyclo[4.2.0]octa-l,3,5-trienyl)-4-methyl-l,2,4-triazin-5-one; and pharmaceutically acceptable salts thereof. ompound according to any one of claims 1 to 7, wherein the compound is selected from

3-(4-Hydroxyindan-5-yl)-4-methyl-6-(6-methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3- c]pyridin- 1 -yl)- 1 ,2,4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-(4- hydroxyindan-5-yl)-4-methyl-l,2,4-triazin-5-one;

6-[(3aR,7aS)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-(4- hydroxyindan-5-yl)-4-methyl-l,2,4-triazin-5-one;

6-[6-(2-Hydroxyethyl)-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-[2- hydroxy-4-(trifluoromethoxy)phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-(2-Hydroxyethyl)-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l- yl]-3-[2-hydroxy -4-(tri fluoromethoxy )phenyl]-4-methyl-l, 2, 4-triazin-5-one;

6-[(3aR,7aS)-6-(2-Hydroxyethyl)-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l- yl]-3-[2-hydroxy -4-(tri fluoromethoxy )phenyl]-4-methyl-l, 2, 4-triazin-5-one; and pharmaceutically acceptable salts thereof. ompound according to any one of claims 1 to 8, wherein the compound is selected from

6-[(3aR,7aS)-6-methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-[2- hydroxy-4-(trifluoromethoxy)phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aR,7aS)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-(4- hydroxy-2,3-dihydrobenzofuran-5-yl)-4-methyl-l,2,4-triazin-5-one;

6-[(3aR,7aS)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-[4- (difluoromethoxy)-2-hydroxy-phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-[2- hydroxy-4-(trifluoromethyl)phenyl]-4-methyl-l,2,4-triazin-5-one; 6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-(2- hydroxy-3-bicyclo[4.2.0]octa-l,3,5-trienyl)-4-methyl-l,2,4-triazin-5-one; and pharmaceutically acceptable salts thereof. A compound according to any of claims 1 to 8, wherein the compound is selected from

6-[(3aS,7aR)-6-methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-[2- hydroxy-4-(trifluoromethoxy)phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-(4- hydroxy-2,3-dihydrobenzofuran-5-yl)-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-[4- (difluoromethoxy)-2-hydroxy-phenyl]-4-methyl-l,2,4-triazin-5-one; and pharmaceutically acceptable salts thereof. A compound according to any of claims 1 to 7, or claim 9, wherein the compound is 6- [(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-(4- hydroxyindan-5-yl)-4-methyl-l,2,4-triazin-5-one, or a pharmaceutically acceptable salt thereof. A compound according to any of claims 1 to 12, wherein the compound is selected from

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-[2- hydroxy-4-(trifluoromethyl)phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-(2- hydroxy-3-bicyclo[4.2.0]octa-l,3,5-trienyl)-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-[2- hydroxy-4-(trifluoromethoxy)phenyl]-4-methyl-l,2,4-triazin-5-one; 6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-(4- hydroxy-2,3-dihydrobenzofuran-5-yl)-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-[4- (difluoromethoxy)-2-hydroxy-phenyl]-4-methyl-l,2,4-triazin-5-one;

6-[(3aS,7aR)-6-Methyl-3,3a,4,5,7,7a-hexahydro-2H-pyrrolo[2,3-c]pyridin-l-yl]-3-(4- hydroxyindan-5-yl)-4-methyl-l,2,4-triazin-5-one; and pharmaceutically acceptable salts thereof.

14. A compound according to any one of claims 1 to 13 for use as a therapeutically active substance.

15. A compound according to any one of claims 1 to 13 for use in the treatment or prevention of a disease, disorder or condition, wherein the disease, disorder or condition is responsive to NLRP3 inhibition.

16. A pharmaceutical composition comprising a compound according to any one of claims 1 to 13 and a therapeutically inert carrier.

17. The use of a compound according to any one of claims 1 to 13 for the treatment or prophylaxis of a disease, disorder or condition, wherein the disease, disorder or condition is responsive to NLRP3 inhibition.

18. A compound according to any one of claims 1 to 13 for the treatment or prophylaxis of a disease, disorder or condition selected from Asthma or COPD.

19. A compound according to any one of claims 1 to 13 for the treatment or prophylaxis of a disease, disorder or condition selected from Parkinson’s Disease or Alzheimer’s Disease.

20. The use of a compound according to any one of claims 1 to 13 in the treatment or prophylaxis of a disease, disorder or condition selected from Asthma or COPD. The use of a compound according to any one of claims 1 to 13 in the treatment or prophylaxis of a disease, disorder or condition selected from Parkinson’s Disease or Alzheimer’s Disease. The use of a compound according to any one of claims 1 to 13 for the preparation of a medicament for the treatment or prophylaxis of a disease, disorder or condition selected from Asthma or COPD. The use of a compound according to any one of claims 1 to 13 for the preparation of a medicament for the treatment or prophylaxis of a disease, disorder or condition selected from Parkinson’s Disease or Alzheimer’s Disease. A method of inhibiting NLRP3, which method comprises administering an effective amount of a compound as claimed in any one of claims 1 to 13 to inhibit NLRP3. A method for the treatment or prophylaxis of a disease, disorder or condition, which method comprises administering an effective amount of a compound according to any one of claims 1 to 13 wherein the disease, disorder or condition is selected from Asthma or COPD. A method for the treatment or prophylaxis of a disease, disorder or condition, which method comprises administering an effective amount of a compound according to any one of claims 1 to 13, wherein the disease, disorder or condition is selected from Parkinson’s Disease or Alzheimer’s Disease.