Novel compounds and uses thereof

Abstract

A compound of Formula I or II or a salt thereof; Wherein R1 and R3 are as herein defined; R3 is selected from C1-6 alkyl, C1-6 fluoroalkyl, Cl, F, I, Br, CN, hydrogen, and C1-6 alkyoxy; R4 is selected

Description

FIELD OF THE INVENTION

[0001] The present invention relates to novel synthetic compounds related to naturally occurring flavonoids such as 7,8-dihydroxyflavone (7,8-DHF). The invention further relates to the method of synthesis, the use of these compounds as research tools and their use as pharmaceuticals. BACKGROUND TO THE INVENTION

[0002] Flavonoids are a large class of plant secondary metabolites, and they are also common polyphenols in the human diet. Studies have shown that flavonoids have various pharmacological activities, such as anti-tumor, anti-inflammatory, and antioxidant properties. The flavonoid, 7,8-dihydroxyflavone (7,8-DHF), is a naturally occurring flavone found in Godmania aesculifolio, Tridax procumbens and Primula helleri leaves. It is known to act as a potent and selective agonist of tropomyosin receptor kinase B (TrkB), which is the main signaling receptor of neurotrophin brain-derived neurotrophic factor (BDNF).

[0003] 7,8-DHF has been shown to have therapeutic efficacy in several animal models including depression, Alzheimer’s disease, cognitive deficits in schizophrenia, Parkinson’s disease, Huntington’s disease, amyotrophic lateral sclerosis, traumatic brain injury, cerebral ischemia, fragile X syndrome and Rett syndrome.

[0004] A derivative of 7,8-DHF, 4-(6-oxo-2-(trifluoromethyl)-3,6-dihydrochromeno[7,8-d]imidazol-8- yl)benzonitrile, also known as CF3CN has been shown to be useful in the treatment of various diseases and conditions including neurodegenerative diseases and movement disorders.

[0005] There is a need for the development of improved flavonoid-like compounds, as well as compositions and therapeutic uses thereof. BRIEF SUMMARY OF THE INVENTION

[0006] At its most general, the present invention relates to novel synthetic flavonoid compounds.

[0007] Compounds were tested in a primary neuronal culture functional assay to determine whether they were able to promote neuronal survival upon insult.

[0008] In a first aspect of the present invention there is provided a compound of Formula I or Formula II or a salt thereof, Formula I Formula II Wherein: is a single or double bond; Ri is selected from the group consisting of: Cl, F, Br, H, OH, O-alkyl (C1-C6), O-fluoroalkyl (C1-C6), alkyl (C1-C6), fluoroalkyl(C1-C6), NH2, NH-alkyl(C1-C6) N-alkyh (C1-C6), cycloalkyl (C3-C6), fluorocycloalkyl (C3-C6), CH2-cycloalkyl (C3-C6), and CHF-cycloalkyl (C3-C6). R2 is selected from the group consisting of: alkyl (C1-C6), fluoroalkyl (C1-C6), Cl, F, I, Br, H, OH, O-alkyl (C1-C6), SMe, CN, cycloalkyl (C3-C6), and fluorocycloalkyl (C3-C6). R3 if present, when Y is selected from N or C, is selected from the group consisting of: H, alkyl(C1-C6), fluoroalkyl(C1-C6); cycloalkyl (C3-C6), alkyl (C1-C6)-OH, alkyl-(C1-C6)-OMe, and alkyl-(C1-C6)-cycloalkyl (C3-C6). R4 is selected from the group consisting of: semi-saturated or saturated cycloalkyls including bridged bicyclic or spirocyclic ring systems, unsaturated, semi-saturated or saturated heterocycloalkyls including bridged bicyclic or spirocyclic ring systems, or aryls, including but not limited to benzene, pyridine, pyrimidine, pyridazine or piperidine. In the instance that R4 is an aryl, monocyclic cycloalkyl or monocyclic heterocycloalkyl then the monocyclic ring may carry a 2, 3 or 4 position substituent from the list: H, CN, F, Cl, Br, OH, alkyl(C1-C6), fluoroalkyl (C1-C6), N-alkyh (C1-C6), NH-alkyl (C1-C6), NH-cycloalkyl (C3-C6) N-cycloalkyl (C3-C6), alkyl (C1-C6)-NH2 , alkyl (C1-C6)-OH , CH2-N-Cycloalkyl (C3-C6), CH2-heterocycloalkyl(C1-C6), CH2N-Alkyl2 (C1-C6), CH2NH-Alkyl (C1-C6), CH2N-cycloalkyl, and CH2N-fluorocycloalkyl. R5 is selected from the group consisting of: alkyl (C1-C6), fluoroalkyl (C1-C6), Cl, F, I, Br, H, OH, OMe, SMe, CN, cycloalkyl (C3-C6), and fluorocycloalkyl (C3-C6). X is independently selected from the group consisting of: N or C. Y is independently selected from the group consisting of: O, N, C, and S.

[0009] In a second aspect of the present invention, there is provided a compound of Formula III, or a salt thereof, Formula III Wherein: ----is a single or double bond; Ri and R2 are independently selected from the group consisting of: OH, OMe, O-alkyl(C1-C6), O-fluoro alkyl(C1-C6), OiPr, SMe, SF5, alkyl(C1-C6), fluoro alkyl(C1-C6), pyrazole, methyl pyrazole, oxazole, imidazole, thiazole, triazole oxadiazole, and thiadiazole; where the heterocycles can be substituted with alkyl(C1-C3). R3 is selected from the group consisting of: alkyl(C1-C6), fluoro alkyl(C1-C6), Cl, F, I, Br, CN, H O-alkyl (C1-C6). R4 is selected from the group consisting of: semi-saturated or saturated cycloalkyls including bridged bicyclic or spirocyclic ring systems, unsaturated, semi-saturated or saturated heterocycloalkyls including bridged bicyclic or spirocyclic ring systems, or aryls, including but not limited to benzene, pyridine, pyrimidine, pyridazine or piperidine. In the instance that R4 is an aryl, monocyclic cycloalkyl or monocyclic heterocycloalkyl then the monocyclic ring may carry a 2, 3 or 4 position substituent from the list: H, CN, F, Cl, Br, OH, alkyl(C1-C6), fluoroalkyl (C1-C6), N-alkyh (C1-C6), NH-alkyl (C1-C6), NH-cycloalkyl (C3-C6) N-cycloalkyl (C3-C6), alkyl 3 (C1-C6)-NH2 , alkyl (C1-C6)-OH , CH2-N-Cycloalkyl (C3-C6), CH2-heterocycloalkyl(C1-C6), CH2N-Alkyl2 (C1-C6), CH2NH-Alkyl (C1-C6), CH2N-cycloalkyl, and CH2N-fluorocycloalkyl. Rs is selected from the group consisting of: alkyl (C1-C6), fluoroalkyl (C1-C6), Cl, F, I, Br, H, OH, OMe, SMe, CN cycloalkyl (C3-C6), and fluorocycloalkyl (C3-C6).

[0010] Preferably the compound of the invention with respect to either Formula I, Formula II or Formula III is as defined by any one of the compounds numbered 1 to 50 in Table 1.

[0011] In a third aspect of the present invention, there is provided a pharmaceutical composition comprising a compound of Formula I, Formula II or Formula III, ora salt thereof, together with one or more ingredients selected from carriers, diluents, excipients, adjuvants, fillers, buffers, binders, disintegrants, preservatives, antioxidants, lubricants, stabilisers, solubilisers, surfactants (e.g., wetting agents), masking agents, colouring agents, flavouring agents, and sweetening agents.

[0012] In a fourth aspect of the present invention, there is provided a compound of Formula I, Formula II, or Formula III or a pharmaceutical composition comprising the compound of Formula I, Formula II, or Formula III, for use as a medicament.

[0013] In a fifth aspect of the invention, there is a method of treatment comprising administering to a subject in need of treatment a therapeutically effective amount of a compound of Formula I, Formula II, or Formula III.

[0014] In a sixth aspect of the invention, there is provided a method of synthesizing the compound of Formula I, Formula II, or Formula III.

[0015] In a seventh aspect of the invention, there is provided an intermediate formed in the method of synthesis of the compound of Formula I, Formula II, or Formula III.

[0016] These and other aspects and embodiments of the invention are described in further detail below. DEFINITIONS

[0017] “Alkyl” refers to substituted or unsubstituted saturated hydrocarbon groups, including straight-chain alkyl and branched-chain alkyl groups. An alkyl group may contain from one to twelve carbon atoms (e.g., C-m2 alkyl), such as one to eight carbon atoms (C1-8 alkyl) or one to six carbon atoms (C1-6 alkyl). Exemplary alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, hexyl, septyl, octyl, nonyl, and decyl. An alkyl group is attached to the rest of the molecule by a single bond. Unless stated otherwise specifically in the specification, an alkyl group is optionally substituted by one or more substituents such as those substituents described herein.

[0018] “Haloalkyl” refers to an alkyl group that is substituted by one or more halogens. Exemplary haloalkyl groups include trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, and 1,2-dibromoethyl.

[0019] The term “cycloalkyl” employed alone or in combination with other terms, refers to a nonaromatic hydrocarbon ring system (monocyclic, bicyclic, or polycyclic), including cyclized alkyl and alkenyl groups. Cycloalkyl groups can include mono- or polycyclic (e.g. having 2, 3, or 4 fused rings) groups and spirocycles. Cycloalkyl groups can have 3, 4, 5, 6, or 7 ring-forming carbons (C3 - C7). In some embodiments, the cycloalkyl group has 3 to 6 ring members, 3 to 5 ring members, or 3 to 4 ring members. In some embodiments, the cycloalkyl group is monocyclic. In some embodiments, the cycloalkyl group is bicyclic or spirocyclic. Ring-forming carbon atoms of a cycloalkyl group can be optionally oxidized to form an oxo or sulfido group. Also included in the definition of cycloalkyl are moieties that have one or more aromatic rings fused (i.e. having a bond in common with) to the cycloalkyl ring, e.g. benzo or thienyl derivatives of cyclopentane, cyclohexane, and the like. A cycloalkyl group containing a fused aromatic ring can be attached through any ring-forming atom including a ring-forming atom of the fused aromatic ring. Examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclohexenyl, and cyclohexadienyl. Examples of polycyclic cycloalkyl groups include, but are not limited to, nonane and adamantane. Spirocyclic cycloalkyl groups include, but are not limited to spiro-, heptane, octane, nonane, decane and undecane.

[0020] The term "heterocycloalkyl” employed alone or in combination with other terms, refers to both aromatic and non-aromatic rings or ring systems, which may optionally contain one or more alkenylene groups as part of the ring structure, which has at least one heteroatom ring member independently selected from nitrogen, sulfur, oxygen, and phosphorus, and which has 3-12 total ring members. Any heterocycloalkyl ring member may be further mono-, or di-, alkylated (C1-C6) if allowed by rules for carbon valency. Included within the term "heterocycloalkyl” are monocyclic 3-, 4-, 5-, 6-, and 7-, membered heterocycloalkyl groups. Heterocycloalkyl groups can include bicyclic (e.g. having two fused or bridged rings) or spirocyclic (e.g. having two ring systems fused by a single shared carbon atom) ring systems containing 3-12 total ring members. In some embodiments, the heterocycloalkyl group is a monocyclic group having 1,2, or 3 heteroatoms independently selected from nitrogen, sulfur, and oxygen. In some embodiments, the heterocycloalkyl group is a poly-, or spirocyclic group having 1,2, or 3 heteroatoms independently selected from nitrogen, sulfur, and oxygen. Ringforming carbon atoms and heteroatoms of a heterocycloalkyl group can be optionally oxidized to form an oxo or sulfido group or other oxidized linkage (e.g. C (O), S (O), C (S) or S (O)2, N-oxide etc.) or a nitrogen atom can be quaternized. Where a heteroatom is selected to be a nitrogen, this heteroatom may be further functionalized to the corresponding N-alkyl(C1-C6), N-fluoroalkyl (C1-C6) or N-CO-Alkyl(C1-C6). The heterocycloalkyl group can be attached through a ring-forming carbon atom ora ring-forming heteroatom. In some embodiments, the heterocycloalkyl group contains 0 to 3 double bonds. In some embodiments, the heterocycloalkyl group contains 0 to 2 double bonds. Also included in the definition of heterocycloalkyl are moieties that have one or more aromatic rings fused (i.e. having a bond in common with) to the heterocycloalkyl ring, e.g. benzo or thienyl derivatives of piperidine, morpholine, azepine, etc. A heterocycloalkyl group containing a fused aromatic ring can be attached through any ring-forming atom including a ring-forming atom of the fused aromatic ring. Examples of monocyclic heterocycloalkyl groups include, but are not limited to oxirane, aziridine, azetidine, pyrrolidine, piperidine, piperizine, pyridine, pyrimidine, pyridazine, pyrazole, pyrrole, imidazole, morpholine, dioxane, pyran, furan, thiophene, thoiazole, and oxazole in any variation of saturation. Examples of polycyclic heterocycloalkyl groups include, but are not limited to, quinoline, isoquinoline, indole, benzofuran, benzimidazole, chromane, chromene, and coumarine. Examples of spirocyclic heterocycloalkyls include, but are not limited to, 1-oxa-8-azaspiro[4.5]decane, 2,2-dimethyl-1-oxa-8-azaspiro[4.5]decane, 1,1-dimethyl-2-oxa-8-azaspiro[4.5]decane 3-oxa-9-azaspiro[5.5]undecane, 2,6-diazaspiro[3.4]octan-5-one

[0021] “Alkenyl” refers to substituted or unsubstituted hydrocarbon groups, including straightchain or branched-chain alkenyl groups containing at least one double bond. An alkenyl group may contain from two to twelve carbon atoms (e.g., C2.12 alkenyl). Exemplary alkenyl groups include ethenyl (i.e., vinyl), prop-1-enyl, but-1-enyl, pent-1-enyl, penta-1,4-dienyl, and the like. Unless stated otherwise specifically in the specification, an alkenyl group is optionally substituted by one or more substituents such as those substituents described herein.

[0022] “Alkynyl” refers to substituted or unsubstituted hydrocarbon groups, including straightchain or branched-chain alkynyl groups containing at least one triple bond. An alkynyl group 6 may contain from two to twelve carbon atoms (e.g., C2-12 alkynyl). Exemplary alkynyl groups include ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like. Unless stated otherwise specifically in the specification, an alkynyl group is optionally substituted by one or more substituents such as those substituents described herein.

[0023] “Aryl” refers to an all carbon monocyclic or fused-ring polycyclic (i.e., rings which share adjacent pairs of carbon atoms) groups having a completely conjugated pi electron system. Examples, without limitation, of aryl groups are phenyl, napthalenyl and anthracenyl. The aryl group may be substituted or unsubstituted.

[0024] “Ester” refers to a functional group -COO and may also be referred to as an “ester link”. Esters are formed by the condensation reaction between an alcohol and a carboxylic acid.

[0025] The term “halo” or, alternatively, “halogen” means fluoro or fluorine, chloro or chlorine, bromo or bromine and iodo or iodine.

[0026] The term “substituted” refers to moieties having substituents replacing a hydrogen on one or more carbons or heteroatoms of the structure. It will be understood that “substitution” or “substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. As used herein, the term “substituted” is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this disclosure, the heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. Substituents can include any substituents described herein, for example, a halogen, a hydroxyl, a carbonyl (such as a carboxyl, an alkoxycarbonyl, a formyl, or an acyl), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an alkoxyl, a phosphoryl, a phosphate, a phosphonate, a phosphinate, an amino, an amido, an amidine, an imine, a cyano, a nitro, an azido, a sulfhydryl, an alkylthio, a sulfate, a sulfonate, a sulfamoyl, a sulfonamido, a sulfonyl, a heterocycle, an aralkyl, a carbocycle, a heterocycle, a cycloalkyl, a heterocycloalkyl, an aromatic and heteroaromatic moiety.

[0027] It will be understood by those skilled in the art that substituents can themselves be substituted, if appropriate. Unless specifically stated as “unsubstituted,” references to chemical moieties herein are understood to include substituted variants. For example, reference to a “heteroaryl” group or moiety implicitly includes both substituted and unsubstituted variants.

[0028] Where substituent groups are specified by their conventional chemical formulae, written from left to right, they equally encompass the chemically identical substituents that would result from writing the structure from right to left, e.g., -CH2O- is equivalent to -OCH2-.

[0029] “Optional” or “optionally” means that the subsequently described event of circumstances may or may not occur, and that the description includes instances where the event or circumstance occurs and instances in which it does not. For example, “optionally substituted aryl” means that the aryl group may or may not be substituted and that the description includes both substituted aryl groups and aryl groups having no substitution.

[0030] Compounds of the present disclosure also include crystalline and amorphous forms of those compounds, pharmaceutically acceptable salts, and active metabolites of these compounds having the same type of activity, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (including anhydrates), conformational polymorphs, and amorphous forms of the compounds, as well as mixtures thereof.

[0031] The compounds described herein may exhibit their natural isotopic abundance, or one or more of the atoms may be artificially enriched in a particular isotope having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number predominantly found in nature. All isotopic variations of the compounds of the present disclosure, whether radioactive or not, are encompassed within the scope of the present disclosure. For example, hydrogen has three naturally occurring isotopes, denoted 1H (protium), 2H (deuterium), and 3H (tritium). Protium is the most abundant isotope of hydrogen in nature. Enriching for deuterium may afford certain therapeutic advantages, such as increased in vivo half-life and / or exposure, or may provide a compound useful for investigating in vivo routes of drug elimination and metabolism. Isotopically-enriched compounds may be prepared by conventional techniques well known to those skilled in the art.

[0032] “Isomers” are different compounds that have the same molecular formula. “Stereoisomers” are isomers that differ only in the way the atoms are arranged in space. “Enantiomers” are a pair of stereoisomers that are non-superimposable mirror images of each 8 other. A 1:1 mixture of a pair of enantiomers is a “racemic” mixture. The term “(±)” is used to designate a racemic mixture where appropriate. “Diastereoisomers” or “diastereomers” are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other. The absolute stereochemistry is specified according to the Cahn-lngold-Prelog R-S system. When a compound is a pure enantiomer, the stereochemistry at each chiral carbon can be specified by either R orS. Resolved compounds whose absolute configuration is unknown can be designated (+) or (-) depending on the direction (dextro- or levorotatory) in which they rotate plane polarized light at the wavelength of the sodium D line. Certain compounds described herein contain one or more asymmetric centers and can thus give rise to enantiomers, diastereomers, and other stereoisomeric forms, the asymmetric centers of which can be defined, in terms of absolute stereochemistry, as (R)- or (S)-. The present chemical entities, pharmaceutical compositions and methods are meant to include all such possible stereoisomers, including racemic mixtures, optically pure forms, mixtures of diastereomers and intermediate mixtures. Optically active (R)- and (S)-isomers can be prepared using chiral synthons or chiral reagents or resolved using conventional techniques. The optical activity of a compound can be analyzed via any suitable method, including but not limited to chiral chromatography and polarimetry, and the degree of predominance of one stereoisomer over the other isomer can be determined.

[0033] Chemical entities having carbon-carbon double bonds or carbon-nitrogen double bonds may exist in Z- or E- form (or cis- or trans- form). Furthermore, some chemical entities may exist in various tautomeric forms. Unless otherwise specified, chemical entities described herein are intended to include all Z-, E- and tautomeric forms as well.

[0034] Isolation and purification of the chemical entities and intermediates described herein can be effected, if desired, by any suitable separation or purification procedure such as, for example, filtration, extraction, crystallization, column chromatography, thin-layer chromatography or thick-layer chromatography, or a combination of these procedures. Specific illustrations of suitable separation and isolation procedures can be had by reference to the examples herein below. However, other equivalent separation or isolation procedures can also be used.

[0035] When stereochemistry is not specified, certain small molecules described herein include, but are not limited to, when possible, their isomers, such as enantiomers and diastereomers, mixtures of enantiomers, including racemates, mixtures of diastereomers, and other mixtures thereof, to the extent they can be made by one of ordinary skill in the art by routine experimentation. In those situations, the single enantiomers or diastereomers, i.e., optically active forms, can be obtained by asymmetric synthesis or by resolution of the racemates or mixtures of diastereomers. Resolution of the racemates or mixtures of diastereomers, if possible, can be accomplished, for example, by conventional methods such as crystallization in the presence of a resolving agent, or chromatography, using, for example, a chiral high-pressure liquid chromatography (HPLC) column. Furthermore, a mixture of two enantiomers enriched in one of the two can be purified to provide further optically enriched form of the major enantiomer by recrystallization and / or trituration. In addition, such certain small molecules include Z- and E- forms (or cis- and trans- forms) of certain small molecules with carbon-carbon double bonds or carbon-nitrogen double bonds. Where certain small molecules described herein exist in various tautomeric forms, the term “certain small molecule” is intended to include all tautomeric forms of the certain small molecule.

[0036] The term “salt” or “pharmaceutically acceptable salt” refers to salts derived from a variety of organic and inorganic counter ions well known in the art. Pharmaceutically acceptable acid addition salts can be formed with inorganic acids and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, specifically such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In embodiments, the pharmaceutically acceptable base addition salt is chosen from ammonium, potassium, sodium, calcium, and magnesium salts.

[0037] The phrase “pharmaceutically acceptable excipient” or “pharmaceutically acceptable carrier” as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. Each carrier 10 must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations.

[0038] As used herein, “treatment” or “treating” refers to an approach for obtaining beneficial or desired results with respect to a disease, disorder, or medical condition including but not limited to a therapeutic benefit and / or a prophylactic benefit. A therapeutic benefit can include, for example, the eradication or amelioration of the underlying disorder being treated. Also, a therapeutic benefit can include, for example, the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the subject, notwithstanding that the subject may still be afflicted with the underlying disorder. In certain embodiments, for prophylactic benefit, the compositions are administered to a subject at risk of developing a particular disease, or to a subject reporting one or more of the physiological symptoms of a disease, even though a diagnosis of this disease may not have been made. DETAILED DESCRIPTION OF THE INVENTION

[0039] Throughout this disclosure, various patents, patent applications and publications are referenced. The disclosures of these patents, patent applications and publications in their entireties are incorporated into this disclosure by reference for all purposes in order to more fully describe the state of the art as known to those skilled therein as of the date of this disclosure. This disclosure will govern in the instance that there is any inconsistency between the patents, patent applications and publications cited and this disclosure.

[0040] Modifying tropoflavin compounds can significantly change their chemical and biological properties. Such chemical functional groups can include a polar moiety, a monosaccharide, disaccharide, a carbohydrate, an amino acid, an acyl group, a diacid group, and other chemical moieties. For biological systems, the addition of such modifying functional groups can significantly alter the resulting biological activity or tissue targeting. For the end use of these compounds, modifications have major impacts on downstream formulations, preparations, pharmacokinetics, pharmacodynamics, and ultimate end uses. Compounds

[0041] In embodiments, the present disclosure provides a compound of Formula I, Formula II, or a salt thereof: Formula I Formula II Wherein: ----is a single or double bond; Ri is selected from the group consisting of: Cl, F, Br, H, OH, O-alkyl (C1-C6), O-fluoroalkyl (C1-C6), alkyl (C1-C6), fluoroalkyl(C1-C6), NH2, NH-alkyl(C1-C6) N-alkyl2 (C1-C6), cycloalkyl (C3-C6), fluorocycloalkyl (C3-C6), CH2-cycloalkyl (C3-C6), and CHF-cycloalkyl (C3-C6); R2 is selected from the group consisting of: alkyl (C1-C6), fluoroalkyl (C1-C6), Cl, F, I, Br, H, OH, O-alkyl (C1-C6), SMe, CN, cycloalkyl (C3-C6), and fluorocycloalkyl (C3-C6); R3 if present when Y is selected from N or C, is selected from the group consisting of: H, alkyl(C1-C6), fluoroalkyl(C1-C6); cycloalkyl (C3-C6), alkyl (C1-C6)-OH, alkyl-(C1-C6)-OMe, and alkyl-(C1-C6)-cycloalkyl (C3-C6); R4 is selected from the group consisting of: semi-saturated or saturated cycloalkyls including bridged bicyclic or spirocyclic ring systems, unsaturated, semi-saturated or saturated heterocycloalkyls including bridged bicyclic or spirocyclic ring systems, or aryls, including but not limited to benzene, pyridine, pyrimidine, pyridazine or piperidine. In the instance that R4 is an aryl, monocyclic cycloalkyl or monocyclic heterocycloalkyl then the monocyclic ring may carry a 2, 3 or 4 position substituent from the list: H, CN, F, Cl, Br, OH, alkyl(C1-C6), acyl, fluoroalkyl (C1-C6), N-alkyl2 (C1-C6), NH-alkyl (C1-C6), NH-cycloalkyl (C3-C6) N-cycloalkyl (C3-C6), alkyl (C1-C6)-NH2 , alkyl (C1-C6)-OH , CH2-N-Cycloalkyl (C3-C6), CH2-heterocycloalkyl(C1-C6), CH2N-Alkyl2 (C1-C6), CH2NH-Alkyl (C1-C6), CH2N-cycloalkyl, and CH2N-fluorocycloalkyl; R5 is selected from the group consisting of: alkyl (C1-C6), fluoroalkyl (C1-C6), Cl, F, I, Br, H, OH, OMe, SMe, CN, cycloalkyl (C3-C6), and fluorocycloalkyl (C3-C6); X is independently selected from the group consisting of: N or C; and Y is independently selected from the group consisting of: O, N, C, and S.

[0042] In embodiments, the present disclosure provides a compound of Formula III or a salt thereof: Formula III Wherein: 3333 is a single or double bond; Ri and R2 are independently selected from the group consisting of: OH, OMe, O-alkyl(C1-C6), O-fluoro alkyl(C1-C6), OiPr, SMe, SFs, alkyl(C1-C6), fluoro alkyl(C1-C6), pyrazole, methyl pyrazole, oxazole, imidazole, thiazole, triazole oxadiazole, and thiadiazole; where the heterocycles can be substituted with alkyl(C1-C3). R3 is selected from the group consisting of: alkyl(C1-C6), fluoro alkyl(C1-C6), Cl, F, I, Br, CN, H O-alkyl (C1-C6). R4 is selected from the group consisting of: semi-saturated or saturated cycloalkyls including bridged bicyclic or spirocyclic ring systems, unsaturated, semi-saturated or saturated heterocycloalkyls including bridged bicyclic or spirocyclic ring systems, or aryls, including but not limited to benzene, pyridine, pyrimidine, pyridazine or piperidine. In the instance that R4 is an aryl, monocyclic cycloalkyl or monocyclic heterocycloalkyl then the monocyclic ring may carry a 2, 3 or 4 position substituent from the list: H, CN, F, Cl, Br, OH, alkyl(C1-C6), fluoroalkyl (C1-C6), N-alkyh (C1-C6), NH-alkyl (C1-C6), NH-cycloalkyl (C3-C6) N-cycloalkyl (C3-C6), alkyl (C1-C6)-NH2, alkyl (C1-C6)-OH , CH2-N-Cycloalkyl (C3-C6), CH2-heterocycloalkyl(C1-C6), CH2N-Alkyl2 (C1-C6), CH2NH-Alkyl (C1-C6), CH2N-cycloalkyl, and CH2N-fluorocycloalkyl. Rs is selected from the group consisting of: alkyl (C1-C6), fluoroalkyl (C1-C6), Cl, F, I, Br, H, OH, OMe, SMe, CN cycloalkyl (C3-C6), and fluorocycloalkyl (C3-C6).

[0043] In embodiments, provided herein is one or more compounds selected from Table 1. The compounds detailed in Table 1, with exception of compound 10 are within the scope of the compounds of Formulas I to III. Such compounds are further exemplified to substantiate the compounds of Formulas I to III.

[0044] In embodiments, provided herein is one or more pharmaceutically acceptable salts of a compound selected from Table 1. Table 1. Compounds Compositions

[0045] The compounds described herein may be formulated as a pharmaceutical composition. A pharmaceutical composition may comprise: (i) a compound of Formula I, Formula II or Formula III, as detailed in Table 1, or a pharmaceutically acceptable salt thereof; and (ii) a pharmaceutically acceptable carrier.

[0046] In embodiments, a pharmaceutically acceptable carrier includes a pharmaceutically acceptable excipient, binder, and / or diluent. In embodiments, suitable pharmaceutically acceptable excipients include, but are not limited to, water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose and polyvinylpyrrolidone. In embodiments, suitable pharmaceutically acceptable carriers include, but are not limited to, inert solid fillers or diluents and sterile aqueous or organic solutions. Sterile phosphate-buffered saline is one example of a pharmaceutically acceptable carrier. Other suitable carriers, diluents, or excipients are well- known to those in the art. (See, e.g., Gennaro (ed.), Remington's Pharmaceutical Sciences (Mack Publishing Company, 19th ed. 1995).) Formulations can further include one or more excipients, preservatives, solubilizers, buffering agents, albumin to prevent protein loss on vial surfaces, etc.

[0047] Pharmaceutical compositions comprising a compound of Formula (I) or Formula (II), as detailed in Table 1 or a pharmaceutically acceptable salt thereof, may also contain one or more additional ingredients including, but not limited to, a mucoadhesive compound, a buffering agent, a plasticizing agent, a stabilizing agent, a taste-masking agent, a flavoring agent, a coloring agent, an antiseptic, an inert filler agent, a preservative, and combinations thereof.

[0048] In embodiments, the formulations may comprise one or more solubilizing agents that increase the solubility of active compounds in the formulation. Suitable solubilizing agents include, for example, complexing agents, surfactants, and the like. Suitable complexing agents include unsubstituted cyclodextrins (such as alpha-cyclodextrin, beta-cyclodextrin) and substituted cyclodextrins, (such as hydroxypropyl beta-cyclodextrin, sulfobutylether-beta-cyclodextrin). Suitable surfactants include polyoxyethylene sorbitan monolaurate (for example, Tween 20), polyoxyethylene sorbitans molooleate (for example, Tween 80), polyethylene glycol (15)-hydroxystearate (for example, Kolliphor® HS 15), PEG-35 castor oil (for example, Kolliphor® EL) and PEG-60 hydrogenated castor oil (for example, Cremophor® RH 60).

[0049] In embodiments, the formulations comprise one or more buffer agents that maintain the pH of the IV solution within a pharmaceutically acceptable range. In certain embodiments, the buffer maintains the pH of the IV solution between about 5 and 9. In specific embodiments, the buffer maintains the pH of the IV solution at about 7.4. Suitable buffers include, for example, citrates, lactate, acetate, maleate, phosphates, and the like. In embodiments, the formulations comprise one or more density modifiers that are used to control the density of the IV formulation. Suitable density modifiers include, for example, dextrose. In embodiments, the formulations comprise one or more isotonicity modifiers that provide a formulation that is iso-osmotic with tissue to prevent pain and irritation when the formulation is administered. Suitable isotonicity modifiers include, for example, electrolytes, monosaccharides, and disaccharides. Examples of isotonicity modifiers include glycerin, dextrose, potassium chloride, and sodium chloride.

[0050] In embodiments, the formulations comprise one or more viscosity enhancers. Suitable viscosity enhancers include, for example, povidone, hydroxyethylcellulose, polyvinyl alcohol, and carbomer (such as, acrylic acid homopolymers and acrylic acid copolymers).

[0051] In embodiments, the formulations comprise one or more preservatives that increase the stability of active compounds in the formulation and / or provide antimicrobial activity. Suitable preservatives include, for example, antimicrobial agents and antioxidants. Examples of antimicrobial agents include benzyl alcohol, methyl paraben, propyl paraben, phenol, cresol, methyl paraben, chlorbutanol, sodium metabisulphite, sodium bisulphite, benzethonium chloride, and benzalkonium chloride. Examples of antioxidants include sodium bisulphite and other sulfurous acid salts, ascorbic acid, salts of ethylenediaminetetraacetic acid (including sodium), alpha tocopherol, butylated hydroxyl hydroxytoluene, and butylated hydroxyanisole.

[0052] A pharmaceutical composition comprising a compound of Formula (I) or Formula (II), as detailed in Table 1, or a pharmaceutically acceptable salt thereof; may be formulated in a dosage form selected from the group consisting of: an oral unit dosage form, an intravenous unit dosage form, an intranasal unit dosage form, a suppository unit dosage form, an intradermal unit dosage form, an intramuscular unit dosage form, an intraperitoneal unit dosage form, a subcutaneous unit dosage form, an epidural unit dosage form, a sublingual unit dosage form, a liquid, a lozenge, a fast disintegrating tablet, a lyophilized preparation, a film, a spray (including a nasal spray, an oral spray, or a topical spray), or a mucoadhesive. The oral unit dosage form may be selected from the group consisting of: tablets, pills, pellets, capsules, powders, lozenges, granules, solutions, suspensions, emulsions, syrups, elixirs, sustained-release formulations, aerosols, and sprays. In embodiments, the modified mesembrine alkaloid is formulated as a liquid, a lozenge, a fast-disintegrating tablet, a lyophilized preparation, a film, a spray, or a mucoadhesive.

[0053] The compounds of Formula (I) or Formula (II), as detailed in Table 1, ora pharmaceutically acceptable salt thereof can be administered to subjects by a variety of administration modes, including, for example, by intramuscular, subcutaneous, intravenous, intra-atrial, intra-articular, parenteral, intranasal, intrapulmonary, transdermal, intrapleural, intrathecal, and oral routes of administration. For prevention and treatment purposes, a compound of Formula (I) or Formula (II), as detailed in Table 1, ora pharmaceutically acceptable salt thereof can be administered to a subject in a single bolus delivery, via continuous delivery (e.g., continuous transdermal delivery) over an extended time period, or in a repeated administration protocol (e.g., on an hourly, daily, weekly, or monthly basis).

[0054] Pharmaceutical compositions comprising a compound of Formula (I) or Formula (II), as detailed in Table 1, or a pharmaceutically acceptable salt thereof can be supplied as a kit comprising a container that comprises the pharmaceutical composition as described herein. A pharmaceutical composition can be provided, for example, in the form of an injectable solution for single or multiple doses, or as a sterile powder that will be reconstituted before injection. Alternatively, such a kit can include a dry-powder disperser, liquid aerosol generator, or nebulizer for administration of a pharmaceutical composition. Such a kit can further comprise written information on indications and usage of the pharmaceutical composition. Therapeutic Use

[0055] The compounds of Formula I, Formula II, or Formula III, as detailed in Table 1, have demonstrated efficacy in an in vitro neuronal survival assay. Therefore, the novel synthetic flavonoid compounds of the invention may be useful in the treatment or prevention of medical conditions. More specifically the activity of these compounds is likely to enable their use in the treatment or prevention of medical conditions associated with neurodegeneration. WORKED EXAMPLES

[0056] The compounds of the present invention were synthesized using the synthetic reaction schema described in Example 1.

[0057] Reaction products can be purified by known methods including silica gel chromatography using various organic solvents such as hexane, dichloromethane, ethyl acetate, methanol and the like or preparative reverse phase high pressure liquid chromatography.

[0058] Preparation of compounds can involve the protection and deprotection of various chemical groups. The need for protection and deprotection, and the selection of appropriate protecting groups can be readily determined by one skilled in the art. The chemistry of protecting groups can be found, for example, in Greene and Wuts, Protective Groups in Organic Synthesis, 44th. Ed., Wiley &Sons, 2006, as well as in Jerry March, Advanced Organic Chemistry, 4th edition, John Wiley &Sons, publisher, New York, 1992 which are incorporated herein by reference in their entirety.

[0059] Example 2 describes the efficacy of these compounds in an in vitro neuronal survival assay. EXAMPLE 1:

[0060] In order to prepare the compounds of Formula I and Formula II as exemplified by the compounds 1 to 50 in Table 1, the following reaction methodologies were employed. Abbreviations: Abbreviation Full name AcOH glacial acetic acid aq aqueous Boc tert-butyloxycarbonyl br broad cone concentrated d doublet DCM dichloromethane DIPEA N,N-diisopropylethylamine DMF N,N-dimethylformamide DMSO dimethyl sulfoxide (ES+) electrospray ionization, positive mode (ES-) Et electrospray ionization, negative mode ethyl EtOAc ethyl acetate HATU 1-[bis(dimethylamino)methylene]-1 H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate HPLC high performance liquid chromatography (reverse phase) h hour(s) m multiplet (M+H)+ protonated molecular ion (M-H)-Me deprotonated molecular ion methyl MeCN acetonitrile MeOH methanol MHz megahertz min minute(s) m / z mass-to-charge ratio NMP N-methylpyrrolidine NMR nuclear magnetic resonance (spectroscopy) Pd(dppf)CI2 [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(ll) Ph phenyl prep HPLC preparative high performance liquid chromatography q quartet rt room temperature s singlet sat saturated sex solid supported cation exchange (resin) t triplet THF tetrahydrofuran THP tetrahydropyranyl General Information Solvents and Reagents

[0061] Organic Solvents and chemical reagents were purchased from commercial sources and used as received unless otherwise stated. Preparation of Intermediates

[0062] Known synthetic intermediates were procured from commercial vendors or were obtained using published literature procedures with reference to original published literature as annotated. Additional intermediates were prepared by the representative synthetic processes described herein. Nuclear Magnetic Resonance Spectroscopy (NMR)

[0063] NMR spectra were recorded using a Bruker 400MHz Avance Neo spectrometer fitted with a Bruker 5mm iProbe, or a Bruker 500MHz Avance III HD spectrometer equipped with a Bruker 5mm SmartProbeTM. Spectra were measured at 298 K, unless indicated otherwise, and were referenced relative to the solvent resonance. The chemical shifts are reported in parts per million. Data were acquired using Bruker TopSpin software and processed using MestreNova software. Mass Spectrometry (MS)

[0064] Compounds were analysed by Ultra High Performance Liquid Chromatography Mass Spectroscopy (UPLC) or Liquid Chromatography Mass Spectrometry (LCMS) as indicated. UPLC (Method 1) Apparatus: Waters HCIass; Binary Solvent Pump, SM-FTN, CMA, PDA, QDa Column: Waters ACQUITY UPLC® BEH C18, 1.7 pm, 2.1 x 30 mm at 40 °C Detection: UV at 210-400 nm unless otherwise indicated, MS by electrospray ionisation Solvents: A: 0.1% Ammonia in water, B: MeCN Gradient: Time %A %B Flow rate (ml / min) 0.00 98 2 0.77 2.50 0 100 0.77 3.00 0 100 0.77 LCMS (Method 1) Apparatus: Agilent 1260; Binary Pump, HiP Sampler, Column Compartment, DAD:, G6150 MS Column: Waters Cortecs C18, 30x2.1 mm, 2.7pm, at 40 °C Detection: UV at 260nm +1- 90nm unless otherwise indicated, MS by electrospray ionisation Solvents: A: 0.1% formic acid in water, B: MeCN Gradient: Time %A %B Flow rate (ml / min) 0.00 98 2 1.35 2.5 0 100 1.35 3 0 100 1.35 General Procedures General Procedure 1: Suzuki coupling with pyrazoleboronic esters

[0065] A N2-purged vial containing the 8-bromoflavone (0.37-0.91 mmol, 1 Eq), pyrazole boronic ester (1.5-2.8 Eq), and Pd(dtbpf)Ch (Pd-118) (5 mol%) was charged with aq. tripotassium phosphate (1.5 molar, 4 Eq) and 1,4-dioxane (4 mL).

[0066] The mixture was stirred at 95 °C for 90 min, then cooled to rt. The mixture was diluted with EtOAc (25 mL), washed with 1:1 water / brine (25 mL) and brine (10 mL), dried over MgSO4, filtered and concentrated. The residue was purified by chromatography or trituration as stated to give the product. General Procedure 2: Triazole displacement

[0067] A vial containing the appropriate triazole bearing chromanone (1 Eq), the amine nucleophile (1.3-2 Eq) and potassium carbonate (2-5 Eq) was charged with NMP (1-2 mL). The mixture was stirred at 80-120 °C for the indicated time.

[0068] The mixture was cooled to rt and either purified immediately if indicated or diluted with EtOAc (10 mL), washed with 1:1 brine / water (2x10 mL) and brine (5 mL), dried over MgSCh, filtered and concentrated. The residue was purified by chromatography or trituration as stated to give the product. General Procedure 3: Demethylation

[0069] A N2-purged flask containing the anisole starting material and pyridine hydrochloride (0.30 g, 2.6 mmol) was stirred at 180 °C for 15 min, then cooled to rt.

[0070] The mixture was diluted with water (4 mL) and sonicated until a fine suspension was obtained. The solid was collected by filtration, rinsed with water (3x1 mL) and MeCN (3x1 mL), then dried to give the phenol. Any further purification was performed as described. General Procedure 4: Purification by High Performance Liquid Chromatography (HPLC)

[0071] Experiments were performed using a 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 Method 1.

[0072] The product was dissolved in a suitable mixture of mobile phases and DMSO to ensure compete solvation. The solution was filtered and injected onto a Waters XBridge BEH C18 ODB prep column, 130A, 5 pm, 30 mm X 100 mm, flow rate 40 mL min-1 eluting with a 0.3% Ammonia in water-MeCN gradient over 17.5 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; 0.5-15.5 min, ramped from 5% MeCN to 35% MeCN; 15.5-15.6 min, ramped from 35% MeCN to 100% MeCN; 15.6-17.5 min, held at 100% MeCN. Method 2.

[0073] The product was dissolved in a suitable mixture of mobile phases and DMSO to ensure compete solvation. The solution was filtered and injected onto a Waters X-Select CSH C18 ODB prep column, 130A, 5 pm, 30 mm X 100 mm, flow rate 40 mL min1 eluting with a 0.1% Formic acid in water-MeCN gradient over 17.5 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; 0.5-15.5 min, ramped from 5% MeCN to 22.5% MeCN; 15.5-15.6 min, ramped from 22.5% MeCN to 100% MeCN; 15.6-17.5 min, held at 100% MeCN.

[0074] The following synthetic pathways were used to prepare compound 1 to 24 of Table 1. Compound 1 8-Bromo-7-hydroxy-2-phenyl-4H-chromen-4-one

[0075] To a solution of 7-hydroxy-2-phenyl-4H-chromen-4-one (5.0 g, 1 Eq, 21 mmol) in DMF (50 mL) at 3 °C (internal) was added a solution of NBS (4.0 g, 1.1 Eq, 22 mmol) in DMF (10 mL). The mixture was stirred at 3 °C for 2 h. The mixture was diluted with water (120 mL) to give a thick suspension. The solid was collected by filtration (4 x 10 mL of water was used to complete the transfer), rinsed with MeCN (4x10 mL) and suspended in isopropanol (150 mL). The slurry was stirred at 80 °C overnight, then left to stand at rt for 1 h. The solid was collected by filtration, rinsed with isopropanol (3x10 mL) and TBME (3x10 mL), then dried to give 8-bromo-7-hydroxy-2-phenyl-4H-chromen-4-one (5.8 g, 15 mmol, 72%) as a white solid.

[0076] 1H NMR (500 MHz, DMSO): 5 11.65 (s, 1H), 8.16-8.11 (m, 2H), 7.89 (d, J= 8.7 Hz, 1H), 7.65-7.55 (m, 3H), 7.11 (d, J =8.8 Hz, 1H), 7.03 (s, 1H).

[0077] MS: The product was analysed by UPLC (Method 1): m / z 317 / 319 [M+H]+ (ES+); 315 / 317 [M-H]' (ES-), at 0.70 min, 80% purity 210-400nm. 8-Bromo-7-methoxy-2-phenyl-4H-chromen-4-one

[0078] To a suspension of 8-bromo-7-hydroxy-2-phenyl-4H-chromen-4-one (1.0 g, 83% Wt, 1 Eq, 2.6 mmol) and potassium carbonate (1.0 g, 2.8 Eq, 7.2 mmol) in DMF (10 mL) was added iodomethane (1.0 g, 0.45 mL, 2.7 Eq, 7.2 mmol). The mixture was stirred at rt for 5 h. The mixture was diluted with water (30 mL). The solid was collected by filtration, rinsed with water (3x5 mL) and dried (0.90 g). Purification by column chromatography using a gradient of 0-75% THF in [1:1 DCM / isohexane] gave 8-bromo-7-methoxy-2-phenyl-4H-chromen-4-one (0.74 g, 2.2 mmol, 85%) as a white solid.

[0079] 1H NMR (500 MHz, DMSO): 5 8.17-8.11 (m, 2H), 8.04 (d, J= 8.9 Hz, 1H), 7.66-7.57 (m, 3H), 7.34 (d, J = 8.9 Hz, 1H), 7.07 (s, 1H), 4.03 (s, 3H).

[0080] MS: The product was analysed by UPLC (Method 1): m / z 331 / 333 [M+H]+(ES+); no ionisation ES-, at 1.58 min, >99% purity 210-400 nm. 7-Methoxy-8-(1-methyl-1H-pyrazol-5-yl)-2-phenyl-4H-chromen-4-one (1)

[0081] 8-Bromo-7-methoxy-2-phenyl-4H-chromen-4-one (300 mg, 1 Eq, 905 pmol) was coupled with 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (290 mg, 1.53 Eq, 1.39 mmol) according to General Procedure 1 (with the exception that the reaction was performed at 80 °C for 1 h). Purification by column chromatography using a gradient of 0- 100% EtOAc in isohexane gave 7-methoxy-8-(1 -methyl- 1H-pyrazol-5-yl)-2-phenyl-4H-chromen-4-one (266 mg, 0.79 mmol, 88%) as a white solid.

[0082] 1H NMR (500 MHz, DMSO): 6 8.19 (d, J = 9.0 Hz, 1H), 7.79-7.73 (m, 2H), 7.63 (d, J = 1.8 Hz, 1H), 7.58-7.52 (m, 1H), 7.52 - 7.45 (m, 2H), 7.42 (d, J =9.1 Hz, 1H), 7.03 (s, 1H), 6.52 (d, J= 1.8 Hz, 1H), 3.95 (s, 3H), 3.62 (s, 3H).

[0083] MS: The product was analysed by UPLC (Method 1): m / z333[M+H]+(ES+); no ionisation ES-, at 1.22 min, >99% purity 210-400nm. Compound 2 8-Bromo-6-chloro-7-hydroxv-2-phenvl-4H-chromen-4-one

[0084] A suspension of 8-bromo-7-hydroxy-2-phenyl-4H-chromen-4-one (contaminated with dibromo derivative) (0.92 g, 86% Wt, 1 Eq, 2.5 mmol) and NCS (0.50 g, 1.5 Eq, 3.7 mmol) in DMF (9 mL) was stirred at 60 °C for 90 min. The mixture was cooled to rt and diluted with TBME (20 mL). The solid was collected by filtration, rinsed with TBME (3x3 mL) and dried to give 8-bromo-6-chloro-7-hydroxy-2-phenyl-4H-chromen-4-one (0.58 g, 1.3 mmol, 52%) as a white solid.

[0085] 1H NMR (500 MHz, DMSO): 6 8.11 (dd, J= 7.7, 2.0 Hz, 2H), 8.08 (s, 1H), 7.64-7.58 (m, 3H), 7.09 (s, 1H).

[0086] MS: The product was analysed by LCMS (Method 1): m / z 351 / 353 / 355 [M+H]+ (ES+); 349 / 351 / 353 [M-H]' (ES-), at 1.51 min, 78% purity 254 nm 8-Bromo-6-chloro-7-methoxv-2-phenyl-4H-chromen-4-one

[0087] To a suspension of 8-bromo-6-chloro-7-hydroxy-2-phenyl-4H-chromen-4-one (0.57 g, 78% Wt, 1 Eq, 1.3 mmol) and potassium carbonate (0.60 g, 3.4 Eq, 4.3 mmol) in DMF (8 mL) was added iodomethane (0.57 g, 0.25 mL, 3.2 Eq, 4.0 mmol). The mixture was stirred at rt overnight. The mixture was diluted with water (25 mL) and stirred at rt for 10 min. The solid was collected by filtration, rinsed with water (3x2 mL) and dried (0.57 g). The crude product was purified by column chromatography using a gradient of 0-30% THF / [1:1 DCM / isohexane]) to give 8-bromo-6-chloro-7-methoxy-2-phenyl-4H-chromen-4-one (0.50 g, 1.1 mmol, 90%) as a white solid.

[0088] 1H NMR (500 MHz, DMSO): 5 8.17 (s, 1H), 8.14-8.10 (m, 2H), 7.67-7.59 (m, 3H), 7.17 (s, 1H), 3.97 (s, 3H).

[0089] MS: The product was analysed by LCMS (Method 1): m / z 365 / 367 / 369 [M+H]+(ES+); no ionisiation ES-, at 1.83 min, 87% purity at 254 nm. 6-Chloro-7-methoxy-2-phenyl-8-(1H-pyrazol-5-yl)-4H-chromen-4-one (2)

[0090] 8-Bromo-6-chloro-7-methoxy-2-phenyl-4H-chromen-4-one (140 mg, 1 Eq, 383 pmol) was coupled with 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (200 mg, 2.69 Eq, 1.03 mmol) according to General Procedure 1. Purification by column chromatography using a gradient of 0-100% [10% MeOH in THF] in isohexane followed by trituration with MeCN (2 mL) gave 6-chloro-7-methoxy-2-phenyl-8-(1H-pyrazol-5-yl)-4H-chromen-4-one (34 mg, 94 pmol, 25%, 98% Purity) as a white solid.

[0091] 1H NMR (500 MHz, DMSO) 5 13.21 (s, 1H), 8.52 (s, 1H), 8.18-8.12 (m, 2H), 7.90 (s, 1H), 7.68 - 7.59 (m, 3H), 7.14 (s, 1H), 6.82 (d, J = 2.3 Hz, 1H), 3.86 (s, 3H).

[0092] MS: The product was analysed by LCMS (Method 1): m / z 353 / 355 [M+H]+(ES+); 351 / 353 [M-H]- (ES-), at 1.53 min, 98% purity at 254 nm. Compound 3 6-Chloro-7-methoxv-8-(1-methvl-1H-pyrazol-5-vl)-2-phenyl-4H-chromen-4-one (3)

[0093] 8-Bromo-6-chloro-7-methoxy-2-phenyl-4H-chromen-4-one (140 mg, 1 Eq, 383 pmol) was coupled with 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (200 mg, 2.51 Eq, 961 pmol) according to General Procedure 1. Purification by column chromatography using a gradient of 0-100% [10% MeOH / EtOAc] in isohexane gave the desired product with one major impurity. Trituration with 25% TBME / isohexane (2 mL) to remove the contaminant gave 6-chloro-7-methoxy-8-(1-methyl-1 H-pyrazol-5-yl)-2-phenyl-4H-chromen-4-one (63 mg, 0.17 mmol, 44%) as a white solid.

[0094] 1H NMR (400 MHz, DMSO): 6 8.19 - 8.11 (m, 2H), 7.89 (s, 1H), 7.69-7.60 (m, 3H), 7.57 (d, J= 1.9 Hz, 1H), 7.20 (s, 1H), 6.50 (d, J = 1.9 Hz, 1H), 3.71 (s, 3H), 3.61 (s, 3H).

[0095] MS: The product was analysed by LCMS (Method 1): m / z 367 / 369 [M+H]+(ES+); No ionisation ES-, at 1.60 min, 98% purity at 254 nm. Compound 4 7-(Benzvloxy)-8-bromo-2-phenyl-4H-chromen-4-one

[0096] To a suspension of 8-bromo-7-hydroxy-2-phenyl-4H-chromen-4-one (1.0 g, 83% Wt, 1 Eq, 2.6 mmol) and potassium carbonate (1.0 g, 2.8 Eq, 7.2 mmol) in DMF (10 mL) was added benzyl bromide (0.72 g, 0.50 mL, 1.6 Eq, 4.2 mmol). The mixture was stirred at rt overnight.

[0097] The mixture was diluted with water (30 mL). The solid was collected by filtration, rinsed with water (3x5 mL) and dried. Purification by column chromatography using a gradient of 0-25% THF in [50% DCM / isohexane] gave 7-(benzyloxy)-8-bromo-2-phenyl-4H-chromen-4-one (0.91 g, 2.2 mmol, 83%) as a white solid.

[0098] 1H NMR (500 MHz, DMSO): 6 8.17-8.11 (m, 2H), 8.03 (d, J= 8.9 Hz, 1H), 7.66-7.56 (m, 3H), 7.56 - 7.48 (m, 2H), 7.47 - 7.40 (m, 3H), 7.40 - 7.34 (m, 1H), 7.08 (s, 1H), 5.41 (s, 2H).

[0099] MS: The product was analysed by LCMS (Method 1): m / z 407 / 409 [M+H]+ (ES+); no ionisation ES-, at 1.94 min, 97% purity at 254 nm. 7-(Benzyloxy)-2-phenyl-8-( 1 H-pyrazol-5-yl)-4H-chromen-4-one

[0100] 7-(Benzyloxy)-8-bromo-2-phenyl-4H-chromen-4-one (150 mg, 1 Eq, 368 pmol) was coupled 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (200 mg, 2.80 Eq, 1.03 mmol) according to General Procedure 1. Purification by column chromatography using a gradient of 0-100% [10% MeOH / EtOAc] in isohexane gave 7-(benzyloxy)-2-phenyl-8-(1H-pyrazol-5-yl)-4H-chromen-4-one (134 mg, 0.33 mmol, 90%) as a white solid.

[0101] 1H NMR (500 MHz, DMSO): 6 13.16 (s, 1H), 8.07 (br-s, 1H), 7.97 (br-s, 1H), 7.94-7.84 (br-m, 1,5H), 7.68 (br-s, 0.5H), 7.58 - 7.53 (m, 1H), 7.53 - 7.47 (m, 2H), 7.47 - 7.40 (m, 3H), 7.40 - 7.35 (m, 2H), 7.35 - 7.28 (m, 1H), 7.02 (s, 1H), 6.61 (s, 1H), 5.34 (s, 2H). Fractional integrals due to pyrazole tautomers.

[0102] MS: The product was analysed by LCMS (Method 1): m / z 395 [M+H]+(ES+); 393 [M-H]' (ES-), at 1.49 min, >98% purity at 254 nm. 7-hydroxy-2-phenyl-8-(1 H-pyrazol-5-yl)-4H-chromen-4-one (4)

[0103] To a solution of 7-(benzyloxy)-2-phenyl-8-(1 H-pyrazol-5-yl)-4H-chromen-4-one (50 mg, 1 Eq, 0.13 mmol) in EtOH (2 mL) and THF (2 mL) was added 5% palladium on carbon (wetted with 50% water, type 87L) (10 mg, 2.5% Wt, 0.019 Eq, 2.3 pmol). The mixture was stirred at rt under 5 bar H2 in a steel pressure reactor for 1 h. Further 5% palladium on carbon (wetted with 50% water, type 87L) (20 mg, 2.5% Wt, 0.037 Eq, 4.7 pmol) was added and the mixture was stirred at 35 °C under 5 bar H2 in a steel pressure reactor for 2 h. The mixture was filtered (Whatman GF / F glass microfibre pad) and the pad was rinsed with MeOH (50 mL). The filtrate was concentrated and the residue was triturated with MeCN (3 mL) and dried. This material was dissolved in 1.5 mLwith 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 C18 ODB prep column, 130A, 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, 2% MeCN; 0.5-10.5 min, ramped from 2% MeCN to 25% MeCN; 10.5-10.6 min, ramped from 25% MeCN to 100% 37 MeCN; 10.6-12.5 min, held at 100% MeCN. The clean fractions were evaporated in a Genevac to give 7-hydroxy-2-phenyl-8-(1H-pyrazol-5-yl)-4H-chromen-4-one (6 mg, 0.02 mmol, 20%) as a white solid.

[0104] 1H NMR (400 MHz, DMSO): 6 13.51 (s, 1H), 12.57 (s, 1H), 8.07-7.95 (m, 3H), 7.94 (d, J = 8.8 Hz, 1H), 7.65 - 7.53 (m, 3H), 7.11 (d, J = 8.8 Hz, 1H), 6.98 - 6.85 (m, 2H).

[0105] MS: The product was analysed by LCMS (Method 1): m / z 305 [M+H]+ (ES+); 303 [M-H]' (ES-), at 1.30 min, 98% purity at 254 nm. Compound 5 7-Hvdroxv-8-(1-methvl-1H-pyrazol-5-vl)-2-phenvl-4H-chromen-4-one (5)

[0106] A N2-purged flask containing 7-methoxy-8-(1-methyl-1H-pyrazol-5-yl)-2-phenyl-4H-chromen-4-one (0.15 g, 1 Eq, 0.45 mmol) and pyridine hydrochloride (0.60 g, 12 Eq, 5.2 mmol) was stirred at 180 °C for 1 h. The mixture was cooled to rt, then diluted with water (10 mL) and sonicated until a fine suspension was obtained. The solid was collected by filtration, rinsed with water (3x1 mL) and MeCN (3x1 mL), then dried (98 mg, 82% LCMS). Purification by column chromatography using a gradient of 0-100% [10% MeOH / THF] in isohexane) gave the product (89 mg), which was triturated with MeCN (2 mL) to give 7-hydroxy-8-(1-methyl-1H-pyrazol-5-yl)-2-phenyl-4H-chromen-4-one (57 mg, 0.17 mmol, 37%, 94% Purity) as an off-white solid. This material was dissolved in 1.2 mL with 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 C18 ODB prep column, 130A, 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 to give 7-hydroxy-8-(1-methyl-1H-pyrazol-5-yl)-2-phenyl-4H-chromen-4-one (14 mg, 44 pmol, 9.6%) as a white solid.

[0107] 1H NMR (500 MHz, DMSO): 6 7.99 (d, J = 8.8 Hz, 1H), 7.78-7.72 (m, 2H), 7.61 (d, J = 1.9 Hz, 1H), 7.56-7.51 (m, 1H), 7.51 -7.44 (m, 2H), 7.11 (d, J= 8.8 Hz, 1H), 6.96 (s, 1H), 6.49 (d, J= 1.9 Hz, 1H), 3.65 (s, 3H).

[0108] MS: The product was analysed by UPLC (Method 1): m / z319[M+H]+ (ES+); 317 [M-H]~ (ES-), at 0.62 min, 98% purity at 210-400 nm. Compound 6 6-Chloro-7-hvdroxv-2-phenvl-8-(1H-pyrazol-5-vl)-4H-chromen-4-one (6) o

[0109] 6-Chloro-7-methoxy-2-phenyl-8-(1H-pyrazol-5-yl)-4H-chromen-4-one (24 mg, 1 Eq, 68 pmol) was demethylated according to General Procedure 3 to give 6-chloro-7-hydroxy-2-phenyl-8-(1 H-pyrazol-5-yl)-4H-chromen-4-one (20 mg, 56 pmol, 82%) as a white solid.

[0110] 1H NMR (500 MHz, DMSO): 6 13.59 (s, 1H), 13.03 (s, 1H), 8.36 (s, 1H), 8.17-8.10 (m, 2H), 8.07 (s, 1H), 7.67-7.58 (m, 3H), 7.17 (d, J= 2.5 Hz, 1H), 7.10 (s, 1H).

[0111] MS: The product was analysed by UPLC (Method 1): m / z 339 / 341 [M+H]+ (ES+); 337 / 339 [M-H]'(ES-), at 0.75 min, 95% purity 254 nm. Compound 7 6-Chloro-7-hvdroxv-8-(1-methvl-1H-pyrazol-5-vl)-2-phenvl-4H-chromen-4-one (7)

[0112] 6-Chloro-7-methoxy-8-(1-methyl-1H-pyrazol-5-yl)-2-phenyl-4H-chromen-4-one (48 mg, 1 Eq, 0.13 mmol) was demethylated according to General Procedure 3. The crude product was further purified by General procedure 4, Method 1. The clean fractions were evaporated in a Genevac to give 6-chloro-7-hydroxy-8-(1-methyl-1H-pyrazol-5-yl)-2-phenyl-4H-chromen-4-one (7 mg, 0.02 mmol, 20%) as a white solid.

[0113] 1H NMR (500 MHz, DMSO): 6 8.13 - 8.06 (m, 2H), 7.63 (s, 1H), 7.62-7.58 (m, 3H), 7.43 (s, 1H), 6.90 (s, 1H), 6.26 (s, 1H), 3.68 (s, 3H).

[0114] MS: The product was analysed by UPLC (Method 1): m / z 353 / 355 [M+H]+ (ES+); 351 / 353 [M-H]' (ES-), at 0.76 min, >98% purity 210-400 nm. Compound 8 8-Bromo-7-hydroxy-4H-chromen-4-one

[0115] To a solution of 7-hydroxy-4H-chromen-4-one (1.50 g, 1 Eq, 9.25 mmol) in DMF (15 mL) in an isopropanol / dry ice bath at -50 °C was added a solution of NBS (1.65 g, 1 Eq, 9.25 mmol) in DMF (3.5 mL) over ca 30 s. The mixture was allowed to warm to 0 °C over a period of 2 h. The mixture was diluted with water (50 mL) and stirred at rt for 5 min. The solid was collected by filtration, rinsed with water (3x10 mL) and dried in vacuo to give 8-bromo-7-hydroxy-4H-chromen-4-one (1.67 g, 5.2 mmol, 56%) as an off-white solid with minor impurities.

[0116] 1H NMR (500 MHz, DMSO): 5 11.62 (s, 1H), 8.30 (d, J = 6.0 Hz, 1H), 7.88 (d, J = 8.8 Hz, 1H), 7.09 (d, J = 8.8 Hz, 1H), 6.31 (d, J= 6.0 Hz, 1H).

[0117] MS: The product was analysed by UPLC (Method 1): m / z 241 / 243 [M+H]+ (ES+); 239 / 241 [M-H]- (ES-), at 0.31 min, 82% purity 210-400nm. 7-(Benzvloxy)-8-bromo-4H-chromen-4-one

[0118] To a suspension of 8-bromo-7-hydroxy-4H-chromen-4-one (1.67 g, 75% Wt, 1 Eq, 5.20 mmol) and potassium carbonate (2.00 g, 2.79 Eq, 14.5 mmol) in DMF (17 mL) was added benzyl bromide (1.73 g, 1.20 mL, 1.94 Eq, 10.1 mmol). The mixture was stirred at 60 °C for 2 h. The mixture was diluted with EtOAc (60 mL), washed with 1:1 water / brine (2 x 50 mL) and brine (30 mL), dried over MgSO4, filtered and concentrated. Purification by column chromatography using a gradient of 0-100% EtOAc in isohexane gave 7-(benzyloxy)-8-bromo-4H-chromen-4-one (1.65 g, 4.9 mmol, 94%) as a tan solid.

[0119] 1H NMR (500 MHz, DMSO): 5 8.35 (d, J = 6.0 Hz, 1H), 8.02 (d, J= 8.9 Hz, 1H), 7.54-7.48 (m, 2H), 7.46 - 7.39 (m, 3H), 7.39 - 7.32 (m, 1H), 6.35 (d, J = 6.0 Hz, 1H), 5.40 (s, 2H).

[0120] MS: The product was analysed by UPLC (Method 1): m / z 331 / 333 [M+H]+ (ES+); no ionisation ES-, at 1.53 min, >98% purity at 254 nm. 7-(Benzyloxy)-8-bromo-2-(1H-1,2,4-triazol-1-yl)-4H-chromen-4-one

[0121] A vial containing 7-(benzyloxy)-8-bromo-4H-chromen-4-one (1.4 g, 1 Eq, 4.2 mmol), 1,2,4-triazole (0.85 g, 2.9 Eq, 12 mmol), iodine (1.6 g, 1.5 Eq, 6.3 mmol) and potassium carbonate (2.9 g, 5.0 Eq, 21 mmol) was charged with DMF (20 mL). The mixture was stirred at 80 °C for 2 h. The mixture was cooled to rt, diluted with 10% Na2S2O3(aq) (20 mL) and water (40 mL), then stirred at rt for 10 min. The solid was collected by filtration, rinsed with water (3x5 mL) and dried to give 7-(benzyloxy)-8-bromo-2-(1H-1,2,4-triazol-1-yl)-4H-chromen-4-one (0.85 g, 2.1 mmol, 49%) as a pale yellow solid.

[0122] 1H NMR (500 MHz, DMSO): 6 9.24 (s, 1H), 8.49 (s, 1H), 8.06 (d, J = 9.0 Hz, 1H), 7.52 (d, J = 7.4 Hz, 2H), 7.48 (d, J = 9.0 Hz, 1H), 7.44 (t, J = 7.5 Hz, 2H), 7.37 (t, J = 7.3 Hz, 1H), 6.72 (s, 1H), 5.44 (s, 2H).

[0123] MS: The product was analysed by LCMS (Method 1): m / z 398 / 400 [M+H]+(ES+); no ionisation ES-, at 1.47 min, >98% purity 254 nm. 7-(benzvloxv)-8-(1-(tetrahydro-2H-pyran-2-vl)-1H-pyrazol-5-vl)-2-(1H-1,2,4-triazol-1-yl)-4H-chromen-4-one

[0124] A nitrogen-purged vial containing 1-(tetrahydro-2H-pyran-2-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (1.8 g, 3 Eq, 6.4 mmol), 7-(benzyloxy)-8-bromo-2-(1H- 1,2,4-triazol-1-yl)-4H-chromen-4-one (0.85 g, 1 Eq, 2.1 mmol) and Pd(dtbpf)CI2 (Pd-118) (70 mg, 0.05 Eq, 0.11 mmol) was charged with 1,4-dioxane (15 mL) and 1.5 M aq. tripotassium phosphate (1.59 g, 5.00 mL, 1.50 molar, 3.5 Eq, 7.50 mmol). The mixture was stirred at 95 °C for 2 h. The mixture was diluted with EtOAc (20 mL), washed with brine (10 mL), dried over MgSO4, filtered and concentrated. Purification by column chromatography using a gradient of 0-100% EtOAc in isohexane gave 7-(benzyloxy)-8-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-5-yl)-2-(1H-1,2,4-triazol-1-yl)-4H-chromen-4-one (0.72 g, 1.5 mmol, 71%) as a white solid.

[0125] 1H NMR (500 MHz, DMSO): 5 8.46 (s, 0.7H), 8.43 (s, 0.7H), 8.41 (s, 0.3H), 8.36 (s, 0.3H), 8.19 (d, J= 8.9 Hz, 0.7H), 8.14 (d, J= 9.0 Hz, 0.3H), 7.74 (d, J= 1.7 Hz, 0.7H), 7.73 (d, J= 1.8 Hz, 0.3H), 7.57 (d, J= 9.0 Hz, 0.7H), 7.44 (d, J= 9.1 Hz, 0.3H), 7.40-7.25 (m, 5H), 6.67 (s, 0.3H), 6.63 (s, 0.7H), 6.60 (d, J= 1.7 Hz, 0.7H), 6.49 (d, J= 1.7 Hz, 0.3H), 5.42-5.35 (m, 1H), 5.35 - 5.29 (m, 1H), 5.15 (dd, J= 9.3, 2.7 Hz, 0.3H), 5.06 (dd, J = 9.5, 2.6 Hz, 0.7H), 3.77 - 3.71 (m, 0.3H), 3.55 - 3.48 (m, 0.7H), 3.37 - 3.32 (m, 0.3H), 3.20 (td, J = 10.9, 3.0 Hz, 0.7H), 2.28-2.14 (m, 1H), 1.90- 1.78 (m, 1H), 1.75- 1.68 (m, 0.3H), 1.61 -1.54 (m, 0.7H), 1.54 - 1.43 (m, 1H), 1.43 - 1.30 (m, 2H).

[0126] MS: The product was analysed by LCMS (Method 1): m / z 386 [M-THP+2H]+ (ES+); no ionisation ES-, at 1.39 min, >99% purity at 254 nm. 7-(Benzvloxv)-2-(2,2-dimethvl-1-oxa-8-azaspiror4.51decan-8-vl)-8-(1-(tetrahvdro-2H-pvran-2-vD-1H-pyrazol-5-yl)-4H-chromen-4-one

[0127] The title compound was prepared according to General Procedure 2 with 7-(benzyloxy)-8-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-5-yl)-2-(1H-1,2,4-triazol-1-yl)-4H-chromen-4-one (100 mg, 1 Eq, 213 pmol), 2,2-dimethyl-1-oxa-8-azaspiro[4.5]decane hydrochloride (88 mg, 2.0 Eq, 0.43 mmol), potassium carbonate (120 mg, 4.08 Eq, 868 pmol) and NMP (1.0 mL) for 1.5 h. The crude product was purified by column chromatography using a gradient of 0-20% MeOH in EtOAc giving the title compound, 7-(Benzyloxy)-2-(2,2-dimethyl-1-oxa-8-azaspiro[4.5]decan-8-yl)-8-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-5-yl)-4H-chromen-4-one (102 mg, 0.18 mmol, 82%) as a colourless glassy foam.

[0128] 1H NMR (500 MHz, DMSO): 5 7.98 (d, J = 8.8 Hz, 0.7H), 7.94 (d, J = 8.9 Hz, 0.3H), 7.66 (d, J = 1.7 Hz, 0.3H), 7.63 (d, J = 1.7 Hz, 0.7H), 7.38 - 7.24 (m, 5.7H), 7.21 (d, J = 8.9 Hz, 0.3H), 6.42 (d, J = 1.7 Hz, 0.7H), 6.39 (d, J = 1.7 Hz, 0.3H), 5.42 (s, 0.7H), 5.42 (s, 0.3H), 5.32 - 5.25 (m, 1H), 5.25-5.18 (m, 1H), 5.03-4.96 (m, 1H), 3.80 - 3.72 (m, 0.3H), 3.70-3.60 (m, 0.7H), 3.39 - 3.32 (m, 1.7H), 3.29 - 3.09 (m, 3.3H), 2.29 - 2.13 (m, 1H), 1.94 - 1.87 (m, 2H), 1.84- 1.73 (m, 3.7H), 1.72- 1.65 (m, 0.3H), 1.61 - 1.36 (m, 7H), 1.19-1.11 (m, 6H).

[0129] MS: The product was analysed by LCMS (Method 1): m / z 570 [M+H]+ (ES+); no ionisation ES-, at 1.76 min, >99% purity at 254 nm. 2-(2,2-Dimethyl-1-oxa-8-azaspiror4.51decan-8-yl)-7-hydroxy-8-(1H-pyrazol-5-yl)-4H-chromen-4-one hydrochloride (8)

[0130] A 2 M HCI solution in MeOH was generated by dropwise addition of acetyl chloride (0.11 g, 0.10 mL, 1.4 mmol) to cold MeOH (0.60 mL). This solution was added to a vial containing 2-(2,2-Dimethyl-1-oxa-8-azaspiro[4.5]decan-8-yl)-7-hydroxy-8-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-5-yl)-4H-chromen-4-one (35 mg, 1 Eq, 73 pmol) at rt. The reaction mixture was stirred at rt for 90 min before being diluted with TBME (2 mL) and stirred at rt for 30 min, the solid was collected by filtration, rinsed with TBME (1 mL) and dried to give 2-(2,2-dimethyl-1-oxa-8-azaspiro[4.5]decan-8-yl)-7-hydroxy-8-(1 H-pyrazol-5-yl)-4H-chromen-4-one as the hydrochloride salt (18 mg, 41 pmol, 57%) as a white solid.

[0131] 1H NMR (500 MHz, DMSO): 5 7.91 (d, J = 2.2 Hz, 1H), 7.87 (d, J= 8.8 Hz, 1H), 7.21 (d, J = 8.8 Hz, 1H), 6.72 (d, J = 2.2 Hz, 1H), 6.28 (s, 1H), 3.89 - 3.78 (m, 2H), 3.57 - 3.48 (m, 2H), 1.90 - 1.84 (m, 2H), 1.84 - 1.79 (m, 2H), 1.77 - 1.63 (m, 4H), 1.20 (s, 6H). Note 3x exchangeable H not resolved.

[0132] MS: The product was analysed by LCMS (Method 1): m / z 396 [M+H]* (ES+); 394 [M-H]~ (ES-), at 1.20 min, >99% purity at 254 nm. Compound 9 7-Hydroxy-8-nitro-2-phenyl-4H-chromen-4-one

[0133] Method A: To a suspension of 7-hydroxy-2-phenyl-4H-chromen-4-one (2.0 g, 1 Eq, 8.4 mmol) in acetic acid (100 mL) was added nitric acid (70% aq.) (5.6 g, 4.0 mL, 70% Wt, 7.5 Eq, 63 mmol). The mixture was stirred at rt for 2 h, then at 40 °C for 1 h, then at 45 °C overnight, then at 50 °C for 90 min. The mixture was stirred at 60 °C for 90 min, UPLC showed 2% conv. to the desired product. Approximately 15-20 mins after this (no changes were made to the conditions), the mixture abruptly became homogeneous and red. The mixture was cooled to rt and poured into water (200 mL) and left to stand at rt for 30 min. The solid was collected by filtration, rinsed with water (3x10 mL) and dried to give 7-hydroxy-8-nitro-2-phenyl-4H-chromen-4-one (1.93 g, 4.8 mmol, 57%)

[0134] 1H NMR (500 MHz, DMSO) 6 12.57 (s, 1H), 8.07 (d, J= 8.9 Hz, 1H), 7.95-7.90 (m, 2H), 7.65-7.52 (m, 3H), 7.19 (d, J = 9.0 Hz, 1H), 7.08 (s, 1H).

[0135] MS: The product was analysed by UPLC (Method 1): m / z284[M+H]+ (ES+); 282 [M-H]' (ES-), at 0.66 min, 72% purity 254 nm.

[0136] Method B: To a solution of 7-hydroxy-2-phenyl-4H-chromen-4-one (1.0 g, 1 Eq, 4.2 mmol) in DMSO (10 mL) at 40 °C was added nitric acid (70% aq.) (2.8 g, 2.0 mL, 70% Wt, 7.5 Eq, 31 mmol). The mixture was stirred at 90 °C for 1 h, then at 95 °C for 50 min. The mixture was cooled to rt, diluted with water (100 mL) and left to stand at rt for 30 min. The solid was collected by filtration, rinsed with water (3x5 mL) and MeCN (5x5 mL) and dried to give 7-hydroxy-8-nitro-2-phenyl-4H-chromen-4-one (0.82 g, 2.6 mmol, 62%) as a yellow solid.

[0137] 1H NMR (500 MHz, DMSO): 6 12.57 (s, 1H), 8.07 (d, J =9.0 Hz, 1H), 7.95-7.90 (m, 2H), 7.65-7.52 (m, 3H), 7.19 (d, J= 9.0 Hz, 1H), 7.07 (s, 1H).

[0138] MS: The product was analysed by UPLC (Method 1): m / z284[M+H]+ (ES+); 282 [M-H]' (ES-), at 0.69 min, 90% purity 254 nm. 7-(Benzyloxy)-8-nitro-2-phenyl-4H-chromen-4-one

[0139] A suspension of 7-hydroxy-8-nitro-2-phenyl-4H-chromen-4-one (2.0 g, 72% Wt, 1 Eq, 5.1 mmol), potassium carbonate (1.5 g, 2.1 Eq, 11 mmol) and benzyl bromide (1.9 g, 1.3 mL, 2.1 Eq, 11 mmol) in DMF (30 mL) was stirred at rt overnight. The mixture was diluted with EtOAc (100 mL), washed with 1:1 water / brine and brine (50 mL), dried over MgSO4, filtered and concentrated.

[0140] Purification by column chromatography using a gradient of 0-30% EtOAc in a mixture of DCM and isohexane [1:1] gave 7-(benzyloxy)-8-nitro-2-phenyl-4H-chromen-4-one (1.5 g, 3.8 mmol, 74%) as a cream solid.

[0141] 'H NMR (500 MHz, DMSO): 6 8.21 (d, J= 9.2 Hz, 1H), 7.95-7.88 (m, 2H), 7.66-7.55 (m, 4H), 7.49-7.41 (m, 4H), 7.40-7.35 (m, 1H), 7.12 (s, 1H), 5.49 (s, 2H).

[0142] MS: The product was analysed by UPLC (Method 1): m / z374[M+H]+ (ES+); No ionisation (ES-), at 1.76 min, 88% purity 210-400nm. 8-Amino-7-hydroxy-2-phenyl-4H-chromen-4-one

[0143] To a solution of 7-(benzyloxy)-8-nitro-2-phenyl-4H-chromen-4-one (1.49 g, 94% Wt, 1 Eq, 3.75 mmol) in a mixture of EtOH (30 mL) and THF (30 mL) was added Pd (type 87L, 5% on C, wetted with 50% water) (0.30 g, 2.5% Wt, 0.019 Eq, 70 pmol). The suspension was stirred in a steel pressure reactor under 5 bar H2 at 25 °C overnight. The mixture was filtered (Whatman GF / F pad), rinsed through with methanol and the filtrate was concentrated. Purification by column chromatography using a gradient of 0-100% of 10% MeOH in THF and isohexane, followed by trituration with TBME (to remove BHT stabiliser from THF) gave 8-amino-7-hydroxy-2-phenyl-4H-chromen-4-one (0.79 g, 3.0 mmol, 79%) as a yellow solid.

[0144] 1H NMR (500 MHz, DMSO): 5 8.26 - 8.20 (m, 2H), 7.63 - 7.52 (m, 3H), 7.20 (d, J = 8.5 Hz, 1H), 6.89 (d, J = 8.5 Hz, 1H), 6.86 (s, 1H).

[0145] MS: The product was analysed by LCMS (Method 1): 3465-83-1-ac, m / z 254 [M+H]+ (ES+); 252 [M-Hp (ES-), at 0.96 min, 93% purity at 254 nm. 8-Phenyl-6H-chromeno[8,7-d1oxazole-2,6(1 H)-dione (9)

[0146] To a suspension of 8-amino-7-hydroxy-2-phenyl-4H-chromen-4-one (150 mg, 1 Eq, 592 pmol) and triethylamine (0.15 g, 0.20 mL, 2.4 Eq, 1.4 mmol) in THF (4.0 mL) was added GDI (110 mg, 1.15 Eq, 678 pmol). The mixture was stirred at rt for 2 h. The mixture was diluted with TBME (4 mL), then the solid was collected by filtration, rinsed with TBME (3 x 1 mL) and dried to give 8-phenyl-6H-chromeno[8,7-d]oxazole-2,6(1H)-dione (104 mg, 0.37 mmol, 62%) as a white solid.

[0147] 1H NMR(500MHz, DMSO): 6 12.92 (s, 1H), 8.31-8.25 (m, 2H), 7.77 (d, J= 8.5 Hz, 1H), 7.67-7.56 (m, 3H), 7.47 (d, J = 8.5 Hz, 1H), 7.11 (s, 1H).

[0148] MS: The product was analysed by LCMS (Method 1): m / z280 (M+H)+ (ES+, weak ionisation); 278 (M-H)- (ES-), at 1.11 min, >99% purity at 254 nm. Compound 10 1-Methvl-8-phenvl-6H-chromenof8,7-d1oxazole-2,6(1 H)-dione (10) O

[0149] To a suspension of 8-phenyl-6H-chromeno[8,7-d]oxazole-2,6(1 H)-dione (40 mg, 1 Eq, 0.14 mmol) and potassium carbonate (60 mg, 3.0 Eq, 0.43 mmol) in DMF (1.0 mL) was added iodomethane (0.1 g, 0.05 mL, 6 Eq, 0.8 mmol). The mixture was stirred at rt for 30 min, then diluted with water (3 mL) and stirred for 5 min. The solid was collected by filtration, rinsed with water (2x2 mL) and MeCN (2x2 mL), then dried to give 1-methyl-8-phenyl-6H-chromeno[8,7-d]oxazole-2,6(1 H)-dione (36 mg, 0.12 mmol, 84%, 98% Purity) as a white solid.

[0150] 1H NMR (500 MHz, DMSO): 5 8.10 - 8.03 (m, 2H), 7.83 (d, J = 8.5 Hz, 1H), 7.68-7.57 (m, 3H), 7.54 (d, J = 8.5 Hz, 1H), 7.08 (s, 1H), 3.80 (s, 3H).

[0151] MS: The product was analysed by LCMS (Method 1): m / z294 (M+H)+ (ES+); no ionisation ES-, at 1.25 min, >98% purity at 254 nm. Compound 11 1-(4-amino-2-hvdroxv-3-nitrophenvl)-3-(4-bromophenyl)prop-2-en-1-one

[0152] A suspension of 1-(4-amino-2-hydroxy-3-nitrophenyl)ethan-1-one (2.50 g, 94% Wt, 1 Eq, 12.0 mmol) and 4-bromobenzaldehyde (3.32 g, 1.5 Eq, 18.0 mmol) in EtOH (40 mL) was heated to 50 °C. Pyrrolidine (953 mg, 1.10 mL, 1.12 Eq, 13.4 mmol) was added in one portion and the reaction mixture was stirred at 50 °C for 4 h, then allowed to cool to rt The precipitate was collected by filtration, washed with EtOH, and dried in vacuo to afford the desired product 1-(4-amino-2-hydroxy-3-nitrophenyl)-3-(4-bromophenyl)prop-2-en-1-one (3.69 g, 10 mmol, 83 %) as a brown free-flowing solid.

[0153] 1H NMR (400 MHz, DMSO): 5 15.81 (s, 1H), 8.20 (d, J = 9.3 Hz, 1H), 7.97 (d, J = 15.5 Hz, 1H), 7.91 -7.82 (m, 2H), 7.78 (d, J = 15.5 Hz, 1H), 7.72-7.64 (m, 2H), 7.56 (s, 2H), 6.42 (d, J = 9.2 Hz, 1H).

[0154] MS: The product was analysed by LCMS (Method 1): m / z 363.0 / 365.0 (M+H)+ (ES+); 361.0 / 363.0 (M-H)’ (ES-), at 1.83 min, 98% purity at 260nm + / - 80nm. 7-amino-2-(4-bromophenyl)-8-nitro-4H-chromen-4-one

[0155] To a stirred suspension of 1-(4-amino-2-hydroxy-3-nitrophenyl)-3-(4-bromophenyl)prop-2-en-1-one (3.69 g, 98% Wt, 1 Eq, 9.96 mmol) in DMSO (4.40 g, 4.00 mL, 5.66 Eq, 56.4 mmol) and toluene (30 mL) was added iodine (591 mg, 0.234 Eq, 2.33 mmol). The reaction mixture was stirred at 110 °C for 4 h and then allowed to cool to rt. The reaction mixture was diluted with water (30 mL) and the precipitate was collected by filtration, washing with EtOH (very slow filtration speed). The solid was sucked dry for 15 min. and then transferred to a flask to dry in vacuo. The desired product 7-amino-2-(4-bromophenyl)-8-nitro-4H-chromen-4-one (3.356 g, 9.1 mmol, 91 %) was afforded as a pale brown solid. The product was used without further purification.

[0156] 1H NMR (400 MHz, DMSO): 5 8.01 - 7.93 (m, 2H), 7.87 (d, J = 9.2 Hz, 1H), 7.84-7.77 (m, 4H), 7.07 (s, 1H), 6.99 (d, J = 9.1 Hz, 1H).

[0157] MS: The product was analysed by LCMS (Method 1): m / z 361.0 / 363.0 (M+H)+ (ES+); 358.8 / 360.8 (M-H) (ES-), at 1.60 min, 98% purity at 260nm + / - 80nm. 7,8-diamino-2-(4-bromophenyl)-4H-chromen-4-one

[0158] To a stirred suspension of 7-amino-2-(4-bromophenyl)-8-nitro-4H-chromen-4-one (3.35 g, 98% Wt, 1 Eq, 9.09 mmol) in DMSO (35 mL), EtOH (35 mL) and water (10 mL) at 80 °C was added sodium dithionite (5.00 g, 2.00 mL, 3.16 Eq, 28.7 mmol). The reaction mixture was stirred at 80 °C for 6.5 h and then allowed to cool to rt overnight. The reaction mixture was poured into ice / water (~200 mL) and the precipitate was collected by filtration. The filter cake was washed with water and then MeCN. The crude product was transferred to a flask and dried in vacuo to afford 7,8-diamino-2-(4-bromophenyl)-4H-chromen-4-one (2.95 g, 8.7 mmol, 96 %) as a pale brown solid.

[0159] 1H NMR (400 MHz, DMSO) 6 8.21 - 8.11 (m, 2H), 7.77 - 7.69 (m, 2H), 7.16 (d, J = 8.4 Hz, 1H), 6.81 (s, 1H), 6.69 (d, J = 8.4 Hz, 1H), 5.60 (br s, 2H), 4.95 (br s, 2H).

[0160] MS: The product was analysed by LCMS (Method 1): m / z 331.0 / 333.0 (M+H)+ (ES+); 375.0 / 377.0 (M+formic acid-H)_ (ES-), at 1.27 min, 98% purity at 260nm + / - 80nm. 8-(4-bromophenyl)-2-(trifluoromethyl)chromenof7,8-d1imidazol-6(1H)-one

[0161] To a stirred suspension of 7,8-diamino-2-(4-bromophenyl)-4H-chromen-4-one (1.028 g, 98% Wt, 1 Eq, 3.042 mmol) in Pyridine (10.0 mL) at 0 °C was added trifluoroacetic anhydride (1.93 g, 1.30 mL, 3.03 Eq, 9.20 mmol) dropwise. The reaction mixture was allowed to warm to rt and stirred for 2 h. The reaction mixture was concentrated in vacuo and the residue was triturated with water. The resultant precipitate was collected by filtration, washing with water, and then dried in vacuo to afford 8-(4-bromophenyl)-2-(trifluoromethyl)chromeno[7,8-d]imidazol-6(1H)-one (1.246 g, 2.8 mmol, 93 %) as a pale brown solid.

[0162] 1H NMR (400 MHz, MeOD): 5 8.18 - 8.14 (m, 2H), 8.12 (d, J = 8.8 Hz, 1H), 7.75 (d, J = 8.7 Hz, 2H), 7.71 (d, J = 8.8 Hz, 1H), 7.04 (s, 1H). 1 proton not observed in MeOD.

[0163] 19F NMR (376 MHz, MeOD) 5 -65.66.

[0164] MS: The product was analysed by LCMS (Method 1): m / z 409.0 / 411.0 (M+H)+ (ES+); 406.8 / 408.8 (M-H)‘ (ES-), at 1.56 min, 98% purity at 260nm +1- 80nm. 8-(4-(pvrrolidin-1-vl)phenvl)-2-(trifluoromethvl)chromenor7,8-d1imidazol-6(1H)-one (11) 0

[0165] A suspension of 8-(4-bromophenyl)-2-(trifluoromethyl)chromeno[7,8-d]imidazol-6(1H)-one (60.0 mg, 98% Wt, 1 Eq, 144 pmol), Pd-177 (12.0 mg, 0.110 Eq, 15.8 pmol), cesium carbonate (143 mg, 3.05 Eq, 439 pmol) and pyrrolidine (17.3 mg, 20.0 pL, 1.69 Eq, 244 pmol) in DMF (1.00 mL) was heated in the microwave at 100 °C for 30 min. The reaction mixture was filtered through cotton wool, washed with DCM, and then concentrated in vacuo. The residue was azeotroped with toluene (3 times) to afford the crude product. The crude product was purified by column chromatography using a greadien of 0-5% MeOH in DCM giving 8-(4-(pyrrolidin-1-yl)phenyl)-2-(trifluoromethyl)chromeno[7,8-d]imidazol-6(1H)-one (28.0 mg, 69 pmol, 48 %) as a yellow solid.

[0166] 1H NMR (400 MHz, DMSO): 6 14.76 (br s, 1H), 8.08 - 7.95 (m, 2H), 7.98 (d, J = 8.8 Hz, 1H), 7.75 - 7.64 (m, 1H), 6.89 (s, 1H), 6.75 - 6.68 (m, 2H), 3.39 - 3.33 (m, 4H), 2.04-1.96 (m, 4H).

[0167] 19F NMR (376 MHz, DMSO): 5 -62.53.

[0168] MS: The product was analysed by LCMS (Method 1): m / z 400.0 (M+H)+ (ES+); 398.0 (M-Hy (ES-), at 1.72 min, 98% purity at 260nm + / - 80nm. Compound 12 8-(4-(piperidin-1-yl)phenvl)-2-(trifluoromethyl)chromenor7,8-d1imidazol-6(1H)-one (12) O

[0169] A suspension of 8-(4-bromophenyl)-2-(trifluoromethyl)chromeno[7,8-d]imidazol-6(1H)-one (60.0 mg, 98% Wt, 1 Eq, 144 pmol), piperidine (21.6 mg, 25.0 pL, 1.76 Eq, 253 pmol), cesium carbonate (143 mg, 3.05 Eq, 439 pmol) and Pd-177 (12.0 mg, 0.110 Eq, 15.8 pmol) in DMF (1.00 mL) was heated in the microwave at 100 °C for 3 h. The reaction mixture was filtered through cotton wool, washing with DCM, and the filtrate was concentrated in vacuo. The residue was azeotroped with toluene (3 times) to afford the crude product.

[0170] The crude product was purified by column chromatography on silica gel (dry load) (12 g cartridge, 0-5% MeOH / DCM). The product containing fractions were concentrated in vacuo and the residue was triturated with EtOH to afford 8-(4-(piperidin-1-yl)phenyl)-2-(trifluoromethyl)chromeno[7,8-d]imidazol-6(1H)-one (14.0 mg, 32 pmol, 22 %) as a yellow solid.

[0171] 1H NMR (400 MHz, DMSO): 6 14.70 (s, 1H), 8.01-7.97 (m, 3H), 7.76-7.64 (m, 1H), 7.10 (d, J = 8.7 Hz, 2H), 6.93 (s, 1H), 3.44-3.35 (m, 4H), 1.67-1.56 (m, 6H).

[0172] 19F NMR (376 MHz, DMSO): 5 -62.60.

[0173] MS: The product was analysed by LCMS (Method 1): m / z 414.2 (M+H)+ (ES+); 412.0 (M-H)- (ES-), at 1.71 min, 96% purity at 260nm + / - 80nm. Compound 13 8-(4-(azetidin-1-yl)phenyl)-2-(trifluoromethyl)chromenor7,8-d1imidazol-6(1H)-one (13) O

[0174] A suspension of 8-(4-bromophenyl)-2-(trifluoromethyl)chromeno[7,8-d]imidazol-6(1H)-one (60.0 mg, 98% Wt, 1 Eq, 144 pmol), Pd-177 (11.0 mg, 0.101 Eq, 14.4 pmol), cesium carbonate (140 mg, 3.0 Eq, 431 pmol) and azetidine (16.9 mg, 20.0 pL, 2.06 Eq, 297 pmol) in DMF (1.00 mL) was heated in the microwave at 80 °C for 1 h. Additional azetidine (84.5 mg, 100 pL, 10.3 Eq, 1.48 mmol) was added and heated in the microwave at 100 °C for 2 h. Additional azetidine (84.5 mg, 100 pL, 10.3 Eq, 1.48 mmol) and Pd-177 (12 mg, 0.109, 15.7 pmol) was added and heated in the microwave at 100 °C for 2 h. The reaction mixture was concentrated in vacuo to afford the crude product. The crude product was purified by column chromatography, using a gradient of 0-5% MeOH in DCM, to afford impure fractions. The product was further purified by column chromatography using a gradient of 0-2.5% MeOH in DCM to afford 8-(4-(azetidin-1-yl)phenyl)-2-(trifluoromethyl)chromeno[7,8-d]imidazol-6(1H)-one (10.0 mg, 25 pmol, 17 %) as a yellow solid.

[0175] 1H NMR (400 MHz, DMSO): 6 14.72 (brs, 1H), 8.09-7.92 (m, 3H), 7.70 (brs, 1H), 6.91 (s, 1H), 6.58 - 6.52 (m, 2H), 3.97 (t, J = 7.3 Hz, 4H), 2.43 - 2.33 (m, 2H).

[0176] 19F NMR (376 MHz, DMSO): 6 -62.55.

[0177] MS: The product was analysed by LCMS (Method 1): m / z 386.0 (M+H)+ (ES+); 384.0 (M-H)’ (ES-), at 1.53 min, 97% purity at 260nm +1- 80nm. Compound 14 7-amino-8-nitro-4H-chromen-4-one

[0178] Sodium hydride (60 wt% in mineral oil) (767 mg, 60% Wt, 4 Eq, 19.2 mmol) was added portionwise to a solution of 1-(4-amino-2-hydroxy-3-nitrophenyl)ethan-1-one (1.00 g, 94% Wt, 1 Eq, 4.79 mmol) in THE (24.0 mL). The resulting suspension was stirred for 5 min before the dropwise addition of ethyl formate (7.10 g, 7.74 mL, 20 Eq, 95.8 mmol). The reaction was heated to 40 °C for 30 min. The reaction was removed from the heating block and sodium hydride (60 wt% in mineral oil) (767 mg, 60% Wt, 4 Eq, 19.2 mmol) was added portionwise (internal temperature ~30 °C). The reaction was heated to 40 °C, then ethyl formate (3.55 g, 3.87 mL, 10 Eq, 47.9 mmol) was added in one portion. The reaction was stirred at 40 °C for 18 h. The reaction mixture was poured into ice (250 mL) and then acidified to pH 1 by addition of cone. HCI. The aqueous phase was extracted with DCM (2 x 250 mL) and the combined organic extracts were concentrated in vacuo. The residue was diluted with THE (10 mL) then vigorously stirred with 6 M aq. HCI (20 mL) for 3 h. The mixture was diluted with DCM (100 mL) and the aqueous phase was basified to pH 12 with 2 M aq. NaOH. Phases were separated and the organic layer was collected and the aqueous phase further extracted with DCM (100 mL). The combined organic extracts were washed with 1:1 water / brine (50 mL) and dried over MgSO4. The solids were filtered off and the solvent was removed in vacuo to afford 7-amino-8-nitro-4H-chromen-4-one (1.25 g, 4.9 mmol, 100 %) as an orange solid.

[0179] 1H NMR (400 MHz, DMSO): 6 8.19 (d, J = 5.9 Hz, 1H), 7.85 (d, J = 9.1 Hz, 1H), 7.60 (s, 2H), 6.96 (d, J = 9.1 Hz, 1H), 6.30 (d, J = 6.0 Hz, 1H).

[0180] MS: The product was analysed by LCMS (Method 1): m / z 207.0 (M+H)+ (ES+); 205.0 (M-H)' (ES-), at 0.73 min, >99% purity at 260nm + / - 80nm. 7,8-diamino-4H-chromen-4-one

[0181] To a stirred suspension of 7-amino-8-nitro-4H-chromen-4-one (860 mg, 80% Wt, 1 Eq, 3.34 mmol) in DMSO (7.50 mL), EtOH (7.50 mL) and water (1.9 mL) at 80 °C was added sodium dithionite (1.74 g, 697 pL, 3 Eq, 10.0 mmol). The reaction was stirred for 8 h before it was allowed to cool overnight. The reaction was diluted with water (100 mL), neutralised with sat. aq. NaHCOs and the resulting suspension was extracted with EtOAc (4 x 100 mL). The combined organics were washed with 1:1 water / brine (100 mL) and brine (50 mL) then dried over MgSO4. The solids were filtered off and the solvent was removed in vacuo to afford an orange solid, contaminated with DMSO. The remaining aqueous phase was further extracted with CHCI3 / IPA (3 x 100 mL), DCM / MeOH (3 x 100 mL) and EtOAc / MeOH (3 x 100 mL) at various pH. The solvents were removed to yield additional product, which was acidified with AcOH and purified by SCX, eluting with 0.7M NH3 in MeOH to yield 59 mg of product. The remaining aqueous phase was concentrated in vacuo to approx. (10 mL). The resulting solids were washed with MeOH (200 mL) and the volume was reduced in vacuo. The resulting solution was acidified with AcOH and re-purified by SCX, eluting with 0.7M NH3 in MeOH to yield an orange solid. The solids were combined, affording 7,8-diamino-4H-chromen-4-one (514 mg, 2.3 mmol, 70 %, 80% Purity) as an orange solid.

[0182] 1H NMR (400 MHz, DMSO): 6 8.07 (d, J = 5.9 Hz, 1H), 7.13 (d, J = 8.4 Hz, 1H), 6.65 (d, J = 8.5 Hz, 1H), 6.06 (d, J = 5.9 Hz, 1H), 5.56 (s, 2H), 4.68 (s, 2H).

[0183] MS: The product was analysed by LCMS (Method 1): m / z 177.0 (M+H)+ (ES+) at 0.26 min, 93% purity at 254nm. 2-(trifluoromethyl)chromeno[7,8-d1imidazol-6(3H)-one

[0184] Trifluoroacetic acid (5.32 g, 3.60 mL, 20 Eq, 46.7 mmol) was slowly added to 7,8-diamino-4H-chromen-4-one (514 mg, 80% Wt, 1 Eq, 2.33 mmol) while cooling over and ice bath. The mixture was stirred at reflux for 2 h, then concentrated in vacuo. The residue was dissolved in EtOAc (20 mL) and the aqueous phase was neutralised with sat. aq. NaHCO3 while stirring. The phases were separated and the organic phase was washed with brine (10 mL), 52 dried over MgSO4, filtered and concentrated. The crude product was purified by column chromatography using a gradient of 0-10% MeOH in DCM) to afford 2-(trifluoromethyl)chromeno[7,8-d]imidazol-6(3H)-one (85 mg, 0.23 mmol, 10 %) as a red solid.

[0185] 1H NMR (400 MHz, DMSO): 6 14.77 (s, 1H), 8.43 (d, J = 6.0 Hz, 1H), 8.01 (d, J = 8.7 Hz, 1H), 7.72 (d, J = 8.7 Hz, 1H), 6.47 (d, J = 5.9 Hz, 1H).

[0186] MS: The product was analysed by LCMS (Method 1): m / z 255.0 (M+H)+ (ES+); 253.0 (M-H)’ (ES-), at 0.85 min, >99% purity at 260nm + / - 80nm. 8-(1H-1.2.4-triazol-1-vl)-2-(trifluoromethyl)chromeno[7.8-d1imidazol-6(3H)-one

[0187] A stirred suspension of 2-(trifluoromethyl)chromeno[7,8-d]imidazol-6(3H)-one (94 mg, 1 Eq, 0.37 mmol), 1H-1,2,4-triazole (77 mg, 3 Eq, 1.1 mmol), iodine (0.14 g, 1.5 Eq, 0.55 mmol) and potassium carbonate (0.26 g, 5 Eq, 1.9 mmol) in DMF (1 mL) was heated to 60 °C for 30 min. The reaction was heated to 80 °C for 2 h, iodine (94 mg, 1 Eq, 0.37 mmol) was added and the reaction was heated for a further 2 h. 1H-1,2,4-triazole (77 mg, 3 Eq, 1.1 mmol) and potassium carbonate (0.26 g, 5 Eq, 1.9 mmol) were added and the reaction was heated to 80 °C for 6 h. The reaction was allowed to cool, diluted with sat. aq. Na2S2O3 (10 mL) and water (20 mL) and extracted with EtOAc (2 x 40 mL). The combined organics were washed with brine (20 mL) and dried over MgSO4. The solids were filtered off and the solvent was removed in vacuo. The crude product was dried onto celite and purified by column chromatography using a gradient of 0-10% MeOH in DCM to afford 8-(1 H-1,2,4-triazol-1 -yl)-2-(trifluoromethyl)chromeno[7,8-d]imidazol-6(3H)-one (18 mg, 55 pmol, 15 %) as a light yellow solid.

[0188] 1H NMR (400 MHz, DMSO, 363K): 6 9.42 (s, 1H), 8.43 (s, 1H), 8.21 (s, 1H), 8.05 (d, J = 8.6 Hz, 1H), 7.82 (d, J = 8.7 Hz, 1H), 6.81 (s, 1H).

[0189] 19F NMR (376 MHz, DMSO): 6 -62.73.

[0190] MS: The product was analysed by LCMS (Method 1): m / z 322.0 (M+H)+ (ES+); 320.0 (M-H)- (ES-), at 0.93 min, >99% purity at 260nm + / - 80nm. 8-(2,2-dimethvl-1-oxa-8-azaspiror4.51decan-8-vl)-2-(trifluoromethvl)chromenor7,8-d1imidazol-6£3H)-gne........(14).

[0191] The title compound was prepared according to General Procedure 2 using Potassium carbonate (38 mg, 5 Eq, 280 pmol) , 8-(1H-1,2,4-triazol-1-yl)-2-(trifluoromethyl)chromeno[7,8-d]imidazol-6(3H)-one (18 mg, 99% Wt, 1 Eq, 55 pmol), 2,2-dimethyl-1-oxa-8-azaspiro[4.5]decane hydrochloride (23 mg, 2 Eq, 110 pmol) and NMP (0.5 mL). The reaction was heated to 120 °C for 24 h.

[0192] The reaction was allowed to cool and the reaction mixture was purified directly by column chromatography using a gradient of 0-6% MeOH in DCM to afford 8-(2,2-dimethyl-1-oxa-8-azaspiro[4.5]decan-8-yl)-2-(trifluoromethyl)chromeno[7,8-d]imidazol-6(3H)-one (7.9 mg, 18 pmol, 32 %) as a beige solid.

[0193] 1H NMR(400MHz, MeOD): 6 8.03 (d, J = 8.7 Hz, 1H), 7.62 (d, J = 8.7 Hz, 1H), 5.69 (s, 1H), 4.01 - 3.93 (m, 2H), 3.78 - 3.67 (m, 2H), 2.03 - 1.97 (m, 2H), 1.97 - 1.90 (m, 2H), 1.81 (t, J = 5.7 Hz, 4H), 1.30 (s, 6H).

[0194] 19F NMR (376 MHz, MeOD): 5 -65.64.

[0195] MS: The product was analysed by LCMS (Method 1): m / z 422.2 (M+H)+ (ES+); 420.0 (M-HX (ES-), at 1.37 min, >99% purity at 260nm + / - 80nm. Compound 15 4-(6-oxo-2-(trifluoromethvl)-3,6-dihydrochromenor7,8-d1imidazol-8-yl)benzaldehyde

[0196] A solution of 8-(4-bromophenyl)-2-(trifluoromethyl)chromeno[7,8-d]imidazol-6(3H)-one (252 mg, 96% Wt, 1 Eq, 591 pmol), Pd(dppf)CI2 (45.0 mg, 0.104 Eq, 61.5 pmol), triethylamine (196 mg, 270 pL, 3.28 Eq, 1.94 mmol) and triethylsilane (218 mg, 300 pL, 3.18 Eq, 1.88 mmol) in DMF (6.00 mL) was heated to 80 °C under 3 bar of CO gas for 16h. The reaction mixture was concentrated in vacuo to afford the crude product. The crude product was purified by column chromatography using a gradient of 0-5% MeOH in DCM to afford 4-(6-oxo-2-(trifluoromethyl)-3,6-dihydrochromeno[7,8-d]imidazol-8-yl)benzaldehyde (146 mg, 0.40 mmol, 68 %) as a pale yellow / brown solid.

[0197] 1H NMR (400 MHz, DMSO): 6 14.80 (brs, 1H), 10.14 (s, 1H), 8.41 -8.36 (m, 2H), 8.18 - 8.11 (m, 2H), 8.06 - 8.01 (m, 1H), 7.79 - 7.74 (m, 1H), 7.34 (s, 1H).

[0198] MS: The product was analysed by LCMS (Method 1): m / z 359.0 (M+H)+ (ES+); 357.0 (M-H) (ES-), at 1.28 min, 98% purity at 260nm +1- 80nm. 8-(4-(azetidin-1-vlmethvl)phenvl)-2-(trifluoromethvl)chromeno(7,8-dlimidazol-6(3H)-one (15) 0

[0199] To a solution of 4-(6-oxo-2-(trifluoromethyl)-3,6-dihydrochromeno[7,8-d]imidazol-8-yl)benzaldehyde (97.0 mg, 1 Eq, 271 pmol) and azetidine (33.9 mg, 40.0 pL, 2.19 Eq, 593 pmol) in THF (2 mL) was added acetic acid (1 drop). The reaction mixture was stirred at rt 30 min. Sodium triacetoxyborohydride (57.4 mg, 1 Eq, 271 pmol) was added in one portion and the reaction mixture was stirred at rt for 1 h. The reaction mixture was carefully quenched with MeOH (~3 mL) and then concentrated in vacuo to afford the crude product. The crude product was purified by column chromatography using a gradient of 0-10% MeOH in DCM, then 1-10% (0.7 M Ammonia / MeOH) in DCM, to afford 8-(4-(azetidin-1-ylmethyl)phenyl)-2-(trifluoromethyl)chromeno[7,8-d]imidazol-6(3H)-one (57.0 mg, 143 pmol, 52%) as a yellow / orange solid. The product was further purified by General procedure 4, Method 2. The clean fractions were evaporated in a Genevacto afford 8-(4-(azetidin-1-ylmethyl)phenyl)-2-(trifluoromethyl)chromeno[7,8-d]imidazol-6(3H)-one, Formic Acid (18.1 mg, 40 pmol, 15 %) as a white solid.

[0200] 1H NMR (400 MHz, MeOD): 6 8.41-8.33 (m, 3H), 8.03 (dd, J = 8.8, 2.5 Hz, 1H), 7.70 (dd, J = 8.7, 2.4 Hz, 1H), 7.65 (dd, J = 8.5, 2.3 Hz, 2H), 7.05 (dt, J = 4.0, 1.1 Hz, 1H), 4.42 (s, 2H), 4.15 (t, J = 8.1 Hz, 4H), 2.52 (p, J = 8.1 Hz, 2H). 1 proton not observed in MeOD.

[0201] 19F NMR (376 MHz, MeOD): 5 -65.35 (d, J = 2.8 Hz).

[0202] MS: The product was analysed by LCMS (Method 1): m / z 400.0 (M+H)+ (ES+); 398.0 (M-H)- (ES-), at 0.77 min, 99% purity at 260nm +1- 80nm. Compound 16 tert-butyl 4-(7-amino-8-nitro-4-oxochroman-2-yl)piperidine-1-carboxylate

[0203] A suspension of 1-(4-amino-2-hydroxy-3-nitrophenyl)ethan-1-one (393 mg, 94% Wt, 1 Eq, 1.88 mmol) and tert-butyl 4-formylpiperidine-1-carboxylate (632 mg, 1.57 Eq, 2.96 mmol) in EtOH (10 mL) was heated to 50 °C before adding pyrrolidine (147 mg, 170 pL, 1.1 Eq, 2.07 mmol) in a single portion and the reaction mixture was stirred at 50 °C for 2 h. A solution of aldehyde (300 mg) in EtOH (5 mL) was added and the reaction mixture was stirred at 50 °C for 1 h. Additional pyrrolidine (147 mg, 170 pL, 1.1 Eq, 2.07 mmol) was added and stirred at 50 °C for 18 h. The reaction mixture was concentrated in vacuo and the residue was partitioned between DCM (100 mL) and sat. aq NaHCOa (50 mL). The organic layer was collected and washed with 50% brine (50 mL), dried over MgSO4and concentrated in vacuo to afford the crude product. The crude product was purified by column chromatography using a gradient of 0-5% MeOH in DCM, to afford tert-butyl 4-(7-amino-8-nitro-4-oxochroman-2-yl)piperidine-1-carboxylate (310 mg, 0.71 mmol, 38 %) as an orange foam.

[0204] 1H NMR (400 MHz, DMSO): 6 7.58 (d, J = 9.0 Hz, 1H), 7.25 (s, 2H), 6.51 (d, J = 9.0 Hz, 1H), 4.43-4.35 (m, 1H), 4.07- 3.88 (m, 2H), 2.81 -2.63 (m, 2H), 1.90- 1.79 (m, 2H), 1.71-1.55 (m, 2H), 1.40 (s, 9H), 1.31 - 1.12 (m, 3H).

[0205] MS: The product was analysed by LCMS (Method 1): m / z 414.2 (M+Na)+ (ES+); 390.2 (M-H)' (ES-), at 1.48 min, 90% purity at 260nm + / - 80nm. tert-butyl 4-(7-amino-8-nitro-4-oxo-4H-chromen-2-vl)piperidine-1-carboxylate

[0206] A solution of tert-butyl 4-(7-amino-8-nitro-4-oxochroman-2-yl)piperidine-1-carboxylate (310 mg, 90% Wt, 1 Eq, 713 pmol) and iodine (452 mg, 2.5 Eq, 1.78 mmol) in pyridine (8 mL) was stirred at 115 °C for 4 h. The reaction mixture was concentrated in vacuo and chased with toluene to remove residuel solvent. The residue was taken up in DCM (100 mL) and washed with, sat. aq sodium thiosulfate (50 mL), sat. aq. NaHCOs (50 mL), 50% brine (50 mL), dried over MgSO4and concentrated in vacuo to afford the crude product. The crude product was purified by column chromatography using a gradient of 0-5% MeOH in DCM to afford tert-butyl 4-(7-amino-8-nitro-4-oxo-4H-chromen-2-yl)piperidine-1-carboxylate (164 mg, 0.42 mmol, 58 %) as a yellow solid.

[0207] 1H NMR (400 MHz, DMSO): 6 7.81 (d, J = 9.1 Hz, 1H), 7.62 (s, 2H), 6.93 (d, J = 9.1 Hz, 1H), 6.17 (s, 1H), 4.03 (d, J = 13.0 Hz, 2H), 2.84 - 2.74 (m, 3H), 1.93-1.85 (m, 2H), 1.52 (qd, J = 12.5, 4.3 Hz, 2H), 1.42 (s, 9H).

[0208] MS: The product was analysed by LCMS (Method 1): m / z 421.2 (M+Na)+ (ES+); 388.0 (M-H)' (ES-), at 1.40 min, 100% purity at 260nm +1- 80nm. 7-amino-8-nitro-2-(piperidin-4-yl)-4H-chromen-4-one hydrochloride

[0209] To a solution of tert-butyl 4-(7-amino-8-nitro-4-oxo-4H-chromen-2-yl)piperidine-1-carboxylate (164 mg, 99% Wt, 1 Eq, 417 pmol) in DCM (4 mL) was added HCI (3 M in CPME) (109 mg, 1.00 mL, 3.00 molar, 7.20 Eq, 3.00 mmol). The resultant suspension was stirred at rt for 18 h. The reaction mixture was concentrated in vacuo to afford the crude product 7-amino-8-nitro-2-(piperidin-4-yl)-4H-chromen-4-one, HCI (143 mg, 0.42 mmol) as a yellow solid. The product was used in the next step without further purification assuming quantitative yield

[0210] 1H NMR (400 MHz, DMSO): 6 9.00 (d, J = 11.2 Hz, 1H), 8.66 (d, J = 11.2 Hz, 1H), 7.82 (d, J = 9.1 Hz, 1H), 7.66 (brs, 2H), 6.96 (d, J = 9.1 Hz, 1H), 6.18 (s, 1H), 3.36 (d, J = 12.6 Hz, 2H), 3.05 - 2.91 (m, 3H), 2.09 (dd, J = 12.3, 3.6 Hz, 2H), 1.85 (qd, J = 12.8, 4.0 Hz, 2H).

[0211] MS: The product was analysed by LCMS (Method 1): m / z 290.2 (M+H)+ (ES+); 288.0 (M-H)' (ES-), at 0.39 min 2-( 1 -acetylpiperidin-4-yl)-7-amino-8-nitro-4H-chromen-4-one

[0212] To a stirred suspension of 7-amino-8-nitro-2-(piperidin-4-yl)-4H-chromen-4-one, HCI (143 mg, 95% Wt, 1 Eq, 417 pmol) and triethylamine (109 mg, 150 pL, 2.58 Eq, 1.08 mmol) in DCM (5 mL) at rt was added a solution of acetyl chloride (34.2 mg, 31.0 pL, 1.05 Eq, 436 pmol) in DCM (0.5 mL) dropwise. The resultant solution was stirred at rt for 1 h. Additional acetyl chloride (11.0 mg, 10.0 pL, 0.337 Eq, 141 pmol) in DCM (0.5 mL) was added and stirred at rt for 1 h. Additional triethylamine (109 mg, 150 pL, 2.58 Eq, 1.08 mmol) and acetyl chloride (11.0 mg, 10.0 pL, 0.337 Eq, 141 pmol) in DCM (0.5 mL) was added and stirred for 1 h. The reaction mixture was concentrated in vacuo and azeotroped with DCM to afford the crude product. The crude product was purified by chromatography on silica gel (12 g cartridge, 0-6% MeOH / DCM, eluted ~4%) to afford 2-(1-acetylpiperidin-4-yl)-7-amino-8-nitro-4H-chromen-4-one (101 mg, 305 pmol, 73.1 %) as a light yellow solid.

[0213] 1H NMR (400 MHz, DMSO): 6 7.81 (d, J = 9.1 Hz, 1H), 7.63 (s, 2H), 6.93 (d, J = 9.1 Hz, 1H), 6.16 (s, 1H), 4.51-4.43 (m, 1H), 3.96-3.88 (m, 1H), 3.12 (td, J = 13.0, 2.7 Hz, 1H), 2.86 (tt, J = 11.7, 3.6 Hz, 1H), 2.61 (td, J = 12.8, 2.8 Hz, 1H), 2.02 (s, 3H), 1.99- 1.87 (m, 2H), 1.60 (qd, J = 12.5, 4.3 Hz, 1H), 1.45 (qd, J = 12.5, 4.3 Hz, 1H).

[0214] MS: The product was analysed by LCMS (Method 1): m / z 332.0 (M+H)+ (ES+); 330.0 (M-H)' (ES-), at 0.88 min, >99% purity at 260nm +1- 80nm. 8-(1-acetvlpiperidin-4-yl)-2-(trifluoromethyl)chromeno[7,8-d1imidazol-6(3H)-one (16)

[0215] To a solution of 2-(1-acetylpiperidin-4-yl)-7-amino-8-nitro-4H-chromen-4-one (46.0 mg, 100% Wt, 1 Eq, 139 pmol) and triethylamine (211 mg, 290 pL, 15 Eq, 2.08 mmol) in EtOH (2 mL), THF (2 mL) and water (0.2 mL) at 60 °C was added sodium dithionite (98.0 mg, 39.2 pL, 4.05 Eq, 563 pmol). The reaction mixture was stirred at 60 °C for 30 min. and then concentrated in vacuo. The residue was diluted with DCM (20 mL) and evaporated in vacuo to afford the crude product as a mixture of 2-(1-acetylpiperidin-4-yl)-7,8-diamino-4H-chromen-4-one and (2-(1 -acetylpiperidin-4-yl)-7-amino-4-oxo-4H-chromen-8-yl)sulfamic acid. The crude product was dissolved in trifluoroacetic acid (2 mL) and stirred at 60 °C for 7 h. The reaction mixture was concentrated in vacuo from DCM to afford the crude product. The crude product was purified by column chromatography using a gradient of 0-10% MeOH in DCM, to afford 8-(1-acetylpiperidin-4-yl)-2-(trifluoromethyl)chromeno[7,8-d]imidazol-6(3H)-one (31.0 mg, 80 pmol, 58 %) as a tan solid, after trituration with diethyl ether.

[0216] 1H NMR (400 MHz, MeOD): 6 8.10 (d, J = 8.8 Hz, 1H), 7.69 (d, J = 8.8 Hz, 1H), 6.39 (s, 1H), 4.77-4.69 (m, 1H), 4.15-4.07 (m, 1H), 3.35-3.26 (m, 1H), 3.10 (tt, J = 12.0, 3.8 Hz, 1H), 2.80 (td, J = 12.9, 2.8 Hz, 1H), 2.17 (s, 5H), 1.97 (qd, J = 12.5, 4.2 Hz, 1H), 1.83 (qd, J = 12.6, 4.4 Hz, 1H).

[0217] 19F NMR (376 MHz, MeOD): 6 -65.72.

[0218] MS: The product was analysed by LCMS (Method 1): m / z 380.2 (M+H)+ (ES+); 378.0 (M-H)' (ES-), at 0.98 min, >98% purity at 260nm + / - 80nm. Compound 17 tert-butyl 6-(4-(6-oxo-2-('trifluoromethvl')-1.6-dihvdrochromeno[7.8-d]imidazol-8-vl)phenvl)-3.4-dihvdropyridine-1(2H)-carboxvlate

[0219] A suspension of 8-(4-bromophenyl)-2-(trifluoromethyl)chromeno[7,8-d]imidazol-6(1H)-one (200 mg, 93% Wt, 1 Eq, 455 pmol), tert-butyl 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydropyridine-1(2H)-carboxylate (169 mg, 1.2 Eq, 546 pmol) and tripotassium phosphate (289 mg, 3 Eq, 1.36 mmol) in DMF (9 mL) and water (1 mL) was sparged with N2 for 5 min. Bis(di-tert-butyl (4-dimethylaminophenyl)phosphine)dichloropalladium(ll) (400 mg, 1.24 Eq, 565 pmol) was added and the mixture was sparged with N2 for 10 min. The reaction mixture was stirred at 90 °C for 20 h. The reaction mixture was concentrated in vacuo and the residue was azeotroped with toluene to afford the crude product. The crude product was purified by column chromatography using a gradient of 0-10% MeOH in DCM. The product containing fractions were combined and concentrated in vacuo to afford impure product. The product was further purified by colomn chromatography using a gradient of 0-100% EtOAc in isohexane to afford tert-butyl 6-(4-(6-oxo-2-(trifluoromethyl)-1,6-dihydrochromeno[7,8-d]imidazol-8-yl)phenyl)-3,4-dihydropyridine-1(2H)-carboxylate (54.0 mg, 53 pmol, 12 %) as a yellow solid with significant impurities. The product was deemed of sufficient quality for further reaction and was not purified further.

[0220] MS: The product was analysed by LCMS (Method 1): m / z 512.2 (M+H)+ (ES+); 510.0 (M-H)- (ES-), at 1.79 min, 50% purity at 260nm + / - 80nm. tert-butyl 2-(4- (6-oxo-2-(trifluoromethyl)-1.6-dihvdrochromeno[7,8-d1imidazol-8-yl)phenyl)piperidine-1-carboxylate

[0221] A suspension of tert-butyl 6-(4-(6-oxo-2-(trifluoromethyl)-1,6-dihydrochromeno[7,8-d]imidazol-8-yl)phenyl)-3,4-dihydropyridine-1(2H)-carboxylate (44.0 mg, 50% Wt, 1 Eq, 43.0 pmol), platinum(IV) oxide (10.0 mg, 1.02 Eq, 44.0 pmol) and palladium (5 wt% on carbon) (20.0 mg, 5% Wt, 0.218 Eq, 9.40 pmol) in EtOAc (5 mL) was stirred under H2 (5 bar.) at 30 °C for 48 h. The reaction mixture was filtered through glass fibre filter paper, washing with EtOAc, and the filtrate was concentrated in vacuo to afford the crude product. The crude product was purified by column chromatography using a gradient of 0-5% MeOH in DCM) to afford tert-butyl 2-(4-(6-oxo-2-(trifluoromethyl)-1,6-dihydrochromeno[7,8-d]imidazol-8-yl)phenyl)piperidine-1-carboxylate (9.00 mg, 13 pmol, 31 %) as a pale yellow solid. MS: The product was analysed by LCMS (Method 1): m / z 514.2 (M+H)+ (ES+); 512.2 (M-H)' (ES-), at 1.84 min, 77% purity at 260nm + / -80nm. 8-(4-(piperidin-2-vl)phenvl)-2-(trifluoromethyl)chromeno[7.8-d1imidazol-6(1H)-one (17)

[0222] To a solution of tert-butyl 2-(4-(6-oxo-2-(trifluoromethyl)-1,6-dihydrochromeno[7,8-d]imidazol-8-yl)phenyl)piperidine-1-carboxylate (9.00 mg, 77% Wt, 1 Eq, 13.5 pmol) in THF (1 mL) was added HCI (3 M in CPME) (55 mg, 0.50 mL, 3.00 molar, 1.1e+2 Eq, 1.5 mmol). The reaction mixture was stirred at rt for 28 h and then concentrated in vacuo to afford the crude product. The crude product was purified by colomn chromatography using a gradient of 0-10% (0.7 M Ammonia / MeOH) in DCM then 20% (0.7 M Ammonia / MeOH) in DCM, to afford 8-(4-(piperidin-2-yl)phenyl)-2-(trifluoromethyl)chromeno[7,8-d]imidazol-6(1H)-one (5.00 mg, 12 pmol, 89 %) as a white solid.

[0223] 1H NMR (400 MHz, MeOD): 5 8.41 (d, J = 8.2 Hz, 2H), 8.16 (d, J = 8.8 Hz, 1H), 7.78-7.69 (m, 3H), 7.12 (s, 1H), 4.40 (dd, J = 12.2, 3.0 Hz, 1H), 3.53 (d, J = 12.6 Hz, 1H), 3.29 - 3.20 (m, 1H), 2.22-2.15 (m, 1H), 2.12 - 1.97 (m, 3H), 1.92- 1.78 (m, 2H).

[0224] 19F NMR (376 MHz, MeOD): 6 -65.62.

[0225] MS: The product was analysed by LCMS (Method 1): m / z 414.2 (M+H)+ (ES+); 412.0 (M-H)- (ES-), at 0.87 min, 99% purity at 260nm + / - 80nm. Compound 18

[0226] 1-(4-amino-5-chloro-2-hydroxy-3-nitrophenyl)ethan-1-one

[0227] NCS (1.04 g, 98% Wt, 1.5 Eq, 7.65 mmol) was added to a solution of 1-(4-amino-2-hydroxy-3-nitrophenyl)ethan-1-one (1.00 g, 1 Eq, 5.10 mmol) in MeCN (30 mL). The reaction was heated to 80 °C for 5 h. The reaction was allowed to cool. The reaction mixture was dried onto celite and purified by column chromatography using a gradient of 0-2% MeOH in DCM to afford 1-(4-amino-5-chloro-2-hydroxy-3-nitrophenyl)ethan-1-one (638 mg, 2.7 mmol, 54 %) as a light yellow solid.

[0228] 1H NMR (400 MHz, DMSO): 6 14.01 (s, 1H), 8.02 (s, 1H), 7.28 (s, 2H), 2.55 (s, 3H).

[0229] MS: The product was analysed by LCMS (Method 1): m / z 231.0 / 233.0 (M+H)+ (ES+); 229.0 / 231.0 (M-H)’ (ES-), at 1.17 min, 99% purity at 260nm + / - 80nm. 4-(3-(4-amino-5-chloro-2-hydroxy-3-nitrophenyl)-3-oxoprop-1-en-1-yl)benzonitrile

[0230] A suspension of 1-(4-amino-5-chloro-2-hydroxy-3-nitrophenyl)ethan-1-one (638 mg, 99% Wt, 1 Eq, 2.74 mmol) and 4-formylbenzonitrile (539 mg, 1.5 Eq, 4.11 mmol) in EtOH (10 mL) was heated to 50 °C before the addition of pyrrolidine (214 mg, 247 pL, 1.1 Eq, 3.01 mmol) in one portion and the reaction mixture was stirred at 50 °C for 2 h and then allowed to cool to rt. The precipitate was collected by filtration, washing with EtOH, and dried in vacuo to afford the desired product 4-(3-(4-amino-5-chloro-2-hydroxy-3-nitrophenyl)-3-oxoprop-1-en-1-yl)benzonitrile (683 mg, 2.0 mmol, 72 %) as a yellow free-flowing solid.

[0231] 1H NMR (400 MHz, DMSO): 5 14.99 (s, 1H), 8.59 (s, 1H), 8.20-8.12 (m, 3H), 7.99-7.92 (m, 2H), 7.87 (d, J = 15.4 Hz, 1H), 7.47 (s, 2H).

[0232] MS: The product was analysed by LCMS (Method 1): m / z 344.0 / 346.0 (M+H)+ (ES+); 342.0 / 344.0 (M-H)' (ES-), at 1.75 min, >99% purity at 260nm +1- 80nm. 4-(7-amino-6-chloro-8-nitro-4-oxo-4H-chromen-2-yl)benzonitrile

[0233] To a stirred suspension of 4-(3-(4-amino-5-chloro-2-hydroxy-3-nitrophenyl)-3-oxoprop-1-en-1-yl)benzonitrile (683 mg, 99% Wt, 1 Eq, 1.97 mmol) in DMSO (1.54 g, 1.40 mL, 10 Eq, 19.7 mmol) and Toluene (7 mL) was added iodine (99.9 mg, 0.2 Eq, 393 pmol). The reaction mixture was stirred at 110 °C for 3 h. The reaction mixture was allowed to cool to room temperature and diluted with water (20 mL). The precipitate was collected by filtration, washing with water (10 mL) and EtOH (10 mL). The solid was dried in vacuo to afford 4-(7-amino-6-chloro-8-nitro-4-oxo-4H-chromen-2-yl)benzonitrile (761 mg, 2.0 mmol, 100 %) as a pale yellow soild. The product was used without further purification.

[0234] 1H NMR (400 MHz, DMSO): 5 8.20-8.12 (m, 2H), 8.12-8.04 (m, 2H), 8.02 (s, 1H), 7.62 (s, 2H), 7.25 (s, 1H).

[0235] MS: The product was analysed by LCMS (Method 1): m / z 342.0 / 344.0 (M+H)+ (ES+); 340.0 / 342.0 (M-H)‘ (ES-), at 1.49 min, >99% purity at 260nm + / - 80nm. 4-(7,8-diamino-6-chloro-4-oxo-4H-chromen-2-yl)benzonitrile

[0236] To a stirred suspension of 4-(7-amino-6-chloro-8-nitro-4-oxo-4H-chromen-2-yl)benzonitrile (761 mg, 90% Wt, 1 Eq, 2.00 mmol) in DMSO (5 mL), EtOH (5 mL) and Water (1 mL) at 80 °C was added sodium dithionite (1.05 g, 419 pL, 3 Eq, 6.01 mmol). After 4 h, the reaction was diluted with an additional aliquot of DMSO (5 mL), EtOH (5 mL) and Water (1 mL). The reaction was heated to 80 °C for a further 6 h. The reaction was allowed to cool then poured into a mixture of ice and water (100 mL). The resulting precipitate was filtered, washed with water (10 mL), then MeCN (5 mL) and dried in vacuo to afford 4-(7,8-diamino-6-chloro-4-oxo-4H-chromen-2-yl)benzonitrile (537 mg, 1.7 mmol, 85 %) as a light orange solid.

[0237] 1H NMR (400 MHz, DMSO): 6 8.47 - 8.39 (m, 2H), 8.05-7.97 (m, 2H), 7.23 (s, 1H), 7.02 (s, 1H), 5.81 (s, 2H), 5.33 (s, 2H).

[0238] MS: The product was analysed by LCMS (Method 1): m / z 312.0 / 314.0 (M+H)+ (ES+); 310.0 / 312.0 (M-H)’ (ES-), at 1.21 min, >99% purity at 260nm +1- 80nm. 4-(4-chloro-6-oxo-2-(trifluoromethvl)-3.6-dihvdrochromeno[7.8-dlimidazol-8-vl)benzonitrile (18)

[0239] TFA (0.36 g, 0.24 mL, 20 Eq, 3.2 mmol) was slowly added to 4-(7,8-diamino-6-chloro-4-oxo-4H-chromen-2-yl)benzonitrile (50 mg, 99% Wt, 1 Eq, 0.16 mmol) while cooling over an ice bath. The mixture was stirred at reflux for 6 h, then concentrated in vacuo. The residue was dissolved in EtOAc (20 mL) and sat. aq. NaHCO3(aq) (10 mL) was slowly added while stirring to avoid excessive gas evolution. The phases were separated and the organic phase was washed with brine (10 mL), dried over MgSCU, filtered and concentrated. The crude product was purified by column chromatography using a gradient of 0-10% MeOH in DOM to afford 4-(4-chloro-6-oxo-2-(trifluoromethyl)-3,6-dihydrochromeno[7,8-d]imidazol-8-yl)benzonitrile (19 mg, 48 pmol, 30 %) as a light yellow solid.

[0240] The product was analysed by LCMS (Method 1): m / z 390.0 / 392.0 (M+H)+ (ES+); 388.0 / 390.0 (M-H)- (ES-), at 1.46 min, >99% purity at 260nm + / - 80nm.

[0241] 1H NMR (500 MHz, DMSO, 298 K) 6 8.46 (b, 2H), 8.16-8.10 (m, 2H), 7.93 (s, 1H), 7.40 (s, 1H).

[0242] 19F NMR (471 MHz, DMSO, 298 K) 0 -62.51.

[0243] 1H NMR (500 MHz, DMSO, 353 K) 6 8.42 (d, J = 8.1 Hz, 2H), 8.11 - 8.06 (m, 2H), 7.95 (s, 1H), 7.29 (s, 1H). Compound 19 4-(7-amino-8-nitro-4-oxo-4H-chromen-2-yl)benzonitrile Synthesized as previously described1 All experimental data was in congruence with literature values. 4-(7,8-diamino-4-oxo-4H-chromen-2-yl)benzonitrile

[0244] To a stirred suspension of 4-(7-amino-8-nitro-4-oxo-4H-chromen-2-yl)benzonitrile (3.0 g, 98% Wt, 1 Eq, 9.57 mmol) in DMSO (15 mL) and EtOH (15 mL) at 80 °C was added sodium dithionite (5.00 g, 2.00 mL, 3 Eq, 28.7 mmol). The reaction mixture was stirred at 80 °C for 30 min. Water (10 mL) was added and the reaction mixture was stirred at 80 °C for 1.5 h. The reaction mixture was diluted with DMSO (10 mL), EtOH (10 mL) and additional sodium dithionite (500 mg, 0.3 eq., 2.87 mmol) was added. The reaction mixture was stirred at 80 °C for 2.5 h. The reaction mixture was poured into a mixture of ice and water (200 mL) and allowed to stand overnight. The resultant precipitate was collected by filtration then washed with water, MeCN and EtOH. The solid was dried in vacuo (40 °C, overnight) to afford 4-(7,8-diamino-4-oxo-4H-chromen-2-yl)benzonitrile (2.778 g, 7.7 mmol, 81 %) as an orange solid.

[0245] 1H NMR(500MHz, DMSO): 5 8.45 - 8.36 (m, 2H), 8.04-7.98 (m, 2H), 7.18 (d, J = 8.5 Hz, 1H), 6.95 (s, 1H), 6.71 (d, J = 8.4 Hz, 1H).

[0246] MS: The product was analysed by LCMS (Method 1): m / z 278.0 (M+H)+ (ES+); 276.0 (M-H)' (ES-), at 0.99 min, 95% purity at 260nm + / - 80nm. 4-(2-(1-fluorocyclopropyl)-6-oxo-3,6-dihydrochromeno|7,8-d1imidazol-8-yl)benzonitrile (19)

[0247] To a suspension of 4-(7,8-diamino-4-oxo-4H-chromen-2-yl)benzonitrile (50.0 mg, 77% Wt, 1 Eq, 139 pmol), 1-fluorocyclopropane-1-carboxylic acid (21.0 mg, 1.45 Eq, 202 pmol) and DIPEA (74.2 mg, 100.0 pL, 4.13 Eq, 574 pmol) in DCM (2 mL) and DMF (2 mL) was added HATU (79.0 mg, 1.50 Eq, 208 pmol). The reaction mixture was stirred at rt for 1 h. Additional HATU (70.0 mg, 1.33 Eq, 184 pmol) was added and stirred for 15 min. The reaction mixture was diluted with DCM (5 mL) and washed with sat. aq NaHCOs (10 mL). Phases were separated, the organic layer was collected and the aqueous layer was extracted with DCM (5 mL). The combined organic extracts were dried using a phase separator and concentrated in vacuo. The residue was azeotroped with toluene to afford the crude product. AcOH (3 mL) was added and the reaction mixture was stirred at 50 °C for 1 h 45 min. The temperature was increased to 85 °C and stirred for 5 h. The reaction mixture was concentrated in vacuo and the residue was azeotroped with toluene to afford the crude product. The crude product was purified by colomn chromatography using a gradient of 0-5% MeOH in DCM to afford 4-(2-(1-fluorocyclopropyl)-6-oxo-3,6-dihydrochromeno[7,8-d]imidazol-8-yl)benzonitrile (34.0 mg, 96 pmol, 69 %) as a beige solid.

[0248] 1H NMR (400 MHz, MeOD): 6 8.49 - 8.41 (m, 2H), 8.02 (d, J = 8.6 Hz, 1H), 7.98-7.93 (m, 2H), 7.64 (d, J = 8.6 Hz, 1H), 7.12 (s, 1H), 1.81 - 1.70 (m, 2H), 1.68-1.58 (m, 2H). 1 proton not observed in MeOD.

[0249] 19F NMR (376 MHz, MeOD): 6 -194.27.

[0250] MS: The product was analysed by LCMS (Method 1): m / z 346.0 (M+H)+ (ES+); 344.0 (M-H)' (ES-), at 1.30 min, 97% purity at 260nm + / - 80nm. Compound 20 4-(2-methyl-6-oxo-1,6-dihvdrochromeno(7,8-d1imidazol-8-yl)benzonitrile (20)

[0251] A suspension of 4-(7,8-diamino-4-oxo-4H-chromen-2-yl)benzonitrile (50.0 mg, 77% Wt, 1 Eq, 139 pmol) and malonic acid (113 mg, 7.82 Eq, 1.09 mmol) in dioxane (2 mL) was heated to 120 °C in the microwave for 5 h. The reaction mixture was diluted with sat. aq. NaHCO3 (5 mL) and extracted with DOM (2x10 mL). The combined organic extracts were dried by phase separator and concentrated in vacuo to afford the crude product. The crude product was purified by column chromatography using a gradient of 0-10% MeOH / DCM) to afford 4-(2-methyl-6-oxo-1,6-dihydrochromeno[7,8-d]imidazol-8-yl)benzonitrile (18.2 mg, 57 pmol, 41 %) as a beige solid. 1H NMR (400 MHz, DMSO): 6 12.84 (s, 1H), 8.38-8.31 (m, 2H), 8.09-8.01 (m, 2H), 7.82 (d, J = 8.5 Hz, 1H), 7.56 (d, J = 8.5 Hz, 1H), 7.11 (s, 1H), 2.63 (s, 3H). MS: The product was analysed by LCMS (Method 1): m / z 302.0 (M+H)+ (ES+); 300.0 (M-H)' (ES-), at 0.97 min, 100% purity at 260nm + / - 80nm. Compound 21

[0252] 4-(2,6-dioxo-1,2,3,6-tetrahydrochromeno[7,8-d]imidazol-8-yl)benzonitrile (21) 0

[0253] To a stirred suspension of 4-(7,8-diamino-4-oxo-4H-chromen-2-yl)benzonitrile (55.0 mg, 77% Wt, 1 Eq, 153 pmol) and cesium carbonate (162 mg, 3.26 Eq, 497 pmol) in THF (4 mL) was added triphosgene (136 mg, 3.00 Eq, 458 pmol) in one portion. The reaction mixture was stirred at rt for 1 h and then carefully quenched with water (4 mL). The resultant suspension was collected by filtration washing with water and MTBE. The solid was dried in vacuo to afford 4-(2,6-dioxo-1,2,3,6-tetrahydrochromeno[7,8-d]imidazol-8-yl)benzonitrile (38.0 mg, 0.12 mmol, pale brown solid.

[0254] 1H NMR(400MHz, DMSO): 6 11.96(brs, 1H), 11.37 (brs, J= 1.7 Hz, 1H), 8.55-8.48 (m, 2H), 8.10-8.02 (m, 2H), 7.66 (d, J = 8.3 Hz, 1H), 7.20 (s, 1H), 7.12 (d, J = 8.4 Hz, 1H).

[0255] MS: The product was analysed by LCMS (Method 1): m / z 304.0 (M+H)+ (ES+); 302.0 (M-H) (ES-), at 0.98 min, 99% purity at 260nm +1- 80nm. Compounds 22 & 23 4-(6-oxo-2-(trifluoromethyl)-3,6-dihydrochromenof7,8-d1imidazol-8-yl)benzonitrile Synthesized as previously described1 All experimental data was in congruence with literature values. 4-(3-methvl-6-oxo-2-(trifluoromethvl)-3,6-dihvdrochromenor7,8-dlimidazol-8-vl)benzonitrile (22) 4-(1-methvl-6-oxo-2-(trifluoromethvl)-1,6-dihvdrochromenor7,8-dlimidazol-8-vl)benzonitrile (23)

[0256] To a stirred suspension of 4-(6-oxo-2-(trifluoromethyl)-1,6-dihydrochromeno[7,8-d]imidazol-8-yl)benzonitrile (107 mg, 98% Wt, 1 Eq, 295 pmol) and potassium carbonate (65.0 mg, 1.59 Eq, 470 pmol) in DMF (3 mL) was added Mel (45.4 mg, 20.0 pL, 1.08 Eq, 320 pmol). The reaction mixture was stirred at rt for 16 h. Additional Mel (45.4 mg, 20.0 pL, 1.08 Eq, 320 pmol) was added and stirred for 4 h. The reaction mixture was diluted with water (10 mL) and the precipitate was collected by filtration, washing with water, to afford a mixture of products (96 mg). The mixture of products (96 mg) were dissolved to 10 mg / mL in DMSO:THF (1:1) with sonication, filtered and was then separated by chiral SFC on a Waters Prep 15 with a PDA detector, 40 °C, 120 bar using a ChiralpaK IH, 10 x 150mm, 5pm, flow rate 15mL / min at 30% MeOH , 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. The samples were then further dried in a vacuum oven at 30 °C / 5mbar overnight to afford the separated isomers. Isomer 1 (22)

[0257] 4-(3-methyl-6-oxo-2-(trifluoromethyl)-3,6-dihydrochromeno[7,8-d]imidazol-8-yl)benzonitrile (28.4 mg, 76 pmol, 26 %) was isolated as a yellow solid.

[0258] 1H NMR (400 MHz, DMSO): 5 8.35 - 8.27 (m, 2H), 8.13-8.09 (m, 2H), 8.08 (d, J = 8.8 Hz, 1H), 7.93 (d, J = 8.9 Hz, 1H), 7.34 (s, 1H), 4.11 - 4.08 (m, 3H). 1

[0259] 9F NMR (376 MHz, DMSO): 6 -61.54.

[0260] MS: The product was analysed by LCMS (Method 1): m / z 370.1 (M+H)+ (ES+); no ionisation (ES ), at 1.41 min, 99% purity 210-400nm. Isomer 2 (23)

[0261] 4-(1-methyl-6-oxo-2-(trifluoromethyl)-1,6-dihydrochromeno[7,8-d]imidazol-8-yl)benzonitrile (30.6 mg, 82 pmol, 28 %) was isolated as a yellow solid.

[0262] 1H NMR (400 MHz, DMSO): 5 8.32 - 8.25 (m, 2H), 8.13-8.05 (m, 2H), 7.97 (d, J = 8.7 Hz, 1H), 7.87 (d, J = 8.7 Hz, 1H), 7.32 (s, 1H), 4.44 (s, 3H).

[0263] 19F NMR (376 MHz, DMSO): 5 -61.25.

[0264] MS: The product was analysed by LCMS (Method 1): m / z 370.1 (M+H)+ (ES+); no ionisation (ES-), at 1.41 min, 99% purity 210-400nm. Compound 24 4-(7,8-diamino-6-methyl-4-oxo-4H-chromen-2-yl)benzonitrile

[0265] A suspension of 4-(7,8-diamino-6-chloro-4-oxo-4H-chromen-2-yl)benzonitrile (202 mg, 99% Wt, 1 Eq, 642 pmol), methylboronic acid (115 mg, 3 Eq, 1.92 mmol), potassium carbonate (197 mg, 2.22 Eq, 1.43 mmol) and Pd-170 (43.2 mg, 0.1 Eq, 64.2 pmol) in dioxane (4 mL) and water (0.4 mL) was stirred at 100 °C under N2 for 16 h. Additional methylboronic acid (115 mg, 3 Eq, 1.92 mmol) and Pd-170 (43.2 mg, 0.1 Eq, 64.2 pmol) were added and stirred at 100 °C for 5 h. The reaction mixture was concentrated in vacuo to afford the crude product. The crude product was purified by column chromatography using a gradient of 0-10% MeOH in DCM to afford 4-(7,8-diamino-6-methyl-4-oxo-4H-chromen-2-yl)benzonitrile (37.0 mg, 0.11 mmol, 18 %) as a yellow solid.

[0266] 1H NMR (400 MHz, DMSO): 6 8.44 - 8.39 (m, 2H), 8.04-7.96 (m, 2H), 7.09 (d, J = 1.0 Hz, 1H), 6.94 (s, 1H), 5.42 (s, 2H), 4.92 (s, 2H), 2.18 (d, J = 0.9 Hz, 3H).

[0267] MS: The product was analysed by LCMS (Method 1): m / z 292.0 (M+H)+ (ES+); 290.0 (M-H)- (ES-), at 1.05 min, 90% purity at 260nm + / - 80nm. 4-(4-methyl-6-oxo-2-(trifluoromethyl)-1.6-dihydrochromeno[7.8-d]imidazol-8-yl)benzonitrile (24) FsC\ HN

[0268] A solution of 4-(7,8-diamino-6-methyl-4-oxo-4H-chromen-2-yl)benzonitrile (37.0 mg, 90% Wt, 1 Eq, 114 pmol) in TFA (2 mL) was stirred at 70 °C for 6 h and then allowed to cool to rt. The reaction mixture was concentrated in vacuo and the residue was azeotroped with DCM to afford the crude product. The crude product was purified by column chromatography using a gradient of 0-3% MeOH in DCM to afford 4-(4-methyl-6-oxo-2-(trifluoromethyl)-1,6- dihydrochromeno[7,8-d]imidazol-8-yl)benzonitrile (34.0 mg, 88 pmol, 77 %) as a pale yellow solid.

[0269] 1H NMR (400 MHz, MeOD): 6 8.47 (d, J = 8.5 Hz, 2H), 8.00-7.93 (m, 2H), 7.88 (s, 1H), 7.15 (s, 1H), 2.70 (d, J= 1.1 Hz, 3H).

[0270] MS: The product was analysed by LCMS (Method 1): m / z 370.0 (M+H)+ (ES+); 368.0 (M-H)- (ES-), at 1.40 min, 96% purity at 260nm +1- 80nm. EXAMPLE 2: EFFECTS OF TEST COMPOUNDS ON SURVIVAL OF MOTOR NEURONS, INTEGRITY OF THE NEURITE NETWORK AND TDP43 MISLOCALISATION IN PRIMARY SOD1 TRANSGENIC SPINAL CORD MOTOR NEURONS FOLLOWING GLUTAMATE INSULT

[0271] The aim of this study was to assess the neuroprotective effects of novel compounds (at several concentrations) in an in vitro model of ALS, based on a primary culture of spinal motor neurons, from SOD1 G93A rat embryos, injured with glutamate. Survival of motor neurons, integrity of the neurite network and TDP43 mislocalisation were evaluated. Methods: Primary culture of spinal motor neurons

[0272] Rat spinal cord motor neurons (MNs) were cultured as described by Boussicault etal., 2020 and Wang etal., 2013. Briefly, pregnant female rats of 14 days gestation were killed using a deep anesthesia with CO2 chamber and a cervical dislocation. Then, embryos were removed from the uterus and immediately placed in ice-cold L15 Leibovitz medium with a 2 % penicillin (10,000 U / mL) and streptomycin (10 mg / mL) solution (PS) and 1 % bovine serum albumin (BSA). Only Tg SOD1G93A embryos were used to prepare the spinal MN culture

[0273] The whole spinal cord was extracted from each embryo and spinal cords from transgenic SOD1G93A embryos were pooled in ice-cold medium of Leibovitz (L15). Spinal cords were then treated for 20 min at 37 °C with a trypsin-EDTA solution at a final concentration of 0.05 % trypsin and 0.02 % Ethylenediaminetetraacetic acid (EDTA). The dissociation was stopped by addition of Dulbecco’s modified Eagle’s medium (DMEM) with 4.5 g / L of glucose, containing DNAse I grade II (final concentration 0.5 mg / ml) and 10 % fetal bovine serum (FBS). Cells were mechanically dissociated by three forced passages through the tip of a 10-mL pipette. Cells were then centrifuged at 515 x g for 10 min at 4 °C. The supernatant was discarded, and the pellet was resuspended in a defined culture medium consisting of Neurobasal medium with a 2 % solution of B27 supplement, 2 mM of L-glutamine, 2 % of PS solution, and 10 ng / mL of brain-derived neurotrophic factor (BDNF). Viable cells were counted in a Neubauer cytometer, using the trypan blue exclusion test. The cells were seeded at a density of 20,000 per well in 96-well plates precoated with poly-L-lysine and were cultured at 37 °C in an air (95 %)-CO2 (5 %) incubator. The medium was changed every other day. Only 60 wells of the 96-well plates were used for the culture: the wells of first and last lines and columns were not used and filled with sterile water to avoid any edge effect. Test compounds and glutamate exposure

[0274] Vehicle: Culture medium (0.1 % dimethyl sulfoxide (DMSO) as final concentration)

[0275] Pre-incubation: On day 13 of culture, test compounds were solubilized in DMSO, then diluted in the culture medium and pre-incubated with primary neurons for 1 h, before glutamate injury. BDNF, solubilized in PBS, was diluted in the culture medium and pre-incubated with primary neurons for 1 h, before glutamate insult. BDNF, was used as a validated positive experimental control.

[0276] Glutamate injury: On day 13 of culture, half of the culture medium (100 pL) was removed from each well and replaced by culture medium containing 10 pM glutamate (so that final concentration of glutamate is 5 pM, except for control condition) and the compounds (2-fold concentrated) for 20 min. After 20 min, glutamate was washed out and fresh culture medium with the compounds or vehicle was added for an additional 24 h period. ORGANISATION OF CULTURE PLATES

[0277] Compounds were tested in 96-well plates (n = 6 culture wells per condition).

[0278] Culture medium was adjusted to 0.1 % DMSO for all the experimental conditions. The stock solution of the test compounds was made at 10 mM in pure DMSO, aliquoted and stored at -80 °C. Next, compounds were submitted to a serial dilution to obtain 1000-fold concentrated stock solutions in DMSO. END POINT EVALUATION Immunostaininq: MAP-2 and TDP43

[0279] 24 hours after glutamate application, the supernatants were collected and stored at -80 °C, and cells were fixed by a cold solution of ethanol (95 %) and acetic acid (5 %) for 5 min at -20 °C. Cell membranes were permeabilized and non-specific binding sites was blocked with a solution of PBS containing 0.1 % of saponin and 1 % FBS for 15 min at room temperature. Cells were incubated for 2 h with the following primary antibodies: a) a mouse monoclonal antibody anti microtubule-associated-protein 2 (MAP-2) at dilution of 1 / 400 in PBS containing 1 % FBS and 0.1 % of saponin. b) a rabbit polyclonal antibody anti-nuclear TAR DNA-binding protein 43 (TDP43) at dilution of 1 / 100 in PBS containing 1 % FBS and 0.1 % of saponin.

[0280] These antibodies were revealed with a Clear Fluor (CF®) 488 goat anti-mouse IgG at the dilution 1 / 400 and a CF® 568 goat anti-rabbit IgG at the dilution 1 / 400, in PBS containing 1 % FBS, 0.1 % saponin, for 1 hour at room temperature. Nuclei were counterstained with the fluorescent dye Hoechst (Sigma Aldrichl / 1000), a marker of total cell survival. Automatic computer analysis

[0281] For each well, 30 pictures (representative of all well area) per well were automatically taken using ImageXpress® (Molecular Devices) with 20x magnification, using the same acquisition parameters. From images, analyses were directly and automatically performed by MetaXpress® (Molecular Devices). The following endpoints were automatically assessed: - Analysis of neuron survival (MAP-2 staining, number of neurons), -Analysis of neurite network (MAP-2 staining, total neurite length in pm), - Analysis of cytoplasmic TDP43 in MAP-2 positive neurons (overlapping between MAP-2 and cytoplasmic TDP43 in pm2).

[0282] According to statistical analysis with an automatized script, 1 to 2 wells per condition can be excluded from analysis due to technical issue, without impacting the quality of results. STATISTICS

[0283] All values are expressed as mean ± SEM (standard error of the mean). Statistical analysis was performed with GraphPad Prism (version 9) using one-way ANOVA followed by Fisher’s LSD test. p< 0.05 was considered significant. Results:

[0284] Table 2 below details the results of the assay. Table 2. Summary of assay data ID# Cone (nM) Neuronal survival (% of control) Neurite network integrity (% of control) Extranuclear TDP43 (% of control) Mean SEM n Sig Mean SEM n Sig Mean SEM n Sig 5 100 71.5 5.9 6 - 75.9 6.7 6 - 156.5 10.7 6 * 1000 74.0 4.4 6 * 82.6 4.4 5 ★ 156.1 6.3 6 * 1 100 63.0 5.1 6 - 65.9 7.1 6 - 165.0 10.3 6 - 1000 83.3 5.5 4 ** 92.4 3.5 4 *** 141.6 4.9 4 ** 10 100 74.3 3.4 6 * 78.3 3.2 6 133.4 2.7 6 1000 71.1 5.2 5 - 70.4 5.6 5 - 132.0 5.7 5 **** 6 100 76.7 2.4 6 *** 80.9 4.9 4 * 107.5 8.9 6 **** 1000 74.3 4.6 5 ** 78.0 1.6 5 ★ 132.1 11.8 5 7 100 69.0 3.6 6 * 68.3 6.7 6 - 130.0 9.7 6 1000 77.4 3.2 6 *** 80.7 7.1 5 127.4 8.8 6 **** 11 10 75.0 5.5 6 * 84.1 5.8 6 *** 155.3 12.4 6 * 100 69.4 4.8 6 - 76.7 5.7 6 * 135.3 12.3 6 *** 12 10 65.5 5.7 6 - 72.7 4.2 6 - 160.2 7.7 6 - 100 75.4 5.4 5 * 75.0 6.9 5 * 167.6 6.4 5 - 13 10 75.4 4.7 6 * 80.7 2.5 6 ** 169.8 5.4 5 - 100 74.4 5.1 6 * 84.4 2.0 6 *** 134.8 9.4 6 *** 18 10 62.0 3.4 6 - 67.1 4.2 6 - 151.5 7.5 6 * 100 67.1 5.2 6 - 68.6 3.4 6 - 141.2 7.5 6 *** 2 10 74.6 3.0 6 ** 79.7 4.8 6 ** 131.6 5.5 6 **** 100 61.1 2.4 6 - 73.0 5.4 6 - 147.5 6.5 6 ** 8 10 63.8 3.6 6 - 68.1 6.3 5 - 145.4 11.0 5 ** 100 63.1 4.1 6 - 76.2 6.2 6 * 140.7 2.3 6 *** 14 10 63.5 4.7 6 - 65.4 4.8 5 - 158.6 4.1 6 - 100 85.0 5.9 5 ** 86.0 5.4 6 *** 128.5 5.1 5 15 10 81.1 5.8 6 ** 82.9 6.1 6 ** 136.5 3.0 6 100 89.8 6.3 6 91.2 4.9 5 *** 121.7 2.1 6 **** 16 10 74.2 5.8 5 - 72.9 4.5 6 - 157.9 6.6 5 - 100 80.7 6.1 6 ** 79.2 4.4 6 * 139.7 5.3 6 **** 20 10 73.5 1.9 4 - 75.3 0.5 4 - 144.4 6.3 4 *** 100 56.5 3.1 5 - 60.3 4.6 5 - 175.2 5.9 5 - 3 10 75.9 2.6 5 ★ 75.6 2.4 6 * 135.8 2.5 5 100 75.8 3.7 6 * 75.7 5.4 6 * 133.8 2.7 6 **** 22 10 67.5 3.6 5 - 65.8 3.1 5 - 159.8 8.6 5 - 100 71.9 1.9 6 * 72.8 2.3 5 * 145.7 3.9 6 ** 23 10 64.5 2.9 6 - 62.4 2.9 6 - 162.5 5.2 6 - 100 66.6 4.0 5 - 64.0 3.0 6 - 147.2 4.3 5 ** 24 10 66.4 5.2 6 - 65.3 2.8 6 - 160.1 5.0 6 - 100 68.2 4.2 6 - 67.1 4.2 5 - 148.4 4.6 6 ** BD NF 50 ng / ml 80.4 1.2 6 ** 82.9 6.1 5 130.3 5.0 4 Note: Significance calculated as a significant change from glutamate insult alone, calculated using One-way ANOVA followed by Fisher's LSD test * = P<0.05 versus glutamate ** = p<0.01 versus glutamate *** = p<0.001 versus glutamate **** = p<0.0001 versus glutamate Conclusion:

[0285] This example demonstrates the effects of the test compounds in an in vitro model of ALS based on primary spinal cord MNs from SOD1 G93A transgenic animals after a glutamatergic stress. BDNF as a validated positive experimental control. The compounds displayed neuroprotective properties with different profiles and as such evidences the use of these compounds as medicaments.

Claims

1. A compound of Formula I or Formula II or a salt thereof,Formula IIWherein:is a single or double bond;Ri is selected from the group consisting of: Cl, F, Br, H, OH, O-alkyl (C1-C6), O-fluoroalkyl (C1-C6), alkyl (C1-C6), fluoroalkyl(C1-C6), NH2, NH-alkyl(C1-C6) N-alkyh (C1-C6), cycloalkyl (C3-C6), fluorocycloalkyl (C3-C6), CH2-cycloalkyl (C3-C6), and CHF-cycloalkyl (C3-C6);R2 is selected from the group consisting of: alkyl (C1-C6), fluoroalkyl (C1-C6), Cl, F, I, Br, H, OH, O-alkyl (C1-C6), SMe, CN, cycloalkyl (C3-C6), and fluorocycloalkyl (C3-C6);R3 if present, when Y is selected from N or C, is selected from the group consisting of: H, alkyl(C1-C6), fluoroalkyl(C1-C6); cycloalkyl (C3-C6), alkyl (C1-C6)-OH, alkyl-(C1-C6)-OMe, and alkyl-(C1-C6)-cycloalkyl (C3-C6);R4 is selected from the group consisting of: semi-saturated or saturated cycloalkyls including bridged bicyclic or spirocyclic ring systems, unsaturated, semi-saturated or saturated heterocycloalkyls including bridged bicyclic or spirocyclic ring systems, or aryls, including but not limited to benzene, pyridine, pyrimidine, pyridazine or piperidine. In the instance that R4 is an aryl, monocyclic cycloalkyl or monocyclic heterocycloalkyl then the monocyclic ring may carry a 2, 3 or 4 position substituent from the list: H, CN, F, Cl, Br, OH, acyl, alkyl(C1-C6), fluoroalkyl (C1-C6), N-alkyl2 (C1-C6), NH-alkyl (C1-C6), NH-cycloalkyl (C3-C6) N-cycloalkyl (C3-C6), alkyl (C1-C6)-NH2 , alkyl (C1-C6)-OH , CH2-N-Cycloalkyl (C3-C6), CH2-heterocycloalkyl(C1-C6), CH2N-Alkyl2 (C1-C6), CH2NH-Alkyl (C1-C6), CH2N-cycloalkyl, and CH2N-fluorocycloalkyl;Rs is selected from the group consisting of: alkyl (C1-C6), fluoroalkyl (C1-C6), Cl, F, I, Br, H, OH, OMe, SMe, CN, cycloalkyl (C3-C6), and fluorocycloalkyl (C3-C6);X is independently selected from the group consisting of: N or C; andY is independently selected from the group consisting of: O, N, C, and S.

2. A compound of Formula III, or a salt thereof, Formula IIIWherein:----is a single or double bond;Ri and R2 are independently selected from the group consisting of: OH, OMe, O-alkyl(C1-C6), O-fluoro alkyl(C1-C6), OiPr, SMe, SF5, alkyl(C1-C6), fluoro alkyl(C1-C6), pyrazole, methyl pyrazole, oxazole, imidazole, thiazole, triazole oxadiazole, and thiadiazole; where the heterocycles can be substituted with alkyl(C1-C3).R3 is selected from the group consisting of: alkyl(C1-C6), fluoro alkyl(C1-C6), Cl, F, I, Br, CN, H O-alkyl (C1-C6).R4 is selected from the group consisting of: semi-saturated or saturated cycloalkyls including bridged bicyclic or spirocyclic ring systems, unsaturated, semi-saturated or saturated heterocycloalkyls including bridged bicyclic or spirocyclic ring systems, or aryls, including but not limited to benzene, pyridine, pyrimidine, pyridazine or piperidine. In the instance that R4 is an aryl, monocyclic cycloalkyl or monocyclic heterocycloalkyl then the monocyclic ring maycarry a 2, 3 or 4 position substituent from the list: H, CN, F, Cl, Br, OH, alkyl(C1-C6), fluoroalkyl (C1-C6), N-alkyh (C1-C6), NH-alkyl (C1-C6), NH-cycloalkyl (C3-C6) N-cycloalkyl (C3-C6), alkyl (C1-C6)-NH2, alkyl (C1-C6)-OH , CH2-N-Cycloalkyl (C3-C6), CH2-heterocycloalkyl(C1-C6), CH2N-Alkyl2 (C1-C6), CH2NH-Alkyl (C1-C6), CH2N-cycloalkyl, and CH2N-fluorocycloalkyl.R5 is selected from the group consisting of: alkyl (C1-C6), fluoroalkyl (C1-C6), Cl, F, I, Br, H, OH, OMe, SMe, CN cycloalkyl (C3-C6), and fluorocycloalkyl (C3-C6).

3. A compound of Formula I, Formula II, or Formula III or as defined by any one of the compounds numbered 1 to 50 in Table 1.

4. A pharmaceutical composition comprising a compound of Formula I, Formula II or Formula III, or a salt thereof, together with one or more ingredients selected from carriers, diluents, excipients, adjuvants, fillers, buffers, binders, disintegrants, preservatives, antioxidants, lubricants, stabilisers, solubilisers, surfactants (e.g., wetting agents), masking agents, colouring agents, flavouring agents, and sweetening agents.

5. A compound of Formula I, Formula II, or Formula III or a pharmaceutical composition comprising the compound of Formula I, Formula II, or Formula III, for use as a medicament.

6. A method of treatment comprising administering to a subject in need of treatment a therapeutically effective amount of a compound of Formula I, Formula II, or Formula III.

7. A method of synthesizing a compound of Formula I, Formula II, or Formula III.

8. An intermediate formed in the method of synthesis of the compound of Formula I,Formula II, or Formula III.