HSD17B13 inhibitor

Abstract

The present invention encompasses a heteroaryl-substituted 2,6-difluorophenol compound of formula (I) (wherein the groups A1 to A3 and Z have the meanings given in the claims and the specification), their use in pharmaceutical compositions containing these compounds, and, in particular, their use as medicaments for preventing the progression of liver diseases to non-alcoholic steatohepatitis, fibrosis and cirrhosis, which are later stages from steatosis. TIFF2025523357000115.tif61124

Description

Technical Field

[0001] The present invention relates to a heteroaryl-substituted 2,6-difluorophenol compound of formula (I).

Chem.

Background Art

[0002] WO2021 / 211974 and WO2022 / 020714 disclose thiophene-carboxamide HSD17B13 inhibitors. WO2022 / 020730 discloses quinazolinone HSD17B13 inhibitors.

[0003] HSD17B13 is a member of the 17b-hydroxysteroid dehydrogenase family of oxidoreductase enzymes and acts on a series of lipid substrates in a group. In humans, the mRNA of HSD17B13 is most highly expressed in the liver and is mainly expressed in hepatocytes. Intracellularly, HSD17B13 is associated with lipid droplets (Su et al, Proc National Acad Sci. 111: 11437-11442, 2014). The physiological function of HSD17B13 is unknown, but multiple substrates including estradiol, retinol and leukotriene B4 have been identified using an in vitro enzyme assay system in which NAD + (nicotinamide adenine dinucleotide, oxidized form) acts as a cosubstrate (Abdul-Husn et al, The New England Journal of Medicine. 378: 1096-1106, 2018).

[0004] Loss-of-function (LoF) gene variants in humans provide evidence for the role of HSD17B13 activity in mediating the risk of certain liver diseases. The presence of a single nucleotide polymorphism (SNP) rs72613567 that encodes a truncated, enzymatically inactive protein has been associated with liver diseases such as hepatic fibrosis or non-alcoholic steatohepatitis (NASH). SNP rs72613567 also reduces the increased risk of liver disease. SNP rs72613567 was found to occur less frequently in liver transplant recipients compared to healthy controls. SNP rs62305723, which encodes an HSD17B13 LoF variant, has been associated with a reduction in the severity of NASH (Ma et al, Hepatology 69: 1504-1519, 2018). SNP rs80182459, which encodes a potential LoF variant, was found to occur less frequently in certain patients with chronic liver disease (Kozlitina et al, The New England Journal of Medicine 379: 1876-1877, 2018). SNP rs6834314, which is highly related to SNP rs72613567, was also found to be associated with fatty liver disease. Small interfering RNA (siRNA) targeting hepatocytes was designed to deplete HSD17B13 in the human liver, and it was found to reduce the activity of serum alanine aminotransferase (ALT), a biomarker of liver injury, in five patients with fatty liver. SUMMARY OF THE INVENTION

[0005] In view of the above data, it is desirable to provide a potent HSD17B13 inhibitor. The "HSD17B13 inhibitor" according to the present invention means a compound that inhibits HSD17B13 in the tests shown in Examples 4 and 6. DETAILED DESCRIPTION OF THE INVENTION

[0006] The compound of formula (I) wherein the groups A1 to A3 and Z have the meanings given below has, in Example 12, a comparative biochemical human IC for HSD17B11 50 As shown by the data, it was not known to act as a selective HSD17B (17β-hydroxysteroid dehydrogenase) inhibitor for HSD17B13. Thus, the compounds according to the invention can be used for the treatment of steatosis such as, for example, non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH). Accordingly, the present invention relates to a compound of formula (I) or a salt thereof.

[0007]

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0008] Those skilled in the art recognize that some heteroaryl groups can be described, for example, in different tautomeric forms, namely pyrazole, triazole, imidazole. Thus, the compounds of the present invention can exist as tautomers. For example, any compound of the present invention containing a pyrazole moiety as a heteroaryl group can exist as the 1H tautomer, or the 2H tautomer, or even a mixture of any amounts of the two tautomers, or the triazole moiety can be the 1H tautomer, 2H tautomer or 4H tautomer, or even a mixture of any amounts of said 1H, 2H or 4H tautomers, namely [Chemical formula] can exist as.

[0009] The present invention includes all possible tautomers of the compounds of the present invention, either as individual tautomers or as any mixture in any proportion of said tautomers.

[0010] In particular, the structure [Chemical formula] can be [Chemical formula] as follows.

[0011] In a group of compounds according to the present invention, the structure [Chemical formula] is selected from the group of structures consisting of [Chemical formula] In another group of compounds of the present invention, the structure

[0012] is [Chemical formula] is [Chemical formula] It is selected from the group of structures consisting of

[0013] In yet another group of the compounds of the present invention, the structure [Chemical formula] is [Chemical formula] selected from the group of structures consisting of Z is preferably [Chemical formula] is. The most preferred structure of Z is [Chemical formula] is.

[0014] The present invention is directed to a compound of formula (I) or a salt thereof that inhibits lipid biosynthesis, and the selective inhibition of HSD17B13 in lipid biosynthesis is a therapeutic benefit including but not limited to the treatment of non-alcoholic steatohepatitis. Accordingly, in another aspect of the present invention, a compound of formula (I) or a pharmaceutically acceptable salt thereof is used as a medicine. The present invention also relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in a method of treating the body of a human or animal.

[0015] Particularly interesting is the use of one or a salt of a compound of formula (I) in the treatment of metabolic disorders. Accordingly, another aspect of the present invention is the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof for the preparation of a pharmaceutical composition comprising at least one compound of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier for the treatment of one of said disorders. Particularly preferred is their use in the preparation of a pharmaceutical composition for modulating a metabolic disorder in the body of a human or animal.

[0016] Also contemplated in the context of the present invention is a method of treating liver disease, metabolic disorders, or cardiovascular disease by using the compounds disclosed herein, or pharmaceutically acceptable salts, solvents or stereoisomers thereof, in combination with additional therapeutic agents. In some embodiments, the additional therapeutic agent is used for the treatment of diabetes or diabetes-related disorders or conditions. In some examples, the additional therapeutic agents include statins, insulin sensitizers, insulin secretagogues, alpha-glucosidase inhibitors, GLP agonists, THR beta agonists, PDE inhibitors, DPP-4 inhibitors (e.g., sitagliptin, vildagliptin, saxagliptin, linagliptin, anagliptin, teneligliptin, alogliptin, gemigliptin or dutogliptin), catecholamines (e.g., epinephrine, norepinephrine or dopamine), peroxisome proliferator-activated receptor (PPAR) gamma agonists (e.g., thiazolidinediones (TZD) [e.g., pioglitazone, rosiglitazone, rivoglitazone or troglitazone], aleglitazar, farigliptazar, muraglitazar or tesaglitazar), peroxisome proliferator-activated receptor (PPAR) alpha agonists, peroxisome proliferator-activated receptor (PPAR) delta agonists, farnesoid X receptor (FXR) agonists (e.g., obeticholic acid), or combinations thereof. In some cases, the statin is an HMG-CoA reductase inhibitor. In other examples, the additional therapeutic agents include fish oil, fibrates, niacin, vitamins such as retinoic acid (e.g., 9-cis retinoic acid), nicotinamide ribonucleoside or analogs thereof, or combinations thereof. In other examples, the additional therapeutic agents include ACC inhibitors, FGF19 and FGF21 mimetics, CCR3 / CCR5 antagonists, or combinations thereof. In some embodiments, the additional therapeutic agent is bivitrol. In some embodiments, the additional therapeutic agent is a statin, such as an HMG-CoA reductase inhibitor, fish oil, fibrate, niacin, or a combination thereof. In other examples, the additional therapeutic agent is a dyslipidemia drug that prevents lipid absorption, such as orlistat. In some embodiments, the additional therapeutic agent is a vitamin, such as retinoic acid or tocopheryl acetate, for the treatment of diabetes and diabetes-related disorders or conditions, such as reducing elevated blood glucose from increased body weight loss and / or food intake. In some embodiments, the additional therapeutic agent is a glucose-lowering agent. In some embodiments, the additional therapeutic agent is an anti-obesity agent. In some embodiments, the additional therapeutic agent is selected from peroxisome proliferator-activated receptor (PPAR) agonists (gamma, dual, or pan), dipeptidyl peptidase (IV) inhibitors, glucagon-like peptide-1 (GLP-1) analogs, insulin or insulin analogs, insulin secretagogues, sodium / glucose cotransporter 2 (SGLT2) inhibitors, glucophage, human amylin analogs, biguanides, alpha-glucosidase inhibitors, meglitinides, thiazolidinediones, and sulfonylureas. In some embodiments, the additional therapeutic agent is a lipid-lowering agent. In some embodiments, the additional therapeutic agent is an antioxidant, a corticosteroid such as budesonide, an anti-tumor necrosis factor (TNF), or a combination thereof. In some embodiments, the additional therapeutic agent is administered concomitantly with the compounds disclosed herein. In some embodiments, the additional therapeutic agent is administered at a lower frequency than the compounds disclosed herein. In some embodiments, the additional therapeutic agent is administered at a higher frequency than the compounds disclosed herein. In some embodiments, the additional therapeutic agent is administered prior to the administration of the compounds disclosed herein. In some embodiments, the additional therapeutic agent is administered after the administration of the compounds disclosed herein.

[0017] In the treatment of mammals, the compounds according to the invention can be administered before, after or simultaneously with at least one other active substance or agent such as a diuretic, antihypertensive, lipid-lowering or antidiabetic agent.

[0018] Formulations Formulations suitable for administering the compounds of the invention will be apparent to those skilled in the art and include, for example, tablets, pills, capsules, suppositories, medicinal candies, lozenges, solutions, elixirs, syrups, sachets, emulsions, inhalants or dispersible powders. Preferred solutions are solutions for injection (s.c., i.v., i.m.) or infusion solutions (injectables). The content of the pharmaceutically active compound needs to be an amount sufficient to achieve the dosage range specified below, for example in the range of 0.10 to 90% by weight, preferably 0.5 to 50% by weight of the total composition. The specified dosage can be given several times a day if necessary.

[0019] Suitable tablets can be obtained, for example, by mixing the active substance of the invention with known excipients such as inert diluents, carriers, disintegrants, adjuvants, surfactants, binders and / or lubricants. Coated tablets can be prepared accordingly by coating a core prepared similarly to a tablet with substances usually used for tablet coating, such as collidon or shellac, gum arabic, talc, titanium dioxide or saccharides. The core may consist of multiple layers in order to achieve delayed release or prevent incompatibilities. Similarly, the tablet coating may consist of multiple layers possibly using the above-mentioned excipients for tablets in order to achieve delayed release. The syrups or elixirs containing the active substances or combinations thereof according to the present invention may additionally contain sweeteners such as saccharine, cyclamate, glycerol or sugar, and flavor enhancers, for example flavoring agents such as vanillin or orange extract. The syrups or elixirs may also contain suspension adjuvants or thickeners such as carboxymethyl cellulose, wetting agents, for example concentrated products of aliphatic alcohols containing ethylene oxide, or preservatives such as p-hydroxybenzoate.

[0020] Solutions for injection and infusion are prepared in the usual manner, for example using emulsifying agents and / or dispersing agents, although stabilizers such as isotonic agents, preservatives such as p-hydroxybenzoate, or alkali metal salts of ethylenediaminetetraacetic acid may be added, and when water is used as the diluent, for example, organic solvents may be used as solvating agents or solubilizing aids and transferred to injection vials or ampoules or infusion bottles. Capsules can be prepared, for example, by mixing the active substance with an inert carrier such as lactose or sorbitol and encapsulating them in gelatin capsules. Suitable suppositories can be prepared, for example, by mixing with carriers provided for this purpose such as neutral fats or polyethylene glycol or their derivatives. Excipients that can be used include pharmaceutically acceptable organic solvents such as water, paraffin (for example, petroleum fractions), vegetable oils (for example, peanut or sesame oil), monofunctional or polyfunctional alcohols (for example, ethanol or glycerol), carriers such as natural mineral powders (for example, kaolin, clay, talc, chalk), synthetic mineral powders (for example, highly dispersed silicic acid and silicates), saccharides (for example, sucrose, lactose and glucose), emulsifying agents (for example, lignin, spent sulfite liquor, methyl cellulose, starch and polyvinylpyrrolidone), and lubricants (for example, magnesium stearate, talc, stearic acid and sodium lauryl sulfate).

[0021] The formulation is administered by a conventional method, preferably by the oral or transdermal route, most preferably by the oral route. For oral administration, tablets, in addition to the carriers described above, of course, contain additives such as sodium citrate, calcium carbonate and dibasic calcium phosphate, together with various additives such as starch, preferably potato starch, gelatin, etc. Lubricants such as magnesium stearate, sodium lauryl sulfate and talc can be used simultaneously for the tableting process. In an aqueous suspension, the active substance can be combined with various flavor enhancers or coloring agents in addition to the excipients described above. For parenteral use, solutions of the active substance with a suitable liquid carrier can be used.

[0022] The total amount of the active ingredient of formula (I) administered is generally in the range of about 0.001 mg / kg to about 200 mg / kg of body weight per day, preferably about 0.01 mg / kg to about 20 mg / kg of body weight per day. Clinically useful dosing schedules range from 1 to 3 times a day to once every 4 weeks. In addition, a "drug holiday" during which the patient is not administered the drug for a specific period may be beneficial to the overall balance between pharmacological effect and tolerance. The unit dose can contain from about 0.5 mg to about 1500 mg of the active ingredient and can be administered more than once a day or less than once a day. The average daily dose for administration by injection, including intravenous, intramuscular, subcutaneous and parenteral injection, and by the use of infusion techniques, is preferably 0.01 - 200 mg / kg of the total body weight. The average daily rectal administration regimen is preferably 0.01 - 200 mg / kg of the total body weight. The average daily vaginal administration regimen is preferably 0.01 - 200 mg / kg of the total body weight. The average daily topical administration regimen is preferably 0.1 - 200 mg administered 1 to 4 times a day. The transdermal concentration is preferably that required to maintain a daily dose of 0.01 - 200 mg / kg. The average daily inhalation administration regimen is preferably 0.01 - 100 mg / kg of the total body weight.

[0023] Depending on an individual's weight, age, route of administration, severity of the disease, individual response to the drug, nature of the formulation, and the time or interval at which the drug is administered (continuous or intermittent treatment with once or multiple daily doses), it may sometimes be necessary to deviate from the amounts specified from time to time. Thus, in some cases, it may be sufficient to use less than the minimum dose given above, and in other cases, it may be necessary not to exceed the upper limit. When administering higher doses, it may be recommended to distribute the higher dose into several lower doses over a day.

[0024] As used herein, "pharmaceutically acceptable salt" refers to a derivative of a disclosed compound in which the parent compound has been modified by making its acidic or basic salt. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral salts or organic acid salts of basic residues such as amines, alkaline salts or organic salts of acidic residues such as carboxylic acids, and the like. For example, such salts include salts derived from benzenesulfonic acid, benzoic acid, citric acid, ethanesulfonic acid, fumaric acid, gentisic acid, hydrobromic acid, hydrochloric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, 4-methyl-benzenesulfonic acid, phosphoric acid, salicylic acid, succinic acid, sulfuric acid, and tartaric acid. Furthermore, pharmaceutically acceptable salts can be formed by cations derived from ammonia, L-arginine, calcium, 2,2'-iminobisethanol, L-lysine, magnesium, N-methyl-D-glucamine, potassium, sodium, and tris(hydroxymethyl)-aminomethane. The pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing basic or acidic moieties by conventional chemical methods. Generally, such salts can be prepared by reacting these compounds in the free acid or base form with a sufficient amount of an appropriate base or acid in water or an organic diluent such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile or a mixture thereof.

[0025] Preparation of the Compounds According to the Invention Overview Unless stated otherwise, all reactions are carried out using methods commonly used in chemical laboratories with commercially available equipment. Starting materials sensitive to air and / or moisture are stored under a protective gas, and the corresponding reactions and operations using them are carried out under a protective gas (nitrogen or argon). The compounds according to the present invention are named using MarvinSketch software (Chemaxon) in accordance with the CAS rules.

[0026] The compounds according to the present invention are prepared by the synthetic methods described below, and the substituents of the general formula have the meanings given above in this specification. These methods are intended to be illustrative of the present invention without limiting the scope of the subject matter and the claimed compounds to these examples. If the preparation of the starting compounds is not described, they are commercially available or known compounds or can be prepared analogously to the methods described in this specification. Substances described in the literature are prepared according to the published synthetic methods.

[0027] Overview Synthesis of Compounds The compounds of the present invention can be prepared as described in the following sections. The schemes and procedures described below illustrate a general synthetic route to the compounds of general formula (I) of the present invention and are not intended to be limiting. Those skilled in the art will understand that the order of the transformations illustrated in the schemes can be modified in various ways. Therefore, the order of the transformations illustrated in the schemes is not intended to be limiting. In addition, interconversions of any substituents can be achieved before and / or after the illustrated transformations. These modifications can be accomplished by introduction of protecting groups, cleavage of protecting groups, exchange of functional groups, reduction or oxidation, halogenation, metalation, substitution, or other reactions known to those skilled in the art. These transformations also include those that introduce functionality that allows further interconversion of substituents. Appropriate protecting groups and their introduction and cleavage are well known to those skilled in the art (see, for example, P.G.M. Wuts and T.W. Greene in “Protective Groups in Organic Synthesis”, 4 th edition, Wiley 2006). Specific examples are described in the following paragraphs. Further, two or more consecutive steps can potentially be carried out in a “one-pot” reaction without work-up between the steps, as is well known to those skilled in the art, for example. The synthesis of the heteroaryl-substituted 2,6-difluorophenol compounds according to the present invention is preferably carried out according to the general synthetic sequence shown in Schemes 1-3.

[0028]

Chemical formula

[0029] Scheme 1: Route for the preparation of compounds of general formulas 8 and 9 (wherein A1, A2, and A3 have the same meanings as given in formula (I) above, X has the meaning of Cl, Br, I, mesylate, or triflate, Y has the meaning of Cl, Br, or I, and R has the meaning of alkyl). Preparation of starting materials (Scheme 1): The bicyclic compounds of general formula 1, 2, 3, 4, 5, 6 or 7 (Scheme 1) are commercially available or described in the literature.

[0030] <<Step 1→2 (Scheme 1)>> Halogenation reaction The conversion of the compound of general formula 1 to the compound of formula 2 is known to those skilled in the art. When Y = Br, the reaction can be carried out using reagents such as bromine, N-bromosuccinimide or copper(II) bromide. When Y = Cl, the reaction can be carried out using reagents such as N-chlorosuccinimide or chlorine. When Y = I, the reaction can be carried out using reagents such as iodine or N-iodosuccinimide.

[0031] <<Step 2→8 (Scheme 1)>> Reduction of ester to alcohol The conversion of the compound of general formula 2 to the alcohol of formula 8 is known to those skilled in the art and can be carried out using reagents such as lithium aluminum tetrahydride, sodium borohydride, calcium borohydride, diisobutylaluminum hydride, etc.

[0032] <<Step 3→8 (Scheme 1)>> Reduction of carboxylic acid to alcohol The conversion of the compound of general formula 3 to the alcohol of formula 8 is known to those skilled in the art and can be carried out using reagents such as lithium aluminum tetrahydride, etc. <<Step 4→8 (Scheme 1)>> Reduction of aldehyde to alcohol The conversion of the compound of general formula 4 to the alcohol of formula 8 is known to those skilled in the art and can be carried out using reagents such as sodium borohydride or lithium aluminum tetrahydride, etc. <<Step 5→6 (Scheme 1)>> Metal-catalyzed methylation reaction The conversion of a compound of general formula 5 to a compound of general formula 6 is known to those skilled in the art and can be carried out, for example, under Suzuki conditions using a reagent such as trimethylboroxine, a palladium catalyst such as (1,1'-bis(diphenylphosphino)ferrocene)palladium(II) dichloride or tetrakis(triphenylphosphine)palladium(0), and a base such as potassium carbonate in an organic solvent such as DMF or dioxane.

[0033] <<Step 7→6 (Scheme 1)>> Halogenation reaction The conversion of a compound of general formula 7 to a compound of general formula 6 is known to those skilled in the art. When Y = Br, the reaction can be carried out using a reagent such as bromine, N-bromosuccinimide or copper(II) bromide. When Y = Cl, the reaction can be carried out using a reagent such as N-chlorosuccinimide or chlorine. When Y = I, the reaction can be carried out using a reagent such as iodine or N-iodosuccinimide. <<Step 6→8 (Scheme 1)>> Hydroxylation of methyl group The conversion of a compound of general formula 6 to the alcohol of formula 8 is known to those skilled in the art and can be carried out using a reagent such as selenium(IV) dioxide or tert-butyl hydroperoxide / manganese triacetate.

[0034] <<Step 8→9 (Scheme 1)>> Conversion of hydroxy to halogen (Br, Cl, I), mesylate or triflate The conversion of alcohol 8 to the halogenated compound of formula 9 (when X = Cl) can be carried out using a chlorination reagent such as thionyl chloride, mesyl chloride / triethylamine, or triphenylphosphine / carbon tetrachloride. When X = Br, reagents such as phosphorus tribromide, trimethylsilyl bromide, hydrogen bromide, carbon tetrabromide / triphenylphosphine, or boron tribromide / triphenylphosphine can be used. When X = I, reagents such as boron trifluoride diethyl ether / potassium iodide, 1H-imidazole / iodine / triphenylphosphine can be used. When X = mesylate, reagents such as mesyl chloride and a base such as triethylamine can be used. When X = triflate, reagents such as trifluoromethanesulfonic anhydride and a base such as pyridine can be used.

[0035] <<Step 7→9 (Scheme 1)>> Radical halogenation of the methyl group The conversion of the compound of general formula 7 to the halide of formula 9 is known to those skilled in the art. Typically, a radical starter such as dibenzoyl peroxide or 2,2'-azobis(isobutyronitrile) and a halogenation reagent are used. When X = Br, the reaction can be carried out using a halogenation reagent such as N-bromosuccinimide. When X = Cl, the reaction can be carried out using a halogenation reagent such as N-chlorosuccinimide.

[0036]

Chemical formula

[0037] Preparation of starting materials (Scheme 2): Compounds of general formula 10, 11, 12, 13 or 14 (Scheme 2) are commercially available or described in the literature. 2,6-Difluorophenol is commercially available.

[0038] <<Step 2,6-difluorophenol → 10 (Scheme 2)>> Introduction of a phenol protecting group The phenol group of 2,6-difluorophenol can be masked using a suitable protecting group R1 that connects to the compound of general formula 10. The reaction conditions for such introduction of a suitable protecting group R1 are known to those skilled in the art (see, for example, Green, Wuts, “Protective groups in organic synthesis” 1999, John Wiley & Sons and references therein). Preferably, benzyl, para-methoxybenzyl and 3,4-methoxybenzyl are used as protecting groups during synthesis. <<Step 10 → 11 (Scheme 2)>> Introduction of a boronic acid or boronic ester The conversion of the compound of general formula 10 to the boronic acid derivative of formula 11 is known to those skilled in the art. When R = H, the reaction can be carried out using reagents such as triisopropyl borate and bases such as n-butyllithium, followed by acid workup of the reaction. When R-R=-C(CH3)2-C(CH3)2-, the reaction can be carried out using reagents such as bis(pinacol) diborane and catalysts such as Pt(N,N'-dicyclohexylimidazol-2-ylidene)(divinyltetramethylsiloxane).

[0039] <<Step 11 → 12 (Scheme 2)>> Removal of the phenol protecting group Removal of the protecting group R1 from the compound of formula 11 leads to the compound of formula 12. Such suitable reaction conditions for the removal of the protecting group R1 are known to those skilled in the art (see, for example, Green, Wuts, “Protective groups in organic synthesis” 1999, John Wiley & Sons and references therein). Preferably, benzyl, paramethoxybenzyl and 3,4-methoxybenzyl are used as protecting groups during the synthesis and they can be removed by hydrogenation. <<Step 13→14 (Scheme 2)>> Introduction of the phenol protecting group The compound of general formula 13 can be masked with a suitable protecting group R1 leading to the compound of general formula 14. Such suitable reaction conditions for the introduction of the protecting group R1 are known to those skilled in the art (see, for example, Green, Wuts, “Protective groups in organic synthesis” 1999, John Wiley & Sons and references therein). Preferably, benzyl, paramethoxybenzyl and 3,4-methoxybenzyl are used as protecting groups during the synthesis.

[0040] <<Step 14→11 (Scheme 2)>> Introduction of the boronic ester The conversion of the compound of general formula 14 to the boronic acid derivative of formula 11 is known to those skilled in the art. When R-R=-C(CH3)2-C(CH3)2-, the reaction can be carried out using reagents such as bis(pinacol)diborane, catalysts such as palladium(II) dichloride [1,1'-bis(diphenylphosphanyl)ferrocene] and bases such as potassium acetate. <<Step 13→12 (Scheme 2)>> Introduction of the boronic ester The conversion of a compound of general formula 13 to a boronic acid derivative of formula 12 is known to those skilled in the art. When R-R=-C(CH3)2-C(CH3)2-, the reaction can be carried out using reagents such as bis(pinacol)diborane, catalysts such as palladium(II) dichloride [1,1'-bis(diphenylphosphanyl)ferrocene], and bases such as potassium acetate.

[0041]

Chem.

[0042] Preparation of starting materials (Scheme 3): The amide compounds of general formula 15 (Scheme 3) can be commercially available, described in the literature or prepared analogously to the literature procedures.

[0043] <<Step 8 + 15 → 16 (Scheme 3)>> Mitsunobu reaction The conversion of a compound of general formula 8 and a compound of general formula 15 to a derivative of formula 16 is known to those skilled in the art. The reaction can be carried out using reagents such as diisopropyl azodicarboxylate / triphenylphosphine, di-tert-butyl azodicarboxylate / triphenylphosphine or diethyl azodicarboxylate / triphenylphosphine under Mitsunobu conditions.

[0044] <<Step 9 + 15 → 16 (Scheme 3)>> Alkylation of amide The conversion of the compounds of general formula 9 and the compounds of general formula 15 to the derivatives of formula 16 is known to those skilled in the art. When X = Cl, Br or I, the reaction can be carried out using a base such as potassium carbonate, cesium carbonate, sodium hydride or LDA in a solvent such as DMF, DMSO or acetonitrile. <<Step 16+11→17(Scheme 3)>> Palladium-catalyzed reaction using boronic acid The heteroaryl halide of formula 16 can be reacted with the boronic acid derivative 11 to obtain the compound of formula 17. The boronic acid derivative can be a boronic acid (R = H) or an ester of boronic acid, for example its isopropyl ester (R = -CH(CH3)2), preferably an ester derived from pinacol, where the boronic acid intermediate forms 2-aryl-4,4,5,5-tetramethyl-1,3,2-dioxoborolane (R-R = -C(CH3)2-C(CH3)2-). The coupling reaction is catalyzed by a palladium catalyst, such as a Pd(0) catalyst such as tetrakis(triphenylphosphine)palladium(0) [Pd(PPh3)4], tris(dibenzylideneacetone)di-palladium(0) [Pd2(dba)3], or a Pd(II) catalyst such as dichlorobis(triphenylphosphine)-palladium(II) [Pd(PPh3)2Cl2], XPhos Pd G2, Pd-Peppsi 2Me-Ipent Cl, palladium(II) acetate and triphenylphosphine, or [1,1'-bis(diphenylphosphino)ferrocene] palladium dichloride.

[0045] The reaction is preferably carried out in a mixture of a solvent such as 1,2-dimethoxymethane, dioxane, DMF, DME, THF, ethanol or isopropanol and water, in the presence of a base such as potassium carbonate, sodium bicarbonate, potassium acetate or potassium phosphate (Overview: D.G. Hall, Boronic Acids, 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim, ISBN 3-527-30991-8 and references in the same document). The reaction is carried out at a temperature in the range of room temperature (i.e., approximately 20 °C) to the boiling point of the corresponding solvent. Furthermore, subsequently, using a pressure tube and a microwave oven, the reaction can be carried out at a temperature above the boiling point. The reaction is preferably terminated after a reaction time of 1 to 36 hours.

[0046] <<Step 17→18 (Scheme 3)>> Removal of the phenol protecting group Removal of the protecting group R1 from the compound of formula 17 leads to the compound of formula 18. Such suitable reaction conditions for the removal of the protecting group R1 are known to those skilled in the art (see, for example, Green, Wuts, “Protective groups in organic synthesis” 1999, John Wiley & Sons and references in the same document). Preferably, benzyl, paramethoxybenzyl and 3,4-methoxybenzyl are used as protecting groups during synthesis and they can be removed by hydrogenation. <<Step 16→19 (Scheme 3)>> Introduction of the boronic ester The conversion of the compound of general formula 16 to the boronic acid derivative of formula 19 is known to those skilled in the art. When R-R=-C(CH3)2-C(CH3)2-, the reaction can be carried out using a reagent such as bis(pinacol)diborane, a catalyst such as palladium(II) dichloride [1,1'-bis(diphenylphosphanyl)ferrocene] and a base such as potassium acetate.

[0047] <<Step 19+13→18 (Scheme 3)>> Palladium-catalyzed reaction using boronic acid The aryl halide of formula 13 can be reacted with the boronic acid derivative 19 to obtain the compound of formula 18. The boronic acid derivative can be a boronic acid (R = H) or an ester of a boronic acid, such as its isopropyl ester (R = -CH(CH3)2), preferably an ester derived from pinacol, where the boronic acid intermediate forms 2-aryl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (R-R = -C(CH3)2-C(CH3)2-). The coupling reaction is catalyzed by a palladium catalyst, such as a Pd(0) catalyst like tetrakis(triphenylphosphine)palladium(0) [Pd(PPh3)4], tris(dibenzylideneacetone)di-palladium(0) [Pd2(dba)3], etc., or a Pd(II) catalyst like dichlorobis(triphenylphosphine)-palladium(II) [Pd(PPh3)2Cl2], XPhos Pd G2, Pd-Peppsi 2Me-Ipent Cl, palladium(II) acetate and triphenylphosphine, etc., or [1,1'-bis(diphenylphosphino)ferrocene] palladium dichloride. The reaction is preferably carried out in a mixture of a solvent such as 1,2-dimethoxymethane, dioxane, DMF, DME, THF, ethanol or isopropanol and water in the presence of a base such as potassium carbonate, sodium bicarbonate, potassium acetate or potassium phosphate (Overview: D.G. Hall, Boronic Acids, 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim, ISBN 3-527-30991-8 and references therein). The reaction is carried out at a temperature in the range of room temperature (i.e., approximately 20 °C) to the boiling point of the corresponding solvent. Furthermore, the reaction can be carried out at a temperature above the boiling point using a pressure tube and a microwave oven. The reaction is preferably terminated after a reaction time of 1 to 36 hours.

[0048] <<Step 16+12→18 (Scheme 3)>> Palladium-catalyzed reaction using boronic acid The heteroaryl halide of Formula 16 can be reacted with the boronic acid derivative 12 to obtain a compound of Formula 18. The boronic acid derivative can be a boronic acid (R = H) or an ester of boronic acid, such as its isopropyl ester (R = -CH(CH3)2), preferably an ester derived from pinacol, where the boronic acid intermediate forms 2-aryl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (R-R = -C(CH3)2-C(CH3)2-). The coupling reaction is catalyzed by a palladium catalyst, such as a Pd(0) catalyst like tetrakis(triphenylphosphine)palladium(0) [Pd(PPh3)4], tris(dibenzylideneacetone)di-palladium(0) [Pd2(dba)3], etc., or a Pd(II) catalyst like dichlorobis(triphenylphosphine)-palladium(II) [Pd(PPh3)2Cl2], XPhos Pd G2, Pd-Peppsi 2Me-Ipent Cl, palladium(II) acetate and triphenylphosphine, etc., or [1,1'-bis(diphenylphosphino)ferrocene] palladium dichloride. The reaction is preferably carried out in a mixture of a solvent such as 1,2-dimethoxymethane, dioxane, DMF, DME, THF, ethanol or isopropanol and water in the presence of a base such as potassium carbonate, sodium bicarbonate, potassium acetate or potassium phosphate (as outlined in D.G. Hall, Boronic Acids, 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim, ISBN 3-527-30991-8 and the references therein). The reaction is carried out at a temperature in the range of room temperature (i.e., approximately 20 °C) to the boiling point of the corresponding solvent. Further, the reaction can be carried out at a temperature above the boiling point using a pressure tube and a microwave oven. The reaction is preferably terminated after a reaction time of 1 to 36 hours.

[0049] <<Step 19 + 14 → 17 (Scheme 3)>> Palladium-catalyzed reaction using boronic acid The aryl halide of formula 14 can be reacted with the boronic acid derivative 19 to obtain the compound of formula 17. The boronic acid derivative can be a boronic acid (R = H) or an ester of a boronic acid, such as its isopropyl ester (R = -CH(CH3)2), preferably an ester derived from pinacol, where the boronic acid intermediate forms 2-aryl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (R-R = -C(CH3)2-C(CH3)2-). The coupling reaction is catalyzed by a palladium catalyst, such as a Pd(0) catalyst like tetrakis(triphenylphosphine)palladium(0) [Pd(PPh3)4], tris(dibenzylideneacetone)di-palladium(0) [Pd2(dba)3], etc., or a Pd(II) catalyst like dichlorobis(triphenylphosphine)-palladium(II) [Pd(PPh3)2Cl2], XPhos Pd G2, Pd-Peppsi 2Me-Ipent Cl, palladium(II) acetate and triphenylphosphine, or [1,1'-bis(diphenylphosphino)ferrocene] palladium dichloride. The reaction is preferably carried out in a mixture of a solvent such as 1,2-dimethoxymethane, dioxane, DMF, DME, THF, ethanol or isopropanol and water in the presence of a base such as potassium carbonate, sodium bicarbonate, potassium acetate or potassium phosphate (Overview: D.G. Hall, Boronic Acids, 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim, ISBN 3-527-30991-8 and references in the same literature). The reaction is carried out at a temperature in the range of room temperature (i.e., approximately 20 °C) to the boiling point of the corresponding solvent. Furthermore, the reaction can be carried out at a temperature above the boiling point using a pressure tube and a microwave oven. The reaction is preferably terminated after a reaction time of 1 to 36 hours.

[0050] General method for preparing the compound of formula (I) The present invention discloses the following method or process for preparing the compound of general formula (I) using compounds X1 and X2. [Chemical formula] The method for obtaining the compound of general formula (I) is characterized as follows. · In the compound of general formula X1, Z, A1, A2 and A3 have the same meanings as defined for the compound of general formula (I), · In the compound of general formula X1, Y can be Br, I or Cl, and it reacts with the compound of general formula X2 using a palladium catalyst and a base; · The reaction proceeds in a solvent or a solvent mixture at a temperature between ambient temperature and the boiling point of the solvent, preferably 50 °C to 120 °C. · The preparation of the compound of general formula (I) can be carried out in an aprotic or protic solvent or a solvent mixture, preferably 1,4-dioxane, tetrahydrofuran, or ethanol / water. · Preferred bases that can be used in the preparation of the compound of general formula (I) are sodium carbonate or cesium carbonate. · Preferred palladium catalysts for the preparation of the compound of general formula (I) are -chloro(2-dicyclohexylphosphino-2’,4’,6’-triisopropyl-1,1’-biphenyl)[2-(2’-amino-1,1’-biphenyl)]palladium(II) (second-generation XPhos precatalyst) or -1,3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)dichloropalladium(II).

[0051] Intermediate compound The present invention also discloses intermediate compounds useful in the preparation of the compound of general formula (I). In particular, the present invention discloses a compound of general formula Ia in which Z is as defined for the compound of general formula (I) above and Y can be Br, I or Cl.

[0052] [Chemical formula] The present invention discloses further intermediate compounds useful in the preparation of compounds of formula (I). In particular, the present invention also discloses compounds of formula (Ib) wherein A1, A2 and A3 are as defined for the compounds of the aforementioned general formula (I).

[0053]

Chemical formula

[0054]

Table 1

[0055] The features and advantages of the present invention will become apparent from the following examples given by way of illustration of the present invention without limiting its scope. Abbreviations

Table 2

Examples

[0056] (Example 1) Analytical HPLC method 1.1 Method A Analysis Column: Water; Sunfire C18 3.0×30mm, 2.5μm; Column Temperature: 60° Equipment: Agilent1200 equipped with DA and MS detectors

[0057]

Table 3

[0058] 1.2 Method B Analysis Column: Water; XBridge C18 3.0×30mm, 2.5μm; Column Temperature: 60° Equipment: Agilent1200 equipped with DA and MS detectors

Table 4

[0059] 1.3 Method C Analysis Column: Water; Sunfire C18 2.1×30mm, 2.5μm; Column Temperature: 60° Equipment: Waters Acquity equipped with DA and MS detectors

Table 5

[0060] 1.4 Method D Analysis Column: Sunfire C18 3.0×30mm, 2.5μm; Column Temperature: 60° Equipment: Waters Acquity, QDa detector

Table 6

[0061] 1.5 Method E Analysis Column: Sunfire C18 3.0×30mm, 2.5μm; Column Temperature: 60° Equipment: Waters Acquity, QDa detector

Table 7

[0062] (Example 2) Preparation of Intermediate 2.1 Intermediate I Intermediate I.1 (General Procedure) 1-[(2-Bromo-1,3-thiazol-5-yl)methyl]-3-methyl-1,2,3,4-tetrahydropyrimidine-2,4-dione [Chemical Structure Diagram]

[0063] 3-Methyl-1,2,3,4-tetrahydropyrimidine-2,4-dione (3.00 g; 23.79 mmol) and 3-bromothiazole-5-methanol (5.08 g; 26.17 mmol) were dissolved in THF (50 mL) and DMF (20 mL). TPP (polymer-bound, 3 mmol / g; 10.50 g; 30.92 mmol) was added, and the reaction mixture was cooled in an ice bath. DTAD (7.12 g; 30.92 mmol) was added. After stirring overnight at RT, the reaction mixture was filtered, and THF was evaporated. The residue was poured into water and extracted several times with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; DCM / MeOH, from 100 / 0 to 95 / 5) to obtain the product. C9H8BrN3O2S (M = 302.2 g / mol) ESI-MS: 302 [M+H] + R t (HPLC): 0.67 min (Method A) Yield: 4.03 g; 56% 1 1H NMR (400 MHz, DMSO-d6) δ ppm: 3.16 (s, 3H), 5.10 (s, 2H), 5.76 (d, J = 7.86 Hz, 1H), 7.73 (s, 1H), 7.84 (d, J = 7.86 Hz, 1H).

[0064] The intermediate compounds I.2 and I.3 shown in the table below were prepared using a procedure similar to that described for intermediate I.1 and appropriate starting materials. As will be understood by those skilled in the art, these similar examples may include variations in common reaction conditions. [Table 8]

[0065] [Table 9]

[0066] Intermediate I.4 1-[(5-Bromo-1,3,4-thiadiazol-2-yl)methyl]-3-methyl-1,2,3,4-tetrahydropyrimidine-2,4-dione [Chemical formula] To a stirred mixture of (5-bromo-1,3,4-thiadiazol-2-yl)methanol (0.20 g; 1.03 mmol) and TEA (0.17 mL; 1.23 mmol) in DCM (5 mL), methanesulfonyl chloride (0.10 mL; 1.23 mmol) was added dropwise. After stirring at RT for 1 h, another portion of methanesulfonyl chloride (0.04 mL) was added and stirring was continued for an additional 1 h. The reaction mixture was diluted with DCM and water. The organic layer was separated through a phase separation cartridge and evaporated. The residue was taken up in DMF (3 mL), and 3-methyl-1,2,3,4-tetrahydropyrimidine-2,4-dione (0.10 g; 0.79 mmol) and K2CO3 (0.28 g; 2.03 mmol) were added. The reaction mixture was stirred overnight at RT, filtered, and purified by column chromatography (reverse phase; Sunfire C18; water / ACN / TFA) to give the product. C8H7BrN4O2S (M = 303.1 g / mol) ESI-MS: 303 / 305(Br)[M+H] + R t(HPLC): 0.71 minutes (Method A) Yield: 0.14 g; 58%

[0067] Intermediate I.5 1-[(5-Bromo-1,2-thiazol-3-yl)methyl]-3-methyl-1,2,3,4-tetrahydropyrimidine-2,4-dione

Chem.

[0068] 3-Methyl-1,2,3,4-tetrahydropyrimidine-2,4-dione (0.04 g; 0.32 mmol), 5-bromo-3-(bromomethyl)-1,2-thiazole (0.09 mL; 0.35 mmol) and K2CO3 (0.10 g; 0.70 mmol) in dry DMF (2 mL) were stirred at RT for 2 h. The reaction mixture was partitioned between EtOAc and water. The organic layer was washed with brine and water, dried over Na2SO4, filtered and concentrated under reduced pressure. C9H8BrN3O2S (M = 302.2 g / mol) ESI-MS: 302 [M+H] + R t (HPLC): 0.71 minutes (Method A) Yield: 0.09 g; 97% 1 1H NMR (400 MHz, DMSO-d6) δ ppm: 3.15 (s, 3 H), 5.07 (s, 2 H), 5.76 (d, J = 7.86 Hz, 1 H), 7.56 (s, 1 H), 7.77 (d, J = 7.86 Hz, 1 H).

[0069] 2.2 Intermediate II [3-(Benzyloxy)-2,4-difluorophenyl]boronic acid

Chem.

[0070] 2-(Benzyloxy)-1,3-difluorobenzene (0.50 g; 2.27 mmol) in THF (10 mL) was cooled to -78 °C. n-BuLi (2.5 mol / L in THF; 1.36 mL; 3.63 mmol) was added dropwise and the mixture was stirred at low temperature for 50 minutes. Triisopropyl borate (0.73 mL; 3.63 mmol) was added dropwise and the mixture was stirred at -78 °C for 10 minutes and then for an additional 30 minutes without cooling. The reaction was quenched with HCl (aqueous solution; 4 mol / L; 5 mL) and stirred for 10 minutes. The mixture was poured into water and extracted several times with EtOAc. The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; DCM / MeOH containing a small amount of AcOH, from 100 / 0 to 96 / 4) to give the product. C 13 H 11 BF2O3 (M = 264.0 g / mol) ESI-MS: 263 [M-H] - R t (HPLC): 0.75 min (Method B) Yield: 0.25 g; 42% 1 1H NMR (400 MHz, DMSO-d6) δ ppm: 5.12 (s, 2 H), 7.05 (ddd, J = 10.27, 8.62, 1.39 Hz, 1 H), 7.25 (dt, J = 8.40, 6.57 Hz, 1 H), 7.30 - 7.56 (m, 5 H), 8.23 (br s, 2 H).

[0071] 2.3 Intermediate III Intermediate III.1 (2,4-Difluoro-3-hydroxyphenyl)boronic acid

Chemical Structure

[0072] Intermediate II.1 (0.10 g; 0.38 mmol) in a mixture of THF and MeOH (3 mL each) was hydrogenated at RT for 3 h using 10% Pd / C (0.02 g) in a parr apparatus. The reaction mixture was concentrated under reduced pressure. The residue was purified by column chromatography (reverse phase; Sunfire C18; water / ACN / TFA) to obtain the product. C6H5BF2O3 (M = 173.9 g / mol) ESI-MS: 173 [M-H] - R t (HPLC): 0.07 min (Method B) Yield: 0.05 g; 77% 1 1H NMR (400 MHz, DMSO-d6) δ ppm: 6.90 - 7.00 (m, 2 H), 8.11 (br s, 2 H), 9.81 (br s, 1 H).

[0073] 2.4 Intermediate IV Intermediate IV.1 (General procedure) 2,4-Difluoro-3-[(4-methoxyphenyl)methoxy]benzaldehyde

Chemical formula

[0074] 1-(Chloromethyl)-4-methoxybenzene (0.19 mL; 1.39 mmol) was added to a mixture of 2,4-difluoro-3-hydroxybenzaldehyde (0.20 g; 1.27 mmol) and K2CO3 (0.27 g; 1.94 mmol) in ACN (5 mL). After stirring at 60 °C for 3 h, the reaction mixture was filtered, diluted with DMF / water / TFA, and purified by column chromatography (reverse phase; Sunfire C18; water / ACN / TFA). ACN was evaporated from the desired fractions, and the product was partitioned between DCM and water. The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. C 15 H 12 F2O3 (M = 278.3 g / mol) ESI-MS: 277 [M-H] - R t (HPLC): 1.12 minutes (Method A) Yield: 0.22 g; 63% 1 H NMR (400 MHz, DMSO-d6) δ ppm: 3.75 (s, 3 H), 5.14 (s, 2 H), 6.93 (d, J = 8.74 Hz, 2 H), 7.26 - 7.39 (m, 3 H), 7.59 (ddd, J = 8.81, 7.41, 6.08 Hz, 1 H), 10.11 (s, 1 H).

[0075] The intermediate compounds IV.2 and IV.4 shown in the following table were prepared using a procedure similar to that described for intermediate IV.1 and appropriate starting materials. As will be understood by those skilled in the art, these similar examples may include variations in general reaction conditions.

Table 10

[0076]

Table 11

[0077] 2.5 Intermediate V Intermediate V.1 ({2,4-Difluoro-3-[(4-methoxyphenyl)methoxy]phenyl}methylidene)hydroxylamine

Chemical formula

[0078] Intermediate IV.1 (0.22 g; 0.79 mmol) and NaOAc (0.08 g; 1.03 mmol) in MeOH (8 mL) and water (3 mL) were stirred at RT. Hydroxylamine hydrochloride (0.04 mL; 1.03 mmol) was added and the reaction mixture was heated to 80 °C for 1.5 h. The reaction mixture was concentrated under reduced pressure and absorbed into water / EtOAc. The aqueous layer was extracted several times with EtOAc. The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. C 15 H 13 F2NO3 (M = 293.3 g / mol) ESI-MS: 294 [M+H] + R t (HPLC): 1.06 min (Method A) Yield: 0.25 g; quantitative 1 1H NMR (400 MHz, DMSO-d6) δ ppm: 3.75 (s, 3 H), 5.10 (s, 2 H), 6.90 - 6.96 (m, 2 H), 7.10 - 7.19 (m, 1 H), 7.31 - 7.37 (m, 2 H), 7.42 (ddd, J = 8.81, 7.79, 6.08 Hz, 1 H), 8.14 (s, 1 H), 11.59 (s, 1 H).

[0079] 2.6 Intermediate VI Intermediate VI.1 1-[(3-{2,4-Difluoro-3-[(4-methoxyphenyl)methoxy]phenyl}-1,2-oxazol-5-yl)methyl]-3-methyl-1,2,3,4-tetrahydropyrimidine-2,4-dione

Chemical Structure

[0080] Intermediate V.1 (0.10 g; 0.34 mmol), 3-methyl-1-(prop-2-yn-1-yl)-1,2,3,4-tetrahydropyrimidine-2,4-dione (0.06 g; 0.34 mmol) and TEA (0.01 mL; 0.03 mmol) in t-BuOH (4 mL) and water (4 mL) were stirred at RT. NaOCl (0.45 mL; 0.58 mmol) was added. After stirring at RT for 2.5 h, EtOAc and water were added. The organic layer was separated, dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (reverse phase; Sunfire C18; water / ACN / TFA) to give the product. C 23 H 19 F2N3O5 (M = 455.4 g / mol) ESI-MS: 456 [M+H] + R t (HPLC): 1.07 min (Method A) Yield: 0.02 g; 12% 1 H NMR (400 MHz, DMSO-d6) δ ppm: 3.17 (s, 3 H), 3.75 (s, 3 H), 5.14 (s, 2 H), 5.20 (s, 2 H), 5.81 (d, J = 7.86 Hz, 1 H), 6.89 (d, J = 2.66 Hz, 1 H), 6.93 (d, J = 8.74 Hz, 2 H), 7.26 (td, J = 9.57, 1.77 Hz, 1 H), 7.35 (d, J = 8.62 Hz, 2 H), 7.56 (ddd, J = 8.90, 7.83, 5.96 Hz, 1 H), 7.86 (d, J = 7.98 Hz, 1 H).

[0081] Intermediate VI.2 (General Procedure) 1-[(5-{2,4-difluoro-3-[(4-methoxyphenyl)methoxy]phenyl}-1,2-oxazol-3-yl)methyl]-3-methyl-1,2,3,4-tetrahydropyrimidine-2,4-dione

Chemical Structure

[0082] Intermediate IX.1 (0.10 g; 0.03 mmol) in DMF (2 mL), 3-methyl-1,2,3,4-tetrahydropyrimidine-2,4-dione (0.01 g; 0.05 mmol) and K2CO3 (0.01 g; 0.08 mmol) were stirred at RT for 1 h and at 50 °C for 30 min. The reaction mixture was filtered and purified by column chromatography (reverse phase; Sunfire C18; water / ACN / TFA) to obtain the product. C 23 H 19 F2N3O5 (M = 455.4 g / mol) ESI-MS: 456 [M+H] + R t (HPLC): 1.07 min (Method A) Yield: 0.01 g; 53%

[0083] The following Intermediate Compound VI.3 was prepared using a procedure similar to that described for Intermediate VI.2, using appropriate starting materials. As will be understood by those skilled in the art, these similar examples may include variations in common reaction conditions.

Table 12

[0084]

Table 13

[0085] 2.7 Intermediate VII Intermediate VII.1 Ethyl 5-(2,4-difluoro-3-hydroxyphenyl)-1,2-oxazole-3-carboxylate

Chemical Structure

[0086] Intermediate VII.2 2,6-Difluoro-3-[5-(hydroxymethyl)-1,3,4-thiadiazol-2-yl]phenol

Chemical Structure

[0087] The reaction was carried out under a nitrogen atmosphere. (5-Bromo-1,3,4-thiadiazol-2-yl)methanol (4.00 g; 20.51 mmol), Intermediate III.1 (4.60 g; 24.61 mmol), and Na2CO3 (5.43 g; 51.27 mmol) were dissolved in EtOH (50 mL) and water (10 mL). Pd-Peppsi 2Me-Ipent Cl (0.86 g; 1.03 mmol) was added, and the mixture was stirred at 80° overnight. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was absorbed in water, and the resulting precipitate was filtered to obtain the product. C9H6F2N2O2S (M = 244.2 g / mol) ESI-MS: 245[M + H] + R t (HPLC): 0.72 min (Method A) Yield: 2.66 g; 53% 1 1H NMR (400 MHz, DMSO-d6) δ ppm: 4.93 (d, J = 4.69 Hz, 2 H), 6.27 (br t, J = 5.39 Hz, 1 H), 7.19 - 7.34 (m, 1 H), 7.65 (ddd, J = 8.84, 7.57, 5.89 Hz, 1 H), 10.75 (s, 1 H).

[0088] 2.8 Intermediate VIII Intermediate VIII.1 (5-{2,4-Difluoro-3-[(4-methoxyphenyl)methoxy]phenyl}-1,2-oxazol-3-yl)methanol

Chemical Structure

[0089] Intermediate IV.2 (0.03 g; 0.07 mmol) in THF (3 mL) was cooled to -20 °C. LiAlH4 (solution in THF; 1 mol / L; 0.05 mL; 0.05 mol / L) was added dropwise and stirred at low temperature for 30 minutes. Two drops of water and NaOH (aqueous solution; 4 mol / L) were added and stirred at RT for an additional 20 minutes. The reaction mixture was filtered, diluted with water, and purified by column chromatography (reverse phase; Sunfire C18; water / ACN / TFA) to obtain the product. C 18 H 15 F2NO4 (M = 347.3 g / mol) ESI-MS: 348 [M+H] + R t (HPLC): 1.04 minutes (Method A) Yield: 0.02 g; 70% 1 1H NMR (400 MHz, DMSO-d6) δ ppm: 3.75 (s, 3 H), 4.57 (d, J = 6.08 Hz, 2 H), 5.16 (s, 2 H), 5.55 (t, J = 6.02 Hz, 1 H), 6.82 (d, J = 3.55 Hz, 1 H), 6.93 (d, J = 8.74 Hz, 2 H), 7.30 (ddd, J = 10.36, 8.90, 1.90 Hz, 1 H), 7.36 (d, J = 8.62 Hz, 2 H), 7.63 (ddd, J = 8.90, 7.83, 5.83 Hz, 1 H).

[0090] 2.9 Intermediate IX Intermediate IX.1 3-(Chloromethyl)-5-{2,4-difluoro-3-[(4-methoxyphenyl)methoxy]phenyl}-1,2-oxazole

Chemical Structure

[0091] Intermediate IX.2 (General procedure) 5-(Chloromethyl)-2-{2,4-difluoro-3-[(4-methoxyphenyl)methoxy]phenyl}-1,3-thiazole

Chemical formula

[0092] Intermediate XI.1 (0.99 g; 2.72 mmol) and TEA (0.60 mL; 4.33 mmol) in DCM (10 mL) were stirred in an ice bath. Methanesulfonyl chloride (0.32 mL; 4.13 mmol) was added dropwise. After stirring at low temperature for 5 min, the reaction mixture was stirred at RT overnight. The reaction mixture was diluted with additional DCM and washed with diluted citric acid. The organic layer was separated using a phase separation cartridge and evaporated. C 18 H 14 ClF2NO2S (M = 381.8 g / mol) ESI-MS: 382 [M+H] + R t (HPLC): 1.22 min (Method A) Yield: 1.01 g; 97% 11H NMR (400 MHz, DMSO-d6) δ ppm: 3.75 (s, 3 H), 5.16 (s, 2 H), 5.18 (s, 2 H), 6.93 (d, J = 8.74 Hz, 2 H), 7.28 (td, J = 9.63, 1.90 Hz, 1 H), 7.36 (d, J = 8.62 Hz, 2 H), 7.89 (ddd, J = 9.00, 8.05, 6.02 Hz, 1 H), 8.03 (d, J = 2.28 Hz, 1 H).

[0093] The following intermediate compound IX.3 was prepared using a procedure similar to that described for intermediate IX.2 and appropriate starting materials. As will be understood by those skilled in the art, these similar examples may include variations in common reaction conditions. [Table 14]

[0094] 2.10 Intermediate X Intermediate X.1 2,4-Difluoro-N’,3-dihydroxybenzene-1-carboximidamide [Chemical formula] 2,4-Difluoro-3-hydroxybenzonitrile (0.03 g; 0.20 mmol) in EtOH (0.5 mL) was stirred at RT. Hydroxylamine hydrochloride (0.03 g; 0.40 mmol) and TEA (0.06 mL; 0.04 mmol) in EtOH (1 mL) were added dropwise. After stirring at RT for 20 h, the reaction mixture was concentrated under reduced pressure and used further as the crude product. C7H6F2N2O2 (M = 188.1 g / mol)

[0095] 2.11 Intermediate XI Intermediate XI.1 (2-{2,4-Difluoro-3-[(4-methoxyphenyl)methoxy]phenyl}-1,3-thiazol-5-yl)methanol [Chemical]

[0096] Intermediate IV.3 (3.00 g; 9.11 mmol), bis(pinacolato)diboron (3.50 g; 13.78 mmol), KOAc (2.30 g; 23.44 mmol) and Pd(dppf)2×CH2Cl2 (0.75 g; 0.92 mmol) in dioxane (50 mL) were stirred at 90 °C overnight. Additional bis(pinacolato)diboron (0.70 g; 2.76 mmol) was added and the reaction mixture was stirred at 100 °C for 2 h. After cooling to RT, Pd(dppf)2×CH2Cl2 (0.38 g; 0.46 mmol), (2-bromo-1,3-thiazol-5-yl)methanol (1.80 g; 9.28 mmol) and Na2CO3 (aqueous solution; 2 mol / L; 15.00 mL; 30.00 mmol) were added and the reaction mixture was stirred for 6 h while refluxing. The mixture was cooled to RT, diluted with EtOAc and filtered through Celite®. The filtrate was concentrated under reduced pressure. The residue was partitioned between EtOAc and water. The organic layer was separated, dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; cyclohexane / EtOAC, 2:1 - 1:1). Et2O was added to the fraction where it evaporated, stirred and filtered. The precipitate was dried in air. C 18 H 15 F2NO3S (M = 363.4 g / mol) ESI-MS: 364 [M+H] + R t (HPLC): 1.07 min (Method A) Yield: 1.10 g; 30% 11H NMR (400 MHz, DMSO-d6) δ ppm: 3.75 (s, 3 H), 4.75 (d, J = 5.70 Hz, 2 H), 5.16 (s, 2 H), 5.64 (t, J = 5.77 Hz, 1 H), 6.93 (d, J = 8.74 Hz, 2 H), 7.26 (td, J = 9.63, 1.77 Hz, 1 H), 7.36 (d, J = 8.62 Hz, 2 H), 7.79 - 7.83 (m, 1 H), 7.86 (ddd, J = 8.93, 8.11, 6.02 Hz, 1 H).

[0097] 2.12 Intermediate XII Intermediate XII.1 (General Procedure) 1-[(2-{2,4-Difluoro-3-[(4-methoxyphenyl)methoxy]phenyl}-1,3-thiazol-5-yl)methyl]-3-ethyl-1,2,3,4-tetrahydropyrimidine-2,4-dione

[0098] [Chemical Structure] Intermediate VI.3 (0.03 g; 0.05 mmol), iodoethane (0.01 mL; 0.16 mmol) and K2CO3 (0.03 g; 0.16 mmol) in DMF (3 mL) were stirred at 50 °C for 2 h. The reaction mixture was diluted with water, filtered and purified by column chromatography (reverse phase; Sunfire C18; water / ACN / TFA) to obtain the product. C 24 H 21 F2N3O4S (M = 485.5 g / mol) R t (HPLC): 1.12 min (Method A) Yield: 0.02 g; 64%

[0099] The following intermediate compounds XII.2 to XII.7 were prepared using a procedure similar to that described for intermediate XII.1 and appropriate starting materials. As will be understood by those skilled in the art, these similar examples may include variations in general reaction conditions.

[0100]

Table 15

[0101]

Table 16

[0102] 2.13 Intermediate XIII Intermediate XIII.1 (General Procedure) 1-[(5-Bromo-1,3,4-thiadiazol-2-yl)methyl]-1,2,3,4-tetrahydropyrimidine-2,4-dione

Chem.

[0103] The reaction was carried out under an argon atmosphere. 1,2,3,4-Tetrahydropyrimidine-2,4-dione (0.50 g; 4.46 mmol) and (E)-(trimethylsilyl N-(trimethylsilyl)ethanimidate (2.75 mL; 11.25 mmol) in ACN (15 mL) were stirred at RT for 5 h. (5-Bromo-1,3,4-thiadiazol-2-yl)methyl methanesulfonat (1.40 g; 5.13 mmol) and tetrabutylazanium iodide (0.17 g; 0.46 mmol) were added. After stirring at 80 °C for 6 h and overnight at RT, 30 mL of water was added slowly. The resulting precipitate was filtered and washed with water, ACN and Et2O. C7H5BrN4O2S (M = 289.1 g / mol) Yield: 0.29 g; 23%

[0104] The following intermediate compounds XIII.2 - XIII.4 were prepared using a procedure similar to that described for intermediate XIII.1 and appropriate starting materials. As will be understood by those skilled in the art, similar examples can include variations in common reaction conditions. [Table 17]

[0105] [Table 18]

[0106] 2.14 Intermediate XIV Intermediate XIV.1 1 - [(5 - Bromo - 1,3,4 - thiadiazol - 2 - yl)methyl] - 6 - methyl - 1,2,3,4 - tetrahydropyrimidine - 2,4 - dione

[0107] [Chemical formula] (5 - Bromo - 1,3,4 - thiadiazol - 2 - yl)methyl methanesulfonate (0.20 g; 0.73 mmol) and 6 - methyl - 1,2,3,4 - tetrahydropyrimidine - 2,4 - dione (0.18 g; 1.46 mmol) were dissolved in DMF (5 mL), and K2CO3 (0.25 g; 1.83 mmol) was added. After stirring at RT for 2 h, iodoethane (0.15 mL; 1.83 mmol) was added and stirring was continued at 50 °C for an additional 1 h. The reaction mixture was purified by column chromatography (reverse phase; Sunfire C18; water / ACN / TFA) to obtain the product. C 10 H 11 BrN4O2S (M = 331.2 g / mol) ESI - MS: 331 [M + H] + R t (HPLC): 0.83 min (Method A) Yield: 0.03 g; 14% 1 1H NMR (400 MHz, DMSO-d6) δ ppm: 1.08 (t, J = 7.03 Hz, 3 H), 2.31 (d, J = 0.63 Hz, 3 H), 3.82 (q, J = 6.97 Hz, 2 H), 5.42 (s, 2 H), 5.71 (q, J = 0.63 Hz, 1 H).

[0108] 2.15 Intermediate XV Intermediate XV.1 1-[(2-Bromo-1,3-thiazol-5-yl)methyl]-1,2,3,4-tetrahydropyrimidine-2,4-dione

[0109]

Chemical Structure

[0110] 2.16 Intermediate XVI Intermediate XVI.1 (General Procedure) 1-[(2-{2,4-Difluoro-3-[(4-methoxyphenyl)methoxy]phenyl}-1,3-thiazol-5-yl)methyl]piperidin-2-one [Chemical Structure Diagram]

[0111] Piperidin-2-one (0.03 g; 0.26 mmol) and sodium hydride (0.01 g; 0.26 mmol) in DMF (2 mL) were stirred at RT for 10 minutes. Intermediate IX.2 (0.04 g; 0.10 mmol) was added. After stirring for 2 h, the mixture was diluted with water and MeOH and purified by column chromatography (reverse phase; Sunfire C18; water / ACN / TFA) to obtain the desired product. C 23 H 22 F2N2O3S (M = 444.5 g / mol) ESI-MS: 445 [M+H] + R t (HPLC): 1.11 minutes (Method A) Yield: 0.02 g; 52%

[0112] The following Intermediate Compounds XVI.2 - XVI.4 shown below were prepared using a procedure similar to that described for Intermediate XVI.1, using appropriate starting materials. As will be understood by those skilled in the art, these similar examples may include variations in general reaction conditions.

[0113] [Table 19]

[0114]

Table 20

[0115] 2.17 Intermediate XVII Intermediate XVII.1 tert-Butyl N-(3-{[(2-{2,4-Difluoro-3-[(4-methoxyphenyl)methoxy]phenyl}-1,3-thiazol-5-yl)methyl]amino}propyl)-N-methylcarbamate

[0116]

Chem.

[0117] 2.18 Intermediate XVIII Intermediate XVIII.1 (General procedure) 1-[(2-Bromo-1,3-thiazol-5-yl)methyl]-2,3-dihydro-1H-indol-2-one

Chem.

[0118] 2.19 Intermediate XIX Intermediate XIX.1 (General procedure) 1-Ethyl-7-methyl-2,3,6,7-tetrahydro-1H-purine-2,6-dione

Chemical Structure

[0119] Ethyl 4-amino-1-methyl-1H-imidazole-5-carboxylate (0.20 g; 1.15 mmol), (bromomethyl)-1,3-thiazole (0.20 g; 1.15 mmol) and isocyanatoethane (0.16 mL; 1.95 mmol) in pyridine (1 mL) were stirred at 70 °C for 2 h, and KOtBu (0.20 g; 1.72 mmol) was added. After stirring overnight at 70 °C, the reaction mixture was quenched with MeOH, concentrated under reduced pressure, absorbed into DMF and water, and purified by column chromatography (reverse phase; Sunfire C18; water / ACN / TFA) to give the desired product. C8H 10 N4O2 (M = 194.2 g / mol) ESI-MS: 195 [M+H] + R t (HPLC): 0.28 min (Method C) Yield: 0.10 g; 43% 1 H NMR (400 MHz, DMSO-d6) δ ppm: 1.10 (t, J = 6.97 Hz, 3 H), 3.81 - 3.90 (m, 5 H), 7.90 (s, 1 H), 11.77 (s, 1 H).

[0120] The following intermediate compounds were prepared using procedures similar to those described for Intermediate XIX.1, using appropriate starting materials. As will be understood by those skilled in the art, these similar examples may include variations in the general reaction conditions. [Table 21]

[0121] [Table 22]

[0122] 2.20 Intermediate XX Intermediate XX.1 (General Procedure) 3-[(5-Bromo-1,3,4-thiadiazol-2-yl)methyl]-1-ethyl-7-methyl-2,3,6,7-tetrahydro-1H-purine-2,6-dione

[0123]

Chem.

[0124] The following intermediate compounds were prepared using procedures similar to those described for Intermediate XX.1, using appropriate starting materials. As will be appreciated by those skilled in the art, these similar examples may include variations in the general reaction conditions.

Table 23

[0125] (Example 3) Preparation of Exemplary Compounds 3.1 General Procedure 1-{[2-(2,4-Difluoro-3-hydroxyphenyl)-1,3-thiazol-5-yl]methyl}-3-methyl-1,2,3,4-tetrahydropyrimidine-2,4-dione (Exemplary Compound 1) [Chemistry]

[0126] To a stirred mixture of Intermediate I.1 (0.40 g; 1.32 mmol), Intermediate III.1 (0.35 g; 2.01 mmol) and Cs2CO3 (1.10 g; 3.38 mmol) in water (1 mL) and EtOH (4 mL), Pd-Peppsi 2Me-IpentCl (0.06 g; 0.07 mmol) was added. After stirring at 100 °C for 1 h, the reaction mixture was filtered through Celite® and washed with EtOH. The filtrate was concentrated under reduced pressure. The residue was taken up in DCM and water. The organic layer was evaporated and the resulting precipitate was filtered off, washed with water, MeOH and Et2O to afford the desired product. C 15 H 11 F2N3O3S (M = 351.3 g / mol) ESI-MS: 352 [M+H] + R t (HPLC): 0.84 min (Method A) Yield: 0.43 g; 92% 1 1H NMR (400 MHz, DMSO-d6) δ ppm: 3.17 (s, 3 H), 5.20 (s, 2 H), 5.77 (d, J = 7.98 Hz, 1 H), 7.18 (td, J = 9.57, 1.90 Hz, 1 H), 7.60 (ddd, J = 8.93, 7.92, 5.96 Hz, 1 H), 7.90 (d, J = 7.86 Hz, 1 H), 8.00 (d, J = 2.28 Hz, 1 H), 10.51 - 10.71 (m, 1 H).

[0127] 3.2 Exemplary Compounds (Ex.) 2 - 14 The following intermediate compounds were prepared using procedures similar to those described above for 3.1, using appropriate starting materials. As will be appreciated by those skilled in the art, these similar examples may include variations in the general reaction conditions.

[0128]

Table 24

[0129]

Table 25

[0130]

Table 26

[0131] 3.3 Exemplary Compound 15 1-{[5-(2,4 - Difluoro - 3 - hydroxyphenyl)-1,3 - thiazol - 2 - yl]methyl}-3 - methyl - 1,2,3,4 - tetrahydropyrimidine - 2,4 - dione

[0132]

Chemical Structure

[0133] 3-Bromo-2,6-difluorophenol (0.04 g; 0.18 mmol), bis(pinacolato)diboron (0.05 g; 0.18 mmol), XPhos Pd G2 (0.01 g; 0.01 mmol), XPhos (0.01 g; 0.01 mmol), and KOAc (0.04 g; 0.36 mmol) in EtOH (2 mL) were stirred in a microwave at 120 °C for 10 minutes. Intermediate I.2 (0.03 g; 0.12 mmol), Na2CO3 (aqueous solution; 2 mol / L; 0.18 mL; 0.36 mmol), and XPhos Pd G2 (0.01 g; 0.01 mmol) were added, and the mixture was stirred in a microwave at 120 °C for an additional 10 minutes. The reaction mixture was diluted with DMF and purified by column chromatography (reverse phase; Sunfire C18; water / ACN / TFA) to obtain the desired product. C 15 H 11 F2N3O3S (M = 351.3 g / mol) ESI-MS: 352 [M+H] + R t (HPLC): 0.61 min (Method D) Yield: 0.01 g; 31% 1 H NMR (400 MHz, DMSO-d6) δ ppm: 3.17 (s, 3 H), 5.29 (s, 2 H), 5.80 (d, J = 7.86 Hz, 1 H), 7.09 - 7.25 (m, 2 H), 7.87 (d, J=7.86 Hz, 1 H), 8.10 (s, 1 H), 10.49 (s, 1 H).

[0134] 3.4 Exemplary Compound 16 1-{[3-(2,4-Difluoro-3-hydroxyphenyl)-1,2-oxazol-5-yl]methyl}-3-methyl-1,2,3,4-tetrahydropyrimidine-2,4-dione

Chemical Structure

[0135] TFA (1.00 mL; 12.96 mmol) and Intermediate VI.1 (0.40 g; 1.32 mmol) in DCM (3 mL) were stirred at RT for 20 min. The reaction mixture was concentrated under reduced pressure. The residue was purified by column chromatography (reverse phase; Sunfire C18; water / ACN / TFA) to afford the desired product. C 15 H 11 F2N3O4 (M = 335.3 g / mol) ESI-MS: 336 [M+H] + R t (HPLC): 0.83 min (Method A) Yield: 0.01 g; 72% 1 H NMR (400 MHz, DMSO-d6) δ ppm: 3.17 (s, 3 H), 5.20 (s, 2 H), 5.82 (d, J = 7.86 Hz, 1 H), 6.89 (d, J = 2.66 Hz, 1 H), 7.10 - 7.22 (m, 1 H), 7.28 (ddd, J = 8.71, 7.57, 5.89 Hz, 1 H), 7.86 (d, J = 7.86 Hz, 1 H), 10.57 (s, 1 H).

[0136] The following exemplary compounds 17 - 22 were prepared using procedures similar to those described above for 3.4 using appropriate starting materials. As will be appreciated by those skilled in the art, these similar examples may include variations in the general reaction conditions.

Table 27

[0137] Analytical data for the compounds described in the table above:

Table 28

[0138] 3.5 Exemplary Compound 23 1-{[3-(2,4-Difluoro-3-hydroxyphenyl)-1,2,4-oxadiazol-5-yl]methyl}-3-methyl-1,2,3,4-tetrahydropyrimidine-2,4-dione [Chemical Structure] A mixture of 2-(3-methyl-2,4-dioxo-1,2,3,4-tetrahydropyrimidin-1-yl)acetic acid (0.04 g; 0.20 mmol), EDC hydrochloride (0.04 g; 0.20 mmol) and HOBT (0.03 g; 0.24 mmol) in DMF (1 mL) was stirred at RT for 15 min. This mixture was added to Intermediate X.1 (0.04 g; 0.20 mmol) in DMF (1 mL). The resulting reaction mixture was stirred at 95 °C for 2 h and then at RT overnight. The reaction mixture was diluted with DMF / water, filtered and purified by column chromatography (reverse phase; Sunfire C18; water / ACN / TFA) to obtain the desired product. C 14 H 10 F2N2O4 (M = 336.3 g / mol) ESI-MS: 337 [M+H] + R t (HPLC): 0.47 min (Method C) Yield: 0.01 g; 6% 1 1H NMR (400 MHz, DMSO-d6) δ ppm: 3.17 (s, 3 H), 5.40 (s, 2 H), 5.87 (d, J = 7.98 Hz, 1 H), 7.22 (td, J = 9.54, 1.71 Hz, 1 H), 7.40 (ddd, J = 8.87, 7.35, 5.96 Hz, 1 H), 7.88 (d, J = 7.98 Hz, 1 H), 10.67 (s, 1 H).

[0139] 3.6 Exemplary Compound 24 1-{[2-(2,4-Difluoro-3-hydroxyphenyl)-1,3-thiazol-5-yl]methyl}-3-methyl-1,3-diazinan-2-one

[0140]

Chem.

[0141] 3.7 Exemplary Compound 25 (General Procedure) 1-{[5-(2,4-Difluoro-3-hydroxyphenyl)-1,3,4-thiadiazol-2-yl]methyl}-3-hydroxy-3-methyl-2,3-dihydro-1H-indol-2-one

Chem.

[0142] Intermediate IX.3 (0.10 g; 0.26 mmol) in DMF (3 mL), 3-hydroxy-3-methyl-2,3-dihydro-1H-indol-2-one (0.04 g; 0.26 mmol) and K2CO3 (0.09 g; 0.65 mmol) were stirred at 70 °C for 20 h. Water was added and the reaction mixture was extracted several times with DCM. The organic layer was separated, dehydrated and concentrated under reduced pressure. The residue was purified by column chromatography (reverse phase; Sunfire C18; water / ACN / TFA) to obtain the intermediate. This intermediate was taken up in DCM (1.5 mL) and TFA (1.00 mL; 49.60 mmol) and stirred overnight at RT. After concentration under reduced pressure, the residue was taken up in ACN and purified by column chromatography (reverse phase; Sunfire C18; water / ACN / TFA) to obtain the desired product. C 18 H 13 F2N3O3S (M = 389.4 g / mol) ESI-MS: 390 [M+H] + R t (HPLC): 0.79 min (Method A) Yield: 0.01 g; 6% 1 H NMR (400 MHz, DMSO-d6) δ ppm: 1.44 (s, 3 H), 5.42 (s, 2 H), 6.09 (br s, 1 H), 7.09 (td, J = 7.45, 0.82 Hz, 1 H), 7.13 (d, J = 7.86 Hz, 1 H), 7.24 (td, J = 9.60, 1.84 Hz, 1 H), 7.27 - 7.35 (m, 1 H), 7.39 (dd, J = 7.35, 0.76 Hz, 1 H), 7.63 (ddd, J = 8.93, 7.54, 5.83 Hz, 1 H), 10.74 (s, 1 H).

[0143] The following exemplary compounds 26 - 30 were prepared using a procedure similar to that described above for 3.7, using appropriate starting materials. As will be understood by those skilled in the art, these similar examples may include variations in common reaction conditions.

Table 29

[0144] Analytical data for the above - mentioned exemplary compound 26

Table 30

[0145] 3.7 Exemplary compound 27 1-{[2-(2,4 - Difluoro - 3 - hydroxyphenyl)-1,3 - thiazol - 5 - yl]methyl}-2,3 - dihydro - 1H - 1,3 - benzodiazol - 2 - one

Chemical formula

[0146] The following compounds 28 - 29 were prepared using a procedure similar to that described above for 3.5 with appropriate starting materials. As will be understood by those skilled in the art, these similar examples may include variations in the general reaction conditions.

[0147] [Table 31]

[0148] Analytical data for the above-mentioned compounds 28 and 29 [Table 32]

[0149] 3.8 Exemplary compound 30 1-{[5-(2,4-Difluoro-3-hydroxyphenyl)-1,3,4-thiadiazol-2-yl]methyl}-3-ethyl-1,2,3,4-tetrahydroquinazoline-2,4-dione [Chemical formula]

[0150] Intermediate XIII.4 (0.07 g; 0.14 mmol) in DMF (1 mL), iodoethane (0.02 mL; 0.21 mmol) and K2CO3 (0.04 g; 0.28 mmol) were stirred at 80 °C for 2 h. The reaction mixture was concentrated under reduced pressure and purified by column chromatography (reverse phase; Sunfire C18; water / ACN / TFA). The residue was taken up in TFA (1.00 mL; 12 - 96 mmol) and DCM (2.5 mL). After stirring overnight at RT, the mixture was purified by column chromatography (reverse phase; Sunfire C18; water / ACN / TFA) to afford the desired compound. C 19 H 14 F2N4O3S (M = 416.4 g / mol) ESI-MS: 417 [M+H] + R t (HPLC): 0.90 min (Method A) Yield: 0.01 g; 16% 1 1H NMR (400 MHz, DMSO-d6) δ ppm: 1.20 (t, J = 7.03 Hz, 3 H), 4.04 (q, J = 7.05 Hz, 2 H), 5.84 (s, 2 H), 7.24 (ddd, J = 10.30, 8.90, 1.84 Hz, 1 H), 7.31 - 7.37 (m, 1 H), 7.59 - 7.64 (m, 1 H), 7.65 (d, J = 8.24 Hz, 1 H), 7.76 - 7.81 (m, 1 H), 8.10 (dd, J = 7.86, 1.52 Hz, 1 H), 10.75 (br s, 1 H).

[0151] (Example 4) Human HSD17B13 - RapidFire MS / MS biochemical assay Estradiol (Sigma, Cat#E8875), NAD (Roche, Cat#10621650001), and recombinant human HSD17B13 (full-length HSD17B13 (Uniprot ID Q7Z5P4-1), C-terminal His-tagged, expressed in mammalian cells and purified to homogeneity) were diluted in assay buffer (100 mM Tris, Sigma, Cat#T2319; sodium chloride, Roth, Cat#3957.2; 0.5 mM EDTA, Invitrogen, Cat#15575020; 0.1% TCEP, Invitrogen, Cat#T2556; 0.05% BSA fraction V (protease- and fatty acid-free), Serva, Cat#11945; 0.001% Tween20, Serva, Cat#37470). Compounds were serially diluted in DMSO (Sigma, Cat#5879) and spotted in a 384-well microplate, PP, V-bottom (Greiner, Cat#781280) plate by Labcyte Echo 55× (1% DMSO during the assay). First, 6 μL / well of recombinant hHSD17B13 (final 1 nM) dilution was added, followed by incubation at RT for 15 min. Next, 6 μL / well of diluted estradiol (final 30 μM) and NAD (final 0.5 mM) were added, mixed, and incubated at RT for 4 h. Following 1 μL of d4-estrone (final 50 nM; Sigma, Cat#489204), 2.4 μL of Girard's reagent P (final 6.5 mM; TCI, Cat#G0030) dissolved in 90% methanol (Sigma, Cat#34860) and 10% formic acid (Merck, Cat#33015) were added to derivatize the samples and stop the enzymatic reaction. Incubation was at RT for 12 - 24 h, after which 70 μL of dH2O was added. Analytical sample processing was performed using a RapidFire autosampler system (Agilent, Waldbronn, Germany) coupled to a triple quadrupole mass spectrometer (Triple Quad 6500, AB Sciex Germany GmbH, Darmstadt, Germany). The liquid sample was aspirated into a 10 μL sample loop by a vacuum pump for 250 ms, and subsequently flushed for 3000 ms with an aqueous mobile phase (99.5% water, 0.49% acetic acid, 0.01% trifluoroacetic acid, flow rate 1.5 mL / min) through a C18 cartridge (Agilent, Waldbronn, Germany). In the solid phase extraction step, the analyte was retained while removing interfering matrices (e.g., buffer components). The analyte was eluted from the cartridge for 3000 ms using an organic mobile phase (49.75% methanol, 49.75% acetonitrile, 0.49% acetic acid, 0.01% trifluoroacetic acid, flow rate 1.25 mL / min), re-eluted, and flashed into the mass spectrometer in MRM mode for detection. The MRM transition for estrone was 404.1 < 157.1 Da (cluster separation potential 27 V, collision energy 43 V), and for the internal standard D4-estrone it was 408.1 < 159.1 Da (cluster separation potential 27 V, collision energy 43 V). The retention time for each MRM transition was 25 ms, and the dwell time between MRMs was 5 ms. The mass spectrometer was operated in positive ionization mode (curtain gas 35 Au, collision gas medium, ion spray voltage 4200 V, temperature 550 °C, ion source gas 1 65 Au, ion source gas 2 80 Au). While performing a backflush to the mass spectrometer, the sample loop and associated tubing were flushed with the organic mobile phase to prevent carryover of the analyte or matrix components to the next sample. The equilibration time of the system was 500 ms. To minimize the effect of carryover, the cleaning station of the RapidFire system was used to perform needle cleaning between samples using pure water (100%) and pure methanol (100%).The solvent delivery device of the RapidFire system consists of two HPLC pumps (G1310A, Agilent, Waldbronn, Germany) that continuously move and operate at a uniform concentration and one binary HPLC pump channel B (G4220A, Agilent, Waldbronn, Germany). MS data processing was performed on a GMSU (Alpharetta, GA, USA), and the peak area ratio of the sample / internal standard was reported for IC50 calculation. The IC50 values were calculated using a four-parameter non-linear regression curve fitting model (developed in-house by Software Megalab). For data evaluation and calculation, the minimum measurement value (without the HSD17B13 enzyme) was set as the 0% control, and the maximum measurement value (including NAD, estrone, and HSD17B13) was set as the 100% control. The IC50 values were calculated using the standard four-parameter logistic regression equation: Y = minimum value + (maximum value - minimum value) / (1 + 10^((LogIC50 - X) × slope + log((maximum value - minimum value) / (50 - minimum value) - 1))).

[0152] (Example 5) Mouse HSD17B13-RapidFire MS / MS Biochemical Assay Estradiol (Sigma, Cat#E8875), NAD (Roche, Cat#10621650001), and recombinant mouse HSD17B13 (U-Protein Express BV, Netherlands) were diluted in assay buffer (100 mM Tris, Sigma, Cat#T2319; sodium chloride, Roth, Cat#3957.2; 0.5 mM EDTA, Invitrogen, Cat#15575020; 0.1% TCEP, Invitrogen, Cat#T2556; 0.05% BSA fraction V (protease- and fatty acid-free), Serva, Cat#11945; 0.001% Tween20, Serva, Cat#37470). Compounds were serially diluted in DMSO (Sigma, Cat#5879) and spotted in a 384-well microplate, PP, V-bottom (Greiner, Cat#781280) plate by Labcyte Echo 55× (1% DMSO during the assay). First, 6 μL / well of the recombinant mouse HSD17B13 (final 50 nM) dilution was added and subsequently incubated at RT for 15 min. Next, 6 μL / well of the diluted estradiol (final 30 μM) and NAD (final 0.5 mM) were added, mixed, and incubated at RT for 3 h. Following 1 μL of d4-estrone (final 50 nM; Sigma, Cat#489204), 2.4 μL of Girard's reagent P (final 6.5 mM; TCI, Cat#G0030) dissolved in 90% methanol (Sigma, Cat#34860) and 10% formic acid (Merck, Cat#33015) were added to derivatize the samples and stop the enzymatic reaction. Incubation was at RT for 12 - 24 h, after which 70 μL of dH2O was added. Analytical sample processing was performed using a RapidFire autosampler system (Agilent, Waldbronn, Germany) coupled to a triple quadrupole mass spectrometer (Triple Quad 6500, AB Sciex Germany GmbH, Darmstadt, Germany). The liquid sample was aspirated into a 10 μL sample loop by a vacuum pump for 250 ms, and subsequently flushed for 3000 ms with an aqueous mobile phase (99.5% water, 0.49% acetic acid, 0.01% trifluoroacetic acid, flow rate 1.5 mL / min) through a C18 cartridge (Agilent, Waldbronn, Germany). In the solid phase extraction step, the analyte was retained while removing interfering matrix (e.g., buffer components). The analyte was eluted from the cartridge for 3000 ms using an organic mobile phase (49.75% methanol, 49.75% acetonitrile, 0.49% acetic acid, 0.01% trifluoroacetic acid, flow rate 1.25 mL / min), and re-eluted and flashed into the mass spectrometer in MRM mode for detection. The MRM transition for estrone was 404.1 < 157.1 Da (cluster separation potential 27 V, collision energy 43 V), and for the internal standard D4-estrone it was 408.1 < 159.1 Da (cluster separation potential 27 V, collision energy 43 V). The dwell time for each MRM transition was 25 ms, and the pause time between MRMs was 5 ms. The mass spectrometer was operated in positive ionization mode (curtain gas 35 Au, collision gas medium, ion spray voltage 4200 V, temperature 550 °C, ion source gas 1 65 Au, ion source gas 2 80 Au). While performing a backflush to the mass spectrometer, the sample loop and associated tubing were flushed with the organic mobile phase to prevent carryover of the analyte or matrix components to the next sample. The equilibration time of the system was 500 ms. To minimize the effect of carryover, the cleaning station of the RapidFire system was used to perform needle cleaning between samples using pure water (100%) and pure methanol (100%).The solvent delivery device of the RapidFire system consists of two HPLC pumps (G1310A, Agilent, Waldbronn, Germany) that continuously move and operate at a uniform concentration, and one binary HPLC pump channel B (G4220A, Agilent, Waldbronn, Germany). MS data processing was performed on a GMSU (Alpharetta, GA, USA), and the peak area ratio of the sample / internal standard was reported for IC50 calculation. The IC50 values were calculated using a four-parameter non-linear regression curve fitting model (developed in-house by Software Megalab). For data evaluation and calculation, the minimum measurement value (without the HSD17B13 enzyme) was set as the 0% control, and the maximum measurement value (including NAD, estrone, and HSD17B13) was set as the 100% control. The IC50 values were calculated using the standard four-parameter logistic regression equation: Y = minimum value + (maximum value - minimum value) / (1 + 10^((LogIC50 - X) × slope + log((maximum value - minimum value) / (50 - minimum value) - 1))).

[0153] (Example 6) Human HSD17B13-RapidFire MS / MS Cell Assay Dilutions of estradiol (Sigma, Cat#E8875) and cells (DDK-tagged clone HEK293 cells stably overexpressing human HSD17B13-Myc, Lakepharma) were prepared in serum-free medium (DMEM, Sigma, Cat#D5796; heat-inactivated 10% FBS, Gibco, Cat#100500; 1× Glutamax, Gibco, Cat#35050-087; 1× sodium pyruvate, Gibyo, Cat#11360070). 25 μL of 0.4 * 10^6 cells / mL dilutions were seeded into 384-well microplates (culture plates, Perkin Elmer, Cat#6007680) 24 h before the compound test. The compound was serially diluted with DMSO (Sigma, Cat#5879), spotted on a pre-seeded cell plate with Labcyte Echo 55× (1% DMSO during the assay), and incubated at 37 °C for 30 min in a humidified incubator (rH = 95%, CO2 = 5%). Then, 25 μL of a 60 μM estradiol dilution was added to the plate and incubated at 37 °C for 3 h in a humidified incubator (rH = 95%, CO2 = 5%). 20 μL of the supernatant was collected, followed by 2.5 μL of d4-estrone (final 50 nM; Sigma, Cat#489204), and 5 μL of Girard's reagent P (final 6.5 mM; TCI, Cat#G0030) and 10% formic acid (Merck, Cat#33015) dissolved in 90% methanol (Sigma, Cat#34860) were added to derivatize the sample. Incubation was at RT for 12 - 24 h, after which 70 μL of dH2O was added. Analytical sample processing was performed using a RapidFire autosampler system (Agilent, Waldbronn, Germany) coupled to a triple quadrupole mass spectrometer (Triple Quad 6500, AB Sciex Germany GmbH, Darmstadt, Germany). The liquid sample was aspirated into a 10 μL sample loop by a vacuum pump for 250 ms, followed by flushing with an aqueous mobile phase (99.5% water, 0.49% acetic acid, 0.01% trifluoroacetic acid, flow rate 1.5 mL / min) through a C18 cartridge (Agilent, Waldbronn, Germany) for 3000 ms. The solid phase extraction step retained the analyte while removing interfering matrices (e.g., buffer components). The analyte was eluted from the cartridge with an organic mobile phase (49.75% methanol, 49.75% acetonitrile, 0.49% acetic acid, 0.01% trifluoroacetic acid, flow rate 1.25 mL / min) for 3000 ms and re-eluted and flashed to the mass spectrometer in MRM mode for detection. The MRM transition for estrone was 404.1 < 157.1 Da (cluster separation potential 27 V, collision energy 43 V), and for the internal standard D4-estrone it was 408.1 < 159.1 Da (cluster separation potential 27 V, collision energy 43 V). The dwell time for each MRM transition was 25 ms, and the pause time between MRMs was 5 ms. The mass spectrometer was operated in positive ionization mode (curtain gas 35 Au, collision gas medium, ion spray voltage 4200 V, temperature 550 °C, ion source gas 1 65 Au, ion source gas 2 80 Au). While performing a backflush to the mass spectrometer, the sample loop and associated tubing were flushed with the organic mobile phase to prevent carryover. MS data processing was performed at GMSU (Alpharetta, GA, USA), and the peak area ratio of the sample / internal standard was reported for IC50 calculation. The IC50 values were calculated using a 4-parameter non-linear regression curve fitting model (developed in-house by Software Megalab). For data evaluation and calculation, the minimum measurements (cells containing estradiol and a 10 μM HSD17B13 inhibitor identified internally) were set as 0% control, and the maximum measurements (cells containing estradiol) were set as 100% control. The IC50 values were calculated using the standard 4-parameter logistic regression equation: Y = minimum value + (maximum value - minimum value) / (1 + 10^((LogIC50 - X) × slope + log((maximum value - minimum value) / (50 - minimum value) - 1))).

[0154] (Example 7) Human HSD17B13 Cell Viability Assay Dilutions of estradiol (Sigma, Cat#E8875) and cells (DDK-tagged clone HEK293 cells stably overexpressing human HSD17B13-Myc, Lakepharma) were prepared in serum-free medium (DMEM, Sigma, Cat#D5796; heat-inactivated 10% FBS, Gibco, Cat#100500; 1× Glutamax, Gibco, Cat#35050-087; 1× sodium pyruvate, Gibyo, Cat#11360070). 25 μL of 0.4 * 10^6 cells / mL dilutions were seeded into 384-well microplates (culture plates, Perkin Elmer, Cat#6007680) 24 h prior to compound testing. Compounds were serially diluted with DMSO (Sigma, Cat#5879) and spotted in the pre-seeded cell plates using Labcyte Echo 55× (1% DMSO during the assay). Incubation was for 30 min at 37 °C in a humidified incubator (rH = 95%, CO2 = 5%). Then, 25 μL of 60 μM estradiol dilutions were added to the plates and incubated for 3 h at 37 °C in a humidified incubator (rH = 95%, CO2 = 5%). 5 μL of CellTiter Glo 2 (Promega, Cat#G9242) was added to the cell plate and incubated at room temperature for 15 minutes, and the fluorescence was measured with PHERAstar FSX (BMG Labtech, Ortenberg, Germany). The IC50 value was calculated using a 4-parameter non-linear regression curve fitting model (developed in-house by Software Megalab). For data evaluation and calculation, the minimum measurement value (without cells, containing estradiol) was set as the 0% control, and the maximum measurement value (with cells and estradiol) was set as the 100% control. The IC50 value was calculated using the standard 4-parameter logistic regression equation: Y = minimum value + (maximum value - minimum value) / (1 + 10^((LogIC50 - X) * Slope + log((maximum value - minimum value) / (50 - minimum value) - 1))).

[0155] (Example 8) Human HSD17B11-RapidFire MS / MS Biochemical Assay Estradiol (Sigma, Cat#E8875), NAD (Roche, Cat#10621650001), and recombinant hHSD17B11 (U-Protein Express BV, Netherlands) were diluted in assay buffer (100 mM Tris, Sigma, Cat#T2319; sodium chloride, Roth, Cat#3957.2; 0.5 mM EDTA, Invitrogen, Cat#15575020; 0.1% TCEP, Invitrogen, Cat#T2556; 0.05% BSA fraction V (protease- and fatty acid-free), Serva, Cat#11945; 0.001% Tween20, Serva, Cat#37470). Compounds were sequentially diluted in DMSO (Sigma, Cat#5879) and spotted with a Labcyte Echo 55× (1% DMSO during the assay) on a 384-well microplate, PP, V-bottom (Greiner, Cat#781280) plate. First, 6 μL / well of the recombinant hHSD17B11 (final 35 nM) dilution was added and subsequently incubated at RT for 15 min. Next, 6 μL / well of the diluted estradiol (final 30 μM) and NAD (final 0.5 mM) were added, mixed, and incubated at RT for 4 h. Following 1 μL of d4-estrone (final 50 nM; Sigma, Cat#489204), 2.4 μL of the Girard reagent P (final 6.5 mM; TCI, Cat#G0030) dissolved in 90% methanol (Sigma, Cat#34860) and 10% formic acid (Merck, Cat#33015) were added to derivatize the samples and stop the enzymatic reaction. Incubation was at RT for 12 - 24 h, after which 70 μL of dH2O was added. Analytical sample processing was performed using a RapidFire autosampler system (Agilent, Waldbronn, Germany) coupled to a triple quadrupole mass spectrometer (Triple Quad 6500, AB Sciex Germany GmbH, Darmstadt, Germany). The liquid sample was aspirated into a 10 μL sample loop by a vacuum pump for 250 ms, followed by flushing with an aqueous mobile phase (99.5% water, 0.49% acetic acid, 0.01% trifluoroacetic acid, flow rate 1.5 mL / min) through a C18 cartridge (Agilent, Waldbronn, Germany) for 3000 ms. In the solid-phase extraction step, the analyte was retained while removing interfering matrices (e.g., buffer components). The analyte was eluted from the cartridge with an organic mobile phase (49.75% methanol, 49.75% acetonitrile, 0.49% acetic acid, 0.01% trifluoroacetic acid, flow rate 1.25 mL / min) for 3000 ms and then re-eluted and flashed into the mass spectrometer in MRM mode for detection. The MRM transition for estrone was 404.1 < 157.1 Da (cluster separation potential 27 V, collision energy 43 V), and for the internal standard D4-estrone it was 408.1 < 159.1 Da (cluster separation potential 27 V, collision energy 43 V). The dwell time for each MRM transition was 25 ms, and the inter-MRM pause time was 5 ms. The mass spectrometer was operated in positive ionization mode (curtain gas 35 Au, collision gas medium, ion spray voltage 4200 V, temperature 550 °C, ion source gas 1 65 Au, ion source gas 2 80 Au). While performing a backflush to the mass spectrometer, the sample loop and associated tubing were flushed with the organic mobile phase to prevent carryover of the analyte or matrix components to the next sample. The equilibration time of the system was 500 ms. To minimize the effect of carryover, the cleaning station of the RapidFire system was used to perform needle cleaning between samples with pure water (100%) and pure methanol (100%).The solvent delivery device of the RapidFire system consisted of two HPLC pumps (G1310A, Agilent, Waldbronn, Germany) that continuously moved and operated at a uniform concentration and one binary HPLC pump channel B (G4220A, Agilent, Waldbronn, Germany). MS data processing was performed on a GMSU (Alpharetta, GA, USA), and the peak area ratio of the analyte / internal standard was reported for IC50 calculation. The IC50 values were calculated using a four-parameter non-linear regression curve fitting model (in-house development by Software Megalab). For data evaluation and calculation, the minimum value (without the HSD17B13 enzyme) was set as the 0% control, and the maximum value (including NAD, estrone, and HSD17B13) was set as the 100% control. The IC50 values were calculated using the standard four-parameter logistic regression equation: Y = minimum value + (maximum value - minimum value) / (1 + 10^((LogIC50 - X) × slope + log((maximum value - minimum value) / (50 - minimum value) - 1))).

[0156] (Example 9) Human HSD17B11 - RapidFire MS / MS Assay Estradiol (Sigma, Cat#E8875), NAD (Roche, Cat#10621650001), and recombinant hHSD17B11 (U-Protein Express BV, Netherlands) were diluted in assay buffer (100 mM Tris, Sigma, Cat#T2319; sodium chloride, Roth, Cat#3957.2; 0.5 mM EDTA, Invitrogen, Cat#15575020; 0.1% TCEP, Invitrogen, Cat#T2556; 0.05% BSA fraction V (protease- and fatty acid-free), Serva, Cat#11945; 0.001% Tween20, Serva, Cat#37470). Compounds were sequentially diluted in DMSO (Sigma, Cat#5879) and spotted with a Labcyte Echo 55× (1% DMSO during the assay) on a 384-well microplate, PP, V-bottom (Greiner, Cat#781280) plate. First, 6 μL / well of the recombinant hHSD17B11 (final 35 nM) dilution was added, followed by incubation at RT for 15 min. Next, 6 μL / well of the diluted estradiol (final 30 μM) and NAD (final 0.5 mM) were added, mixed, and incubated at RT for 4 h. Following 1 μL of d4-estrone (final 50 nM; Sigma, Cat#489204), 2.4 μL of Girard's reagent P (final 6.5 mM; TCI, Cat#G0030) dissolved in 90% methanol (Sigma, Cat#34860) and 10% formic acid (Merck, Cat#33015) were added to derivatize the samples and stop the enzymatic reaction. Incubation was at RT for 12 - 24 h, after which 70 μL of dH2O was added. Analytical sample processing was performed using a RapidFire autosampler system (Agilent, Waldbronn, Germany) coupled to a triple quadrupole mass spectrometer (Triple Quad 6500, AB Sciex Germany GmbH, Darmstadt, Germany). The liquid sample was aspirated into a 10 μL sample loop by a vacuum pump for 250 ms, and subsequently flushed for 3000 ms with an aqueous mobile phase (99.5% water, 0.49% acetic acid, 0.01% trifluoroacetic acid, flow rate 1.5 mL / min) through a C18 cartridge (Agilent, Waldbronn, Germany). In the solid phase extraction step, the analyte was retained while removing interfering matrices (e.g., buffer components). The analyte was eluted from the cartridge for 3000 ms using an organic mobile phase (49.75% methanol, 49.75% acetonitrile, 0.49% acetic acid, 0.01% trifluoroacetic acid, flow rate 1.25 mL / min), and re-eluted and flashed into the mass spectrometer in MRM mode for detection. The MRM transition for estrone was 404.1 < 157.1 Da (cluster separation potential 27 V, collision energy 43 V), and for the internal standard D4-estrone it was 408.1 < 159.1 Da (cluster separation potential 27 V, collision energy 43 V). The dwell time for each MRM transition was 25 ms, and the pause time between MRMs was 5 ms. The mass spectrometer was operated in positive ionization mode (curtain gas 35 Au, collision gas medium, ion spray voltage 4200 V, temperature 550 °C, ion source gas 1 65 Au, ion source gas 2 80 Au). While performing a backflush to the mass spectrometer, the sample loop and associated tubing were flushed with the organic mobile phase to prevent carryover of the analyte or matrix components to the next sample. The equilibration time of the system was 500 ms. To minimize the effect of carryover, the cleaning station of the RapidFire system was used to perform needle cleaning between samples using pure water (100%) and pure methanol (100%).The solvent delivery device of the RapidFire system consists of two HPLC pumps (G1310A, Agilent, Waldbronn, Germany) that continuously move and operate at a uniform concentration and one binary HPLC pump channel B (G4220A, Agilent, Waldbronn, Germany). MS data processing was performed on a GMSU (Alpharetta, GA, USA), and the peak area ratio of the analyte / internal standard was reported for IC50 calculation. The IC50 values were calculated using a four-parameter non-linear regression curve fitting model (developed in-house by Software Megalab). For data evaluation and calculation, the minimum (excluding the HSD17B13 enzyme) measurement value was set as the 0% control, and the maximum (including NAD, estrone, and HSD17B13) measurement value was set as the 100% control. The IC50 values were calculated using the standard four-parameter logistic regression equation: Y = minimum value + (maximum value - minimum value) / (1 + 10^((LogIC50 - X) × slope + log((maximum value - minimum value) / (50 - minimum value) - 1))).

[0157] (Example 10) Pharmacokinetic in vitro assay of metabolic stability in liver microsomes The metabolic degradation of the test compound was assayed at 37 °C using pooled liver microsomes. The final culture volume of 60 μl at each time point contained TRIS buffer (0.1 M) at pH 7.6 at RT, magnesium chloride (5 mM), microsomal protein (0.5 - 2 mg / ml), and the test compound at a final concentration of 1 μM. Following a short pre-incubation at 37 °C, the reaction was initiated by the addition of reduced beta-nicotinamide adenine dinucleotide phosphate (NADPH, 1 mM) and stopped at different time points by transferring an aliquot to the solvent. The quenched culture was pelleted by centrifugation (10000 g, 5 min). An aliquot of the supernatant was assayed for the amount of remaining parent compound by LC-MS / MS. The half-life (t1 / 2 INVITRO) was determined by the slope of the semi-logarithmic plot of the concentration-time profile. The intrinsic clearance (CL_INTRINSIC) was calculated by taking into account the amount of protein in the culture. CL_INTRINSIC [μl / min / mg protein] = (Ln 2 / (half-life [min] * protein content [mg / ml])) * 1000

[0158] (Example 11) Pharmacokinetic in vitro assay for metabolic stability in human hepatocytes (HHEP assay) An assay in human hepatocytes was performed to evaluate the metabolic stability of the compound. The metabolic degradation of the test compound was assayed in a human hepatocyte suspension. After recovery from cryopreservation, human hepatocytes were diluted with DMEM (supplemented with 3.5 μg / 500 ml of glucagon, 2.5 mg / 500 ml of insulin, 3.75 mg / 500 ml of hydrocortisone, 5% or 50% human serum, or in the absence of serum), and a final cell density of 1.0×10 6 cells / ml or 4.0×10 6 cells / ml was obtained. After a 30-minute pre-incubation in a cell culture incubator (37 °C, 10% CO2), the test compound solution was added to the hepatocyte suspension, resulting in a final test compound concentration of 1 μM and a final DMSO concentration of 0.05%. The cell suspension was incubated at 37 °C (cell culture incubator, horizontal shaker), and samples were removed from the culture at 0, 0.5, 1, 2, 4, and 6 hours. The samples were quenched with acetonitrile (containing an internal standard) and pelleted by centrifugation. The supernatant was transferred to a 96-deep well plate and prepared for analysis of the decrease in the parent compound by HPLC-MS / MS. The percentage of the remaining test compound was calculated using the peak area ratio at each incubation time point (test compound / internal standard) relative to the peak area ratio at time zero. The log-transformed data was plotted against the incubation time, and the absolute value of the slope obtained by linear regression analysis was used to evaluate the in vitro half-life (T 1 / 2 ). The in vitro intrinsic clearance (Cl int ) was calculated from the in vitro T 1 / 2 and applied to the following equation using 120 × 10 6 hepatocytes per gram of liver weight, human liver weight per 25.7 g / kg of body weight, and in vitro incubation parameters, and scaled up to the whole liver: CL_INTRINSIC_INVIVO [ml / min / kg] = (CL_INTRINSIC [μl / min / 10 6 cells] × hepatocytes per gram of weight [10 6 hepatocytes / g] × liver factor [g / kg of body weight]) / 1000 The in vivo blood clearance (CL) of the liver was predicted according to a well-stirred liver model, taking into account an average hepatic blood flow (QH) of 20.7 ml / min / kg. CL [ml / min / kg] = CL_INTRINSIC_INVIVO [ml / min / kg] × hepatic blood flow [ml / min / kg] / (CL_INTRINSIC_INVIVO [ml / min / kg] + hepatic blood flow [ml / min / kg]) The results were expressed as a percentage of hepatic blood flow. QH [%] = CL [ml / min / kg] / hepatic blood flow [ml / min / kg])

[0159] (Example 12) Biological data of exemplary compounds 1 to 30

Table 33

Claims

1. A compound of formula (I) or a salt thereof. 【Chemical Formula 1】 (Wherein, a) Z- * is 【Chemical 2】 【Chem.】 selected from the group consisting of, b) 【Chemical Formula 3】 the structure of 【Chemical Formula 4】 selected from the group of structures consisting of)

2. 【Fig. 5】 the structure of ​ selected from the group of structures consisting of, the compound according to claim 1.

3. Z- * is [Chemical Formula 7] which is, the compound according to claim 1.

4. A compound of formula (I) or a salt thereof, selected from the group 01 - 30 of the compound of formula (I), according to claim 1.

5. A salt of the compound according to any one of claims 1 to 4 for use as a medicament.

6. A medicament prepared using the compound according to any one of claims 1 to 4 or a salt thereof.

7. A method for preparing the compound of claim 1 having the general formula (I), the method comprising reacting a compound X1 (wherein Y can be Br, I or Cl) with a compound X2 to obtain the compound (I). [Chemical Formula 8] (Wherein, in the compounds of X1 and (I), Z, A1, A2 and A3 have the same meaning as defined in claim 1)

8. The method according to claim 7, which occurs at a temperature between ambient temperature and the boiling point of the solvent, preferably at a temperature of 50 °C to 120 °C, in an aprotic or protic solvent or a solvent mixture.

9. A pharmaceutical composition comprising at least one compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients.

10. The pharmaceutical composition according to claim 9, comprising a therapeutically effective amount of the compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof in the range of 0.1 to 90% by mass of the composition, preferably in the range of 0.5 to 50% by mass of the composition.

11. A compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 10 for use in the treatment or prevention of steatosis such as non - alcoholic fatty liver disease (NAFLD) or non - alcoholic steatohepatitis (NASH).

12. A compound according to any one of claims 1 to 5 for use in the treatment and / or prevention of non - alcoholic steatohepatitis, which is administered before, after, or simultaneously with at least one other active substance.

13. A compound having formula (Ia), useful as an intermediate compound for the preparation of the compound of formula (I) according to claim 1, wherein Z is as defined in claim 1 and Y can be Br, I or Cl. 【Chemical Formula 9】

14. A compound having formula (Ib), which is useful as an intermediate compound for the preparation of a compound of formula (I) according to claim 1, wherein A1, A2 and A3 are as defined in claim 1. 【Chemical Formula 10】

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