Pyrrolo[2,3-b]pyridine-3-carboxamide composition and method for improving hearing loss

Abstract

Disclosed are N-(3-substituted thiazole-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamides and N-(3-substituted oxazole-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamides of formula (I) and formula (II). These compounds activate Yap and inhibit Lats kinase. Therefore, they are useful for treating hearing loss.

Description

[0001] Statement of Government Authority

[0002] This invention was made with government support under Grant No. T32GM007739 awarded by the National Institutes of Health. The government has certain rights in this invention.

[0003] Background of the Invention Technical Field

[0004] The present application generally relates to N-(3-substituted thiazole-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamides and N-(3-substituted oxazole-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamides, which inhibit laminin kinase and thereby increase Yap activity. The compounds can be used to induce the proliferation of supporting cells in the inner ear and are therefore potentially useful for treating hearing loss. Background Art

[0005] Initiated in response to injury, regeneration is a complex process that can restore the structure and function of damaged tissues. Some adult mammalian tissues retain a regenerative capacity that gradually decreases during development. Regeneration occurs through the activation and expansion of resident stem cells, such as in the epithelium of the skin and intestine, or through cell dedifferentiation and proliferation, such as in the liver. In other cases, such as central nervous and myocardial tissues, cells exhibit little or no potential for regeneration after injury.

[0006] Given its fundamental roles in development, proliferation, stem cell maintenance, and dedifferentiation, Hippo signaling is an inviting target for driving regeneration. The regenerative potential of the Hippo pathway is enriched in many organs, including the heart, retina, liver, and intestine. Hippo signaling limits the size of the developing mouse utricle, a sensory organ in the vestibular portion of the inner ear, and the Yap-Tead complex is active and necessary for the proliferation and regeneration of the neonatal utricle. Hippo signaling limits the size of the developing mouse utricle, a sensory organ in the vestibular portion of the inner ear, and the Yap-Tead complex is active and necessary for the proliferation and regeneration of the neonatal utricle. These observations suggest that chemical activation of YAP signaling may lead to support cell proliferation in adult tissues, a key missing step in the regeneration of the mammalian inner ear.

[0007] To identify activators of YAP, we performed a small molecule screen in cultured cells. We identified a compound that we found to act as an inhibitor of LATS kinase. To test our initial hypothesis, we treated utricles removed from adult mice with this compound and found that a few days of exposure caused supporting cells to reenter the cell cycle, a critical step for robust hair cell regeneration. Summary of the Invention

[0008] The present invention relates to N-(3-substituted oxazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamides, pharmaceutical compositions, and methods for inhibiting LATS or activating Yap to stimulate target cell (particularly hair cell) regeneration.

[0009] In a first aspect, the present invention relates to compounds of formula I:

[0010]

[0011] in:

[0012] R 1 Selected from (C1-C6) alkyl, carboxyl, (C3-C7) carbon monocyclic, (C9-C 11 ) carbon bicyclic group, heteromonocyclic group and heterobicyclic group, wherein the (C1-C6) alkyl, (C3-C7) carbon monocyclic group, (C9-C 11 )Carbobicyclic, heteromonocyclic and heterobicyclic groups may be optionally selected from halogen, cyano, hydroxy, nitro, amino, acetoxy, carboxyl, (C1-C7)alkyl, halo(C1-C6)alkyl, (C1-C3)alkoxy, halo(C1-C3)alkoxy, (C1-C6)acyl, (C1-C3)alkoxy(C1-C3)alkyl, hydroxy(C1-C3)alkyl, heteroaryl, phenylsulfonyl, (C1-C3)alkoxycarbonyl, aminocarbonyl, (C1-C3)alkyl substituted with 1 to 3 substituents selected from the group consisting of aminocarbonyl, di(C1-C3)alkylaminocarbonyl, (C1-C3)alkylamino, di(C1-C3)alkylamino, amino(C1-C3)alkyl, (C1-C3)alkylamino(C1-C3)alkyl(C1-C3)dialkylamino(C1-C3)alkyl, (C1-C3)alkylthio, (C1-C3)alkylsulfonylamino, (C1-C3)alkylsulfinyl, (C1-C3)alkylsulfonyl, phenoxy, and benzyloxy;

[0013] R 2 Selected from hydrogen, halogen, (C1-C7) hydrocarbon, halo (C1-C6) alkyl, (C1-C6) acyl, hydroxy (C1-C3) alkyl, -C(=O)O(C1-C6) alkyl, -C(=O)NR 20 R 21 and (C1-C6)oxaalkyl;

[0014] R 3 is selected from hydrogen, halogen, (C1-C6)alkyl, halogenated (C1-C6)alkyl, (C1-C6)acyl and (C1-C3)alkoxy;

[0015] R 4is selected from hydrogen, halogen, (C1-C6)alkyl, halogenated (C1-C6)alkyl, (C1-C6)acyl and (C1-C3)alkoxy;

[0016] R 10 is independently selected in each instance from hydrogen and methyl;

[0017] R 20 is selected from hydrogen and (C1-C6)alkyl;

[0018] R 21 is selected from hydroxy, (C1-C6)alkyl, (C1-C6)oxaalkyl, amino(C1-C6)alkyl, (C1-C3)alkylamino(C1-C6)alkyl, di(C1-C3)alkylamino(C1-C6)alkyl, and -(CH2) m -Het, wherein Het is an aliphatic mono- or bicyclic heterocycle, optionally substituted by substituents selected from hydroxy, amino, acetoxy, carboxyl, (C1-C7)alkyl, halo(C1-C6)alkyl, (C1-C3)alkoxy, halo(C1-C3)alkoxy, (C1-C6)acyl, (C1-C3)alkoxy(C1-C3)alkyl, hydroxy(C1-C3)alkyl, aminocarbonyl, (C1-C3)alkylaminocarbonyl, di(C1-C3)alkylaminocarbonyl, (C1-C3)alkylamino and di(C1-C3)alkylamino;

[0019] Or, together with the nitrogen to which they are attached, R 20 and R 21 Forming aliphatic heterocycles;

[0020] n is 0, 1, or 2;

[0021] m is 0, 1, or 2; and

[0022] X is S; or, when n is 1 and R 1 When X is an optionally substituted phenyl group, X may alternatively be O; provided that when R 1 When phenyl, X is sulfur and n is 1, R 2 、R 3 、R 4 and R 10 At least one of them is not hydrogen.

[0023] In another aspect, the present invention relates to a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound as described herein.

[0024] In another aspect, the invention relates to a method for activating YAP in a cell expressing YAP, comprising exposing the cell to a compound as described herein.

[0025] In another aspect, the invention relates to a method for LATS inhibition in a cell population expressing LATS comprising exposing the cell population to a compound as described herein.

[0026] In another aspect, the invention relates to a method for LATS inhibition in a cell population comprising exposing the cell population to a compound as described herein.

[0027] In another aspect, the invention relates to a method for stimulating hair cell regeneration comprising exposing a supporting-cell population to a compound as described herein. DETAILED DESCRIPTION

[0028] It has been found that the compound of formula I

[0029]

[0030] Can be used to inhibit LATS or activate Yap, and thus is a potential therapeutic agent for stimulating the regeneration of target cells (especially hair cells). Such compounds can be used to treat hearing loss. Genus I can be divided into two subgenera.

[0031] In the first subgenus, X is sulfur and the compound is thiazol-2(3H)-ylidene-1H-pyrrolo[2,3-b]pyridine-3-carboxamide of formula II:

[0032]

[0033] In the second subgenus, X is oxygen and the compound is oxazol-2(3H)-ylidene-1H-pyrrolo[2,3-b]pyridine-3-carboxamide of formula III:

[0034]

[0035] Where n is 1, and R 1 is an optionally substituted phenyl group.

[0036] In some embodiments of Formulas I and II, n may be 0. In other embodiments of Formulas I-III, n may be 1. In these embodiments, R 10 It may be hydrogen.

[0037] In some embodiments, R 1 is an optionally substituted (C1-C4)alkyl, carboxyl, phenyl, cyclohexyl, 5-membered heterocyclyl, 6-membered heterocyclyl or heterobicyclyl. 1 Can be methyl, ethyl, aminobutyl and carboxyethyl. In other embodiments, R 1 is an optionally substituted cyclohexyl group, or R 1is an optionally substituted phenyl group, or R 1 is an optionally substituted heterocyclic group, such as pyridyl, pyrazolyl, piperidinyl, tetrahydropyranyl, tetrahydrofuranyl or tetrahydroisoquinolinyl. 1 When R is an optionally substituted phenyl group, it may carry one or two substituents independently selected from halogen, cyano, hydroxy, amino, carboxyl, (C1-C6)alkyl, trifluoromethyl, methoxy, acetyl, formyl, hydroxy(C1-C3)alkyl, methoxycarbonyl [-C(=O)OCH3], formylamino [-C(=O)NH2], methanesulfonylamino and amino(C1-C3)alkyl. 1 When pyridyl, pyrazolyl, piperidinyl, tetrahydropyranyl or tetrahydroisoquinolinyl, each heterocyclic ring may be optionally substituted with one or two substituents independently selected from amino, hydroxy and (C1-C6)alkyl.

[0038] In some embodiments, R 1 is selected from carboxyl and optionally substituted (C1-C4) alkyl, phenyl, cyclohexyl, 5-membered heterocyclyl, 6-membered heterocyclyl and heterobicyclyl. In some embodiments, these R 1 is selected from the group consisting of methyl, ethyl, aminobutyl and carboxyethyl. 1 In some embodiments, these R 1 It is an optionally substituted heterocyclic radical. These heterocycles can be pyridyl, pyrazolyl, piperidyl, tetrahydropyranyl, tetrahydrofuranyl and tetrahydroisoquinolyl, each of which is optionally substituted. The optional substituents may include one or two substituents independently selected from amino, hydroxyl and (C1-C6) alkyl.

[0039] In some embodiments, R 1 is an optionally substituted phenyl group. The phenyl group may be substituted with one or two substituents independently selected from halogen, cyano, hydroxy, amino, carboxyl, (C1-C6) alkyl, trifluoromethyl, methoxy, acetyl, formyl, hydroxy (C1-C3) alkyl, methoxycarbonyl, formamido, methylsulfonylamino, and amino (C1-C3) alkyl. In some of these embodiments, R 1 is phenyl substituted in the ortho position and n is 0; in other embodiments, R 1 is an optionally substituted phenyl group and n is 1.

[0040] In some embodiments, R 2 Selected from -C(=O)O(C1-C6)alkyl, -C(=O)NR 20 R 21 and (C1-C6)oxaalkyl. In some embodiments, R 20 is selected from hydrogen and methyl, and R 21selected from hydrogen, methyl, (C1-C6) oxaalkyl, dimethylamino (C1-C6) alkyl and -(CH2) m -Het. In other embodiments, R 20 and R 21 Together with the nitrogen to which they are attached, they form a 4-7 membered aliphatic heterocyclic ring. Exemplary aliphatic heterocyclic rings include piperidine, piperazine, morpholine, pyrrolidine, azetidine, azepine, and the like.

[0041] In some embodiments, R 2 is selected from hydrogen, methyl, ethyl, propyl, cyclopropyl, hydroxymethyl and trifluoromethyl.

[0042] In some embodiments, R 3 and R 4 is selected from hydrogen, chlorine and methyl.

[0043] It should be understood that in various embodiments, the pharmaceutical compositions of the present invention contain one or more pharmaceutically acceptable excipients, including but not limited to one or more binders, fillers, buffers, stabilizers, surfactants, wetting agents, lubricants, diluents, disintegrants, viscosity enhancers or reducing agents, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, glidants, processing aids, colorants, sweeteners, taste masking agents, flavorings, flavorings, diluents, polishing agents, polymer matrix systems, plasticizers and other known additives to provide an elegant presentation of the drug or adjuvant in the preparation of a drug or drug product containing the composition of the present invention. Examples of carriers and excipients are well known to those skilled in the art and are described in detail in Ansel, Howard C, et al., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, Alfonso R, et al., Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C. Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005.

[0044] In various embodiments, non-limiting examples of excipients include, but are not limited to, corn starch, potato starch or other starches, gelatin, natural and synthetic gums such as gum arabic, sodium alginate, alginic acid, other alginates, powdered tragacanth gum, guar gum, cellulose and its derivatives (e.g., ethylcellulose, cellulose acetate, carboxymethylcellulose calcium, carboxymethylcellulose sodium), polyvinyl pyrrolidone, methylcellulose, pregelatinized starch, hydroxypropyl methylcellulose (e.g., Nos. 2208, 2906, 2910), hydroxypropyl cellulose, titanium dioxide, talc, calcium carbonate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, dextrin, kaolin, silicic acid, Sorbitol, starch, pregelatinized starch, agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, potato or tapioca starch, other starch, pregelatinized starch, other starch, clay, other algal toxins, other cellulose, gum, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oils (e.g., peanut, cottonseed, sunflower, sesame, olive, corn, and soybean), zinc stearate, ethyl oleate, lauryl esters, agar, syloid silica (AEROSIL 200, manufactured by W.R. Grace Co. of Baltimore, Maryland, USA), condensation aerosol of synthetic silica (sold by Degussa Corporation of Plano, Texas, USA), CAB-O-SIL (a fumed silica product sold by Cabot Corporation of Boston, Massachusetts, USA), colorants, and mixtures thereof.

[0045] The terms "subject" or "subject in need thereof" are used interchangeably herein. These terms refer to a patient who has been diagnosed with the underlying disease to be treated. Typically, the patient will be a human. The subject may currently be experiencing symptoms associated with the condition or may have experienced symptoms in the past. Additionally, a "subject in need thereof" may be a patient at risk for a particular disease, or a patient reporting risk of one or more physiological systems for a disease, even though a diagnosis of the disease may not have been made.

[0046] As used herein, the terms "treatment" or "treating" are used interchangeably. These terms refer to an approach for obtaining beneficial or desired results, including, but not limited to, a therapeutic benefit. A therapeutic benefit includes eradication or amelioration of the underlying condition being treated; it also includes eradication or amelioration of one or more symptoms associated with the underlying condition, such that an improvement is observed in the patient, although the patient may still be suffering from the underlying condition.

[0047] As used herein, the term "optionally substituted" is used interchangeably with "unsubstituted or substituted". The term "substituted" refers to the replacement of one or more hydrogen atoms in a specified group with a specified group. For example, substituted aryl, heterocyclyl, etc. refers to a group in which one or more H atoms in each residue are replaced by halogen, haloalkyl, alkyl, (C1-C8) hydrocarbon, acyl, alkoxyalkyl, hydroxy lower alkyl, carbonyl, phenyl, heteroaryl, benzenesulfonyl, hydroxy, lower alkoxy, haloalkoxy, oxaalkyl, carboxyl, alkoxycarbonyl [i.e., -C(=O)O-alkyl], carboxamido [i.e., -C(=O)NH2], alkylaminocarbonyl [i.e., -C(=O)NH-alkyl], cyano, acetoxy, nitro, amino, alkylamino, dialkylamino , dialkylaminoalkyl, dialkylaminoalkoxy, heterocyclylalkoxy, arylalkyl, (cycloalkyl)alkyl, heterocyclyl, heterocyclylalkyl, alkylaminoalkyl, heterocyclylaminoalkyl, heterocyclylalkylaminoalkyl, cycloalkylaminoalkyl, cycloalkylalkylaminoalkyl, arylaminoalkyl and arylalkylaminoalkyl, mercapto, alkylthio, alkylsulfinyl, benzyl, heterocyclyl, phenoxy, benzyloxy, heteroaryloxy, aminosulfonyl, amidino, guanidino, urea, -SO2alkyl, -SO2NH2, -SO2NHalkyl substituted aryl or heterocyclyl. Preferred substituents are halogen, cyano, hydroxy, nitro, amino, acetoxy, carboxyl, (C1-C7)alkyl, halo(C1-C6)alkyl, (C1-C3)alkoxy, halo(C1-C3)alkoxy, (C1-C6)acyl, (C1-C3)alkoxy(C1-C3)alkyl, hydroxy(C1-C3)alkyl, heteroaryl, phenylsulfonyl, (C1-C3)alkoxycarbonyl [i.e., -C(=O)O(C1-C3)alkyl], carboxamido [i.e., -C(=O)NH2], (C1-C6)acyl, (C1-C3)alkoxy(C1-C3)alkyl, hydroxy(C1-C3)alkyl, heteroaryl, phenylsulfonyl, (C1-C3)alkoxycarbonyl [i.e., -C(=O)O(C1-C3)alkyl], formamido [i.e., -C(=O)NH2], (C1-C6)acyl, (C1-C3)alkoxy(C1-C3)alkyl, hydroxy(C1-C3)alkyl, heteroaryl, (C1-C3)alkylamino, (C1-C3)alkylamino, (C1-C3)alkylamino, (C1-C3)alkylamino, (C1-C3)alkylamino, (C1-C3)alkylamino, (C1-C3)alkylamino, (C1-C3)alkylamino, (C1-C3)alkylamino, (C1-C3)alkylamino, (C1-C3)alkylamino, (C1-C3)alkylamino, (C1-C3)alkylamino, (C1-C3)alkylthio, (C1-C3)alkylsulfonyl, (C1-C3)alkylsulfinyl, (C1-C3)alkylsulfonyl, phenoxy and benzyloxy.

[0048] Unless otherwise indicated, an alkyl group is a straight or branched hydrocarbon group. Unless otherwise indicated, an unsubstituted alkyl group has 1 to 20 carbon atoms (e.g., 1 to 6 carbon atoms). Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, and tert-butyl.

[0049] Hydrocarbon or hydrocarbyl (as substituent) comprises alkyl, cycloalkyl, polycycloalkyl, alkenyl, alkynyl, aryl and combination thereof.Examples include cyclopropylmethyl, benzyl, phenethyl, cyclohexylmethyl, camphoryl and naphthylethyl.Hydrocarbon refers to any substituent consisting of hydrogen and carbon as the only elemental components.Cycloalkyl is a subset of hydrocarbyl and includes cyclic hydrocarbyl groups of 3 to 8 carbon atoms.Examples of cycloalkyl include C-propyl, C-butyl, C-pentyl, norbornyl etc.

[0050] Unless otherwise indicated, the term "carbocycle" is a ring system in which all of the ring atoms are carbon, not in any oxidation state. Thus, (C3-C8)carbocycle refers to both non-aromatic and aromatic systems, including systems such as cyclopropane, benzene, and cyclohexene; (C8-C 12 ) Carbocyclic refers to systems such as norbornane, decane, indene, and naphthalene. Unless otherwise limited, carbocyclic refers to monocyclic, bicyclic, and polycyclic rings.

[0051] Oxyalkyl refers to an alkyl residue in which one or more carbons (and their associated hydrogens) have been replaced by oxygen. Examples include methoxypropoxy, 3,6,9-triazoloyl, and the like. The term "oxaalkyl" is understood in the art [see Naming and Indexing of Chemical Substances for Chemical Abstracts, published by the American Chemical Society, 196, but not limited to 127(a)], i.e., it refers to a compound in which an oxygen is bonded to its adjacent atom by a single bond (forming an ether linkage); it does not refer to a doubly bonded oxygen, such as is found in a carbonyl group. Alkoxy or alkoxy is a subset of oxaalkyl, which refers to a group having a straight chain or branched configuration of 1 to 8 carbon atoms, which is attached to the parent structure through an oxygen. Examples include methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy, cyclohexyloxy, and the like. Lower alkoxy refers to a group containing 1-4 carbon atoms. For the purposes of this application, alkoxy and lower alkoxy include methylenedioxy and ethylenedioxy.

[0052] Unless otherwise indicated, acyl refers to formyl and groups having 1, 2, 3, 4, 5, 6, 7, and 8 carbon atoms in straight, branched, cyclic configurations, saturated, unsaturated, aromatic, and combinations thereof, attached to the parent structure via a carbonyl functional group. As long as the point of attachment to the parent remains at the carbonyl, one or more carbons in the acyl residue may be substituted with nitrogen, oxygen, or sulfur. Examples include formyl, acetyl, benzoyl, propionyl, isobutyryl, tert-butyloxycarbonyl, benzyloxycarbonyl, and the like. Lower acyl refers to groups containing one to four carbon atoms. When referred to as a substituent, the double bonded oxygen itself is referred to as "oxo."

[0053] Aryl and heteroaryl refer to (i) phenyl (or benzene) or a monocyclic 5- or 6-membered heteroaromatic ring containing 1-4 heteroatoms selected from O, N or S; (ii) a bicyclic 9- or 10-membered aromatic or heteroaromatic ring system containing 0-4 heteroatoms selected from O, N or S; (iii) a tricyclic 13- or 14-membered aromatic or heteroaromatic ring system containing 0-5 heteroatoms selected from O, N or S. Aromatic 6- to 14-membered carbocyclic rings include, for example, benzene, naphthalene, indane, tetralin and fluorene, and 5- to 10-membered aromatic heterocyclic rings include, for example, imidazole, pyridine, indole, thiophene, benzopyrone, thiazole, furan, benzimidazole, quinoline, isoquinoline, quinoxaline, pyrimidine, pyrazine, tetrazole and pyrazole. As used herein, aryl and heteroaryl refer to residues in which one or more rings are aromatic, but not all rings are required to be so.

[0054] Arylalkyl refers to a substituent in which an aryl residue is attached to the parent structure via an alkyl group. Examples include benzyl, phenethyl, and the like. Heteroarylalkyl refers to a substituent in which a heteroaryl residue is attached to the parent structure via an alkyl group. In one embodiment, the alkyl group of an arylalkyl or heteroarylalkyl group is an alkyl group having 1 to 6 carbon atoms. Examples include, for example, pyridylmethyl, pyrimidinylethyl, and the like.

[0055] Heterocycle refers to a cycloalkyl or aryl carbocyclic residue in which 1-4 carbon atoms are substituted with heteroatoms selected from N, O, and S. Nitrogen and sulfur heteroatoms may be optionally oxidized, and nitrogen heteroatoms may be optionally quaternized. Unless otherwise indicated, heterocycles may be non-aromatic (i.e., aliphatic) or aromatic. Examples of heterocycles include pyrrolidine, pyrazole, pyrrole, indole, quinoline, isoquinoline, tetrahydroisoquinoline, benzofuran, benzodioxane, benzodioxolane (commonly referred to as methylenedioxyphenyl when appearing as a substituent), tetrazole, morpholine, thiazole, pyridine, pyridazine, pyrimidine, thiophene, furan, oxazole, oxazoline, isoxazole, dioxane, tetrahydrofuran, etc. It should be noted that heteroaryl is a subset of heterocycles in which heterocycles are aromatic. Examples of heteroaromatic rings include furan, benzofuran, isobenzofuran, pyrrole, indole, isoindole, thiophene, benzothiophene, imidazole, benzimidazole, purine, pyrazole, indazole, oxazole, benzoxazole, isoxazole, benzisoxazole, thiazole, benzothiazole, triazole, tetrazole, pyridine, quinoline, isoquinoline, pyrazine, quinoxaline, acridine, pyrimidine, quinazoline, pyridazine, cinnoline, phthalazine, and triazine. Examples of heterocyclyl residues further include piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazepinyl, azepanyl, 4-piperidinyl, pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyrazinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinylquinuclidine, isothiazolidinyl, benzimidazolyl, thiadiazolyl, benzopyranyl, benzothiazolyl, tetrahydrofuranyl, tetrahydropyranyl, thienyl, benzothienyl, thiamorphinyl, thiamorphinyl sulfoxide, thiamorphinyl sulfone, oxadiazolyl, triazolyl and tetrahydroquinolinyl.

[0056] An oxygen heterocycle is a heterocycle containing at least one oxygen in the ring; it may contain additional oxygen, as well as other heteroatoms. A sulfur heterocycle is a heterocycle containing at least one sulfur in the ring; it may contain additional sulfur, as well as other heteroatoms. Oxyheteroaryls are a subset of oxygen heterocycles; examples include furan and oxazole. Sulfur heteroaryls are a subset of sulfur heterocycles; examples include thiophene and thiazine. Azoheterocycles are heterocycles containing at least one nitrogen in the ring; it may contain additional nitrogen, as well as other heteroatoms. Aliphatic nitrogen-containing heterocycles include piperidine, piperazine, morpholine, pyrrolidine, thiomorpholine, azetidine, azapine, and olanzapine. Azoaryls are a subset of nitrogen heterocycles; examples include pyridine, pyrrole, and thiazole.

[0057] As used herein and as will be understood by those skilled in the art, recitation of a "compound" is intended to include salts of the compound unless expressly further limited. Thus, for example, recitation of a "compound of Formula I" as described above would include T32 as the free base and its salts:

[0058]

[0059] wherein Z is any counterion. In one embodiment, the term "compound of Formula I" refers to the compound or a pharmaceutically acceptable salt thereof. The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic acid or base, including inorganic acids and bases and organic acids and bases. When the compound of the present invention is basic, as described above in this paragraph, the salt can be prepared from a pharmaceutically acceptable non-toxic acid, including inorganic acids and organic acids. Suitable pharmaceutically acceptable acid addition salts of the compounds of the present invention include acetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid (benzenesulfonate), benzoic acid, boric acid, butyric acid, camphor, camphorsulfonic acid, carbonic acid, citric acid, ethanedisulfonic acid, ethanesulfonic acid, ethylenediaminetetraacetic acid, formic acid, fumaric acid, glucoheptanoic acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, hydroiodic acid, hydroxynaphthoic acid, isethionic acid, lactic acid, lactobionic acid, laurylsulfonic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalenesulfonic acid, nitric acid, oleic acid, palmitic acid, pantothenic acid, phosphoric acid, pivalic acid, polygalacturonic acid, salicylic acid, stearic acid, succinic acid, sulfuric acid, tannic acid, tartaric acid, teoclatic acid, p-toluenesulfonic acid, and the like. When the compound contains an acidic side chain, for example, as in T34, when R 1When is COOH, pharmaceutically acceptable base addition salts suitable for the compounds of the present invention include, but are not limited to, organic salts prepared from aluminum, calcium, lithium, magnesium, potassium, sodium, and zinc, or from lysine, arginine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine), and procaine. Other pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium cations and carboxylate, sulfonate, and phosphonate anions attached to alkyl groups having 1 to 20 carbon atoms.

[0060] Treatment

[0061] The compositions described herein can be administered by a variety of routes, such as topical administration to the middle or inner ear (e.g., administration to or through the oval window, round window, or semicircular canals (e.g., horizontal canals), or by intratympanic or intratympanic injection), intravenous, parenteral, intradermal, transdermal, intramuscular, intranasal, subcutaneous, transdermal, intratracheal, intraperitoneal, intraarterial, intravascular, inhalation, infusion, lavage, and oral administration. The most appropriate route of administration in any given case will depend on the specific composition being administered, the patient, the method of pharmaceutical preparation, the method of administration (e.g., time of administration and route of administration), the patient's age, weight, sex, severity of the disease being treated, the patient's diet, and the patient's excretion rate. The composition can be administered once or more than once (e.g., once a year, twice a year, three times a year, every two months, every month, or every two weeks).

[0062] A subject that can be treated as described herein is a subject having or at risk of developing hearing loss and / or vestibular dysfunction (eg, a subject having or at risk of developing hearing loss, vestibular dysfunction, or both). The compositions and methods described herein can be used to treat subjects who have or are at risk of developing cochlear hair cell damage (e.g., damage associated with auditory trauma, disease or infection, head trauma, ototoxic drugs, or aging), subjects who have or are at risk of developing vestibular hair cell damage (e.g., damage associated with disease or infection, head trauma, ototoxic drugs, or aging), subjects who have or are at risk of developing sensorineural hearing loss, deafness, or auditory neuropathy, subjects who have or are at risk of developing vestibular dysfunction (e.g., dizziness, vertigo, loss of balance, bilateral vestibulopathy, tremor, or balance disorders), subjects who have tinnitus (e.g., tinnitus alone or tinnitus associated with sensorineural hearing loss or vestibular dysfunction), subjects who have a genetic mutation associated with hearing loss and / or vestibular dysfunction, or subjects who have a family history of hereditary hearing loss, deafness, auditory neuropathy, or vestibular dysfunction. In some embodiments, the subject suffers from hearing loss and / or vestibular dysfunction associated with or caused by hair cell (e.g., cochlear or vestibular hair cell) loss or is at risk of development. The methods described herein may include screening the subject's known one or more mutations in genes associated with hearing loss and / or vestibular dysfunction before treating with a composition as described herein or administering a composition as described herein. The subject's genetic mutations can be screened using standard methods known to those skilled in the art (e.g., genetic testing). The methods described herein may also include assessing the subject's hearing and / or vestibular function before treating with a composition as described herein or administering. Hearing can be assessed using standard tests, such as audiometry, auditory brainstem response (ABR), electrocochlear glottis (ECOG), and otoacoustic emissions. Vestibular function can be assessed using standard tests, such as eye movement testing (e.g., electronic nystagmus (ENG) or video nystagmus (VNG)), vestibulo-ocular reflex (VOR) testing (e.g., head impulse testing (Haimagyi-Curthoys test), which can be performed at the bedside or using video head impulse testing (VHIT) or caloric reflex testing), posturography, rotary chair testing, ECOG, vestibular evoked myogenic potentials (VEMPs), and specialized clinical balance tests, such as those described in Mancini and Horak, Eur J Phys Rehabil Med, 46:239 (2010). These tests can also be used to assess hearing and / or vestibular function in a subject following treatment or administration with a composition described herein.The compositions and methods described herein can also be administered as a prophylactic treatment to patients at risk for developing hearing loss and / or vestibular dysfunction, such as patients with a family history of hearing loss or vestibular dysfunction (e.g., hereditary hearing loss or vestibular dysfunction), patients who carry a genetic mutation associated with hearing loss or vestibular dysfunction but have not yet manifested hearing loss or vestibular dysfunction, or patients exposed to risk factors for acquiring hearing loss (e.g., acoustic trauma, disease or infection, head trauma, ototoxic drugs, or aging) or vestibular dysfunction (e.g., disease or infection, head trauma, ototoxic drugs, or aging).

[0063] The compositions and methods described herein can be used to induce or increase hair cell regeneration (e.g., cochlear and / or vestibular hair cell regeneration) in a subject. Subjects who may benefit from compositions that induce or increase hair cell regeneration include subjects with hearing loss or vestibular dysfunction due to hair cell loss (e.g., hair cell loss associated with trauma (e.g., acoustic trauma or head trauma), disease or infection, ototoxic drugs, or aging), as well as abnormal hair cells (e.g., hair cells that do not function properly compared to normal hair cells), damaged hair cells (e.g., hair cell damage associated with trauma (e.g., acoustic trauma or head trauma), disease or infection, ototoxic drugs, or aging), or a decrease in the number of hair cells due to genetic mutations or congenital abnormalities.

[0064] The compositions and methods described herein can also be used to prevent or reduce hearing loss and / or vestibular dysfunction caused by hair cell damage or death (e.g., cochlear hair cell and / or vestibular hair cell damage or death) induced by ototoxic drugs in subjects who have received ototoxic drug treatment, or who are currently receiving or are about to begin receiving ototoxic drug treatment. Ototoxic drugs are toxic to inner ear cells and can cause sensorineural hearing loss, vestibular dysfunction (e.g., vertigo, dizziness, imbalance, bilateral vestibulopathy, tremor or balance disorder), tinnitus, or a combination of these conditions. Drugs that have been found to have ototoxicity include aminoglycoside antibiotics (e.g., gentamicin, neomycin, streptomycin, tobramycin, kanamycin, vancomycin, and amikacin), puromycins, antitumor drugs (e.g., platinum-containing chemotherapeutic agents, such as cisplatin, carboplatin, and oxaliplatin), loop diuretics (e.g., ethacrynic acid and furosemide), salicylates (e.g., aspirin, especially high doses), and quinine. In some embodiments, the methods and compositions described herein can be used to treat bilateral vestibulopathy or tremor. Bilateral vestibulopathy and tremor can be induced by aminoglycosides (e.g., the methods and compositions described herein can be used to promote or increase hair cell regeneration in a subject having or at risk for developing aminoglycoside-induced bilateral vestibulopathy or tremor).

[0065] Treatment can include administering a composition containing a compound described herein in various unit doses. Each unit dose typically contains a predetermined amount of the therapeutic composition. The amount to be administered, as well as the specific route of administration and formulation, are within the skill of those skilled in the art of clinical practice. The unit dose need not be administered as a single injection, but can include continuous infusion over a set time period. Administration can be performed using a syringe pump to control the infusion rate to minimize damage to the inner ear (e.g., cochlea and / or vestibular system).

[0066] Compositions as described herein are enough to improve hearing, improve vestibular function (for example, improve balance or reduce dizziness or vertigo), reduce tinnitus, treat bilateral vestibulopathy, treat tremor, treat balance disorder, increase or induce the amount of hair cell regeneration (for example, cochlear and / or vestibular hair cell regeneration), increase hair cell number, activate YAP, and / or inhibit LATS.Hearing can be assessed using standard audiometry (for example, audiometry, ABR, electrocochlear electrogram (ECOG) and otoacoustic emissions), and can be improved compared to the audiometry obtained before treatment.Standard tests for balance and vertigo (for example, eye movement test (for example, ENG or VNG), posturography, VOR test (for example, head impulse test (Halmagyi-Curthoys test, for example, VHIT) or caloric reflex)), swivel chair test, ECOG, VEMP and special clinical balance test) can be used to assess vestibular function, and may be improved compared to the measured value obtained before treatment.In some embodiments, compositions are administered with an amount sufficient to improve the ability of the subject to understand language. The compositions described herein can also be administered in an amount sufficient to slow or prevent the development or progression of sensorineural hearing loss and / or vestibular dysfunction (e.g., in subjects who carry a genetic mutation associated with hearing loss or vestibular dysfunction, who have a family history of hearing loss or vestibular dysfunction (e.g., hereditary hearing loss or vestibular dysfunction), or who have been exposed to risk factors associated with hearing loss or vestibular dysfunction (e.g., ototoxic drugs, head trauma, disease or infection, or auditory trauma) but do not exhibit hearing loss or vestibular dysfunction (e.g., vertigo, dizziness, or imbalance), or in subjects who exhibit mild to moderate hearing loss or vestibular dysfunction). For example, these effects may occur within 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 15 weeks, 20 weeks, 25 weeks, or more after administration of the compositions described herein. Depending on the dosage and route of administration used for treatment, the patient may be evaluated 1 month, 2 months, 3 months, 4 months, 5 months, 6 months or longer after administration of the composition. Based on the results of the evaluation, the patient may receive additional treatment.

[0067] Preparation of compounds

[0068] The following abbreviations are used in the synthetic routes: THF (tetrahydrofuran), MeOH (methanol), DCM (dichloromethane), DMF (N,N-dimethylformamide), ACN (acetonitrile), EtOH (ethanol), EtOAc (ethyl acetate), IPA (2-propanol), DMSO (dimethyl sulfoxide), MTBE (tert-butyl methyl ester), TEA (triethylamine), DIPEA (N,N-diisopropylethylamine), TMEDA (tetramethylethylenediamine), DMAP (N,N-dimethyl Pyridin-4-amine), EDCI (N-(3-dimethylaminopropyl)-N'-ethylcarbodiamine hydrochloride), HOBt (1-hydroxybenzotriazole hydrate), HBTU (2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate), T3P (propanephosphonic anhydride), TBAI (tetrabutylammonium iodide), LAH (lithium aluminum hydride), XPhos (2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl), and TFA (trifluoroacetic acid).

[0069] Preparative HPLC purification refers to the use of a water / acetonitrile gradient, with or without additives such as HCl, formic acid, TFA, or NH 4 HCO 3 , using an appropriate hydrophobic stationary phase.

[0070] The compounds of the present invention can be prepared as illustrated in general Schemes I-IV and in more detail below in Schemes 1-69. Detailed descriptions of the syntheses of intermediates and exemplary compounds are also disclosed below.

[0071]

[0072] As shown in Scheme 1, it is known to those skilled in the art that compounds of Formula (Ia) containing an imine group, when treated with compounds of Formula (Ib) wherein X is chloro, bromo, or -OH under coupling conditions, will provide compounds of Formula I. Typical conditions for the reaction of compounds of Formula (Ib) wherein X is chloro and compounds of Formula (Ia) include, but are not limited to, stirring an equimolar mixture of the compounds in a solvent such as chloroform or dichloromethane in the presence of a base such as, but not limited to, triethylamine and N,N-dimethylpyridin-4-amine (DMAP) at 5-25° C. for 1-12 hours. The acid coupling conditions for the compound of formula (Ib) wherein X is -OH and the compound of formula (Ia) include, but are not limited to, stirring an equimolar mixture of the compound with a coupling agent in a solvent in the presence of a base, such as, but not limited to, N-(3-dimethylaminopropyl)-N'-ethylcarbodiamine hydrochloride (EDCI), 1-hydroxybenzotriazole hydrate (HOBt), 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU), propanephosphonic anhydride (T3P), wherein the base includes, but is not limited to, N,N-diisopropylethylamine (DIPEA), and the solvent includes, but is not limited to, DMF, EtOAc, and pyridine. A typical reaction can be carried out at 25-110°C for 1-12 hours.

[0073]

[0074] As shown in Scheme II, compounds of formula (IIa) can be converted to compounds of formula I, which are representative compounds of the present invention. Typical conditions include, but are not limited to, treating the compound of formula (IIa) with potassium carbonate (K2CO3) in DMF at 25°C, followed by the addition of a reagent such as R1-Y, wherein R1 is as defined in formula I and Y is chlorine, bromine, iodine, methanesulfonic acid, or toluenesulfonic acid. Typical reactions can be performed in a microwave reactor at 100°C to facilitate alkylation.

[0075]

[0076] Compounds of formula (Ia) in Scheme I can be prepared according to the methods outlined in Scheme III. Treatment of a compound of formula (IIIa) with potassium carbonate (K2CO3) in DMF at 25°C followed by addition of a reagent such as R1-Y, wherein R1 is as defined in formula I and Y is chloro, bromo, iodo, mesyl or tosylate, will provide compounds of formula (IIIb / Ia). A typical reaction can be carried out at 100°C in a microwave reactor to facilitate alkylation.

[0077]

[0078]

[0079] Scheme IV describes another alternative method for preparing compounds of formula (Ia) in Scheme I. When the compound of formula (IVa) is treated with ethyl N-(thiomethylene)urethane and tetramethylethylenediamine (TMEDA), the compound of formula (IVb) is provided. When the compound of formula (IVb) is treated with a compound of formula (IVc) in acetonitrile in the presence of cesium carbonate (Cs2CO3), the compound of formula (IVd) is provided. When the compound of formula (IVd) is treated with sulfonyl chloride (SOCl2) in the presence of N,N-diisopropylethylamine (DIPEA), the compound of formula (IVe) is provided. When the compound of formula (IVe) is treated with sodium hydroxide (NaOH), the compound of formula (IVf / Ia) is provided.

[0080] The acids Ib used in Scheme I are described in Table A. For known acids, the CAS number is shown in the reference column. Synthetic schemes for all other acids are described in the application.

[0081] Table A

[0082]

[0083]

[0084] Intermediates Ia / IIIb used in Scheme I and prepared using the methods outlined in Scheme III are described in Table B.

[0085] Table B

[0086]

[0087]

[0088]

[0089] Intermediates Ia / IVf used in Scheme I and prepared using the method outlined in Scheme IV are described in Table C.

[0090] Table C

[0091]

[0092]

[0093]

[0094]

[0095] The preparation of the intermediates in Table D is described in Schemes 51-52.

[0096] Table D

[0097]

[0098] Preparation of intermediate B1

[0099]

[0100] To a solution of 2-thiazolamine (10.00 g, 99.86 mmol, 1.0 equivalent) in DMF (200 mL) was added chloromethylbenzene (18.96 g, 149.8 mmol, 1.5 equivalents). The reaction mixture was heated to 80 ° C and stirred for 12 hours. The reaction mixture was concentrated under reduced pressure. The resulting residue was diluted with EtOAc (200 mL). An aqueous NaOH solution (100 mL, 40 w%) was added at 0 ° C and stirred at 0 ° C for 1 hour. The mixture was extracted with EtOAc (3 × 100 mL). The combined organic layers were washed with brine (3 × 100 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography (petroleum ether / EtOAc=5: 1 to 1: 1) to obtain 3-benzylthiazole-2 (3H)-imine B1 (13.00 g, crude product) as a yellow oil.

[0101] Alternative method for preparing intermediate B1

[0102]

[0103] N-(Benzylaminomethylthioyl)urethane

[0104] At 25 ℃ under nitrogen atmosphere, to benzylamine (5.00g, 46.7mmol, 1.0 equivalent) and O-ethyl carbon isothiocyanate (6.43g, 49.0mmol, 1.05 equivalent) in ethyl acetate (40mL), add TMEDA (0.54g, 4.67mmol, 0.1 equivalent).Mixture was stirred 6 hours at 25 ℃, then concentrating under reduced pressure.Resistates is ground twice from the solution of petroleum ether and ethyl acetate (v / v=10 / 1, 55mL), obtain 10.0g N-(benzylaminomethylthioyl) ethyl carbamate as yellow solid. 1 H NMR (400MHz, DMSO-d6) δ11.04 (br.s, 1H), 10.22 (br.t, J = 5.6Hz, 1H), 7.39-7. 23(m,5H), 4.81(d,J=5.6Hz,2H), 4.14(q,J=7.2Hz,2H), 1.21(t,J=7.2Hz,3H).

[0105] (Z)-Ethyl((benzylamino)((2-oxoethyl)thio)methylene)carbamate

[0106] To a mixture of N-(benzylaminomethylthio)ethyl carbamate (4.00 g, 16.8 mmol, 1.0 equivalent) and cesium carbonate (13.7 g, 42.0 mmol, 2.5 equivalents) in acetonitrile (50 mL) was added 2-chloroacetaldehyde (13.2 g, 67.2 mmol, 4.0 equivalents). The reaction mixture was stirred at 25 ° C for 12 hours, poured into ice water (20 mL), and then extracted with MTBE (2×200 mL). The combined organic phase was washed with brine (2×20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate=100:1 to 3:1) to give 5.00 g of (Z)-ethyl ((benzylamino)((2-oxoethyl)thio)methylene)carbamate as a yellow solid. LCMS (m / z[M+H] + ):280.9.

[0107] (Z)-(3-Benzylthiazol-2(3H)-ylidene)carbamic acid ethyl ester

[0108] To a mixture of (Z)-ethyl ((benzylamino)((2-oxoethyl)thio)methylene)carbamate (5.00 g, 12.4 mmol, 1.0 equiv) and diisopropylethylamine (4.81 g, 37.2 mmol, 3.0 equiv) in tetrahydrofuran (50 mL) was added thionyl chloride (1.48 g, 12.4 mmol, 1.0 equiv). The mixture was stirred at 25 ° C for 16 hours, then poured into ice water (20 mL) and extracted with ethyl acetate (3×100 mL). The combined organic phases were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 100: 1 to 4: 1) to give 2.80 g of (Z)-ethyl (3-benzylthiazole-2(3H)-ylidene)carbamate as a yellow solid. 1 H NMR (400MHz, CD3OD) δ7.36-7.28(m,5H),7.22(d,J=4.8Hz,1H),6.84(d,J=4.8Hz,1H),5.31(s,2H),4.20(q,J=7.2Hz,2H),1.30(t,J=7.2Hz,3H). LCMS(m / z[M+H] + ):262.9.

[0109] Intermediate B1

[0110] A mixture of (Z)-ethyl(3-benzylthiazol-2(3H)-ylidene)carbamate (2.80 g, 9.06 mmol, 1.0 equiv) and sodium hydroxide (7.25 g, 181 mmol, 20 equiv) in ethanol (20 mL) was stirred at 50° C. for 1.5 hours. The mixture was poured into ice water (20 mL) and extracted with ethyl acetate (5×100 mL). The combined organic phases were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford 1.60 g of 3-benzylthiazol-2(3H)-imine B1 as a yellow gum.

[0111] Preparation of intermediate B2

[0112]

[0113] To a solution of 2-thiazolamine (1.00 g, 9.99 mmol, 1.0 equivalent) in EtOH (10 mL) was added MeI (1.84 g, 13.0 mmol, 808 μL, 1.3 equivalents). The reaction mixture was stirred at 90 ° C for 12 hours. The suspension was filtered, the solid was washed with EtOH (60 mL) and dried under reduced pressure. The solid was collected and diluted with 40% NaOH aqueous solution (10 mL) and extracted with EtOAc (3×10 mL). The combined organic layer was washed with 40% NaOH aqueous solution (3×10 mL) and brine (3×10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue to give 160 mg of 3-methyl-thiazol-2-amine B2.

[0114] Preparation of intermediate B3

[0115]

[0116] A mixture of 2-thiazolamine (3.00 g, 30.0 mmol, 1.0 equiv) and ethyl iodide (6.07 g, 38.9 mmol, 1.3 equiv) in EtOH (30 mL) was degassed and purged with N 2 three times and stirred at 90° C. under N 2 atmosphere for 12 hours. The reaction mixture was diluted with 40% aqueous NaOH (50 mL) and extracted with EtOAc (2×30 mL). The combined organic layers were washed with 40% aqueous NaOH (3×20 mL) and brine (3×20 mL), dried over anhydrous Na 2 SO 4 , filtered and concentrated under reduced pressure to give 2.80 g of 3-ethylthiazole-2(3H)-imine B3 as a dark red oil.

[0117] Preparation of intermediate B4

[0118]

[0119] 2-thiazolamine (120 mg, 1.20 mmol, 1.0 equiv), tert-butyl (4-bromobutan-2-yl)carbamate (302 mg, 1.20 mmol, 1.0 equiv) and DIPEA (170 mg, 1.32 mmol, 230 μL, 1.1 equiv) were stirred in a 5 mL sealed tube at 90° C. for 4 hours. The reaction mixture was diluted with MeOH (1 mL) and the mixture was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1 to 0 / 1, then ethyl acetate / ammonium hydroxide (25%) = 50 / 1) to give 30 mg of tert-butyl (4-(2-iminothiazol-3(2H)-yl)butan-2-yl)carbamate B4 as a yellow oil.

[0120] Preparation of intermediate B5

[0121]

[0122] A mixture of 2-thiazolamine (1.00 g, 10.0 mmol, 1.0 equiv) and 4-(bromomethyl)benzonitrile (1.96 g, 10.0 mmol, 1.0 equiv) in i-PrOH (20 mL) was stirred at 60° C. for 12 h. The reaction mixture was filtered to collect the solid, which was washed with 20 mL of EtOAc and dried under vacuum to give 2.1 g of 2-amino-3-(4-cyanobenzyl)thiazol-3-ium B5 as a yellow solid.

[0123] Preparation of intermediate B6

[0124]

[0125] To a solution of 2-thiazolamine (1.00g, 9.99mmol, 1.0 equivalent) and (bromomethyl) cyclohexane (1.77g, 9.99mmol, 1.39mL, 1.0 equivalent) in DMF (10mL) is added TBAI (1.84g, 4.99mmol, 0.5 equivalent) at room temperature. The reaction mixture is then stirred at 85°C for 12 hours under N2. Another 1.0 equivalent of (bromomethyl) cyclohexane is added to the mixture and the reaction mixture is stirred at 100°C for another 24 hours under N2. The reaction mixture is diluted with saturated NaHCO3 aqueous solution (100mL) and extracted with EtOAc (3x50mL). The combined organic layer is washed with salt water (3x50mL), through Na2SO4 drying, filtered and the filtrate is concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate / MeOH=0:1:0 to 0:1:1) to give 428 mg of 3-(cyclohexylmethyl)thiazol-2(3H)-imine B6 as a yellow solid.

[0126] Preparation of intermediate B7

[0127]

[0128] 5-(Chloromethyl)-1-methyl-1H-pyrazole

[0129] At 25 ℃, to (1-methyl-1H-pyrazole-5-yl) methanol (200mg, 1.78mmol, 1.0 equivalent) in DCM (4mL) stirred solution SOCl2 (0.32mL).Then the reaction mixture was stirred at 25 ℃ for 2 hours under N2.The reaction mixture was diluted with water (20mL) and adjusted to pH 8 with a saturated aqueous solution of NaHCO3.The solution was extracted with EtOAc (3×20mL), and the combined organic layers were washed with brine (3×10mL), dried over Na2SO4, and filtered.The filtrate was concentrated under reduced pressure to obtain 200mg of 5-(chloromethyl)-1-methyl-1H-pyrazole crude product as a yellow oil. 1 H NMR: (400MHz, CHLOROFORM-d) δ7.41 (d, J = 1.8 Hz, 1H), 6.28 (d, J = 1.8 Hz, 1H), 4.62 (s, 2H), 3.93 (s, 3H).

[0130] Intermediate B7

[0131] To a solution of 5-(chloromethyl)-1-methyl-1H-pyrazole (120 mg, 919 μmol, 1.0 equivalent) in i-PrOH (1 mL) was added 2-thiazolamine (74 mg, 735 μmol, 0.8 equivalent) at room temperature, followed by stirring at 60 ° C under N2 for 12 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate / MeOH=1:1:0 to 0:1:0 to 0:1:9) to give 76 mg of 3-((1-methyl-1H-pyrazol-5-yl)methyl)thiazole-2(3H)-imine B7 as a yellow solid.

[0132] Preparation of intermediate B8

[0133]

[0134] To a solution of ethyl 2-aminothiazole-4-carboxylate (2.00 g, 11.6 mmol, 1.0 equiv) in i-PrOH (20 mL) was added (bromomethyl)benzene (1.38 mL, 11.6 mmol, 1.0 equiv) at 25 ° C. The mixture was heated to 80 ° C and stirred for 6 hours. The reaction mixture was cooled to room temperature and allowed to stand for 3 days. The precipitate was collected by filtration and the filter cake was dried under high vacuum to give 0.80 g of ethyl 3-benzyl-2-imino-2,3-dihydrothiazole-4-carboxylate B8 as a white solid.

[0135] Preparation of intermediate B9

[0136]

[0137] To a solution of 5-methyl-2-thiazolamine (1.00 g, 8.76 mmol, 1.0 equiv) in acetone (15 mL) at 25°C was added bromotoluene (1.65 g, 9.63 mmol, 1.1 equiv). The reaction mixture was heated to 70°C and stirred for 5 hours. The reaction mixture was cooled to 25°C and adjusted to pH > 7.0 by adding aqueous NaOH (2N). The reaction mixture was filtered and the filter cake was washed with acetone (5 mL), MTBE (10 mL), and dried under reduced pressure to give 1.1 g of crude 3-benzyl-5-methylthiazole-2(3H)-imine B9 as a brown solid. The structure was confirmed by NOE.

[0138] Preparation of intermediate B10

[0139]

[0140] A mixture of 2-thiazolamine (100 mg, 998 μmol, 1.0 equivalent), 2-(bromomethyl)pyridine (278 mg, 1.10 mmol, 1.1 equivalent, HBr) and KCO (152 mg, 1.10 mmol, 1.1 equivalent) in acetone (5 mL) was degassed and purged with N 3 times, and the reaction mixture was then stirred at 60 ° C under an N atmosphere for 5 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC to give 100 mg of compound 3-(pyridin-2-ylmethyl)thiazole-2(3H)-imine B10 as a white solid.

[0141] Preparation of intermediate B11

[0142]

[0143] To a solution of 2-thiazolamine (0.30 g, 3.0 mmol, 1.0 equiv) in i-PrOH (5 mL) was added 2-(bromomethyl)benzonitrile (587 mg, 3.00 mmol, 1.0 equiv). The reaction mixture was stirred at 25° C. for 12 hours, then heated to 60° C. and stirred for an additional 4 hours. The reaction mixture was directly concentrated under reduced pressure to give a residue, which was purified by silica gel flash column chromatography (ethyl acetate / MeOH) to give 0.41 g of 2-((2-iminothiazol-3(2H)-yl)methyl)benzonitrile B11 as a yellow solid.

[0144] Preparation of intermediate B12

[0145]

[0146] To a solution of 2-bromoethylbenzene (3.70 g, 20.0 mmol, 1.0 equiv) in i-PrOH (35 mL) was added 2-thiazolamine (2.00 g, 20.0 mmol, 1.0 equiv). The reaction mixture was stirred at 80° C. for 12 hours. The reaction mixture was cooled to room temperature and then directly concentrated to give a residue, which was purified by reverse phase column chromatography to give 2.00 g of compound 3-phenethylthiazole-2(3H)-imine B12 as a light yellow solid.

[0147] Preparation of intermediate B13

[0148]

[0149] Methyl (tetrahydro-2H-pyran-4-yl) 4-methylbenzenesulfonate

[0150] To a solution of (tetrahydro-2H-pyran-4-yl)methanol (2.00 g, 17.2 mmol, 1.0 equiv) in 2-methyltetrahydrofuran (20 mL) was added NaOH (4.13 g, 51.6 mmol, 50% purity, 3.0 equiv) followed by the dropwise addition of a solution of 4-methylbenzenesulfonyl chloride (5.91 g, 31.0 mmol, 1.8 equiv) in 2-methyltetrahydrofuran (6 mL) at 0°C. The reaction mixture was then stirred at 25°C for 12 hours. The reaction mixture was diluted with aqueous HCl (6 M) and 3.0 mL of cyclohexane was added. The solution was stirred at 0°C for 0.5 hours but no solid precipitated. The mixture was then extracted with EtOAc (3×60 mL), the combined organic layers were washed with brine (100 mL), dried over Na 2 SO 4 , filtered, and the filtrate was concentrated under reduced pressure to give 4.2 g of crude (tetrahydro-2H)-pyran-4-yl)-4-methylbenzenesulfonic acid methyl ester as a white solid. LCMS (m / z [M+H] + ):271.1.

[0151] Intermediate B13

[0152] To a solution of (tetrahydro-2H-pyran-4-yl)methyl 4-methylbenzenesulfonate (2.70 g, 9.99 mmol, 1.0 equiv) and 2-thiazolamine (1.00 g, 9.99 mmol, 1.0 equiv) in DMF (10 mL) was added TBAI (1.84 g, 4.99 mmol, 0.5 equiv) at room temperature. The reaction mixture was stirred at 80° C. under N2 for 12 hours. The reaction mixture was diluted with water (80 mL) and extracted with EtOAc (3×60 mL). The combined organic layers were discarded and the aqueous phase was adjusted to pH=9 with NaOH (50% purity). The aqueous solution was extracted with EtOAc (3×50 mL), and the combined organic layers were washed with brine (100 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to provide 500 mg of 3-((tetrahydro-2H-pyran-4-yl)methyl)thiazol-2(3H)-imine B13 as a yellow solid.

[0153] Preparation of intermediate B14

[0154]

[0155] To a solution of 2-thiazolamine (0.60 g, 5.99 mmol, 1.0 equivalent) in i-PrOH (10 mL) was added methyl 3-(bromomethyl)benzoate (1.37 g, 5.99 mmol, 1.0 equivalent). The reaction mixture was stirred at 60 ° C for 12 hours. The reaction mixture was cooled to 25 ° C and diluted with MTBE. The suspension was decanted and dried under reduced pressure to give 1.5 g of crude product methyl 3-((2-iminothiazol-3(2H)-yl)methyl)benzoate B14 as a brown solid.

[0156] Preparation of intermediate B15

[0157]

[0158] A mixture (5 mL) of 2-thiazolamine (100 mg, 1.00 mmol, 1.0 equivalent), 2-(bromomethyl)pyridine (278 mg, 1.10 mmol, 1.1 equivalents, HBr) and KCO (152 mg, 1.10 mmol, 1.1 equivalents) in acetone was degassed and purged with N3 three times. The reaction mixture was stirred at 60 ° C under an N2 atmosphere for 5 hours. The resulting mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC to give 100 mg of 3-(pyridin-2-ylmethyl)thiazole-2(3H)-imine B15 as a white solid.

[0159] Preparation of intermediate B16

[0160]

[0161] To a solution of 2-thiazolamine (1.00 g, 9.99 mmol, 1.0 equiv) in EtOAc (20 mL) was added 3-(bromomethyl)benzonitrile (1.96 g, 9.99 mmol, 1.0 equiv) at 25°C. The reaction mixture was then stirred at 25°C for 12 hours. The reaction mixture was diluted with EtOAc (100 mL) and then NaOH (100 mL, 40%) was added at 0°C. The resulting mixture was stirred at 0°C for 1 hour. The solution was extracted with EtOAc (3×100 mL). The combined organic layers were washed with brine (3×100 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether / EtOAc=5:1 to EtOAc:NH3·H2O (25%)=100:1) to give 500 mg of 3-((2-iminothiazol-3(2H))-yl)methyl)benzonitrile B16 as a yellow oil.

[0162] Preparation of intermediate B17

[0163]

[0164] 4-(Chloromethyl)-1-methyl-1H-pyrazole

[0165] To a stirred solution of (1-methyl-1H-pyrazol-4-yl)methanol (500 mg, 4.46 mmol, 1.0 eq) in DCM (1.6 mL) was added SOCl (2.39 g, 20.1 mmol, 1.46 mL, 4.5 eq) dropwise at 25° C. The reaction mixture was stirred at 25° C. under N for 2 hours. The reaction mixture was concentrated under reduced pressure to give 500 mg of 4-(chloromethyl)-1-methyl-1H-pyrazole as a white solid. 1 H NMR (400MHz, DMSO-d6) δ7.79(s,1H),7.48(s,1H),4.68(s,2H),3.81(s,3H).

[0166] Intermediate B17

[0167] To a solution of 4-(chloromethyl)-1-methyl-1H-pyrazole (450 mg, 3.45 mmol, 1.0 equiv) in DMF (4.5 mL) was added 2-thiazolamine (345 mg, 3.45 mmol, 1.0 equiv), followed by the addition of K2CO3 (953 mg, 6.89 mmol, 2.0 equiv) at room temperature. The mixture was then stirred at 80 ° C for 12 hours under N2. The reaction mixture was diluted with water (30 mL) and extracted with EtOAc (3×20 mL), the organic layer was washed with brine (30 mL), dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate / MeOH=1:1:0 to 0:1:1) to give 50 mg of 3-((1-methyl-1H-pyrazol-4-yl)methyl)thiazole-2(3H)-imine B17 as a yellow solid.

[0168] Preparation of intermediate B18

[0169]

[0170] To a solution of 2-oxazolamine (0.50 g, 5.95 mmol, 1.0 equiv) in acetone (10 mL) was added bromomethylbenzene (1.12 g, 6.54 mmol, 777 μL, 1.1 equiv), and the mixture was stirred at 60° C. for 5 h. The reaction mixture was concentrated under reduced pressure to afford 0.70 g of 3-benzyloxazole-2(3H)-imine B18 as a yellow gelatinous oil.

[0171] Preparation of intermediate B19

[0172]

[0173]

[0174] To a solution of 4-methyl-2-oxazolamine (0.400 g, 3.26 mmol, 1.0 equiv) in i-PrOH (2 mL) was added (bromomethyl)benzene (0.387 mL 3.26 mmol, 1.0 equiv) dropwise at 20°C. The reaction mixture was heated to 80°C and stirred for 3 hours. The mixture was concentrated under reduced pressure and the residue was purified by preparative HPLC to give 0.20 g of 3-benzyl-4-methyloxazole-2(3H)-imine 2,2,2-trifluoroacetic acid B19 as a yellow oil.

[0175] Preparation of intermediate B20

[0176]

[0177] To a solution of ethyl 2-imino-3H-oxazole-4-carboxylate (1.35 g, 8.65 mmol) in isopropanol (14 mL) was added bromotoluene (1.03 mL, 8.65 mmol) at 25 ° C, and the mixture was stirred at 80 ° C for 12 hours. The reaction mixture was cooled to room temperature and the precipitate was collected by filtration. The filter cake was dried under reduced pressure to obtain 1.20 g of ethyl 3-benzyl-2-imino-2,3-dihydrooxazole-4-carboxylate B20 as a white solid.

[0178] Preparation of intermediate C1

[0179]

[0180] N-(phenylaminomethylthio)urethane

[0181] To a solution of aniline (2.00 g, 21.48 mmol, 1.0 equivalent) in EtOAc (20 mL) was added N-(thiomethylene) ethyl carbamate (2.82 g, 21.48 mmol, 1.0 equivalent) and TMEDA (250 mg, 2.15 mmol, 0.1 equivalent). The reaction mixture was stirred at 25 ° C for 5 hours. The reaction mixture was directly concentrated under reduced pressure to give a yellow solid. The solid was redissolved in EtOH (10 mL) and stirred for 0.5 hours and then filtered. The filter cake was washed with EtOH (2 × 5 mL) and dried under vacuum to give 3.70 g of N-(phenylaminomethylthioyl) ethyl carbamate as a white solid.

[0182] (Z)-Ethyl (((2-oxoethyl)thio)(phenylamino)methylene)carbamate

[0183] To a solution of ethyl N-(phenylaminomethylthio)carbamate (2.70 g, 12.0 mmol, 1.0 equiv) and CsCO (6.67 g, 20.5 mmol, 1.7 equiv) in CHCN (40 mL) was added 2-chloroacetaldehyde (2.95 g, 15.0 mmol, 1.3 equiv), maintaining the temperature below 25°C. After addition, the reaction mixture was stirred at 25°C for 12 hours. The reaction mixture was diluted with MTBE (80 mL) at 20°C, washed with saturated aqueous NaHCO (75 mL), brine (75 mL), dried over NaSO, and filtered. The filtrate was concentrated under reduced pressure to give 0.32 g of crude product (Z)-ethyl (((2-oxoethyl)thio)(phenylamino)methylene)carbamate as a brown oil.

[0184] (Z)-(3-phenylthiazol-2(3H)-ylidene)carbamic acid ethyl ester

[0185] To a solution of (Z)-ethyl(((2-oxoethyl)thio)(phenylamino)methylene)carbamate (3.20 g, 12.0 mmol, 1.0 equiv) in THF (40 mL) was added DIPEA (4.66 g, 36.0 mmol, 3.0 equiv) and SOCl2 (1.43 g, 12.0 mmol, 1.0 equiv) at 0°C. The reaction mixture was stirred at below 15°C for 4 hours. The reaction mixture was concentrated under reduced pressure. The residue was redissolved in MTBE (100 mL) and adjusted to pH 8.0 by the addition of saturated aqueous NaHCO3. The organic layer was washed with brine, dried over Na2SO4, and concentrated in vacuo. The residue was purified by flash silica gel chromatography (ethyl acetate / petroleum ether) to give 1.40 g of (Z)-ethyl(3-phenylthiazol-2(3H)-ylidene)carbamate as a yellow solid.

[0186] Intermediate C1

[0187] To a solution of (Z)-ethyl(3-phenylthiazol-2(3H)-ylidene)carbamate (0.80 g, 3.22 mmol, 1.0 equiv) in EtOH (10 mL) was added 6N NaOH (10 mL, 60.0 mmol, 19 equiv). The reaction mixture was stirred at 90°C for 4 hours. The reaction mixture was diluted with water (25 mL) and extracted with EtOAc (2 x 25 mL). The combined organic layers were dried over Na2SO4 and concentrated to give 0.60 g of crude 3-phenylthiazol-2-imine C1 as a brown solid. The crude product was purified by reverse phase MPLC to give 0.23 g of pure 3-phenylthiazol-2(3H)-imine C1 as a brown oil.

[0188] Preparation of intermediate C2

[0189]

[0190] Ethyl N-[(2-bromopropane-4-pyridine)aminomethylthio]carbamate

[0191] 2-Bromopyridin-4-amine (5.00g, 28.9mmol, 1.0 equivalent), N-(thiomethylene) ethyl carbamate (3.79g, 28.9mmol, 3.41mL, 1.0 equivalent) and TMEDA (336mg, 2.89mmol, 436uL, 0.1 equivalent) are degassed in EtOAc (50mL) and purged 3 times with N at room temperature, and then the reaction mixture is stirred at 25 DEG C under N atmosphere for 12h. The mixture is directly concentrated under reduced pressure to obtain a yellow solid. The solid is redissolved in EtOH (30mL) and stirred for 10 minutes and then filtered. The filter cake is washed with EtOH (2×5mL) and then concentrated under reduced pressure to obtain 4.2g of N-[(2-bromo-4-pyridyl)aminomethylthio] ethyl carbamate as a light yellow solid.

[0192] (Z)-Ethyl(((2-bromopyridin-4-yl)amino)((2-oxoethyl)thio)methylene)carbamate

[0193] To a solution of ethyl N-[(2-bromo-4-pyridinyl)aminomethylthio]carbamate (1.50 g, 4.93 mmol, 1.0 equiv) and Cs2CO3 (2.73 g, 8.38 mmol, 1.7 equiv) in MeCN (30 mL) was added 2-chloroacetaldehyde (484 mg, 6.16 mmol, 397 μL, 1.2 equiv) at room temperature. After addition, the reaction mixture was stirred at 25 ° C for 12 hours. The mixture was diluted with water (50 mL) and extracted with ethyl acetate (3×30 mL). The combined organic phases were washed with brine (30 mL), dried over anhydrous Na2SO4, filtered and concentrated in vacuo. (Z)-ethyl (((2-bromopyridin-4-yl)amino)((2-oxoethyl)thio)methylene)carbamate (1.60 g, crude) was obtained as a brown oil.

[0194] (Z)-Ethyl (3-(2-bromopyridin-4-yl)thiazol-2(3H)ylidene)carbamate

[0195] To a solution of (Z)-ethyl (((2-bromopyridin-4-yl)amino)((2-oxoethyl)thio)methylene)carbamate (1.60 g, 4.62 mmol, 1.0 equiv) in THF (20 mL) was added DIPEA (1.79 g, 13.9 mmol, 2.42 mL, 3.0 equiv) and SOCl (550 mg, 4.62 mmol, 335 μL, 1.0 equiv) at 0°C. The reaction mixture was stirred below 20°C for 12 h. The mixture was quenched with saturated aqueous NaHCO (50 mL) and extracted with dichloromethane (3 x 20 mL). The combined organic layers were washed with brine, dried over NaSO, and concentrated to afford 1.40 g of (Z)-ethyl (3-(2-bromopyridin-4-yl)thiazol-2(3H)-ylidene)carbamate as a brown oil.

[0196] (Z)-Ethyl (3-(2-((tert-butoxycarbonyl)amino)pyridin-4-yl)thiazol-2(3H)-ylidene)carbamate

[0197] (Z)-ethyl (3-(2-bromopyridin-4-yl)thiazol-2(3H)-ylidene)carbamate (700 mg, 2.13 mmol, 1.0 equiv), NH2Boc (750 mg, 6.40 mmol, 3.0 equiv), K3PO4 (1.81 g, 8.53 mmol, 4.0 equiv), Pd2(dba)3 (195 mg, 213 μmol, 0.1 equiv) and Xantphos (123 mg, 213 μmol, 0.1 equiv) were degassed and purged with N2 three times in dioxane (10 mL), and the reaction mixture was stirred at 110 ° C. under N2 atmosphere for 12 hours. The mixture was poured into water (30 mL) and extracted with ethyl acetate (3×20 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50 / 1 to 1 / 1) to give 140 mg of (Z)-ethyl (3-(2-((tert-butoxycarbonyl)amino)pyridin-4-yl)thiazol-2(3H)-ylidene)carbamate as a yellow solid. 1 H NMR (400MHz, METHANOL-d4) δ8.37(d,J=5.50Hz,1H),8.09(s,1H),7.43(d,J=4.75Hz,1H),7.29(dd,J=5 .50,1.75Hz,1H),7.01(d,J=4.88Hz,1H),4.18(q,J=7.13Hz,2H),1.54(s,9H),1.26(t,J=7.07Hz,3H).

[0198] Intermediate C2

[0199] To a solution of (Z)-ethyl (3-(2-((tert-butoxycarbonyl)amino)pyridin-4-yl)thiazol-2(3H)-ylidene)carbamate (110 mg, 302 μmol, 1.0 equiv) in EtOH (4 mL) was added NaOH (6 M, 4.0 mL, 80 equiv). The reaction mixture was stirred at 60 °C for 2 h. The mixture was diluted with water (30 mL) and extracted with THF (2 x 20 mL). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to afford the intermediate C2 tert-butyl (4-(2-iminothiazol-3(2H)-yl)pyridin-2-yl)carbamate (100 mg, crude) as a brown solid.

[0200] Preparation of intermediate C3

[0201]

[0202] N-(o-Tolylaminomethylthio)urethane

[0203] To a solution of o-toluidine (2.00 g, 18.7 mmol, 1.0 equivalent) in ethyl acetate (20 mL) was added TMEDA (0.217 g, 1.87 mmol, 0.1 equivalent) and O-ethyl carboisothiocyanate (2.45 g, 18.7 mmol, 1.0 equivalent) at 25°C. The mixture was stirred at 25°C for 2 hours and then evaporated under reduced pressure. The residue was triturated in ethanol (10 mL) to give 3.90 g of ethyl N-(o-tolylaminomethylthioyl)urethane as a white solid. 1 H NMR(400MHz, CDCl3)δ11.18(br.s,1H),8.36(br.s,1H),7.71-7.58(m,1H),7 .38-7.20(m,3H),4.31(q,J=7.2Hz,2H),2.34(s,3H),1.37(t,J=7.2Hz,3H). LCMS(m / z[M+H] + ):239.1.

[0204] (Z)-Ethyl (((2-oxoethyl)thio)(o-tolylamino)methylene)carbamate

[0205] To a solution of ethyl N-(o-tolylaminomethylthio)carbamate (3.90 g, 16.3 mmol, 1.0 equiv) and Cs2CO3 (9.05 g, 27.8 mmol, 1.7 equiv) in acetonitrile (40 mL) was added 2-chloroacetaldehyde (3.85 g, 19.6 mmol, 40% aqueous solution, 1.2 equiv) maintaining the temperature below 25°C. After addition, the mixture was stirred at 25°C for 12 hours, then diluted with MTBE (100 mL), washed with saturated aqueous NaHCO3 (100 ml), then with brine (50 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to give 5.80 g of crude (Z)-ethyl (((2-oxoethyl)thio)(o-tolylamino)methylene)carbamate as a brown gum. 1 H NMR (400MHz, CDCl3) δ7.40-7.27(m,5H),5.39(d,J=5.2Hz,1H),4.16(q,J=7.2Hz,2H),3 .65(dd,J=6.0,12.4Hz,1H),3.28(d,J=12.4Hz,1H),2.20(s,3H),1.26(t,J=7.2Hz,3H). LCMS(m / z[M+H] + ):281.1.

[0206] (Z)-Ethyl (3-(o-tolyl)thiazole-2(3H) substituent) carbamate

[0207] To a solution of (Z)-ethyl (((2-oxoethyl)thio)(o-tolylamino)methylene)carbamate (5.80 g, 12.1 mmol, 1.0 equiv) and DIEA (6.32 mL, 36.3 mmol, 3.0 equiv) in THF was added dropwise SOCl2 (0.878 mL, 12.1 mmol, 1.0 equiv) (60 mL) at 0 ° C. The mixture was stirred at 0 ° C for 4 hours, then quenched with water (100 mL) and extracted with ethyl acetate (30 mL×3). The combined organic layers were dried over sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=10:1 to 3:1) to give 2.70 g of (Z)-ethyl (3-(o-tolyl)thiazole-2(3H)-)dimethylene)carbamate as a yellow oil. 1H NMR (400MHz, CDCl3) δ7.41-7.28(m,3H),7.25-7.19(m,1H),6.85(d,J=4.8Hz,1H) ,6.69(d,J=4.8Hz,1H),4.21(q,J=7.2Hz,2H),2.14(s,3H),1.29(t,J=7.2Hz,3H). LCMS(m / z[M+H] + ):263.1.

[0208] Intermediate C3

[0209] A solution (6 mL) of (Z)-ethyl(3-(o-tolyl)thiazole-2(3H)-)dimethylene)carbamate (0.500 g, 1.79 mmol, 1.0 equiv) and NaOH (1.43 g, 35.8 mmol, 20 equiv) in ethanol was stirred at 50 ° C for 1 hour. The mixture was cooled to room temperature and diluted in ethyl acetate (20 mL), washed with water (20 mL), then washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to give 0.23 g of 3-(o-tolyl)thiazole-2(3H)-imine C3 as a yellow oil.

[0210] Preparation of intermediate C4

[0211]

[0212] Ethyl N-[(2-methoxyphenyl)aminomethylthio]carbamate

[0213] To a solution of 2-methoxyaniline (2.00 g, 16.2 mmol, 1.0 equiv) in ethyl acetate (20 mL) was added O-ethyl carboisothiocyanate (1.92 mL, 16.2 mmol, 1.0 equiv) and tetramethylethylenediamine (0.245 mL, 1.62 mmol, 0.1 equiv) at 25°C. The mixture was stirred at this temperature for 5 hours. The mixture was concentrated under reduced pressure to remove the ethyl acetate. The residue was triturated twice with ethanol (5 mL) to give 3.40 g of ethyl N-[(2-methoxyphenyl)aminomethylthio]carbamate as an off-white solid. 1H NMR (400MHz, DMSO-d6) δ11.90(br.s,1H),11.25(br.s,1H),8.48(dd,J=8.0,1.2Hz,1H),7.19(dt,J=8.4,1.6Hz,1H) ,7.10(dd,J=8.0,1.2Hz,1H),6.96(dt,J=8.0,1.2Hz,1H),4.22(q,J=7.2Hz,2H),3.85(s,3H),1.25(t,J=7.2Hz,3H).

[0214] Ethyl (Z)-(((2-methoxyphenyl)amino)((2-oxoethyl)thio)methylene)carbamate

[0215] To a suspension of N-[(2-methoxyphenyl)aminomethylthio]ethyl carbamate (3.40 g, 13.4 mmol, 1.0 equiv) and cesium carbonate (7.41 g, 22.7 mmol, 1.7 equiv) in acetonitrile (40 mL) was added 2-chloroacetaldehyde (2.69 mL, 16.7 mmol, 1.2 equiv) at 25°C. The resulting mixture was stirred at this temperature for 12 hours. Additional 2-chloroacetaldehyde (1.08 mL, 6.68 mmol, 0.5 equiv) was added dropwise to the mixture at 25°C and stirred for an additional 2 hours. The mixture was poured into water (150 mL) and extracted with MTBE (100 mL x 3). The combined organic layers were washed with brine (75 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give 4.60 g of (Z)-ethyl(((2-methoxyphenyl)amino)((2-oxoethyl)thio)methylene)carbamate as a brown oil. 1 H NMR (400MHz, DMSO-d6): δ7.37(dt,J=8.4,1.6Hz,1H),7.19(dd,J=7.6,1.6Hz,1H),7.13(dd,J=8.4,0.8Hz,1H),7.00(dt,J=7.6,1.2Hz,1H ), 6.75 (d, J = 7.2Hz, 1H), 5.46 (dt, J = 7.6, 2.4Hz, 1H), 3.93 (q, J = 7.2Hz, 2H), 3.77 (s, 3H), 3.59 (dd, J = 12.4, 6.4Hz, 1H), 1.12-1.10 (m, 3H). LCMS(m / z[M+H] + ):296.8.

[0216] (Z)-Ethyl (3-(2-methoxyphenyl)thiazol-2(3H)-ylidene)carbamate

[0217] To a solution of (Z)-ethyl(((2-methoxyphenyl)amino)((2-oxoethyl)thio)methylene)carbamate (2.60 g, 6.10 mmol, 1.0 equiv) in THF (20 mL) was added diisopropylethylamine (3.19 mL, 18.3 mmol, 3.0 equiv) followed by SOCl2 (0.442 mL, 6.10 mmol, 1.0 equiv) at 0°C. The mixture was stirred at below 15°C for 4 hours. The mixture was added to saturated aqueous sodium bicarbonate solution (20 mL) and stirred for 5 minutes, and extracted with ethyl acetate (100 mL×3). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 5 / 1) to give 1.00 g of (Z)-ethyl (3-(2-methoxyphenyl)thiazol-2(3H)-propylidene)carbamate as an off-white solid. 1 H NMR (400MHz, DMSO-d6): δ7.51-7.47(m,1H),7.35(dd,J=6.8,1.6Hz,1H),7.32(d,J=4.8Hz,1H),7.25(dd,J=8.4, 0.8Hz, 1H), 7.10-7.06 (m, 1H), 6.99 (d, J = 4.8Hz, 1H), 3.98 (q, J = 7.2Hz, 2H), 3.76 (s, 3H), 1.13 (t, J = 7.2Hz, 3H). LCMS(m / z[M+H] + ):278.8.

[0218] Intermediate C4

[0219] To a solution of ethyl (3-(2-methoxyphenyl)thiazol-2(3H)-ylidene)carbamate (0.500 g, 1.80 mmol, 1.0 equiv) in ethanol (5 mL) was added sodium hydroxide (1.44 g, 35.9 mmol, 20 equiv) in one portion at 25°C, and the mixture was stirred for 1 hour. The mixture was poured into water (20 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with brine (25 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford 0.350 g of 3-(2-methoxyphenyl)thiazol-2(3H)-imine C4 as a brown oil.

[0220] Preparation of intermediate C5

[0221]

[0222] Ethyl N-[(2-chlorophenyl)aminomethylthio]carbamate

[0223] To a solution of 2-chloroaniline (2.20 g, 17.3 mmol, 1.0 equiv) and O-ethylcarbothiocyanate (2.26 g, 17.3 mmol, 1.0 equiv) in ethyl acetate (20 mL) was added tetramethylethylenediamine (0.200 g, 1.72 mmol, 0.1 equiv) at 0°C. The resulting mixture was stirred at 25°C for 2 hours and then concentrated under reduced pressure. The residue was triturated with methanol (30 mL) to give 3.00 g of ethyl N-[(2-chlorophenyl)aminomethylthio]carbamate as a white solid. 1 H NMR (400MHz, DMSO-d6) δ11.60(s,1H),11.48(s,1H),7.94(dd,J=8.0,1.6Hz,1H),7.56(dd,J=8.0 ,1.6Hz,1H),7.40-7.36(m,1H),7.33-7.29(m,1H),4.23(q,J=7.2Hz,2H),1.26(t,J=7.2Hz,3H).

[0224] Ethyl (Z)-(((2-chlorophenyl)amino)((2-oxoethyl)thio)methylene)carbamate

[0225] To a suspension of ethyl N-[(2-chlorophenyl)aminomethylthio]carbamate (3.00 g, 11.6 mmol, 1.0 equiv) and Cs2CO3 (6.42 g, 19.7 mmol, 12 equiv) in acetonitrile (40 mL) was added 2-chloroacetaldehyde (2.84 g, 14.5 mmol, 1.2 equiv), maintaining the temperature below 25°C. The reaction mixture was stirred at 25°C for 12 hours. The mixture was poured into water (100 mL) and extracted with methyl TBME (150 mL x 2). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 3.80 g of crude (Z)-ethyl (((2-chlorophenyl)amino)((2-oxoethyl)thio)methylene)carbamate as a brown gum. 1 H NMR (400MHz, DMSO-d6) δ7.66-7.58(m,1H),7.49-7.35(m,3H),6.99(d,J=6.4Hz,1H),5.53-5 .42(m,1H),4.00-3.90(m,2H),3.70-3.65(m,1H),3.23-3.15(m,1H),1.11(t,J=7.2Hz,3H).

[0226] (Z)-(3-(2-chlorophenyl)thiazol-2(3H)-ylidene)carbamic acid ethyl ester

[0227] To a solution of (Z)-ethyl (((2-chlorophenyl)amino)((2-oxoethyl)thio)methylene)carbamate (1.80 g, 5.98 mmol, 1.0 equiv) in THF (20 mL) was added diisopropylethylamine (2.32 g, 17.9 mmol, 3.0 equiv) and thionyl chloride (0.710 g, 5.98 mmol, 1.0 equiv) at 0°C. The mixture was stirred at 15°C for 4 hours, then poured into water (100 mL) and extracted with ethyl acetate (150 mL x 4). The combined organic phases were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 0 to 10 / 1) to give 1.101 g of (Z)-ethyl (3-(2-chlorophenyl)thiazole-2(3H)-propylene)carbamate as a yellow solid. 1 HNMR (400MHz, DMSO-d6) δ7.73-7.71(m,1H),7.60-7.53(m,3H),7.45(d,J=4. 8Hz, 1H), 7.09 (d, J = 4.8Hz, 1H), 4.00 (q, J = 6.8Hz, 2H), 1.14 (t, J = 7.2Hz, 3H). LCMS(m / z[M+H] + ):282.9.

[0228] Intermediate C5

[0229] To a solution of (Z)-ethyl(3-(2-chlorophenyl)thiazol-2(3H)-ylidene)carbamate (0.80 g, 2.75 mmol, 1.0 equiv) in ethanol (10 mL) was added sodium hydroxide (2.34 g, 58.4 mmol, 21 equiv) at 25°C. The mixture was heated to 50°C and stirred for 1 hour, then poured into water (80 mL) and extracted with ethyl acetate (120 mL x 4). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give 0.54 g of 3-(2-chlorophenyl)thiazol-2(3H)-imine C5 as a yellow solid.

[0230] Preparation of intermediate C6

[0231]

[0232] N-(m-Tolylaminomethylthio)urethane

[0233] To a solution of m-toluidine (2.00 g, 18.6 mmol, 1.0 equiv) in ethyl acetate (20 mL) was added O-ethyl carboisothiocyanate (2.21 mL, 18.6 mmol, 1.0 equiv) and tetramethylethylenediamine (0.282 mL, 1.87 mmol, 0.1 equiv). The mixture was stirred at 25 ° C for 5 hours. The mixture was concentrated under reduced pressure to remove the ethyl acetate. The residue was triturated twice with ethanol (5 mL) to give 3.00 g of ethyl N-(m-tolylaminomethylthioyl)carbamate as a white solid. 1 H NMR (400MHz, DMSO-d6) δ11.51(br.s,1H),11.23(br.s,1H),7.43-7.38(m,2H),7.26(t,J=7.6 Hz, 1H), 7.05 (d, J = 7.6Hz, 1H), 4.20 (q, J = 8.0, 7.2Hz, 2H), 2.30 (s, 3H), 1.25 (t, J = 6.8Hz, 3H).

[0234] Ethyl (Z)-(((2-oxoethyl)thio)(m-toluamino)methylene)carbamate

[0235] To a suspension of ethyl N-(m-tolylaminomethylthio)carbamate (3.00 g, 12.6 mmol, 1.0 equiv) and cesium carbonate (6.97 g, 21.4 mmol, 1.7 equiv) in acetonitrile (40 mL) was added 2-chloroacetaldehyde (2.53 mL, 15.7 mmol, 1.25 equiv) dropwise at 25°C and the mixture was stirred for 12 hours. The mixture was poured into water (150 mL) and extracted with methyl tert-butyl ether (100 mL x 3). The combined organic layers were washed with brine (75 mL), dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give 4.00 g of (Z)-ethyl (((2-oxoethyl)thio)(m-tolylamino)methylene)carbamate as a brown oil. 1 H NMR (400MHz, DMSO-d6) δ7.34-7.30(m,1H),7.16-7.13(m,3H),6.85(d,J=7.2Hz,1H) ,5.60(dt,J=7.2,1.6Hz,1H),3.96(q,J=7.2Hz,2H),2.32(s,3H),1.14-1.10(m,3H). LCMS(m / z[M+H] + ):280.9.

[0236] Ethyl (Z)-(3-(m-tolyl)thiazol-2(3H)-ylidene)carbamate

[0237] To a solution of (Z)-ethyl (((2-oxoethyl)thio)(m-tolylamino)methylene)carbamate (2.00 g, 5.58 mmol, 1.0 equivalent) in THF (20 mL) at 0° C. was added diisopropylethylamine (2.92 mL, 16.8 mmol, 3.0 equivalents) followed by SOCl 2 (0.405 mL, 5.58 mmol, 1.0 equivalents). The mixture was stirred for 4 hours at a temperature below 15° C. The mixture was poured into a saturated aqueous sodium bicarbonate solution to adjust the pH to 8 and then extracted with ethyl acetate (100 mL x 3). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate=10 / 1) to give 1.00 g of (Z)-ethyl (3-(m-tolyl)thiazole-2(3H)-ylidene)carbamate as a yellow oil. 1 H NMR(400MHz,DMSO-d6)δ7.49(d,J=4.8Hz,1H),7.42(t,J=8.0Hz,1H),7.30-7.28(m ,3H),7.05(d,J=4.8Hz,1H),4.04-3.99(m,2H),2.37(s,3H),1.15(t,J=7.2Hz,3H). LCMS(m / z[M+H] + ):262.8.

[0238] Intermediate C6

[0239] To a solution of (Z)-(3-(m-tolyl)thiazol-2(3H)-ylidene)carbamic acid ethyl ester (0.200 g, 0.762 mmol, 1.0 equiv) in ethanol (2 mL) was added sodium hydroxide (0.610 g, 15.2 mmol, 20 equiv) in one portion at 25°C, and the mixture was then heated to 50°C and stirred for 1 hour. The mixture was added to water (50 mL) and extracted with ethyl acetate (50 mL x 4). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford 140 mg of 3-(m-tolyl)thiazol-2(3H)-imine C6 as a brown oil.

[0240] Preparation of intermediate C7

[0241]

[0242] Ethyl N-[(3-methoxyphenyl)aminomethylthio]carbamate

[0243] To a solution of 3-methoxyaniline (2.00 g, 16.2 mmol, 1.0 equivalent) in ethyl acetate (10 mL) was added TMEDA (0.189 g, 1.62 mmol, 0.1 equivalent) and O-ethyl carbonyl isothiocyanate (2.13 g, 16.2 mmol, 1.0 equivalent) at 25 ° C. The mixture was stirred for 2 hours at 25 ° C. and then evaporated under reduced pressure. Residue was crystallized from petroleum ether / ethyl acetate (v / v=5:1, 10 mL) to give 3.70 g of N-[(3-methoxyphenyl)aminomethylthio]ethyl carbamate as a yellow solid. 1 HNMR (400MHz, CDCl3) δ11.48(br.s,1H),8.11(br.s,1H),7.41(t,J=2.0Hz,1H),7.30(t,J=8.0Hz,1H),7.14(dd, J=1.6,8.0Hz,1H),6.82(ddd,J=0.8,2.4,8.4Hz,1H),4.30(q,J=7.2Hz,2H),3.83(s,3H),1.36(t,J=7.2Hz,3H). LCMS(m / z[M+H] + ):255.1.

[0244] Ethyl (Z)-(((3-methoxyphenyl)amino)((2-oxoethyl)thio)methylene)carbamate

[0245] To a solution of ethyl N-[(3-methoxyphenyl)aminomethylthio]carbamate (3.70 g, 14.5 mmol, 1.0 equiv) and CsCO (8.01 g, 24.6 mmol, 1.7 equiv) in acetonitrile (40 mL) was added 2-chloroacetaldehyde (3.40 g, 17.3 mmol, 40% aqueous solution, 1.2 equiv), maintaining the temperature below 25° C. The mixture was stirred at 25° C. for 12 hours, then diluted with MTBE (100 mL) and washed with saturated aqueous NaHCO (100 mL) and brine (50 mL), dried over NaSO, and filtered. The filtrate was concentrated under reduced pressure to give 5.00 g of (Z)-ethyl (((3-methoxyphenyl)amino)((2-oxoethyl)thio)methylene)carbamate as a yellow oil. 1H NMR (400MHz, CDCl3) δ7.33(t,J=8.0Hz,1H),7.04-6.95(m,2H),6.87(ddd,J=0.8,2.4,8.4Hz,1H),5.61(d,J=5.2Hz,1H) ,4.18(q,J=7.2Hz,2H),3.81(s,3H),3.60(dd,J=5.6,12.0Hz,1H),3.22(dd,J=1.2,12.4Hz,1H),1.28(t,J=7.2Hz,3H). LCMS(m / z 297.1[M+H] + ):297.1.

[0246] (Z)-(3-(3-methoxyphenyl)thiazol-2(3H)-ylidene)carbamic acid ethyl ester

[0247] To a solution of (Z)-ethyl (((3-methoxyphenyl)amino)((2-oxoethyl)thio)methylene)carbamate (5.00 g, 11.3 mmol, 1.0 equiv) and DIEA (4.37 g, 33.8 mmol, 3.0 equiv) in THF (60 mL) was added dropwise SOCl2 (0.818 mL, 11.3 mmol, 1.0 equiv) at 0 ° C. The mixture was stirred at 0 ° C for 4 hours, then quenched with water (100 mL) and extracted with ethyl acetate (30 mL×3). The combined organic layers were dried over sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate=10: 1 to 3: 1) to give 2.70 g of (Z)-ethyl (3-(3-methoxyphenyl)thiazole-2(3H)-dimethylene)carbamate as a yellow oil. 1 H NMR(400MHz, CDCl3)δ7.38(t,J=8.0Hz,1H),7.07-6.98(m,3H),6.94(dd,J=2.0,8.0Hz ,1H),6.66(d,J=4.8Hz,1H),4.22(q,J=7.2Hz,2H),3.83(s,3H),1.30(t,J=7.2Hz,3H). LCMS(m / z[M+H] + ):279.1.

[0248] Intermediate C7

[0249] A solution (6 mL) of (Z)-ethyl (3-(3-methoxyphenyl)thiazole-2(3H)-ylidene)carbamate (0.500 g, 1.70 mmol, 1.0 equiv) and NaOH (1.36 g, 34.1 mmol, 20 equiv) in ethanol was stirred at 50 ° C for 1 hour. The mixture was cooled to room temperature and diluted in ethyl acetate (20 mL), washed with water (20 mL), then washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane: methanol = 20: 1) to give 230 mg of 3-(3-methoxyphenyl)thiazole-2(3H)-imine C7 as a yellow oil.

[0250] Preparation of intermediate C8

[0251]

[0252] Ethyl N-[(3-chlorophenyl)aminomethylthio]carbamate

[0253] To a solution of 3-chloroaniline (2.00 g, 15.7 mmol, 1.0 equivalent) in ethyl acetate (20 mL) was added O-ethyl carboisothiocyanate (1.85 mL, 15.7 mmol, 1.0 equivalent) and tetramethylethylenediamine (0.237 mL, 1.57 mmol, 0.1 equivalent). The mixture was stirred at 25 ° C for 5 hours. The mixture was concentrated under reduced pressure. The residue was triturated twice with petroleum ether / ethyl acetate (v / v=10 / 1, 11 mL) to give 3.20 g of ethyl N-[(3-chlorophenyl)aminomethylthio]carbamate as a yellow solid. 1 HNMR (400MHz, DMSO-d6): δ11.55(br.s,1H),11.35(br.s,1H),7.84(s,1H),7.49-7.47(m,1 H), 7.41 (t, J = 7.6Hz, 1H), 7.32-7.30 (m, 1H), 4.21 (q, J = 7.6Hz, 2H), 1.25 (t, J = 7.2Hz, 3H).

[0254] Ethyl (Z)-(((3-chlorophenyl)amino)((2-oxoethyl)thio)methylene)carbamate

[0255] To a suspension of N-[(3-chlorophenyl)aminomethylthio]ethyl carbamate (3.20 g, 12.4 mmol, 1.0 equiv) and cesium carbonate (6.85 g, 21.0 mmol, 1.7 equiv) in acetonitrile (40 mL) was added 2-chloroacetaldehyde (2.49 mL, 15.4 mmol, 1.25 equiv) dropwise at 25 ° C. The resulting mixture was stirred at this temperature for 12 hours. The mixture was poured into water (150 mL) and extracted with MTBE (100 mL x 3) at 25 ° C. The combined organic layers were washed with brine (75 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 3.80 g of (Z)-ethyl (((3-chlorophenyl)amino)((2-oxoethyl)thio)methylene)carbamate as a brown oil. 1 H NMR (400MHz, DMSO-d6): δ7.50-7.46(m,2H),7.43-7.40(m,1H),7.38-7.36(m,1H),6.95(br.s,1H),5 .69-5.68(m,1H),3.98(t,J=7.6Hz,2H),3.68-3.55(m,1H),3.10-3.05(m,1H),1.14(t,J=7.6Hz,3H). LCMS(m / z[M+H] + ):300.8. (Z)-(3-(3-chlorophenyl)thiazol-2(3H)-ylidene)carbamic acid ethyl ester

[0256] To a solution of (Z)-ethyl(((3-chlorophenyl)amino)((2-oxoethyl)thio)methylene)carbamate (1.80 g, 3.68 mmol, 1.0 equiv) in THF (20 mL) was added diisopropylethylamine (1.92 mL, 11.0 mmol, 3.0 equiv) and then SOCl2 (0.267 mL, 3.68 mmol, 1.0 equiv) at 0°C under a nitrogen atmosphere. The mixture was stirred for 4 hours below 15°C. SOCl2 (0.134 mL, 1.84 mmol, 0.5 equiv) was added dropwise to the mixture at 0°C, and the reaction mixture was stirred for 2 hours at 15°C. The mixture was added to a saturated aqueous sodium bicarbonate solution to adjust pH=8 and extracted with ethyl acetate (100 mL x 3). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 5 / 1) to give 0.95 g of (Z)-ethyl (3-(3-chlorophenyl)thiazole-2(3H)-dimethylene)carbamate as an off-white solid. 1H NMR (400MHz, DMSO-d6): δ7.70 (s, 1H), 7.58-7.54 (m, 3H), 7.53-7.51 (m, 1H), 7.07 (d, J = 4.8Hz, 1H), 4.03 (q, J = 7.2Hz, 2H), 1.16 (t, J = 7.2Hz, 3H). LCMS(m / z[M+H] + ):282.8.

[0257] Intermediate C8

[0258] To a solution of (Z)-ethyl(3-(3-chlorophenyl)thiazol-2(3H)-ylidene)carbamate (0.200 g, 0.707 mmol, 1.0 equiv) in ethanol (2 mL) was added sodium hydroxide (0.566 g, 14.2 mmol, 20 equiv) in one portion at 25°C. The mixture was stirred at this temperature for 1 hour. The mixture was added to water (50 mL) and extracted with ethyl acetate (50 mL x 4). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford 130 mg of 3-(3-chlorophenyl)thiazol-2(3H)-imine C8 as a brown solid.

[0259] Preparation of intermediate C9

[0260]

[0261] To a solution of p-toluidine (2.00 g, 18.7 mmol, 1.0 equivalent) in ethyl acetate (10 mL) was added TMEDA (0.217 g, 1.87 mmol, 0.1 equivalent) and O-ethyl carboisothiocyanate (2.45 g, 18.7 mmol, 1.0 equivalent) at 25°C. The mixture was stirred at 25°C for 2 hours and then evaporated under reduced pressure. The residue was triturated in ethanol (10 mL) to give 3.70 g of ethyl N-(p-tolylaminomethylthio)urethane as a yellow solid. 1 H NMR(400MHz,CDCl3)δ11.35(br s,1H),8.21(br s, 1H), 7.48 (d, J = 8.4Hz, 2H), 7.21 (d, J = 8.0Hz, 2H), 4.29 (q, J = 7.2Hz, 2H), 2.37 (s, 3H), 1.35 (t, J = 7.2Hz, 3H). LCMS(m / z[M+H] + ):239.1.

[0262] (Z)-(((2-Oxoethyl)thio)(p-tolylamino)methylene)carbamate

[0263] To a solution of ethyl N-(p-tolylaminomethylthio)carbamate (3.70 g, 15.5 mmol, 10 equiv) and Cs2CO3 (8.59 g, 26.4 mmol, 1.7 equiv) in acetonitrile (40 mL) was added 2-chloroacetaldehyde (3.65 g, 18.6 mmol, 40% aqueous solution, 1.2 equiv) maintaining the temperature below 25°C. After addition, the mixture was stirred at 25°C for 12 hours, then diluted with MTBE (100 mL), washed with saturated aqueous NaHCO3 (100 ml), then with brine (50 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to give 5.30 g of crude (Z)-ethyl (((2-oxoethyl)thio)(p-tolylamino)methylene)carbamate as a yellow oil. 1 H NMR (400MHz, CDCl3) δ7.39-7.18(m,5H),5.56(dd,J=0.5,5.6Hz,1H),4.16(q,J=7.2Hz,2H),3 .57(dd,J=5.6,12.4Hz,1H),3.19(dd,J=1.2,12.0Hz,1H),2.35(s,3H),1.26(t,J=7.2Hz,3H). LCMS(m / z[M+H] + ):281.1.

[0264] Ethyl (Z)-(3-(p-Tolyl)thiazol-2(3H)-ylidene)carbamate

[0265] To a solution of (Z)-ethyl (((2-oxyethyl)thio) (p-tolylamino) methylene) carbamate (5.30 g, 10.8 mmol, 1.0 equiv) and DIEA (4.19 g, 32.4 mmol, 3.0 equiv) in THF was added dropwise SOCl2 (0.783 mL, 10.8 mmol, 1.0 equiv) (60 mL) at 0 ° C. The mixture was stirred at 0 ° C for 4 hours, then quenched with water (100 mL) and extracted with ethyl acetate (30 mL × 3). The combined organic layers were dried over sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10: 1 to 3: 1) to give 2.20 g of (Z)-ethyl (3-(p-tolyl)thiazole-2(3H)-)dimethylene) carbamate as a yellow oil. 1H NMR (400MHz, CDCl3) δ7.26 (d, J = 8.4Hz, 2H), 7.21 (d, J = 8.4Hz, 2H), 6.93 (d, J = 4.8Hz, 1H), 6.58 (d, J = 4.8Hz, 1H), 4.14 (q, J = 7.2Hz, 2H), 2.32 (s, 3H), 1.22 (t, J = 7.2Hz, 3H). LCMS o(m / z[M+H] + ):263.1.

[0266] Intermediate C9

[0267] A solution of (Z)-(3-(p-tolyl)thiazole-2(3H)-ylidene)carbamic acid ethyl ester (0.500 g, 1.86 mmol, 1.0 equiv) and NaOH (1.49 g, 37.2 mmol, 20 equiv) in ethanol (6 mL) was stirred at 50 ° C for 1 hour. The mixture was cooled to room temperature and diluted in ethyl acetate (20 mL), washed with water (20 mL), then washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane / methanol=20:1) to give 230 mg of 3-(p-tolyl)thiazole-2(3H)-imine C9 as a yellow oil.

[0268] Preparation of intermediate C10

[0269]

[0270] Ethyl N-[(4-methoxyphenyl)aminomethylthio]carbamate

[0271] To a solution of 4-methoxyaniline (2.00 g, 16.2 mmol, 1.0 equiv) and O-ethylcarboisothiocyanate (2.13 g, 16.2 mmol, 1.0 equiv) in ethyl acetate (20 mL) was added tetramethylethylenediamine (0.189 g, 1.62 mmol, 0.1 equiv) at 25° C., and the mixture was stirred for 2 hours at 25° C. The mixture was concentrated in vacuo, the residue was triturated with ethanol (8 mL), and the precipitate was dried in vacuo to give 3.50 g of ethyl N-[(4-methoxyphenyl)aminomethylthio]carbamate as a white solid. 1 H NMR (400MHz, CDCl3): δ11.37(br.s,1H),11.18(br.s,1H),7.44(d,J=8.8Hz,2H), 6.94 (d, J = 8.8 Hz, 2H), 4.20 (q, J = 7.2 Hz, 2H), 3.76 (s, 1H), 1.26 (t, J = 6.8 Hz, 3H).

[0272] Ethyl (Z)-(((4-methoxyphenyl)amino)((2-oxoethyl)thio)methylene)carbamate

[0273] To a solution of ethyl N-[(4-methoxyphenyl)aminomethylthio]carbamate (3.50 g, 13.7 mmol, 1.0 equiv) and cesium carbonate (7.62 g, 23.4 mmol, 1.7 equiv) in acetonitrile (40 mL) was added 2-chloroacetaldehyde (3.38 g, 17.2 mmol, 1.25 equiv), maintaining the temperature below 25°C. After addition, the mixture was stirred at 25°C for 12 hours. The mixture was poured into water (150 mL) and extracted with TBME (150 mL x 2). The combined organic layers were washed with brine (200 mL), dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give 3.80 g of (Z)-ethyl (((4-methoxyphenyl)amino)((2-oxoethyl)thio)methylene)carbamate as a brown gum. 1H NMR (400MHz, DMSO-d6): δ7.28-7.20(m,2H),7.02-6.95(m,2H),6.92(d,J=7.2Hz, 1H), 5.59-5.54 (m, 1H), 3.95 (q, J = 7.2Hz, 2H), 3.78 (s, 3H), 1.12 (t, J = 7.2Hz, 3H). LCMS(m / z[M+H] + ):297.0.

[0274] Ethyl (Z)-(3-(4-methoxyphenyl)thiazol-2(3H)-ylidene)carbamate

[0275] To a solution of (Z)-ethyl(((4-methoxyphenyl)amino)((2-oxoethyl)thio)methylene)carbamate (1.80 g, 6.07 mmol, 1.0 equiv) in tetrahydrofuran (20 mL) was added DIEA (2.36 g, 18.2 mmol, 3.0 equiv) at 0°C followed by thionyl chloride (0.723 g, 6.07 mmol, 1.0 equiv). After addition, the mixture was warmed to 15°C and stirred for 4 hours. The mixture was directly concentrated under reduced pressure. The residue was dissolved in ethyl acetate (100 mL) and adjusted to pH = 8.0 by addition of saturated aqueous sodium bicarbonate solution. The organic layer was separated and washed with brine (20 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 5 / 1) to give 1.20 g of (Z)-ethyl (3-(4-methoxyphenyl)thiazole-2(3H)-propylidene)carbamate as a yellow solid. 1H NMR (400MHz, CDCl3) δ7.47(d,J=4.8Hz,1H),7.41(d,J=5.2Hz,2H),7.07(d,J=4.8Hz, 2H), 7.04 (d, J = 4.8Hz, 1H), 4.01 (q, J = 6.8Hz, 2H), 3.82 (s, 3H), 1.16 (t, J = 7.2Hz, 3H).

[0276] Intermediate C10

[0277] To a solution of ethyl (3-(4-methoxyphenyl)thiazole-2(3H)-ylidene)carbamate (0.200 g, 0.718 mmol, 1.0 equiv) in ethanol (2 mL) was added sodium hydroxide (0.575 g, 14.4 mmol, 20 equiv). The mixture was stirred at 50° C. for 2 hours. The mixture was diluted with water (30 mL) and extracted with ethyl acetate (15 mL×2). The combined organic layers were dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane / methanol=10 / 1) to give 100 mg of 3-(4-methoxyphenyl)thiazole-2(3H)-imine C10 as a yellow oil.

[0278] Preparation of intermediate C11

[0279]

[0280] Ethyl N-[(4-chlorophenyl)aminomethylthio]carbamate

[0281] To a solution of 4-chloroaniline (2.00 g, 15.7 mmol, 1.0 equiv) and O-ethylcarboisothiocyanate (2.06 g, 15.7 mmol, 1.0 equiv) in ethyl acetate (20 mL) was added tetramethylethylenediamine (0.182 g, 1.57 mmol, 0.1 equiv) at 25°C, and the resulting mixture was stirred at 25°C for 2 hours. The mixture was concentrated under reduced pressure. The residue was triturated with ethanol (8 mL) to give 3.00 g of ethyl N-[(4-chlorophenyl)aminomethylthio]carbamate as a white solid. 1 H NMR (400MHz, CDCl3-d): δ11.51 (br.s, 1H), 11.31 (br.s, 1H), 7.62 (d, J = 8.8Hz, 2H), 7.44 (d, J = 8.8Hz, 2H), 4.21 (q, J = 7.2Hz, 2H), 1.26 (t, J = 6.8Hz, 3H).

[0282] Ethyl (Z)-(((4-chlorophenyl)amino)((2-oxoethyl)thio)methylene)carbamate

[0283] To a suspension of ethyl N-[(4-chlorophenyl)aminomethylthio]carbamate (3.00 g, 11.60 mmol, 1.0 equiv) and cesium carbonate (6.42 g, 19.7 mmol, 1.7 equiv) in acetonitrile (40 mL) was added 2-chloroacetaldehyde (2.84 g, 14.5 mmol, 1.25 equiv) dropwise, maintaining the temperature below 25°C. After addition, the mixture was stirred at 25°C for 12 hours. The mixture was diluted with tert-butyl methyl ether (150 mL x 2), washed with brine (50 mL), dried over sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give 3.50 g of (Z)-ethyl (((4-chlorophenyl)amino)((2-oxoethyl)thio)methylene)carbamate as a brown gum. 1 H NMR (400MHz, DMSO-d6) δ7.57-7.46(m,2H),7.43-7.33(m,2H),6.92(d,J=7.2Hz,1H),5.70-5.61(m,1H),3.98(q,J=6.8Hz,2H),1.13(t,J=7.2Hz,3H). LCMS(m / z[M+H] + ):301.0.

[0284] (Z)-(3-(4-chlorophenyl)thiazol-2(3H)-ylidene)carbamic acid ethyl ester

[0285] To a solution of (Z)-ethyl(((4-chlorophenyl)amino)((2-oxoethyl)thio)methylene)carbamate (1.80 g, 5.78 mmol, 1.0 equiv) in tetrahydrofuran (20 mL) was added N-ethyl-N-isopropylpropan-2-amine (2.24 g, 17.3 mmol, 3.0 equiv) and thionyl chloride (0.687 g, 5.78 mmol, 1.0 equiv) at 0°C. The mixture was stirred at 25°C for 4 hours. The mixture was directly concentrated under reduced pressure. The residue was dissolved in ethyl acetate (100 mL) and adjusted to pH = 8.0 by adding saturated aqueous sodium bicarbonate solution. The organic layer was separated and washed with brine (20 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 5 / 1) to give 1.10 g of (Z)-ethyl (3-(4-chlorophenyl)thiazol-2(3H)-propylidene)carbamate as a yellow solid. 1 H NMR (400MHz, CDCl3-d): δ7.65-7.55 (m, 4H), 7.54 (d, J = 4.4Hz, 1H), 7.07 (d, J = 4.4Hz, 1H), 4.03 (q, J = 7.2Hz, 2H), 1.16 (t, J = 7.2Hz, 3H).

[0286] Intermediate C11

[0287] To a solution of (Z)-ethyl(3-(4-chlorophenyl)thiazol-2(3H)-ylidene)carbamate (0.200 g, 0.707 mmol, 1.0 equiv) in ethanol (2 mL) was added sodium hydroxide (0.569 g, 14.2 mmol, 20 equiv) at 25°C. The reaction mixture was then heated to 50°C and stirred for 1.5 hours. The mixture was diluted with water (80 mL) and extracted with ethyl acetate (50 mL x 2). The combined organic layers were dried over sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to afford 120 mg of 3-(4-chlorophenyl)thiazol-2(3H)-imine C11 as a yellow oil.

[0288] Preparation of intermediate C12

[0289]

[0290] Ethyl N-(3-pyridylaminomethylthio)carbamate

[0291] To a solution of pyridin-3-amine (1.00 g, 10.6 mmol, 1.0 equiv) and O-ethyl carboisothiocyanate (1.39 g, 10.6 mmol, 1.0 equiv) in ethyl acetate (10 mL) was added tetramethylethylenediamine (0.123 g, 1.06 mmol, 0.1 equiv) at 25° C., and the mixture was stirred at that temperature for 1 hour. The mixture was concentrated, the residue was triturated with ethanol (10 mL), and the precipitate was collected by filtration and dried under reduced pressure to give 1.80 g of ethyl N-(3-pyridylcarbamoylthio)carbamate as a white solid. 1 HNMR (400MHz, CDCl3) δ11.56(br.s,1H),8.70(d,J=2.4Hz,1H),8.52(dd,J=4.8Hz,1.6Hz,1H),8. 30(br.s,1H),8.28-8.23(m,1H),7.39-7.33(m,1H),4.33(q,J=7.2Hz,2H),1.38(t,J=7.2Hz,3H).

[0292] (Z)-Ethyl(((2-oxoethyl)thio)(pyridin-3-ylamino)methylene)carbamate

[0293] To a suspension of ethyl N-(3-pyridylaminomethylthio)carbamate (0.60 g, 2.66 mmol, 1.0 equiv) and cesium carbonate (1.48 g, 4.53 mmol, 1.7 equiv) in acetonitrile (10 mL) was added 2-chloroacetaldehyde (1.05 g, 5.33 mmol, 2.0 equiv) dropwise at 0°C, maintaining the temperature at 25°C. After addition, the mixture was stirred at this temperature for 12 hours. The mixture was poured into water (100 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 0.40 g of (Z)-ethyl (((2-oxoethyl)thio)(pyridin-3-ylamino)methylene)carbamate as a white solid. 1 H NMR (400MHz, DMSO-d6) δ8.58(d,J=2.0Hz,1H),8.52(dd,J=1.6Hz,4.8Hz,1H),7.83-7.80(m,1H),7.50(q,J=4.8Hz,1H),7.00(d,J =7.2Hz, 1H), 5.72 (t, J = 6.0Hz, 1H), 4.04-3.96 (m, 2H), 3.66-3.61 (m, 1H), 3.15 (dd, J = 12.4Hz, 1.2Hz, 1H), 1.13 (t, J = 6.8Hz, 3H). LCMS(m / z[M+H] + ):268.1.

[0294] (Z)-Ethyl (3-(pyridin-3-yl)thiazol-2(3H)ylidene)carbamate

[0295] To a solution of (Z)-ethyl (((2-oxoethyl)thio)(pyridin-3-ylamino)methylene)carbamate (0.570 g, 2.13 mmol, 1.0 equiv) in tetrahydrofuran (6 mL) was added diisopropylethylamine (0.830 g, 6.40 mmol, 3.0 equiv) and thionyl chloride (0.250 g, 2.13 mmol, 1.0 equiv). The mixture was stirred at 25 ° C for 4 hours. The mixture was directly concentrated under reduced pressure. The residue was dissolved in ethyl acetate (100 mL) and adjusted to pH = 8.0 by saturated sodium bicarbonate aqueous solution. The organic layer was separated and washed with brine (20 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure to give 0.40 g of ethyl (Z)-(3-(pyridin-3-yl)thiazol-2(3H))-ylidene)carbamate as a yellow solid. 1H NMR (400MHz, DMSO-d6) δ8.77(d,J=2.4Hz,1H),8.65(dd,J=4.8Hz,1.6Hz,1H),8.07-8.00(m,1H),7.63( d,J=4.8Hz,1H),7.63-7.59(m,1H),7.12(d,J=4.8Hz,1H),4.03(q,J=7.2Hz,2H),1.15(t,J=7.2Hz,3H).

[0296] Intermediate C12

[0297] To a solution of ethyl (3-(pyridin-3-yl)thiazol-2(3H)-ylidene)carbamate (0.200 g, 0.800 mmol, 1.0 equiv) in ethanol (2 mL) was added sodium hydroxide (0.640 g, 16.1 mmol, 20 equiv) at 25° C., and the mixture was heated to 50° C. and stirred for 1 hour. The mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL×2). The combined organic layers were dried over sodium sulfate, filtered, and concentrated to give 140 mg of 3-(pyridin-3-yl)thiazol-2(3H)-imine C12 as a brown oil. 1 H NMR(400MHz,DMSO-d6,400MHz)δ8.80(d,J=2.0Hz,1H),8.45(d,J=4.0Hz,1H),8.28(br.s,1 H), 8.06 (d, J = 7.6Hz, 1H), 7.51-7.43 (m, 1H), 7.12 (d, J = 5.2Hz, 1H), 6.25 (d, J = 5.2Hz, 1H).

[0298] Preparation of intermediate C13

[0299]

[0300] Ethyl N-(2-pyridylaminomethylthio)carbamate

[0301] To a solution of pyridine-2-amine (1.00 g, 10.6 mmol, 1.0 equivalent) in ethyl acetate (10 mL) was added O-ethyl carbonyl isothiocyanate (1.26 mL, 10.6 mmol, 1.0 equivalent) and N1, N1, N2, N2-tetramethylethane-1,2-diamine (0.160 mL, 1.06 mmol, 0.1 equivalent) at 25 ° C. The mixture was then stirred at this temperature for 5 hours. The mixture was concentrated under reduced pressure. The residue was triturated 3 times with ethanol (10 mL) to give 0.95 g of ethyl N-(2-pyridylaminomethylthio)carbamate as an off-white solid. 1H NMR (400MHz, CDCl3) δ12.06(br.s,1H),8.80(d,J=8.0Hz,1H),8.41(d,J=4.0Hz,1H),8.07(b r.s,1H),7.79-7.74(m,1H),7.16-7.13(m,1H),4.31(q,J=7.2Hz,2H),1.35(t,J=7.2Hz,3H). LCMS(m / z[M+H] + ):225.8.

[0302] Ethyl (Z)-(((2-oxoethyl)thio)(pyridin-2-ylamino)methylene)carbamate

[0303] To a suspension of ethyl N-(2-pyridylaminomethylthio)carbamate (0.800 g, 3.55 mmol, 1.0 equiv) and cesium carbonate (1.97 g, 6.04 mmol, 1.7 equiv) in acetonitrile (8 mL) was added 2-chloroacetaldehyde (0.714 mL, 4.44 mmol, 1.25 equiv) dropwise at 25°C. After addition, the mixture was stirred for 5 hours. 2-Chloroacetaldehyde (0.285 mL, 1.78 mmol, 0.5 equiv) was then added to the mixture and stirred for an additional 2 hours. The mixture was poured into water (20 mL) and extracted with ethyl acetate (70 mL x 3). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give (Z)-ethyl (((2-oxoethyl)thio)(pyridin-2-ylamino)methylene)carbamate (1.00 g) as a brown oil. 1 H NMR (400MHz, CDCl3): δ8.34-8.32(m,1H),8.06(d,J=8.4Hz,1H),7.93-7.79(m,1H),7.17-7.14(m,1H),6.19( d,J=6.4Hz,1H),5.44(s,1H),4.28-4.22(m,2H),3.51-3.47(m,1H),3.28-3.24(m,1H),1.33(t,J=7.2Hz,3H).

[0304] (Z)-Ethyl (3-(pyridin-2-yl)thiazol-2(3H)-ylidene)carbamate

[0305] To a solution of (Z)-ethyl (((2-oxoethyl)thio)(pyridin-2-ylamino)methylene)carbamate (0.500 g, 1.87 mmol, 1.0 equiv) in THF (5 mL) was added N-ethyl-N-isopropylpropan-2-amine (0.977 mL, 5.61 mmol, 3.0 equiv) at 0°C, followed by SOCl2 (136 μL, 1.87 mmol, 1.0 equiv). The mixture was stirred at below 15°C for 4 hours. The mixture was poured into water (10 mL) and extracted with ethyl acetate (40 mL×3). The combined organic layers were washed with brine (25 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1 to 10 / 1) to give 210 mg of (Z)-ethyl (3-(pyridin-2-yl)thiazol-2(3H))-ylidene)carbamate as a brown solid. 1 H NMR (400MHz, CDCl3) δ8.51-8.49(m,1H),8.42(d,J=8.4Hz,1H),7.92-7.88(m,1H),7.80(d,J=5.2Hz,1H) ,7.31-7.28(m,1H),0.65(d,J=5.2Hz,1H),4.28(q,J=7.2Hz,2H),1.36(t,J=7.2Hz,3H).LCMS(m / z[M+H] + ):249.8.

[0306] Intermediate C13

[0307] To a solution of sodium hydroxide (0.674 g, 16.8 mmol, 20.0 equiv) in ethanol (3 mL) was added (Z)-ethyl (3-(pyridin-2-yl)thiazol-2(3H)-ylidene)carbamate (0.210 g, 0.842 mmol, 1.0 equiv) at 25 °C. The mixture was then heated to 50 °C and stirred for 1 hour. The mixture was poured into water (15 mL) and stirred for 3 minutes and extracted with ethyl acetate (40 mL x 3). The combined organic phases were washed with brine (15 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give 130 mg of 3-(pyridin-2-yl)thiazol-2(3H)-imine C13 as a brown oil. 1 H NMR (400MHz, DMSO-d6) δ8.82 (s, 1H), 8.62-8.60 (m, 1H), 8.41 (d, J = 4.0Hz, 1H ),7.89-7.86(m,1H),7.60(s,1H),7.24-7.21(m,1H),7.26(d,J=5.2Hz,1H). LCMS(m / z[M+H] +):177.9.

[0308] Preparation of intermediate C14

[0309]

[0310] Ethyl N-(tetrahydropyran-4-ylaminomethylthio)carbamate

[0311] To the mixture of tetrahydro-2H-pyrans-4-amine (0.800g, 7.91mmol, 1.0 equivalent) and O-ethyl carbon isothiocyanate (1.09g, 8.30mmol, 1.05 equivalent) in ethyl acetate (10mL), add TMEDA (92mg, 0.791mmol, 0.1 equivalent). The mixture was stirred at 25 ℃ for 6 hours, then concentrated under reduced pressure. The solution of residue with petroleum ether / ethanol (v / v=20 / 1,21mL) was ground twice to obtain 1.80g of N-(tetrahydropyrans-4-ylaminomethylthio) ethyl carbamate, which was a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ10.99(br.s,1H),9.83(d,J=7.2Hz,1H),4.40-4.26(m,1H),4.12(q,J=7.2Hz,2 H),3.87-3.73(m,2H),3.48-3.36(m,2H),1.97-1.86(m,2H),1.59-1.45(m,2H),1.21(t,J=7.2Hz,3H).

[0312] (Z)-Ethyl(((2-oxoethyl)thio)((tetrahydro-2H-pyran-4-yl)amino)methylene)carbamate

[0313] To a mixture of ethyl N-(tetrahydropyran-4-ylaminomethylthio)carbamate (0.800 g, 3.44 mmol, 1.0 equiv) and cesium carbonate (1.91 g, 5.85 mmol, 1.7 equiv) in acetonitrile (10 mL) was added 2-chloroacetaldehyde (2.09 g, 10.66 mmol, 3.1 equiv). The resulting mixture was stirred at 25 ° C for 9 hours, then poured into ice water (10 mL) and extracted with ethyl acetate (100 mL × 2). The combined organic phase was washed with brine (20 mL × 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 100 / 1 to 0 / 1) to give 0.30 g of (Z)-ethyl (((2-oxoethyl)thio)((tetrahydro-2H-)pyran-4-yl)amino)methylene)carbamate as a yellow solid. LCMS (m / z [M+H] + ):274.9.

[0314] (Z)-Ethyl (3-(tetrahydro-2H-pyran-4-yl)thiazol-2(3H)-ylidene)carbamate

[0315] A mixture of (Z)-ethyl (((2-oxoethyl)thio)((tetrahydro-2H-pyran-4-yl)amino)methylene)carbamate (0.30 g, 0.52 mmol, 1.0 equiv) and diisopropylethylamine (133 mg, 1.03 mmol, 2.0 equiv) in tetrahydrofuran (10 mL) was added thionyl chloride (61 mg, 0.52 mmol, 1.0 equiv). The mixture was stirred at 25° C. for 2 hours, then poured into ice water (20 mL) and extracted with ethyl acetate (100 mL×3). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 100 / 1 to 0 / 1) to give 120 mg of (Z)-ethyl (3-(tetrahydro-2H-pyran-4-yl)thiazol-2(3H)-ylidene)carbamate as a yellow solid. 1 H NMR (400MHz, CD3OD) δ7.41(d,J=4.8Hz,1H),6.87(d,J=4.8Hz,1H),5.02-4.91(m,1H),4.20(q, J=7.2Hz,2H),4.09-4.04(m,2H),3.62-3.58(m,2H),1.99-1.89(m,4H),1.31(t,J=7.2Hz,3H). LCMS(m / z[M+H] + ):256.9.

[0316] Intermediate C14

[0317] A mixture of (Z)-ethyl (3-(tetrahydro-2H-pyran-4-yl)thiazol-2(3H)-ylidene)carbamate (0.120 g, 0.468 mmol, 1.0 equiv) and sodium hydroxide (0.374 g, 9.36 mmol, 20 equiv) in ethanol (1 mL) was stirred at 50° C. for 1 hour. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane / methanol = 100 / 1 to 10 / 1) to give 80 mg of 3-(tetrahydro-2H-pyran-4-yl)thiazol-2(3H)-imine C14 as a yellow solid.

[0318] Preparation of intermediate C15

[0319]

[0320] N-(phenylaminomethylthio)urethane

[0321] To a solution of aniline (1.96 mL, 21.5 mmol, 1.0 equiv) in ethyl acetate (20 mL) was added O-ethyl carboisothiocyanate (2.54 mL, 21.5 mmol, 1.0 equiv) and tetramethylethylenediamine (0.324 mL, 2.15 mmol, 0.1 equiv). The mixture was stirred at 25°C for 4 hours and then concentrated under reduced pressure. The residue was triturated with petroleum ether (8 mL) to give 4.20 g of ethyl N-(phenylaminomethylthio)carbamate as a yellow oil. 1 H NMR(400MHz,DMSO-d6)δ11.55(br.s,1H),11.25(br.s,1H),7.61-7.59(m,2H),7. 41-7.37(m,2H),7.26-7.22(m,1H),4.21(q,J=7.2Hz,2H),1.26(t,J=7.2Hz,3H).

[0322] (Z)-Ethyl (((2-oxopropyl)thio)(phenylamino)methylene)carbamate

[0323] To a solution of N-(phenylaminomethylthio)carbamate (4.20 g, 18.7 mmol, 1.0 equivalent) and cesium carbonate (10.4 g, 31.8 mmol, 1.7 equivalent) in acetonitrile (50 mL) was added 1-chloropropane-2-one (2.17 g, 23.4 mmol, 1.25 equivalents), keeping the temperature below 25 ° C. After addition, the mixture was stirred at 25 ° C for 2 hours, then diluted with MTBE (120 mL) and washed with saturated sodium bicarbonate aqueous solution (100 mL) and brine (100 mL), filtered with anhydrous sodium sulfate. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50: 1 to 4: 1) to give 1.80 g of (Z)-ethyl (((2-oxopropyl)thio) (phenylamino) methylene) carbamate as a brown oil. 1 H NMR (400MHz, CDCl3) δ7.46-7.42(m,2H),7.40-7.36(m,1H),7.28-7.27(m,1H),7.27-7.26(m,1H),4.14 (q,J=7.2.0Hz,2H),3.47(d,J=12.0Hz,1H),3.65(d,J=12.0Hz,1H),1.47(s,3H),1.26(t,J=7.6Hz,4H). LCMS(m / z[M+H] + ):280.9.

[0324] (Z)-(4-Methyl-3-phenylthiazol-2(3H)-ylidene)carbamic acid ethyl ester

[0325] To a solution of (Z)-ethyl (((2-oxopropyl)thio)(phenylamino)methylene)carbamate (0.600 g, 2.09 mmol, 1.0 equiv) in THF (6 mL) was added diisopropylethylamine (0.810 g, 6.27 mmol, 3.0 equiv) and thionyl chloride (0.167 mL, 2.30 mmol, 1.1 equiv) at 0 ° C. The mixture was stirred at 20 ° C for 4 hours and then concentrated under reduced pressure. The residue was diluted with saturated aqueous sodium bicarbonate solution (40 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with brine (60 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by reverse phase HPLC to give 0.32 g of (Z)-ethyl (4-methyl-3-phenylthiazole-2(3H)-ylidene)carbamate as a yellow oil. 1 H NMR (400MHz, CDCl3) δ7.68 (s, 3H), 7.45 (s, 2H), 6.92 (s, 1H), 4.34 (q, J = 7.2Hz, 2H), 2.08 (s, 3H), 1.34 (t, J = 7.2Hz, 3H). LCMS(m / z[M+H] + ):263.3.

[0326] Intermediate C15

[0327] To a solution of (Z)-ethyl(4-methyl-3-phenylthiazol-2(3H)-ylidene)carbamate (0.240 g, 0.891 mmol, 1.0 equiv) in ethanol (4 mL) was added sodium hydroxide (0.713 g, 17.8 mmol, 20 equiv) at 25°C. The mixture was heated to 60°C and stirred for 1 hour, then diluted with water (40 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford 180 mg of 4-methyl-3-phenylthiazol-2(3H)-imine C15 as a yellow solid.

[0328] Preparation of intermediate C16

[0329]

[0330] N-(Benzylaminomethylthioyl)urethane

[0331] To a solution of benzylamine (5.00g, 46.7mmol, 1.0 equivalent) and O-ethyl carbon isothiocyanate (6.43g, 49.0mmol, 1.05 equivalent) in ethyl acetate (40mL) under 25 ℃ of nitrogen atmospheres, add TMEDA (0.542g, 4.67mmol, 1.0 equivalent). The mixture was stirred 6 hours at 25 ℃, then concentrated under reduced pressure. Residue was ground twice from a solution of petroleum and ethyl acetate (v / v=10 / 1,55mL) to obtain 10.0g of N-(benzylaminomethylthioyl) ethyl carbamate, which was a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ11.04 (br.s, 1H), 10.22 (br.t, J = 5.6Hz, 1H), 7.39-7. 23(m,5H), 4.81(d,J=5.6Hz,2H), 4.14(q,J=7.2Hz,2H), 1.21(t,J=7.2Hz,3H).

[0332] (Z)-Ethyl((benzylamino)((2-oxobutyl)thio)methylene)carbamate

[0333] To a suspension of N-(benzylaminomethylthioyl)urethane (3.00 g, 12.6 mmol, 1.0 equiv) and Cs2CO3 (6.97 g, 21.4 mmol, 1.7 equiv) in acetonitrile (45 mL) was added 1-bromobutane-2-one (2.38 g, 15.7 mmol, 1.25 equiv), maintaining the temperature below 25°C. After addition, the mixture was stirred at 25°C for 5 hours, then poured into water (80 mL) and extracted with ethyl acetate (120 mL×3). The combined organic layers were washed with brine (100 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate=1:0 to 5:1) to give 2.50 g of (Z)-ethyl ((benzylamino)((2-oxobutyl)thio)methylene)carbamate as a yellow oil. 1 HNMR (400MHz, CDCl3) δ7.32-7.27(m,3H),7.25-7.22(m,2H),4.96(d,J=15.6Hz,1H),4.71(d,J=15.6Hz,1H),4.18(q,J=7.2Hz,2H ), 3.28 (d, J = 12.0Hz, 1H), 3.11 (d, J = 12.0Hz, 1H), 1.97-1.92 (m, 1H), 1.67-1.62 (m, 1H), 1.31-1.28 (m, 3H), 0.88 (t, J = 7.6Hz, 3H). LCMS(m / z[M+H] + ):308.9.

[0334] (Z)-Ethyl(3-benzyl-4-ethylthiazol-2(3H)-ylidene)carbamate

[0335] To a solution of (Z)-ethyl((benzylamino)((2-oxobutyl)thio)methylene)carbamate (2.50 g, 8.11 mmol, 1.0 equiv) in THF (38 mL) was added diisopropylethylamine (3.14 g, 24.3 mmol, 3.0 equiv) and SOCl2 (0.964 g, 8.11 mmol, 1.0 equiv) at 0°C. The mixture was stirred at 25°C for 16 hours, then poured into water (80 mL) and extracted with ethyl acetate (100 mL×3). The combined organic layers were washed with brine (100 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give 2.20 g of (Z)-ethyl(3-benzyl-4-ethylthiazol-2(3H)-ylidene)carbamate as a dark brown oil. 1 H NMR (400MHz, DMSO-d6) δ7.37-7.33(m,2H),7.29-7.27(m,1H),7.09(d,J=7.2Hz,2H),6.61(s,1H),5.36-5. 34(m,2H),4.07-4.03(m,2H),2.46(dd,J=2.0Hz,7.6Hz,2H),1.17(t,J=7.2Hz,3H),1.11(t,J=7.2Hz,3H). LCMS(m / z[M+H] + ):291.1.

[0336] Intermediate C16

[0337] To a solution of (Z)-ethyl(3-benzyl-4-ethylthiazole-2(3H)-ylidene)carbamate (1.00 g, 3.44 mmol, 1.0 equiv) in ethanol (10 mL) was added 6N NaOH (2.75 g, 68.9 mmol, 20 equiv) at 25 ° C. The mixture was heated to 50 ° C and stirred for 1 hour, then poured into water (80 mL) and extracted with ethyl acetate (100 mL×3). The combined organic layers were washed with brine (100 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate=1:0 to 5:1) to give 0.49 g of 3-benzyl-4-ethylthiazole-2(3H)-imine C16 as a dark brown oil.

[0338] Preparation of intermediate C17

[0339]

[0340] (Z)-Ethyl((benzylamino)((3-methyl-2-oxobutyl)thio)methylene)carbamate

[0341] To a mixture of N-(benzylaminomethylthio)ethyl carbamate (0.500 g, 2.10 mmol, 1.0 equiv) and cesium carbonate (1.16 g, 3.57 mmol, 1.7 equiv) in acetonitrile (5 mL) was added 1-bromo-3-methylbutane-2-one (0.519 g, 3.15 mmol, 1.5 equiv) at 25 ° C. The mixture was stirred for 5 hours, then poured into water (10 mL) and extracted with MTBE (100 mL x 2). The combined organic phases were washed with brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give 0.67 g of crude (Z)-ethyl ((benzylamino)((3-methyl-2-oxobutyl)thio)methylene)carbamate as a yellow solid. LCMS (m / z[M+H] + ):323.0.

[0342] Ethyl (Z)-ethyl (3-benzyl-4-isopropylthiazol-2(3H)-propylene) carbamate

[0343] To a solution of (Z)-ethyl((benzylamino)((3-methyl-2-oxobutyl)thio)methylene)carbamate (0.670 g, 2.08 mmol, 1.0 equiv) and diisopropylethylamine (1.09 mL, 6.23 mmol, 3.0 equiv) in THF (10 mL) was added thionyl chloride (0.247 g, 2.08 mmol, 1.0 equiv). The mixture was stirred at 25 °C for 16 hours, then poured into water (20 mL) and extracted with ethyl acetate (100 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 100 / 1 to 4 / 1) to give 0.53 g of ethyl (Z)-ethyl (3-benzyl-4-isopropylthiazol-2(3H)-propylene)carbamate as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ7.38-7.24(m,3H),7.04(d,J=7.2Hz,2H),6.70(s,1H),5.39(s,2H ), 4.04 (q, J = 7.2Hz, 2H), 2.87-2.77 (m, 1H), 1.16 (t, J = 7.2Hz, 3H), 1.08 (d, J = 6.8Hz, 6H). LCMS(m / z:305.1[M+H] + ):305.1.

[0344] Intermediate C17

[0345] A mixture of (Z)-ethyl (3-benzyl-4-isopropylthiazole-2 (3H) -ylidene) carbamate (0.300 g, 0.830 mmol) and sodium hydroxide (0.664 g, 16.61 mmol) in ethanol (5 mL) was stirred at 50 ° C for 1 hour. The mixture was poured into water (10 mL) and extracted with ethyl acetate (100 mL × 3). The combined organic layer was washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane / methanol = 100 / 1 to 10 / 1) to give 150 mg of 3-benzyl-4-isopropylthiazole-2 (3H) -imine C17 as a yellow solid.

[0346] Preparation of intermediate C18

[0347]

[0348] (Z)-Ethyl((benzylamino)((2-cyclopropyl-2-oxoethyl)thio)methylene)carbamate

[0349] To a mixture of N-(benzylaminomethylthio)ethyl carbamate (0.500 g, 2.10 mmol, 1.0 equiv) and cesium carbonate (1.16 g, 3.57 mmol, 1.7 equiv) in acetonitrile (5 mL) was added 2-bromo-1-cyclopropyl-ethanone (0.513 g, 3.15 mmol, 1.5 equiv). The resulting mixture was stirred at 25 ° C for 5 hours, then poured into ice water (10 mL) and extracted with MTBE (100 mL x 2). The combined organic layer was washed with brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give (Z)-ethyl ((benzylamino)((2-cyclopropyl-2-oxoethyl)thio)methylene)carbamate (0.670 g, crude) as a yellow solid. LCMS (m / z [M + H] + ):321.1.

[0350] (Z)-Ethyl(3-benzyl-4-cyclopropylthiazol-2(3H)-ylidene)carbamate

[0351] To a mixture of (Z)-ethyl((benzylamino)((2-cyclopropyl-2-oxoethyl)thio)methylene)carbamate (0.670 g, 2.08 mmol, 1.0 equiv) and diisopropylethylamine (0.810 g, 6.27 mmol, 3.0 equiv) in tetrahydrofuran (10 mL) was added thionyl chloride (0.248 g, 2.09 mmol, 1.2 equiv). The mixture was stirred at 25° C. for 16 hours, then poured into ice water (20 mL) and extracted with ethyl acetate (100 mL×3). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 100 / 1 to 4 / 1) to give 0.25 g of ethyl (Z)-ethyl (3-benzyl-4-cyclopropylthiazole-2(3H)-dimethylene)carbamate as a yellow gum. 1 H NMR (400MHz, DMSO-d6): δ7.40-7.24(m,3H),7.14(d,J=7.2Hz,2H),6.59(s,1H),5.48(s,2H),4.03 (q,J=7.2Hz,2H),1.69-1.60(m,1H),1.17(t,J=7.2Hz,3H),0.82-0.75(m,2H),0.60-0.54(m,2H). LCMS(m / z[M+H] + ):305.1.

[0352] Intermediate C18

[0353] A mixture of (Z)-ethyl(3-benzyl-4-cyclopropylthiazol-2(3H)-ylidene)carbamate (0.200 g, 0.661 mmol, 1.0 equiv) and 6N sodium hydroxide (0.529 g, 13.23 mmol, 20 equiv) in ethanol (4 mL) was stirred at 50° C. for 1 hour. The mixture was poured into ice water (10 mL) and extracted with ethyl acetate (100 mL×3). The combined organic layers were washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford 160 mg of 3-benzyl-4-cyclopropylthiazol-2(3H)-imine C18 as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ7.57(br.s,1H),7.36-7.30(m,2H),7.27-7.20(m,3H),5. 64(s,1H),5.04(s,2H),1.48-1.36(m,1H),0.72-0.64(m,2H),0.52-0.42(m,2H). LCMS(m / z[M+H] + ):233.2.

[0354] Preparation of intermediate C19

[0355]

[0356] (Z)-Ethyl((benzylamino)((3,3,3-trifluoro-2-oxopropyl)thio)methylene)carbamate

[0357] To a suspension of N-(benzylaminomethylthio)ethyl carbamate (0.500 g, 2.10 mmol, 1.0 equivalent) and cesium carbonate (1.16 g, 3.57 mmol, 1.7 equivalents) in acetonitrile (8 mL) was added 3-bromo-1,1,1-trifluoropropane-2-one (0.654 mL, 6.30 mmol, 3.0 equivalents) and the temperature was kept below 25 ° C. After addition, the mixture was stirred at 25 ° C for 12 hours, then poured into water (40 mL) and extracted with MTBE (30 mL x 3). The combined organic layer was washed with brine (50 mL), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50:1: to 10:1) to give 0.75 g of (Z)-ethyl ((benzylamino)((3,3,3-trifluoro-2-oxopropyl)thio)methylene)carbamate as a yellow oil. 1 H NMR (400MHz, CDCl3) δ7.35-7.28(m,5H),5.37(d,J=15.6Hz,1H),4.62(d,J=16.0Hz,1H),4.27-4.18 (m,2H),3.79(br.s,1H),3.69(d,J=12.8Hz,1H),3.25(dd,J=12.0Hz,0.8Hz,1H),1.35-1.31(m,3H). 19 F NMR (400MHz, CDCl3) δ-79.04 (s, 3F). LCMS(m / z[M+H] + ):348.9.

[0358] (Z)-Ethyl(3-benzyl-4-(trifluoromethyl)thiazol-2(3H)-ylidene)carbamate

[0359] To a solution of (Z)-ethyl((benzylamino)((3,3,3-trifluoro-2-oxopropyl)thio)methylene)carbamate (0.550 g, 1.44 mmol, 1.0 equiv) in THF (20 mL) was added diisopropylethylamine (0.754 mL, 4.33 mmol, 3.0 equiv) and thionyl chloride (0.115 mL, 1.59 mmol, 1.1 equiv) at 0°C. The mixture was heated to 60°C and stirred for 2 hours, then added to saturated aqueous sodium bicarbonate solution (40 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layers were washed with brine (60 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50:1 to 10:1) to give 0.40 g of (Z)-ethyl (3-benzyl-4-(trifluoromethyl)thiazol-2(3H))-ylidene)carbamate as a brown oil. 1 H NMR (400MHz, CDCl3): δ7.34-7.28(m,3H), 7.18-7.13(m,3H), 5.48(s,2H), 4.26(q,J=7.2Hz,2H), 1.34(t,J=6.8Hz,3H). LCMS(m / z[M+H] + ):330.9.

[0360] Intermediate C19

[0361] To a solution of (Z)-ethyl (3-benzyl-4-(trifluoromethyl)thiazole-2(3H)-ylidene)carbamate (0.300 g, 0.884 mmol, 1.0 equiv) in ethanol (5 mL) was added sodium hydroxide (0.707 g, 17.7 mmol, 20 equiv) at 25 ° C. The mixture was heated to 60 ° C and stirred for 1 hour, then poured into water (40 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic layer was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate=20: 1) to give 150 mg of 3-benzyl-4-(trifluoromethyl)thiazole-2(3H)-imine C19 as a brown oil.

[0362] Preparation of intermediate C20

[0363]

[0364] Ethyl N-[(3-cyanophenyl)aminomethylthio]carbamate

[0365] To a stirred mixture of 3-aminobenzonitrile (2.50 g, 21.16 mmol, 1.0 equivalent) in EtOAc (25 mL) was added O-ethyl carbonyl isothiocyanate (2.78 g, 21.16 mmol, 1.0 equivalent) and TMEDA (246 mg, 2.12 mmol, 319 μL, 0.1 equivalent) at 25 ° C. The reaction mixture was then stirred at 25 ° C for 5 hours under N2. The reaction mixture was concentrated under reduced pressure. EtOH (50 mL) was added to the mixture and the resulting mixture was stirred at 25 ° C for 0.5 h. The mixture was filtered and the filter cake was dried under reduced pressure to obtain 2.80 g of yellow solid N-[(3-cyanophenyl)aminomethylthio]ethyl carbamate.

[0366] Ethyl (Z)-(((3-cyanophenyl)amino)((2-oxoethyl)thio)methylene)carbamate

[0367] To a mixture of N-[(3-cyanophenyl)aminomethylthio]ethyl carbamate (2.80 g, 11.2 mmol, 1.0 equivalent) and CsCO (6.22 g, 19.1 mmol, 1.7 equivalent) in MeCN (40 mL) was added 2-chloroacetaldehyde (2.76 g, 14.0 mmol, 2.26 mL, 1.2 equivalent) at 25 ° C. The reaction mixture was stirred at 25 ° C for 12 hours under N2. Another 1.0 equivalent of 2-chloroacetaldehyde was added and the reaction mixture was stirred at 25 ° C for another 12 hours. The reaction mixture was diluted with MTBE (100 mL) and washed with NaHCO solution (2x50 mL). The organic layer was washed with brine (50 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to give 2.80 g of (Z)-ethyl(((3-cyanophenyl)amino)((2-oxoethyl)thio)methylene)carbamate as a brown solid.

[0368] (Z)-(3-(3-cyanophenyl)thiazol-2(3H)-ylidene)carbamic acid ethyl ester

[0369] To a mixture of (Z)-ethyl (((3-cyanophenyl)amino)((2-oxoethyl)thio)methylene)carbamate (2.80 g, 9.61 mmol, 1.0 equiv) in THF (30 mL) was added DIPEA (3.73 g, 28.8 mmol, 5.02 mL, 3.0 equiv) and SOCl (1.14 g, 9.61 mmol, 697 μL, 1.0 equiv) at 0°C. The reaction mixture was then stirred at 25°C under N for 15 hours. The reaction mixture was concentrated under reduced pressure. The residue was dissolved in MTBE (200 mL) and the mixture was adjusted to pH = 8 with a saturated solution. A saturated aqueous solution of NaHCO was added, and the organic layer was discarded. The aqueous phase was extracted with EtOAc (3 × 100 mL), and the organic layer was washed with brine (100 mL), dried over NaSO, filtered, and the filtrate was concentrated under reduced pressure. MTBE (10 mL) was added to the mixture and the mixture was stirred for 0.5 hours at 25° C. The mixture was then filtered and the filter cake was dried under reduced pressure to give 1.30 g of ethyl (Z)-ethyl(3-(3-cyanophenyl)thiazol-2(3H)-ylidene)carbamate as a brown solid. 1 H NMR: (400MHz, DMSO-d6) δ8.12(s,1H),7.99-7.91(m,2H),7.79-7.74(m,1H),7.61 (d, J=4.8Hz, 1H), 7.11 (d, J=4.8Hz, 1H), 4.09-3.98 (m, 2H), 1.17 (t, J=7.1Hz, 3H).

[0370] Intermediate C20

[0371] To a mixture of (Z)-ethyl(3-(3-cyanophenyl)thiazole-2(3H)-ylidene)carbamate (400 mg, 1.46 mmol, 1.0 equiv) in EtOH (4 mL) was added NaOH (6 M, 4.00 mL, 16.4 equiv) at 25°C. The reaction mixture was then stirred at 25°C for 12 hours and at 60°C for 2 hours under N2. The pH of the reaction mixture was adjusted to 7 with concentrated hydrochloric acid. The resulting solution was then filtered and the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC to give 130 mg of 3-(2-iminothiazole-3(2H)-yl)benzamide C20 as a brown solid.

[0372] Preparation of intermediate C21

[0373]

[0374] tert-Butyl 7-nitro-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0375] To a solution of 7-nitro-1,2,3,4-tetrahydroisoquinoline (4.00 g, 22.4 mmol, 1.0 equiv) in DCM (40 mL) was added TEA (4.54 g, 44.9 mmol, 6.25 mL, 2.0 equiv) and di-tert-butyl dicarbonate (5.39 g, 24.7 mmol, 5.67 mL, 1.1 equiv) at 25 ° C. The reaction mixture was stirred at 25 ° C for 2 hours. The reaction mixture was diluted with water (100 mL) and extracted with EtOAc (4×200 mL). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (eluent: 0-17% ethyl acetate in petroleum ether gradient, 100 mL / min) to provide 5.30 g of tert-butyl 7-nitro-3,4-dihydroisoquinoline-2(1H)-carboxylate as a yellow oil. 1 H NMR (400MHz, DMSO-d6) δ8.12(s,1H),8.02(dd,J=1.9,8.4Hz,1H),7.45(d,J=8.4Hz,1H),4.63(br s,2H),3.64-3.52(m,2H),2.97-2.80(m,2H),1.43(s,9H).

[0376] tert-Butyl 7-amino-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0377] To a solution of tert-butyl 7-nitro-3,4-dihydroisoquinoline-2(1H)-carboxylate (5.9 g, 21.2 mmol, 1.0 equiv) in MeOH (50 mL) was added Pd / C (2.5 g, 10% purity) at 25° C. under N . The suspension was degassed under vacuum and purged with H 3 times. The reaction mixture was stirred at 25° C. under H (15 psi) for 12 hours. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give tert-butyl 7-amino-3,4-dihydroisoquinoline-2(1H)-carboxylate (4.5 g, crude) as a red oil.

[0378] tert-Butyl 7-(3-(ethoxycarbonyl)thioureido)-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0379] To a mixture of tert-butyl 7-amino-3,4-dihydroisoquinoline-2(1H)-carboxylate (1.00 g, 4.03 mmol, 1.0 equiv) in EtOAc (10 mL) was added O-ethyl carbonyl isothiocyanate (528 mg, 4.03 mmol, 1.0 equiv) and TMEDA (47 mg, 0.40 mmol, 61 μL, 0.1 equiv) at 25 ° C. The reaction mixture was then stirred at 25 ° C for 5 hours under N2. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was dissolved in EtOH (10 mL) and the mixture was stirred at 25 ° C for 0.5 hours. The mixture was filtered and the filter cake was dried under reduced pressure to give 1.20 g of tert-butyl 7-(3-(ethoxycarbonyl)thiourea)-3,4-dihydroisoquinoline-2(1H)-carboxylate as a white solid. LCMS (m / z [M + Na] + ):402.2.

[0380] (Z)-tert-Butyl 7-((((ethoxycarbonyl)imino)((2-oxoethyl)thio)methyl)amino)-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0381] To a mixture of tert-butyl 7-(3-(ethoxycarbonyl)thioureido)-3,4-dihydroisoquinoline-2(1H)-carboxylate (1.80 g, 4.74 mmol, 1.0 equiv) in MeCN (20 mL) was added CsCO (2.63 g, 8.06 mmol, 1.7 equiv) and 2-chloroacetaldehyde (1.16 g, 5.93 mmol, 954 μL, 1.25 equiv) at 25° C. The reaction mixture was then stirred at 25° C. under N for 12 hours. The reaction mixture was diluted with MTBE (100 mL) and the mixture was washed with saturated NaHCO solution (2×50 mL). The organic layer was washed with brine (50 mL), dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure to give (Z)-tert-butyl 7-((((ethoxycarbonyl)imino)((2-oxoethyl)thio)methyl)amino)-3,4-dihydroisoquinoline-2(1H)-carboxylate (2.4 g, crude) as a brown oil.

[0382] tert-Butyl 7-[(2Z)-2-ethoxycarbonyliminothiazol-3-yl]-3,4-dihydro-1H-isoquinoline-2-carboxylate

[0383] To a mixture of (Z)-tert-butyl 7-((((ethoxycarbonyl)imino)((2-oxoethyl)thio)methyl)amino)-3,4-dihydroisoquinoline-2(1H)-carboxylate in THF (30 mL) (2.40 g, 5.69 mmol, 1.0 equiv) was added DIPEA (2.21 g, 17.1 mmol, 2.98 mL, 3.0 equiv) and SOCl2 (677 mg, 5.69 mmol, 413 μL, 1.0 equiv) at 0°C. The reaction mixture was then stirred at 25°C under N2 for 12 hours. The reaction mixture was diluted with water (100 mL) and extracted with EtOAc (3×50 mL). The organic layer was washed with brine (100 mL), dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography ( 20g Silica Flash Column, eluent: 0-40% ethyl acetate / petroleum ether gradient @ 75 mL / min) to afford 1.0 g of (Z)-tert-butyl 7-((((ethoxycarbonyl)imino)((2-oxoethyl)thio)methyl)amino)-3,4-dihydroisoquinoline-2(1H)-carboxylate as a brown oil.

[0384] Intermediate C21

[0385] To a mixture of (Z)-tert-butyl 7-((((ethoxycarbonyl)imino)((2-oxoethyl)thio)methyl)amino)-3,4-dihydroisoquinoline-2(1H)-carboxylate (800 mg, 1.98 mmol, 1.0 equiv) in EtOH (8 mL) was added NaOH (6 M, 8.00 mL, 24 equiv) at room temperature. The reaction mixture was then stirred at 60 °C under N2 for 2 hours. The reaction mixture was filtered, the filter cake was washed with EtOAc (3 x 20 mL), and the filtrate was concentrated under reduced pressure. The residue was purified by reverse phase column to give 170 mg of tert-butyl 7-(2-iminothiazol-3(2H)-yl)-3,4-dihydroisoquinoline-2(1H)-carboxylate C21 as a yellow solid.

[0386] Preparation of intermediate C22

[0387]

[0388] Ethyl N-[(2-bromophenyl)aminomethylthio]carbamate

[0389] To a solution of 2-bromoaniline (10.00 g, 58.1 mmol, 1.0 equivalent) in EtOAc (100 mL) was added N-(thiomethylene) ethyl carbamate (7.62 g, 58.1 mmol, 1.0 equivalent) and TMEDA (675 mg, 5.81 mmol, 877 μL, 0.1 equivalent) at room temperature. The mixture was stirred at 20 ° C for 12 hours. The reaction mixture was concentrated under reduced pressure. The residue was ground with EtOH (50 mL) for 0.5 hours. The mixture was filtered and the filter cake was washed with EtOH (2×30 mL). The filter cake was collected to give 16.0 g of N-[(2-bromophenyl)aminomethylthio] ethyl carbamate as a white solid.

[0390] Ethyl (Z)-(((2-bromophenyl)amino)((2-oxoethyl)thio)methylene)carbamate

[0391] To a solution of ethyl N-[(2-bromophenyl)aminomethylthio]carbamate (16.0 g, 52.8 mmol, 1.0 equiv) in ACN (200 mL) was added CsCO (31.0 g, 95.0 mmol, 1.8 equiv) and 2-chloroacetaldehyde (20.7 g, 106 mmol, 17.0 mL, 2.0 equiv) at room temperature. The mixture was stirred at 20°C for 12 hours. The reaction mixture was quenched by the addition of water (500 mL) and extracted with ethyl acetate (3 x 500 mL). The combined organic layers were dried over NaSO, filtered, and concentrated under reduced pressure to afford 24.0 g of (Z)-ethyl (((2-bromophenyl)amino)((2-oxoethyl)thio)methylene)carbamate as a light yellow solid.

[0392] Ethyl (Z)-(3-(2-bromophenyl)thiazol-2(3H)-ylidene)carbamate

[0393] To a solution of (Z)-ethyl (((2-bromophenyl)amino)((2-oxoethyl)thio)methylene)carbamate (24.0 g, 69.5 mmol, 1.0 equiv) in THF (240 mL) was added DIPEA (27.0 g, 209 mmol, 36.3 mL, 3.0 equiv) and SOCl (8.27 g, 69.5 mmol, 5.0 mL, 1.0 equiv) at 0 ° C. The mixture was stirred at 15 ° C for 4 hours. The reaction mixture was quenched by adding saturated NaHCO solution (400 mL) and extracted with ethyl acetate (3x400 mL). The combined organic layers were dried over Na SO, filtered and concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1 to 1 / 1) to give 13.43 g of (Z)-(3-(2-bromophenyl)thiazole-2(3H)-ylidene)carbamic acid ethyl ester. LCMS: (m / z[M+H+ ]):329.0.

[0394] Intermediate C22

[0395] To a solution of ethyl (Z)-(3-(2-bromophenyl)thiazol-2(3H)-ylidene)carbamate (2.00 g, 6.11 mmol, 1.0 equiv) in EtOH (20 mL) was added 6N NaOH (4.89 g, 122 mmol, 20 equiv) at room temperature. The mixture was stirred at 50 ° C for 1.5 hours under N2. The reaction mixture was quenched by the addition of water (40 mL) and extracted with ethyl acetate (3×40 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give 1.56 g of 3-(2-bromophenyl)thiazol-2(3H)-imine C22 as a yellow oil. LCMS (m / z [M+H] + ):255.1.

[0396] Preparation of intermediate C23

[0397]

[0398] tert-Butyl 2-aminobenzylcarbamate

[0399] To a solution of 2-(aminomethyl)aniline (5.00 g, 40.9 mmol, 1.0 equivalent) in DCM (50 mL) was added a solution of BOC2O (8.93 g, 40.9 mmol, 9.40 mL, 1.0 equivalent). The reaction mixture was stirred at 25 ° C for 12 hours under an N2 atmosphere. The reaction mixture was extracted with H2O (25 mL). The combined organic layer was washed with salt water (25 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain 8.36 g of 2-aminobenzylcarbamate-)methyl] tert-butyl carbamate, a light yellow solid. 1 H NMR (400MHz, DMSO-d6) δ7.18(s,1H),7.19-7.18(m,1H),7.05-7.00(m,1H),7.02(dt,J=1.3,7.6Hz,1H),6.95(d,J=7.3H z,1H),6.65-6.53(m,2H),4.19-4.14(m,1H),4.19-4.11(m,1H),4.16(d,J=6.4Hz,2H),1.36-1.36(m,1H),1.37(s,9H). LCMS(m / z[M+H] + ):166.9.

[0400] Ethyl N-[[2-[(tert-Butoxycarbonylamino)methyl]phenyl]aminomethylthio]carbamate

[0401] To a solution of tert-butyl 2-aminobenzylaminomethyl]methyl]carbamate (5.00 g, 22.5 mmol, 1.0 equivalent) in EtOAc (50 mL) was added TMEDA (2.61 g, 22.5 mmol, 3.39 mL, 1.0 equivalent) and N-(thiomethylene)carbamate (4.43 g, 33.7 mmol, 1.5 equivalents). The reaction mixture was stirred at 25 ° C for 5 hours. The mixture was directly concentrated under reduced pressure to obtain a yellow oil. The oil was dissolved in MTBE (20 mL) and stirred for 0.5 hours, then filtered. The filter cake was washed with MTBE (2 × 10 mL) and dried in vacuo to obtain 7.75 g of ethyl N-[[2-[(tert-butoxycarbonylamino)methyl]phenyl]aminomethylthio]carbamate as a yellow solid. 1 H NMR(400MHz,DMSO-d6)δ11.18(br s,1H),11.21-11.15(m,1H),11.24-11.09(m,1H),11.24-11.09(m,1H),7. 35-7.31(m,1H),7.29-7.19(m,3H),7.12-7.03(m,1H),7.12-7.02(m,1H),7 .13-7.00(m,1H),4.62(s,1H),4.65-4.55(m,1H),4.29-4.12(m,2H),4.10- 4.02(m,2H),1.37(s,9H),1.28-1.19(m,1H),1.23(td,J=7.2,16.0Hz,3H). LCMS (m / z [M+H] + ):298.1.

[0402] (Z)-Ethyl(((2-(((tert-butoxycarbonyl)amino)methyl)phenyl)amino)((2-oxoethyl)thio)methylene)carbamate

[0403] To a solution of ethyl N-[[2-[(tert-butoxycarbonylamino)methyl]phenyl]aminomethylthio]carbamate (3.70 g, 10.5 mmol, 1.0 equiv) in MeCN (50 mL) was added CsCO (5.80 g, 17.8 mmol, 1.7 equiv) followed by 2-chloroacetaldehyde (2.57 g, 13.1 mmol, 2.10 mL, 1.25 equiv) at 10 ° C. The reaction mixture was stirred at 20 ° C for 12 hours. The reaction mixture was filtered and concentrated under reduced pressure. The residue was diluted with water (50 mL) and extracted with EtOAc (3×20 mL). The combined organic layers were washed with brine (50 mL), dried over NaSO, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 0 to 1 / 1) to give 3.0 g of (Z)-ethyl (((2-((tert-butoxycarbonyl)amino)methyl))phenyl)amino)((2-oxoethyl)thio)methylene)carbamate as a yellow solid. LCMS (m / z [M+H] + ):396.1.

[0404] (Z)-Ethyl (3-(2-(((tert-butoxycarbonyl)amino)methyl)phenyl)thiazole-2(3H)-methylenecarbamate

[0405] To (Z)-ethyl (((2-(((tert-butoxycarbonyl)amino)methyl)phenyl)amino)((2-oxoethyl)thio)methylene)carbamate (2.00 g, 5.06 mmol, 1.0 equiv) in THF (40 mL) was added DIPEA (1.96 g, 15.2 mmol, 2.64 mL, 3.0 equiv) and SOCl (602 mg, 5.06 mmol, 367 μL, 1.0 equiv) at 0°C. The reaction mixture was stirred at 15°C under N atmosphere for 1 hour. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with saturated aqueous NaHCO (25 mL) and extracted with MTBE (3x25 mL). The combined organic layers were washed with brine (25 mL), dried over NaSO, filtered and concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 0 to 2 / 1) to give 1.06 g of (Z)-ethyl (3-(2-((tert-butoxycarbonyl)amino))methyl)phenyl)thiazol-2(3H)-ylidene)carbamate as a yellow solid. 1H NMR (400MHz, DMSO-d6) δ7.55-7.47(m,1H),7.42(br s,1H),7.46-7.39(m,1H),7.37(d,J=4.6Hz,1H),7.28(d,J=7.7Hz,1H),7.24(br t,J=5.6Hz,1H),7.08(d,J=4.9Hz,1H),4.07-3.93(m,2H),3.88(br d,J=6.0Hz,2H),1.40-1.21(m,9H),1.20-1.09(m,3H). LCMS(m / z[M+H] + ):378.3.

[0406] Intermediate C23

[0407] To a solution of (Z)-ethyl(3-(2-(((tert-butoxycarbonyl)amino)methyl)phenyl)thiazol-2(3H)-ylidene)carbamate (340 mg, 901 μmol, 1.0 equiv) in EtOH (3.4 mL) was added NaOH (831 mg, 20.8 mmol, 23 equiv). The reaction mixture was stirred at 50 °C under N2 atmosphere for 1.5 hours. The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to afford 133 mg of tert-butyl 2-(2-iminothiazol-3(2H)-yl)benzylcarbamate C23 as a light yellow solid.

[0408] Preparation of intermediate C24

[0409]

[0410] tert-Butyl 3-aminobenzylcarbamate

[0411] At 25 ℃, to 3-(aminomethyl)aniline (5.00g, 40.9mmol, 1.0 equivalent) MeCN (50mL) solution, Boc was added dropwise o (8.13g, 37.2mmol, 8.56mL, 0.9 equivalent) MeCN (50mL) solution.Then the reaction mixture was stirred at 25 ℃ for 12 hours under N2.The reaction mixture was diluted with ethyl acetate (100mL) and filtered through a layer of silica gel, and then the layer of silica gel was washed with ethyl acetate (500mL).The filtrate was concentrated under reduced pressure to obtain 9.4g yellow oily tert-butyl 3-aminobenzylcarbamate. 1H NMR (400MHz, DMSO-d6) δ7.23 (br t, J = 5.9Hz, 1H), 6.95 (t, J = 7.6Hz, 1H), 6.48-6.38 (m, 3H), 4.99 (s, 2H), 4.00 (br d, J = 6.1Hz, 2H), 1.42 (s, 9H).

[0412] Ethyl N-[[3-[(tert-Butoxycarbonylamino)methyl]phenyl]aminomethylthio]carbamate

[0413] To a solution of tert-butyl 3-aminobenzylcarbamate (5.00 g, 22.5 mmol, 1.0 equivalent) in EtOAc (50 mL) was added O-ethyl carbonyl isothiocyanate (2.95 g, 22.5 mmol, 1.0 equivalent) and TMEDA (261 mg, 2.25 mmol, 339 μL, 0.1 equivalent) at 25 ° C. The reaction mixture was stirred at 25 ° C for 12 hours under N2. The reaction mixture was concentrated under reduced pressure. The residue was diluted with MTBE (50 mL) and stirred at 25 ° C for 0.5 h. 4.3 g of N-[[3-[(tert-butoxycarbonylamino)methyl]phenyl]aminomethylthio]ethyl carbamate was collected by filtration as a white solid. 1 H NMR(400MHz,DMSO-d6)δ11.55(s,1H),11.22(br s,1H),7.53(br d,J=7.8Hz,1H),7.46-7.38(m,2H),7.33(t,J=7.8Hz,1H),7.11(d,J=7.6Hz,1H),4.22(q,J=7.1Hz,2H),4.13(br d,J=6.1Hz,2H),1.40(s,9H),1.27(t,J=7.1Hz,3H).

[0414] (Z)-Ethyl(((3-(((tert-butoxycarbonyl)amino)methyl)phenyl)amino)((2-oxoethyl)thio)methylene)carbamate

[0415] To ethyl N-[[3-[(tert-butoxycarbonylamino)methyl]phenyl]aminomethylthio]carbamate (4.30 g, 12.2 mmol, 1.0 equiv) and Cs2CO3 (6.74 g, 20.7 mmol, 1.7 equiv) in MeCN (44 mL) was added 2-chloroacetaldehyde (2.98 g, 15.2 mmol, 2.45 mL, 1.25 equiv), maintaining the temperature below 25° C. After the addition, the reaction mixture was stirred at 25° C. under N2 for 12 h. The reaction mixture was diluted with MTBE (80 mL), washed with saturated NaHCO3 (75 mL) solution, brine (75 mL), dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to provide 4.7 g of (Z)-ethyl (((3-(((tert-butoxycarbonyl)amino)methyl)phenyl)amino)((2-oxoethyl)thio)methylene)carbamate) as a brown solid.

[0416] Ethyl (Z)-(3-(3-(((tert-Butoxycarbonyl)amino)methyl)phenyl)thiazol-2(3H)-ylidene)carbamate

[0417] To a solution of (Z)-ethyl (((3-(((tert-butoxycarbonyl)amino)methyl)phenyl)amino)((2-oxoethyl)thio)methylene)carbamate (4.70 g, 11.9 mmol, 1.0 equiv) in THF (47 mL) was added DIPEA (4.61 g, 35.6 mmol, 6.21 mL, 3.0 equiv) and SOCl2 (1.41 g, 11.9 mmol, 862 μL, 1.0 equiv) at 0°C. The reaction mixture was stirred at 25°C under N2 for 1 hour. The reaction mixture was concentrated under reduced pressure. The residue was dissolved in EtOAc (20 mL) and diluted with a saturated aqueous solution of NaHCO3 (60 mL). And extracted with EtOAc (3x80 mL). The combined organic layers were washed with brine (80 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography, eluting with a 0-40% ethyl acetate in petroleum ether gradient at 75 mL / min, to afford 2.4 g of (Z)-ethyl (3-(3-((tert-butoxycarbonyl))amino)methyl)phenyl)thiazol-2(3H)-ylidene)carbamate as a white solid. 1 H NMR (400MHz, DMSO-d6) δ7.53-7.47(m,2H),7.41-7.33(m,3H),7.08(d,J=4.8Hz,1H),4.21(br d,J=6.1Hz,2H),4.06-4.00(m,2H),1.40(s,9H),1.16(t,J=7.1Hz,3H).

[0418] Intermediate C24

[0419] To a solution of (Z)-ethyl (3-(3-(((tert-butoxycarbonyl)amino)methyl)phenyl)thiazole-2(3H)-ylidene)carbamate (1.00 g, 2.65 mmol, 1.0 equiv) in EtOH (10 mL) was added 6N NaOH (2.40 g, 60.0 mmol, 23.0 equiv) at room temperature. The reaction mixture was stirred at 60 ° C under N2 for 5 hours. The reaction mixture was adjusted to pH 7 with an aqueous HCl (6 M) solution. The neutralized solution was then filtered and the filter cake was washed with EtOH (200 mL). The filtrate was concentrated under reduced pressure. The crude product was purified by reverse phase column to give 300 mg of tert-butyl 3-(2-iminothiazole-3(2H)-yl)benzylcarbamate C24 as a yellow solid.

[0420] Preparation of intermediate C25

[0421]

[0422] Ethyl N-[(4-bromophenyl)aminomethylthio]carbamate

[0423] To a solution of 4-bromoaniline (5.00 g, 29.1 mmol, 1.0 equiv) in EtOAc (50 mL) was added N-(thiomethylene) ethyl carbamate (3.81 g, 29.1 mmol, 1.0 equiv) and TMEDA (338 mg, 2.91 mmol, 439 μL, 0.1 equiv) at room temperature. The reaction mixture was stirred at 20 ° C for 12 hours. The reaction mixture was concentrated under reduced pressure to obtain a residue. The residue was triturated with EtOH (25 mL) for 0.5 hours. The mixture was filtered, and the filter cake was washed with EtOH (2×25 mL) and dried to give 7.5 g of N-[(4-bromophenyl)aminomethylthio] ethyl carbamate as a white solid.

[0424] Ethyl (Z)-(((4-bromophenyl)amino)((2-oxoethyl)thio)methylene)carbamate

[0425] To a solution of N-[(4-bromophenyl)aminomethylthio]ethyl carbamate (7.50 g, 24.7 mmol, 1.0 equivalent) in CH3CN (100 mL) was added Cs2CO3 (13.70 g, 42.0 mmol, 1.7 equivalents) and 2-chloroacetaldehyde (6.07 g, 30.9 mmol, 4.97 mL, 1.25 equivalents) at room temperature. The reaction mixture was stirred at 20 ° C for 12 hours under N2. Cs2CO3 (8.06 g, 24.7 mmol, 1.0 equivalents) and 2-chloroacetaldehyde (3.64 g, 18.6 mmol, 2.98 mL, 0.75 equivalents) were added to the reaction solution. The reaction mixture was stirred at 20 ° C for 4 hours under N2. The reaction mixture was quenched by adding water (300 mL) and extracted with ethyl acetate (3×300 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give 10 g of (Z)-ethyl(((4-bromophenyl)amino)((2-oxoethyl)thio)methylene)carbamate as a light yellow solid. LCMS (m / z [M+H] + ):345.0.

[0426] (Z)-Ethyl (3-(4-bromophenyl)thiazol-2(3H)-ylidene)carbamate

[0427] To a solution of (Z)-ethyl (((4-bromophenyl) amino)((2-oxoethyl) thio) methylene) carbamate (10.00 g, 28.97 mmol, 1.0 equivalent) in THF (120 mL) was added DIPEA (11.23 g, 86.90 mmol, 15.14 mL, 3.0 equivalents) and SOCl (3.45 g, 28.97 mmol, 2.10 mL, 1.0 equivalents) at 0 ° C. The reaction mixture was stirred at 15 ° C for 4 hours. The reaction mixture was quenched by adding saturated NaHCO solution (200 mL) and extracted with ethyl acetate (3x200 mL). The combined organic layer was dried over Na SO, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1 to 1 / 1) to give 3.60 g of compound (Z)-ethyl (3-(4-bromophenyl)thiazol-2(3H))-ylidene)carbamate as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ7.75 (d, J = 8.63Hz, 2H), 7.47-7.56 (m, 3H), 7.08 (d, J = 4.88Hz, 1H), 4.03 (q, J = 7.09Hz, 2H), 1.16 (t, J = 7.07Hz, 3H).

[0428] Intermediate C25

[0429] To a solution of (Z)-ethyl (3-(4-bromophenyl)thiazole-2(3H)-ylidene)carbamate (1.00 g, 3.06 mmol, 1.0 equiv) in EtOH (10 mL) was added 6N NaOH (2.44 g, 61.1 mmol, 20 equiv). The reaction mixture was stirred at 50 ° C under N for 1.5 hours. The reaction was combined with the previous batch (with 100 mg of starting material). The reaction mixture was quenched by the addition of water (100 mL) and extracted with ethyl acetate (3×100 mL). The combined organic layers were dried over Na SO , filtered and concentrated under reduced pressure to give 750 mg of 3-(4-bromophenyl)thiazole-2(3H)-imine C25 as a yellow oil.

[0430] Preparation of intermediate C26

[0431]

[0432] Ethyl N-((cyclohexylmethyl)aminomethylthio)urethane

[0433] To a solution of cyclohexylmethylamine (2.00 g, 17.7 mmol, 1.0 equiv) in ethyl acetate (20 mL) was added O-ethyl carboisothiocyanate (2.09 mL, 17.7 mmol, 1.0 equiv) and tetramethylethylenediamine (0.267 mL, 1.77 mmol, 0.1 equiv) at 25°C. The mixture was then stirred at this temperature for 5 hours. The mixture was concentrated under reduced pressure and the residue was triturated twice with ethanol (10 mL) to give 3.50 g of ethyl N-((cyclohexylmethyl)aminomethylthio)carbamate as an off-white solid. 1 H NMR (400MHz, DMSO-d6) δ10.94(br.s,1H),9.92(br.t,J=5.2Hz,1H),4.14(q,J=7.2Hz,2H) ,3.42-3.39(m,2H),2.83(s,1H),1.69-1.60(m,6H),1.22-1.10(m,5H),0.98-0.90(m,2H). LCMS(m / z[M+H] + ):244.9.

[0434] Ethyl (Z)-(3-(cyclohexylmethyl)-4-methylthiazol-2(3H)-ylidene)carbamate

[0435] To a solution of N-((cyclohexylmethyl)aminomethylthio)ethyl carbamate (1.00 g, 4.09 mmol, 1.0 equivalent) and cesium carbonate (2.27 g, 6.96 mmol, 1.7 equivalent) in acetonitrile (10 mL) was added 1-chloroacetone (0.568 g, 6.14 mmol, 1.5 equivalents) at 25 ° C. After addition, it was stirred at this temperature for 2 hours. The mixture was poured into water (30 mL) and extracted with ethyl acetate (50 mL × 3). The combined organic layer was washed with brine (25 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 5 / 1) to give 0.80 g of (Z)-(3-(cyclohexylmethyl)-4-methylthiazole-2(3H))-ylidene)ethyl carbamate as a yellow oil. 1 H NMR(400MHz,DMSO-d6)δ6.44(s,1H),4.01-3.99(m,2H),3.22-3.09(m,2H),3.08-3.05(m,1H ),1.65-1.61(m,6H),1.48(s,3H),1.15-1.14(m,3H),1.12-1.10(m,2H),0.90-0.88(m,2H). LCMS(m / z[M+H] + ):282.9.

[0436] Intermediate C26

[0437] To a solution of (Z)-(3-(cyclohexylmethyl)-4-methylthiazole-2(3H)-ylidene)carbamic acid ethyl ester (0.350 g, 1.08 mmol, 1.0 equiv) in ethanol (3 mL) was added 6N sodium hydroxide (0.862 g, 21.5 mmol, 20 equiv) in one portion at 25°C. The reaction mixture was heated to 50°C and stirred for 1 hour. The mixture was added to water (15 mL) and extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with brine (25 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 0 / 1) to give 150 mg of 3-(cyclohexylmethyl)-4-methylthiazole-2(3H)-imine C26 as a yellow oil.

[0438] Preparation of intermediate C27

[0439]

[0440] N-(Cyclohexylaminomethylthio)ethyl carbamate

[0441] To the mixture of cyclohexylamine (0.800g, 8.07mmol, 1.0 equivalent) and O-ethyl carbonylthiocyanate (1.11g, 8.47mmol, 1.05 equivalent) in ethyl acetate (10mL) at 25 ℃, add TMEDA (93mg, 0.806mmol, 1.0 equivalent). Stir the mixture at this temperature for 6 hours, then concentrate under reduced pressure. Residue is passed through silica gel column chromatography (column height: 250mm, diameter: 100mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate=100 / 1 to 3 / 1), obtain 1.80g of N-(cyclohexylaminomethylthio) ethyl carbamate, be yellow jelly. 1 H NMR(400MHz,CD3Cl)δ9.64(br.s,1H),7.96(br.s,1H),4.29-4.22(m,2H),2.09-2.01 (m,2H),1.77-1.68(m,2H),1.62-1.63(m,1H),1.49-1.33(m,4H),1.33-1.19(m,5H). LCMS(m / z[M+H] + ):231.1.

[0442] (Z)-Ethyl((cyclohexylamino)((2-oxopropyl)thio)methylene)carbamate

[0443] To a mixture of ethyl N-(cyclohexylaminomethylthio)carbamate (0.800 g, 3.47 mmol, 1.0 equiv) and cesium carbonate (1.92 g, 5.90 mmol, 1.7 equiv) in acetonitrile (10 mL) was added 1-chloropropane-2-one (0.353 g, 3.82 mmol, 1.1 equiv) at 25 ° C. The resulting mixture was stirred for 4 hours, then poured into ice water (10 mL) and extracted with MTBE (100 mL×2). The combined organic phases were washed with brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give 900 mg of crude (Z)-ethyl ((cyclohexylamino)((2-oxopropyl)thio))methylene)carbamate as a yellow solid, which was used directly in the next step without purification. LCMS (m / z[M+H] + ):287.1.

[0444] (Z)-(3-cyclohexyl-4-methylthiazol-2(3H)-ylidene)carbamic acid ethyl ester

[0445] To (Z)-ethyl((cyclohexylamino)((2-oxopropyl)thio)methylene)carbamate (0.900 g, 3.14 mmol, 1.0 equiv) and diisopropylethylamine (0.812 g, 6.29 mmol, 2.2 equiv) in tetrahydrofuran (20 mL) was added thionyl chloride (0.224 g, 1.89 mmol, 0.6 equiv) at 0°C. The mixture was stirred at 25°C for 2 hours, then poured into ice water (20 mL) and extracted with ethyl acetate (100 mL x 3). The combined organic phases were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 100 / 1 to 4 / 1) to give 0.50 g of ethyl (Z)-(3-cyclohexyl-4-methylthiazol-2(3H)-ylidene)carbamate as a yellow solid. 1 HNMR (400MHz, DMSO-d6) δ6.54(br.s,1H),4.09-4.03(m,2H),2.29-2.25(m,3H),1.82-1.62(m,6H),1.36-1.23(m,3H),1.21-1.19(m,5H). LCMS(m / z[M+H] + ):268.9.

[0446] Intermediate C27

[0447] To a mixture of (Z)-(3-cyclohexyl-4-methylthiazol-2(3H)-ylidene)carbamic acid ethyl ester (0.200 g, 0.745 mmol, 1.0 equiv) in ethanol (4 mL) was added NaOH (0.596 g, 14.90 mmol, 20 equiv) at 25°C. The mixture was heated to 50°C and stirred for 1 hour, then poured into ice water (50 mL) and extracted with ethyl acetate (50 mL x 5). The combined organic phases were washed with brine (10 x 2 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford 120 mg of 3-cyclohexyl-4-methylthiazol-2(3H)-imine C27 as a yellow solid.

[0448] Preparation of intermediate C28

[0449]

[0450] Ethyl N-[(2-chlorophenyl)aminomethylthio]carbamate

[0451] At 0 ° C, to a solution of 2-chloroaniline (8.26 mL, 78.4 mmol) in ethyl acetate (100 mL) was slowly added carbon isothiocyanate (10.3 g, 78.4 mmol) and tetramethylethylenediamine (1.18 mL, 7.84 mmol), and the mixture was stirred at 50 ° C for 5 hours. The reaction mixture was cooled to room temperature and quenched dropwise with 100 mL of methanol. The precipitated solid was collected by filtration and then ground with methanol (100 mL) for 10 minutes at 25 ° C to give 16.5 g of N-[(2-chlorophenyl)aminomethylthio]ethyl carbamate as a white solid. 1 H NMR(DMSO-d6,400MHz)δ11.60(br.s,1H),11.47(br.s,1H),7.94(d,J=8.0Hz,1H),7.57-7.5 5(m,1H),7.41-7.36(m,1H),7.33-7.29(m,1H),4.23(q,J=7.2Hz,2H),1.27(t,J=7.2Hz,3H). LCMS(m / z[M+H] + ):258.9.

[0452] Methyl (Z)-3-((N-(2-chlorophenyl)-N'-(ethoxycarbonyl)carbamoyl)thio)-2-oxopropanoate

[0453] To N-[(2-chlorophenyl) aminomethylthio] ethyl carbamate (5.00g, 19.3mmol) and cesium carbonate (7.56g, 23.2mmol) in acetonitrile (100mL) was added 3-bromo-2-oxopropanoic acid methyl ester (2.67mL, 25.1mmol) at 25 DEG C, then the mixture was stirred at 25 DEG C for 2 hours. The reaction mixture was quenched with water (100mL) at 25 DEG C, and extracted with ethyl acetate (200mL×3). The combined organic layer was washed with brine (200mL x 2), dried over anhydrous NaSO and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 2 / 1) to give 2.80 g of methyl (Z)-3-((N-(2-chlorophenyl)-N'-(ethoxycarbonyl)carbamoyl)thio)-2-oxopropanoate as a brown oil. 1H NMR(CDCl3,400MHz)δ7.53-7.49(m,1H),7.47-7.44(m,1H),7.35-7.31(m,2H),4.90(br.s,1H),4. 16(q,J=7.2Hz,2H),4.00(d,J=11.6Hz,1H),3.75(s,3H),3.47-3.42(m,1H),1.26(t,J=7.2Hz,3H). LCMS(m / z[M+H] + ):358.9.

[0454] (Z)-3-(2-chlorophenyl)-2-((ethoxycarbonyl)imino)-2,3-dihydrothiazole-4-carboxylic acid methyl ester

[0455] To a mixture of (Z)-3-((N-(2-chlorophenyl)-N'-(ethoxycarbonyl)carbamoyl)thio)-2-oxopropanoic acid methyl ester (2.80 g, 7.80 mmol) and DIPEA (2.04 mL, 11.7 mmol) in THF (15 mL) was added sulfonyl chloride (0.85 mL, 11.7 mmol) at 0 ° C. The mixture was stirred at 25 ° C for 2 hours under a nitrogen atmosphere. The reaction mixture was quenched with water (100 mL) at 25 ° C. and extracted with ethyl acetate (200 mL×3). The combined organic layers were washed with brine (200 mL x 2), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to give 2.20 g of (Z)-3-(2-chlorophenyl)-2-((ethoxycarbonyl)imino)-2,3-dihydrothiazole-4-carboxylic acid methyl ester as a yellow oil, which was used in the next step without further purification. 1 HNMR(CDCl3,400MHz)δ7.64(s,1H),7.54-7.51(m,1H),7.46-7.41(m,2H),7 .40-7.37(m,1H),4.20(q,J=7.2Hz,2H),3.74(s,3H),1.29(t,J=7.2Hz,3H). LCMS(m / z[M+H] + ):340.9.

[0456] Ethyl (Z)-(3-(2-chlorophenyl)-4-(methylcarbamoyl)thiazol-2(3H)-ylidene)carbamate

[0457] To a mixture of (Z)-methyl 3-(2-chlorophenyl)-2-((ethoxycarbonyl)imino)-2,3-dihydrothiazole-4-carboxylate (1.5 g, 4.40 mmol) in THF (15 mL) was added methylamine (2 M, 22 mL) and 3,4,6,7,8,9-hexahydro-2H-pyrimido[1,2-a]pyrimidine (0.306 g, 2.20 mmol) dropwise at 25 ° C. The mixture was stirred at 60 ° C for 2 hours under a nitrogen atmosphere, then slowly poured into ice water (50 mL) and subsequently extracted with ethyl acetate (100 mL × 3). The combined organic layer was washed with brine (200 mL x 2), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to give 1.55 g of (Z)-ethyl (3-(2-chlorophenyl)-4-(methylcarbamoyl)thiazol-2(3H)-ylidene)carbamate as a yellow solid. 1 H NMR(DMSO-d6,400MHz)δ8.63(br.d,J=4.8Hz,1H),7.59(d,J=7.6Hz,1H),7.54(s,1H),7.4 9-7.43 (m, 3H), 4.02 (q, J = 7.2Hz, 2H), 2.57 (d, J = 4.8Hz, 3H), 1.19-1.04 (t, J = 7.2Hz, 3H). LCMS(m / z[M+H] + ):339.9.

[0458] Intermediate C28

[0459] To a solution of ethyl (Z)-(3-(2-chlorophenyl)-4-(methylcarbamoyl)thiazol-2(3H)-ylidene)carbamate (0.360 g, 1.06 mmol) in DMF (7 mL) was added bromotrimethylsilane (0.687 mL, 5.30 mmol) at 25°C. The reaction mixture was stirred at 80°C for 1 hour and then slowly poured into ice water (20 mL). The mixture was neutralized with saturated aqueous sodium bicarbonate. The mixture was directly purified by reverse phase flash distillation (0.1% NH3·H2O / MeCN / water) to give 80 mg of 3-(2-chlorophenyl)-2-imino-N-methyl-2,3-dihydrothiazole-4-carboxamide C28 as a yellow solid.

[0460] Preparation of intermediate C29

[0461]

[0462] Preparation of Intermediate C29 Ethyl (Z)-(3-(2-chlorophenyl)-4-(dimethylcarbamoyl)thiazol-2(3H)-ylidene)carbamate

[0463] To a mixture of (Z)-3-(2-chlorophenyl)-2-((ethoxycarbonyl)imino)-2,3-dihydrothiazole-4-carboxylic acid methyl ester (0.380 g, 1.12 mmol) and dimethylamine (2 M, 5.6 mL) in THF (1 mL) was added 3,4,6,7,8,9-hexahydro-2H-pyrimido[1,2-a]pyrimidine (0.078 g, 0.560 mmol) in a single portion at 25°C under a nitrogen atmosphere. The reaction mixture was stirred at 60°C for 2 hours and then carefully poured into 100 mL of ice water. The aqueous phase was extracted with ethyl acetate (50 mL x 3) and then washed with brine (10 mL). The organic layer was dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure to give a residue which was purified by reverse phase flash (0.1% trifluoroacetic acid / MeCN / water) to give 140 mg of ethyl (Z)-[3-(2-chlorophenyl)-4-(dimethylcarbamoyl)thiazol-2-ylidene]carbamate as a yellow solid. 1 H NMR (DMSO-d6, 400MHz) δ7.63-7.61 (m, 1H), 7.52-7.43 (m, 3H), 7.36 (s, 1H), 4.04 (m, 2H), 3.11 (s, 3H), 2.76 (s, 3H), 1.14 (t, J = 7.2Hz, 3H). LCMS(m / z[M+H] + ):353.9.

[0464] Intermediate C29

[0465] To a solution of (Z)-[3-(2-chlorophenyl)-4-(dimethylcarbamoyl)thiazole-2-methylene]carbamic acid ethyl ester (0.390 g, 1.10 mmol) in ethanol (4 mL) was added solid sodium hydroxide (0.960 g, 24.0 mmol). The mixture was stirred at 50 °C for 2 hours. The crude product was directly purified by reverse phase flash distillation without any workup (0.1% ammonium hydroxide / MeCN / water) to afford 90 mg of 3-(2-chlorophenyl)-2-imino-N,N-dimethyl-thiazole-4-carboxamide C29 as a yellow solid.

[0466] Preparation of intermediate D1

[0467]

[0468] 2-Phenylacetyl chloride

[0469] To a solution of 2-phenylacetic acid (5.00 g, 36.7 mmol, 4.63 mL, 1.0 equiv) in DCM (50 mL) was added DMF (268 mg, 3.67 mmol, 283 μL, 0.1 equiv) at 0°C, and SOCl2 (8.74 g, 73.4 mmol, 5.33 mL, 2.0 equiv) was added to the mixture. The reaction mixture was stirred at 25°C for 1 hour. The reaction mixture was concentrated under reduced pressure to remove the solvent. Compound 2-phenylacetyl chloride (5.68 g, crude product) was obtained as a colorless oil.

[0470] 1-diazo-3-phenylpropan-2-one

[0471] To a solution of 2-phenylacetyl chloride (5.68 g, 36.7 mmol, 4.90 mL, 1.0 eq) in MeCN (50 mL) was added TMSCHN2 (2 M, 36.74 mL, 2 eq) at 0° C. The reaction mixture was then stirred at 25° C. for 2 hours. The resulting yellow solution was used directly in the next step without further purification.

[0472] 1-Bromo-3-phenylpropan-2-one

[0473] To a MeCN solution of 1-diazo-3-phenylpropan-2-one was added HBr (13.5 g, 55.1 mmol, 9.06 mL, 1.5 equivalents) (33% in AcOH). The mixture was stirred at 25 ° C for 12 hours. The mixture was poured into water (50 mL) and extracted with EtOAc (3 × 30 mL). The organic phase was washed with brine (50 mL), dried over Na SO , filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate=0 / 1 to 6 / 1) to obtain 2.30 g of 1-bromo-3-phenylpropan-2-one as a red oil.

[0474] Intermediate D1

[0475] To a solution of thiourea (715 mg, 9.39 mmol, 1.0 equivalent) in refluxing EtOH (20 mL) was added 1-bromo-3-phenylpropan-2-one (2.00 g, 9.39 mmol, 1.0 equivalent) and pyridine (742 mg, 9.39 mmol, 758 μL, 1.0 equivalent). The reaction mixture was stirred at 70 ° C for 4 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate=50 / 1 to 1 / 1) to give 0.6 g of crude product. 490 mg of crude product was purified on preparative HPLC to give 160 mg of 4-benzylthiazole-2(3H)-imine D1 as a white solid. 1H NMR: (400 MHz, DMSO-d6) δ 7.31-7.12 (m, 5H), 6.83 (s, 2H), 6.12 (s, 1H), 3.71 (s, 2H). The structure was confirmed by HMBC and HSQC.

[0476] Preparation of intermediate D3

[0477]

[0478] To a solution of benzylthiourea (1.00 g, 6.02 mmol, 1.0 equiv) in acetone (15 mL) was added HCl (1.20 g, 12.0 mmol, 1.18 mL, 36% purity, 2.0 equiv) and DMSO (939 mg, 12.0 mmol, 939 μL, 2.0 equiv) at 40°C. The reaction mixture was stirred for 48 hours. The reaction mixture was directly concentrated under reduced pressure. The residue was purified by reverse phase MPLC to give 0.80 g of 3-benzyl-4-methylthiazole-2(3H)-imine D3 as a white solid.

[0479] Preparation of acid D

[0480]

[0481] 4-Methyl-1H-pyrrolo[2,3-b]pyridine

[0482] To a solution of 4-chloro-1H-pyrrolo[2,3-b]pyridine (10.00 g, 65.54 mmol, 1.0 equiv) and Pd(dppf)Cl2 (1.20 g, 1.64 mmol, 0.025 equiv) in toluene (200 mL) was added MeMgBr (3 M, 109 mL, 5.0 equiv) dropwise at 25 °C. After addition, the reaction mixture was stirred at 80 °C for 12 hours, then cooled to 25 °C, quenched at 0 °C by addition of ice water (300 mL), and extracted with EtOAc (2 x 200 mL). The combined organic layers were washed with saturated aqueous NH4Cl solution (400 mL), brine (200 mL), dried over Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by flash chromatography on silica gel (eluting with a 0-100% ethyl acetate / petroleum ether gradient at 100 mL / min) to afford 7.00 g of 4-methyl-1H-pyrrolo[2,3-b]pyridine as a white solid. LCMS (m / z [M+H] + ):133.1.

[0483] 2,2,2-Trichloro-1-(4-methyl-1H-pyrrolo[2,3-b]pyridin-3-yl)ethanone

[0484] To a solution of 4-methyl-1H-pyrrolo[2,3-b]pyridine (1.00 g, 7.57 mmol, 1.0 equiv) in DCM (20 mL) was added AlCl3 (2.52 g, 18.9 mmol, 2.5 equiv) at 25 ° C. The reaction mixture was stirred at 25 ° C for 10 minutes, and then 2,2,2-trichloroacetyl chloride (2.06 g, 11.4 mmol, 1.5 equiv) was added. The reaction mixture was stirred at 25 ° C for more than 3 hours and 50 minutes. The mixture was poured into ice water (30 mL) and the precipitate was filtered. The filtrate was extracted with DCM (2×30 mL) and concentrated under reduced pressure to give 2.0 g of crude 2,2,2-trichloro-1-(4-methyl-1H-pyrrolo[2,3-b]pyridin-3-yl)ethanone as a light yellow solid.

[0485] 4-Methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid (Acid D)

[0486] A mixture of 2,2,2-trichloro-1-(4-methyl-1H-pyrrolo[2,3-b]pyridin-3-yl)ethanone (2.00 g, 7.21 mmol, 1.0 equiv) in aqueous NaOH (3 M, 30 mL, 12.5 equiv) was stirred at 25°C under N2 atmosphere for 3 hours. The mixture was cooled to 0°C and adjusted to pH <6.0 by adding concentrated aqueous HCl. The precipitate was collected by filtration, and the filter cake was washed with water (2 x 5 mL) and dried under vacuum to give 1.2 g of crude 4-methyl-1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid (acid D) as an off-white solid.

[0487] Preparation of Example T01

[0488]

[0489] To a mixture of 3-benzylthiazole-2(3H)-imine B1 (1.20 g, 6.31 mmol, 1.0 equiv), 1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid A (0.818 g, 5.05 mmol, 0.8 equiv), HOBt (1.28 g, 9.46 mmol, 1.5 equiv) and diisopropylethylamine (2.45 g, 18.9 mmol, 3.0 equiv) in dimethylformamide (10 mL) was added EDCI (1.81 g, 9.46 mmol, 1.5 equiv). The mixture was stirred at 25° C. for 1.5 hours and then poured into ice water (200 mL). The precipitate was collected by filtration, and the filter cake was washed with acetonitrile (3×30 mL), and then triturated with acetonitrile / methanol = 5:1 (5×30 mL). Then, reverse phase HPLC was performed to obtain 352 mg of (Z)-N-(3-benzothiazole-2(3H) substituent)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T01 from methanol (2×250 mL) as a white solid. 1 H-NMR(DMSO-d6,400MHz)δ12.18(br.s,1H),8.53-8.50(m,1H),8.25-8.10(m,2H),7.60(d, J=4.8Hz,1H),7.42-7.40(m,5H),7.36(d,J=7.2Hz,1H),6.96(d,J=4.8Hz,1H),5.52(s,2H). LCMS(m / z[M+H] + )=335.1.

[0490] The following Examples in Table E were prepared using Intermediate Bl and the appropriate acid from Table A in a manner similar to that shown in Scheme 54 above.

[0491] Table E

[0492]

[0493] The Examples in Table F were prepared in a manner analogous to that shown in Scheme 54 above using the appropriate intermediates in Table B / C and acid A in Table A.

[0494] Table F

[0495]

[0496]

[0497]

[0498]

[0499]

[0500]

[0501]

[0502] The preparation of the other examples in Table G is shown in Schemes 55-71.

[0503] Table G

[0504]

[0505]

[0506]

[0507]

[0508]

[0509] Preparation of Examples T37 and T38

[0510]

[0511] Example T37

[0512] To a solution of 1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid A (199 mg, 1.23 mmol, 1.0 equiv) in DCM (10 mL) was added oxalyl chloride (COCl) 2 (156 mg, 1.23 mmol, 107 μL, 1.0 equiv) and DMF (1 μL, 12.3 μmol, 0.01 equiv) at 0 ° C. The reaction mixture was stirred at 15 ° C for 2 hours, and then TEA (187 mg, 1.84 mmol, 1.5 equiv) and 3-methylthiazol-2-imine B2 (140 mg, 1.23 mmol, 1.0 equiv) were added. The resulting mixture was stirred at 15 ° C for 12 hours. The reaction mixture was quenched by adding H2O (20 mL) and extracted with EtOAc (3 × 15 mL). The combined organic layer was washed with brine (3 × 10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by preparative HPLC to give 21 mg of (Z)-N-(3-methylthiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T37 as a white solid.

[0513] Example T38

[0514] To a solution of 1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid A (379 mg, 2.34 mmol, 1.0 equiv) in DCM (10 mL) was added oxalyl chloride (COCl) 2 (297 mg, 2.34 mmol, 1.0 equiv) and DMF (2 μL, 23.4 μmol, 0.01 equiv) at 0 ° C. The reaction mixture was stirred at 15 ° C for 2 hours, and then TEA (355 mg, 3.51 mmol, 1.5 equiv) and 3-ethylthiazole-2-imine B3 (300 mg, 2.34 mmol, 1.0 equiv) were added. The resulting mixture was stirred at 15 ° C for 12 hours. The reaction mixture was quenched by adding water (30 mL) and extracted with EtOAc (3 × 20 mL). The combined organic layer was washed with brine (2 × 20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by preparative HPLC to afford 24 mg of (Z)-N-(3-ethylthiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T38 as a white solid.

[0515] Preparation of Example T39

[0516]

[0517] (Z)-N-(Thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide

[0518] To a solution of 1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid A (0.60 g, 3.70 mmol, 1.0 equiv) in pyridine (5.88 g, 74.3 mmol, 20 equiv) was added HBTU (2.81 g, 7.40 mmol, 2.0 equiv) and thiazole-2-amine (556 mg, 5.55 mmol, 1.5 equiv). The reaction mixture was heated to 100 ° C and stirred at 100 ° C for 12 hours. The reaction mixture was cooled to 0 ° C and poured into ice water (15 mL). The solid was collected by filtration. The filter cake was washed with water (2×5 mL) and dried under reduced pressure to give a crude product. The crude product was recrystallized in MeOH / EtOH (4 mL / 4 mL) by refluxing at 60 ° C for 0.5 hours, then cooled to 25 ° C and filtered. The solid was washed with cooled EtOH (5 mL) and dried in vacuo to afford 0.41 g of (Z)-N-(thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide as a brown solid.

[0519] (Z)-N-(3-(1-phenylethyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T39

[0520] To a solution of (Z)-N-(thiazole-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (0.20 g, 819 umol, 1.0 equiv) in DMF (3 mL) was added KCO (226 mg, 1.64 mmol, 2.0 equiv) and 1-chloroethylbenzene (173 mg, 1.23 mmol, 1.5 equiv). The reaction mixture was stirred at 25 ° C for 12 hours. The reaction mixture was heated to 60 ° C for 18 hours. The reaction mixture was cooled to room temperature and filtered to provide a yellow solution in DMF. The yellow solution was purified by preparative HPLC to give 25 mg of (Z)-N-(3-(1-phenylethyl)thiazole-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T39 as a light yellow solid.

[0521] Preparation of Example T41

[0522]

[0523] (Z)-tert-Butyl(4-(2-((1H-pyrrolo[2,3-b]pyridine-3-carbonyl)imino)thiazol-3(2H)-yl)butan-2-yl)carbamate

[0524] To a solution of 1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid A (30 mg, 184 μmol, 1.0 equiv), HOBt (37 mg, 276 μmol, 1.5 equiv), EDCI (53 mg, 276 μmol, 1.5 equiv) and DIPEA (71 mg, 553 μmol, 96 μL, 3.0 equiv) in DMF (1 mL) was added tert-butyl (4-(2-iminothiazol-3(2H)-yl)butan-2-yl)carbamate B4 (50 mg, 184 μmol, 1.0 equiv). The reaction mixture was stirred at 25 ° C for 12 hours. The reaction mixture was diluted with 20 mL of water and extracted with EtOAc (3×10 mL). The combined organic layers were washed with 20 mL of brine, dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by preparative TLC on silica gel (petroleum ether / ethyl acetate) to give 40 mg of (Z)-tert-butyl(4-(2-((1H-pyrrolo[2,3-b]pyridine-3-carbonyl)imino)thiazol-3(2H)-yl)butan-2-yl)carbamate as a white solid.

[0525] (Z)-N-(3-(3-aminobutyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T41

[0526] A mixture of (Z)-tert-butyl(4-(2-((1H-pyrrolo[2,3-b]pyridine-3-carbonyl)imino)thiazol-3(2H)-yl)butan-2-yl)carbamate (40 mg, 96 μmol, 1.0 equiv) in HCl / EtOAc (4 M, 2 mL) was stirred at 25 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC to afford 28 mg of (Z)-N-(3-(3-aminobutyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T41 as a white solid.

[0527] Preparation of Examples T42 and T43

[0528]

[0529] (Z)-N-(3-(2-cyanobenzyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T42

[0530] To a solution of 1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid A (369 mg, 2.28 mmol, 1.0 eq) in DMF (6 mL) was added EDCI (524 mg, 2.73 mmol, 1.2 eq), HOBt (369 mg, 2.73 mmol, 1.2 eq), TEA (691 mg, 6.83 mmol, 3.0 eq) and 2-((2-iminothiazol-3(2H)-yl)methyl)benzonitrile B11 (0.49 g, 2.28 mmol, 1.0 eq). The reaction mixture was stirred at 25° C. for 12 h. The reaction mixture was quenched by adding water (15 mL) at 0°C, and the precipitate was collected by filtration and washed with water (2 x 5 mL), and dried under vacuum to give 0.60 g of crude (Z)-N-(3-(2-cyanobenzyl)thiazol-2(3H)-substituent)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T42 as a gray solid. 180 mg of crude T42 was further purified by preparative HPLC to give 92 mg of pure (Z)-N-(3-(2-cyanobenzyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T42 as a light yellow solid.

[0531] (Z)-N-(3-(2-(aminomethyl)benzyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T43

[0532] To a solution of LAH (48 mg, 1.25 mmol, 3.0 equiv) in THF (4 mL) was added (Z)-N-(3-(2-cyanobenzyl)thiazole-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T42 (0.15 g, 417 μmol, 1.0 equiv) in portions at 0 ° C. After addition, the reaction mixture was slowly warmed to 25 ° C and stirred at 25 ° C for 4 hours. The reaction mixture was quenched by adding HCl aqueous solution (2N, 0.5 mL) at 0 ° C and stirred for 0.5 hours. The mixture was diluted with THF / DMF (20 mL / 4 mL) and filtered. The filter cake was washed with MeOH (2x5 mL). The combined filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC to afford 8.3 mg of (Z)-N-(3-(2-(aminomethyl)benzyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T43 as an off-white solid.

[0533] Preparation of Examples T44 and T45

[0534]

[0535] (Z)-N-(3-(3-cyanobenzyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T44

[0536] To a solution of 1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid A (264 mg, 1.63 mmol, 1.0 equiv), HOBt (330 mg, 2.44 mmol, 1.5 equiv), EDCI (468 mg, 2.44 mmol, 1.5 equiv), and DIPEA (630 mg, 4.88 mmol, 850 μL, 3.0 equiv) in DMF (8 mL) was added 3-((2-iminothiazol-3(2H)-yl)methyl)benzonitrile B17 (350 mg, 1.63 mmol, 1.0 equiv) at 25°C. The reaction mixture was then stirred at 25°C for 12 hours. The reaction mixture was diluted with 50 mL of water and extracted with EtOAc (3 x 30 mL). The combined organic layers were washed with 50 mL of brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was diluted with MeOH (20 mL) and stirred for 20 minutes; the suspension was filtered to collect the solid. The collected solid was washed with MeOH (10 mL) and then dried under vacuum to give 500 mg of (Z)-N-(3-(3-cyanobenzyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T44 as a yellow solid. 100 mg of the crude product was purified on preparative HPLC to give 62 mg of pure (Z)-N-(3-(3-cyanobenzyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T44 as a white solid.

[0537] (Z)-N-(3-(3-(Aminomethyl)benzyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T45

[0538] To a suspension of (Z)-N-(3-(3-cyanobenzyl)thiazole-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T44 (200 mg, 556 μmol) in THF (5 mL) was added LAH (63 mg, 1.67 mmol, 3.0 equiv) at 0°C under a nitrogen atmosphere. The reaction mixture was then stirred at 25°C for 3 hours. The reaction mixture was quenched by the addition of water (5 mL) and acidified to pH = 3 with HCl (4N), and the mixture became clear. The mixture was directly purified by preparative HPLC to give 76 mg of (Z)-N-(3-(3-(aminomethyl)benzyl)thiazole-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T45 as a white solid.

[0539] Preparation of Examples T46, T47, and T48

[0540]

[0541] (Z)-N-(3-(4-cyanobenzyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T46

[0542] To a solution of 1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid A (1.09 g, 6.75 mmol, 1.0 equiv), HOBt (1.37 g, 10.1 mmol, 1.5 equiv), EDCI (1.94 g, 10.1 mmol, 1.5 equiv), and DIPEA (2.62 g, 20.3 mmol, 3.5 mL, 3.0 equiv) in DMF (20 mL) was added 4-((2-iminothiazol-3(2H)-yl)methyl)benzonitrile B5 (2.00 g, 6.80 mmol, 1.0 equiv) at 25°C. The reaction mixture was stirred at 25°C for 12 hours. The reaction mixture was diluted with 150 mL of water and extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with 200 mL of brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a residue. The residue was diluted with MeOH (50 mL) and stirred for 20 minutes; the suspension was filtered to collect the solid. The filter cake was washed with MeOH (20 mL) and dried under vacuum to afford 1.8 g of (Z)-N-(3-(4-cyanobenzyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T46 as a yellow solid. 200 mg of crude T46 was purified by preparative HPLC to afford 132 mg of pure (Z)-N-(3-(4-cyanobenzyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T46 as a white solid.

[0543] (Z)-N-(3-(4-(Aminomethyl)benzyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T47 (Z)-N-(3-(4-Formylbenzyl)thiazol-2(3H)-substituted)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T48

[0544] To a mixture of (Z)-N-(3-(4-cyanobenzyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T46 (300 mg, 835 μmol, 1.0 equiv) in THF (7 mL) was added LAH (95 mg, 2.5 mmol, 3.0 equiv) under N2 at 0°C. The mixture was stirred at 25°C for 3 hours. The reaction mixture was quenched by the addition of water (5 mL) and acidified to pH = 3 with 4N HCl. The mixture was purified by preparative HPLC to give 108 mg of (Z)-N-(3-(4-(aminomethyl)benzyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T47 as a white solid and 31 mg of (Z)-N-(3-(4-formylbenzyl)thiazol-2(3H)substituent)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T48 as a white solid.

[0545] Preparation of Examples T54 and T64

[0546] To a solution of the appropriate imine (1.0 equiv) in pyridine was added HBTU (2.0 equiv) and 1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid A (1.0 equiv). The reaction mixture was stirred at 100°C for 12 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC to afford T54 and T64.

[0547] Preparation of Example T51

[0548]

[0549] N-(Thiazol-2-yl)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide

[0550] To a solution of 1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid A (0.60 g, 3.70 mmol, 1.0 equiv) in pyridine (6 mL) was added HBTU (2.81 g, 7.40 mmol, 2.0 equiv) and 2-thiazolamine (556 mg, 5.55 mmol, 1.5 equiv) at 0°C. The reaction mixture was slowly heated to 100°C and stirred at 100°C for 12 hours. The reaction mixture was poured into ice water (12 mL). The solid was collected by filtration and washed with H2O (5×2 mL) and dried under reduced pressure. The crude product was recrystallized by refluxing in MeOH / EtOH (4 mL / 4 mL) at 70°C for 0.5 hours, then cooled to room temperature and filtered to give 0.41 g of N-(thiazol-2-yl)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide as a brown solid.

[0551] Methyl 3-[(2Z)-2-(1H-pyrrolo[2,3-b]pyridine-3-carbonylimino)thiazol-3-yl]propanoate

[0552] To a solution of N-(thiazol-2-yl)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (0.29 g, 1.19 mmol, 1.0 equiv) in DMF (5 mL) was added KCO (656 mg, 4.75 mmol, 4.0 equiv) and methyl 3-bromopropionate (595 mg, 3.56 mmol, 3.0 equiv). After addition, the reaction mixture was transferred to a sealed microwave vial and stirred in a microwave reactor at 100 ° C for 2 hours. The reaction mixture was quenched by the addition of H O (10 mL) at 0 ° C and extracted with EtOAc (2 × 10 mL). The combined organic layers were washed with water (20 mL), brine (20 mL), dried over Na SO and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by preparative TLC on silica gel (ethanol / ethyl acetate = 1:1) to obtain 0.30 g of methyl 3-[(2Z)-2-(1H-pyrrolo[2,3-b]pyridine-3-carbonylimino)thiazol-3-yl]propanoate as a brown solid. A portion of the residue (0.10 g) was further purified twice by preparative TLC on silica gel (petroleum ether / ethyl acetate = 2:1) to obtain 20 mg of methyl 3-[(2Z)-2-(1H-pyrrolo[2,3-b]pyridine-3-carbonylimino)thiazol-3-yl]propanoate as a white solid.

[0553] Example T51

[0554] To a solution of methyl 3-[(2Z)-2-(1H-pyrrolo[2,3-b]pyridine-3-carbonylimino)thiazol-3-yl]propanoate (0.20 g, 605 μmol, 1.0 equiv) in dioxane (4 mL) / HO (1 mL) was added LiOH·HO (127 mg, 3.03 mmol, 5.0 equiv). The reaction mixture was stirred at 25 ° C for 12 hours. The mixture was concentrated. The residue was diluted with water (5 mL) and adjusted to pH < 5.0 by adding HCl (concentrated). The suspension was filtered. The filter cake was washed with water (5 mL) and dried under reduced pressure. The residue was purified by preparative HPLC to give 52 mg of (Z)-3-(2-((1H-pyrrolo[2,3-b]pyridine-3-carbonyl)imino)thiazol-3(2H)-yl)propanoic acid as a white solid.

[0555] Preparation of Example T52

[0556]

[0557]

[0558] A mixture of N-(thiazol-2-yl)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (100 mg, 409.38 μmol, 1.0 equiv), 4-(bromomethyl)pyridine (104 mg, 409.38 μmol, 1.0 equiv, HBr), and KCO (170 mg, 1.23 mmol, 3.0 equiv) in DMF (1 mL) was degassed and purged with N three times, and then the reaction mixture was stirred at 30°C under N atmosphere for 12 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC to give 37 mg of (Z)-N-(3-(pyridin-4-ylmethyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T52 as a white solid.

[0559] Preparation of Example T53

[0560]

[0561] A mixture of N-(thiazol-2-yl)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (100 mg, 409 μmol, 1.0 equiv), 3-(bromomethyl)pyridine (104 mg, 409 μmol, 1.0 equiv, HBr), and KCO (170 mg, 1.23 mmol, 3.0 equiv) was degassed in DMF (1 mL) and purged with N three times, and then the reaction mixture was stirred at 30°C under N for 12 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC to afford 32 mg of (Z)-N-(3-(pyridin-3-ylmethyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T53 as a white solid.

[0562] Preparation of Example T57

[0563]

[0564] To a solution of N-(thiazol-2-yl)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (200 mg, 819 μmol, 1.0 equiv) in DMF (2 mL) was added K2CO3 (226 mg, 1.64 mmol, 2.0 equiv) followed by the addition of bromocyclohexane (2.67 g, 16.4 mmol, 2.02 mL, 20 equiv) at room temperature. The reaction mixture was then stirred at 100 ° C for 12 hours. An additional 10 equivalents of bromocyclohexane were added to the reaction mixture, and the mixture was then stirred at 100 ° C for another 12 hours under N2. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (50 mL × 3), and the combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC to give 7.3 mg of (Z)-N-(3-cyclohexylthiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T57 as a white solid. The structure was confirmed by NOE.

[0565] Preparation of Example T61

[0566]

[0567] tert-Butyl (Z)-3-(2-((1H-pyrrolo[2,3-b]pyridine-3)-carbonyl)imino)thiazol-3(2H)-yl)benzylcarbamate

[0568] To a solution of 1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid A (80 mg, 491 μmol, 1.0 equiv) in DMF (2 mL) was added EDCI (141 mg, 737 μmol, 1.5 equiv), HOBt (100 mg, 737 μmol, 1.5 equiv), DIPEA (190 mg, 1.47 mmol, 257 μL, 3.0 equiv) and tert-butyl 3-(2-iminothiazol-3(2H)-yl)benzylcarbamate C24 (150 mg, 491 μmol, 1.0 equiv) at 25 ° C. The reaction mixture was then stirred under N2 for 2 hours at 25 ° C. The reaction mixture was diluted with water (5 mL) and a solid was precipitated. The solid was collected by filtration, washed with water (5 mL) and the residue was dried under pressure to give 170 mg of tert-butyl (Z)-3-(2-((1H-pyrrolo[2,3-b]pyridine-3)-carbonyl)imino)thiazol-3(2H)-yl)benzylcarbamate as a white solid.

[0569] (Z)-N-(3-(3-(aminomethyl)phenyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T61

[0570] To a solution of (Z)-3-(2-((1H-pyrrolo[2,3-b]pyridine-3)-carbonyl)imino)thiazole-3(2H)-yl)benzylcarbamic acid tert-butyl ester (170 mg, 378 μmol, 1.0 equivalent) in EtOAc (1 mL) was added HCl / EtOAc (378 μmol, 3 mL, 1.0 equivalent) at 25 ° C. The solution was then stirred at 25 ° C for 1 hour under N2. The reaction mixture was filtered and the filter cake was washed with EtOAc (30 mL). The filter cake was concentrated under reduced pressure and purified by preparative HPLC to give 52 mg of (Z)-N-(3-(3-(aminomethyl)phenyl)thiazole-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T61 as a white solid.

[0571] Preparation of Example T62

[0572]

[0573] tert-Butyl (Z)-7-(2-((1H-pyrrolo[2,3-b]pyridine-3-carbonyl)imino)thiazol-3(2H)-yl)-3,4-dihydroisoquinoline-2(1H)-carboxylate

[0574] To a mixture of 1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid A (80 mg, 493 μmol, 1.0 eq) in DMF (3 mL) was added HOBt (100 mg, 740 μmol, 1.5 eq), EDCI (142 mg, 740 μmol, 1.5 eq), DIPEA (191 mg, 1.48 mmol, 258 μL, 3.0 eq) and tert-butyl 7-(2-iminothiazol-3(2H)-yl)-3,4-dihydroisoquinoline-2(1H)-carboxylate C21 (164 mg, 493 μmol, 1.0 eq) at 25°C. The reaction mixture was then stirred at 25°C under N2 for 12 hours. The reaction mixture was diluted with water (20 mL) and filtered to collect the solid. The collected solid was dried under reduced pressure to obtain 200 mg of tert-butyl (Z)-7-(2-((1H-pyrrolo[2,3-b]pyridine-3-carbonyl)imino)thiazol-3(2H)-yl)-3,4-dihydroisoquinoline-2(1H)-carboxylate as a brown solid. The crude product was used directly in the next step.

[0575] (Z)-N-(3-(1,2,3,4-tetrahydroisoquinolin-7-yl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T62

[0576] To a mixture of (Z)-tert-butyl 7-(2-((1H-pyrrolo[2,3-b]pyridine-3-carbonyl)imino)thiazole-3(2H)-yl)-3,4-dihydroisoquinoline-2(1H)-carboxylate (170 mg, 357 μmol, 1.0 equiv) in EtOAc (1 mL) was added HCl / EtOAc (4 M, 3 mL, 33 equiv) at 25°C. The reaction mixture was then stirred at 25°C under N2 for 2 hours. The solid was collected by filtration and purified by preparative HPLC to give 88 mg of (Z)-N-(3-(1,2,3,4-tetrahydroisoquinolin-7-yl)thiazole-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T62 as a white solid.

[0577] Preparation of Example T72

[0578]

[0579] (Z)-2-((1H-pyrrolo[2,3-b]pyridine-3-carbonyl)imino)-3-benzyl-2,3-dihydrothiazole-4-carboxylic acid ethyl ester

[0580] To a solution of 1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid (0.238 g, 1.47 mmol, 1.0 equiv) in DMF (5 mL) was added EDCI (0.422 g, 2.20 mmol, 1.5 equiv), HOBT (0.298 g, 2.20 mmol, 1.5 equiv), diisopropylethylamine (0.758 mL, 4.41 mmol, 3.0 equiv) and ethyl 3-benzyl-2-imino-2,3-dihydrothiazole-4-carboxylate (0.400 g, 1.47 mmol, 1.0 equiv) at 25°C. The mixture was then stirred at 25°C for 1 hour. The reaction mixture was then poured into ice water (15 mL) to precipitate a solid which was collected by filtration. The filter cake was triturated three times with ethanol (15 mL) at 25 °C to afford 0.40 g of ethyl (Z)-2-((1H-pyrrolo[2,3-b]pyridine-3-carbonyl)imino)-3-benzyl-2,3-dihydrothiazole-4-carboxylate as a white solid. 1H NMR (400MHz, DMSO-d6): δ12.27(br.s,1H),8.45-8.42(m,1H),8.26(dd,J=4.8,1.2Hz,1H),8.21-8.20(m,1H),7.99(s,1H), 7.35-7.31(m,2H),7.27-7.2(m,3H),7.14(dd,J=8.0,4.8Hz,1H),5.93(s,2H),4.27(q,J=7.2Hz,2H),1.25(t,J=7.2Hz,3H). LCMS(m / z[M+H] + ):407.0.

[0581] (Z)-N-(3-Benzyl-4-(hydroxymethyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T72

[0582] To a suspension of (Z)-2-((1H-pyrrolo[2,3-b]pyridine-3-carbonyl)imino)-3-benzyl-2,3-dihydrothiazole-4-carboxylic acid ethyl ester (0.200 g, 0.407 mmol) in THF (5 mL) was added dropwise a solution of lithium borohydride in THF (2M, 0.4 mL, 0.8 mmol) under a nitrogen atmosphere and 0 ° C. After the addition, the reaction mixture was warmed to 25 ° C and stirred for 1 hour. The mixture was then added dropwise to water (5 mL) at 25 ° C, and the mixture was then added to a saturated aqueous sodium carbonate solution (5 mL). The aqueous phase was extracted with ethyl acetate (50 mL×3). The combined organic layers were washed with brine (15 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by preparative HPLC to afford 32 mg of (Z)-N-(3-benzyl-4-(hydroxymethyl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T72 as a white solid.

[0583] Preparation of Example T75

[0584]

[0585] tert-Butyl (Z)-4-(2-((1H-pyrrolo[2,3-b]pyridine-3-carbonyl)imino)thiazol-3(2H)-yl)piperidine-1-carboxylate

[0586] To a DMF (1 mL) solution of (Z)-N-(thiazole-2(3H) substituent)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide was added KCO (57 mg, 246 μmol, 3.0 equivalents) and tert-butyl 4-bromopiperidine-1-carboxylate (216 mg, 819 μmol, 10.0 equivalents). The reaction mixture was transferred to a microwave tube. The sealed tube was heated at 100 ° C. for 2 hours in a microwave reactor. The reaction mixture was quenched by adding water (5 mL) to form a brown precipitate. The solid was collected by filtration and washed with water (2 mL). The filtrate was dried under reduced pressure to give the crude product (Z)-4-(2-((1H-pyrrolo[2,3-b]pyridine-3-carbonyl)imino)thiazole-3(2H)-yl)piperidine-1-carboxylate (30 mg, crude) as a brown solid.

[0587] (Z)-N-(3-(Piperidin-4-yl)thiazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide hydrochloride T75

[0588] To a solution of (Z)-4-(2-((1H-pyrrolo[2,3-b]pyridine-3-carbonyl)imino)thiazole-3(2H)-yl)piperidine-1-carboxylic acid tert-butyl ester (0.03 g, 70 umol, 1.0 equiv) in EtOAc (1 mL) was slowly added dropwise HCl / EtOAc (4 M, 1.2 mL, 68.4 equiv). The reaction mixture was stirred at 25 ° C for 2 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC to give 5 mg of (Z)-N-(3-(piperidin-4-yl)thiazole-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide hydrochloride T75 as a white solid.

[0589] Preparation of Example T77

[0590]

[0591] To a solution of 3-benzyl-4-methyloxazole-2(3H)-imine 2,2,2-trifluoroacetic acid B19 (0.100 g, 0.285 mmol, 1.0 equiv) and triethylamine (0.198 mL, 1.43 mmol, 5.0 equiv) in chloroform (1 mL) was added a solution of N,N-dimethylpyridin-4-amine (3 mg, 0.03 mmol, 0.1 equiv) and 1H-pyrrolo[2,3-b]pyridine-3-carbonyl chloride hydrochloride A (62 mg, 0.28 mmol, 1.0 equiv) in chloroform (2 mL) at 0°C. The reaction mixture was warmed to 25°C and stirred for 1 hour. The mixture was concentrated under reduced pressure, and the residue was dissolved in methanol (2 mL) and then poured into water (10 mL) to precipitate a solid. The precipitate was collected by filtration and purified by preparative HPLC to give 8 mg of (Z)-N-(3-benzyl-4-methyloxazol-2(3H)-ylidene)-1H-pyrrolo[2,3-b]pyridine-3-carboxamide T77 as a white solid.

[0592] Preparation of Example T78

[0593]

[0594] (Z)-2-((1H-pyrrolo[2,3-b]pyridine-3-carbonyl)imino)-3-benzyl-2,3-dihydrooxazole-4-carboxylic acid ethyl ester

[0595] To a solution of ethyl 3-benzyl-2-imino-2,3-dihydrooxazole-4-carboxylate B20 (0.450 g, 1.83 mmol) and 1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid (acid A) (0.356 g, 2.19 mmol) in DMF (6 mL) was added DIPEA (1.59 mL, 9.14 mmol), EDCI (0.525 g, 2.74 mmol) and HOBt (0.247 g, 1.83 mmol) at 25 ° C. The reaction mixture was stirred at 50 ° C for 2 hours. The reaction was quenched with water (50 mL) and extracted with ethyl acetate (30 mL × 3). The combined organic layers were washed with saturated sodium bicarbonate aqueous solution (50 mL x 3) and brine (50 mL x 3), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure. The crude product was purified by reverse phase HPLC (0.1% NH3·H2O conditions) to give 0.220 g of (Z)-ethyl 2-((1H-pyrrolo[2,3-b]pyridine-3-carbonyl)imino)-3-benzyl-2,3-dihydrooxazole-4-carboxylate as a yellow solid. 1H NMR(DMSO-d6,400MHz)δ12.18(br.s,1H),8.52(s,1H),8.34(dd,J=1.6,6.4Hz,1H),8.24(dd,J=1.6,3.2Hz,1H),8.07(d,J=2.0Hz, 1H),7.37-7.36(m,4H),7.33-7.29(m,1H),7.10(dd,J=3.2,4.8Hz,1H),5.37(s,2H),4.28(q,J=7.2Hz,2H),1.25(t,J=7.2Hz,3H). LCMS(m / z[M+H] + ):391.0.

[0596] (Z)-N-(3-Benzyl-4-(hydroxymethyl)oxazole-2(3H)-substituted)-1-pyrrolo[2,3-b]pyridine-3-carboxamide T78

[0597] To a solution of (Z)-2-((1H-pyrrolo[2,3-b]pyridine-3-carbonyl)imino)-3-benzyl-2,3-dihydrooxazole-4-carboxylic acid ethyl ester (0.100 g, 0.231 mmol) in THF (3.0 mL) was added dropwise a solution of lithium borohydride (0.348 mL, 2M) in THF at 0°C under a nitrogen atmosphere. The reaction mixture was stirred at 0°C for 1 hour. The mixture was poured into water (20 mL) at 25°C, and then a saturated aqueous ammonium chloride solution (20 mL) was added. The aqueous phase was extracted with ethyl acetate (30 mL×3). The combined organic phases were washed with brine (50 mL), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure. The mixture was purified by preparative HPLC (column: Phenomenex Gemini-NX C18 75×30 mm×3 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 17%-47%, 8 minutes) to give 9 mg of (Z)-N-(3-benzyl-4-(hydroxymethyl)oxazole-2(3H) substituent)-1-pyrrolo[2,3-b]pyridine-3-carboxamide T78 as a white solid.

[0598] Preparation of Examples T79 and T80

[0599]

[0600] (2Z)-3-Benzyl-2-(1H-pyrrolo[2,3-b]pyridine-3-carbonylimino)thiazole-4-carboxylic acid ethyl ester T79

[0601] To a solution of 1H-pyrrolo[2,3-b]pyridine-3-carboxylic acid (acid A) (4.62 g, 28.4 mmol) in DMF (100 mL) at 25°C was added HOBt (3.08 g, 22.8 mmol), EDCI (8.20 g, 42.8 mmol), DIPEA (14.9 mL, 85.6 mmol), and ethyl 3-benzyl-2-imino-thiazole-4-carboxylate hydrobromide B8 (10.0 g, 28.5 mmol). The resulting mixture was stirred at 25°C for 1 hour. The reaction was poured into ice water (200 mL). The precipitate was collected by filtration, and the filter cake was triturated with ethanol (150 mL) to afford 10.0 g of ethyl (2Z)-3-benzyl-2-(1H-pyrrolo[2,3-b]pyridine-3-carbonylimino)thiazole-4-carboxylate T79 as a white solid.

[0602] (Z)-3-Benzyl-N-[2-(dimethylamino)ethyl]-2-(1H-pyrrolo[2,3-b]pyridine-3-carbonylimino)thiazole-4-carboxamide T80

[0603] To a solution of ethyl (2Z)-3-benzyl-2-(1H-pyrrolo[2,3-b]pyridine-3-carbonylimino)thiazole-4-carboxylate T79 (0.050 g, 0.123 mmol) and N',N'-dimethylethane-1,2-diamine (0.108 g, 1.23 mmol, 0.134 mL) in THF (1 mL) was added 3,4,6,7,8,9-hexahydro-2H-pyrimido[1,2-a]pyrimidine (0.009 g, 0.062 mmol). The mixture was stirred at 60° C. for 1 hour. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (Phenomenex Waters Xbridge 150*25mm*5um; mobile phase: [water (10mM NH4HCO3)-acetonitrile]; B%: 24%-54%, 10 minutes) to give 54 mg of (Z)-3-benzyl-N-[2-(dimethylamino)ethyl]-2-(1H-pyrrolo[2,3-b]pyridine-3-carbonylimino)thiazole-4-carboxamide T80 as a white solid.

[0604] General approach

[0605] LCMS conditions

[0606]

[0607] 10uM IVKA assay method:

[0608] In vitro kinase assays (HTRF KinEASE-STK S1, CisBio 62ST1PEB) were optimized to the linear reaction range of the enzyme Lats1 (Carna 01-123). As shown below, 10 μM STK1 substrate and 10 μM ATP or 2 mM ATP were used for the reaction. The DMSO concentration was maintained at 0.5% in all experiments, and compounds dissolved in DMSO were added to the reaction using a Janus 384MDT (PerkinElmer) equipped with a 50 nL Pintool (V&P Scientific, Inc.). Unless otherwise stated, the enzymes, substrates, ATP, and test compounds of the present invention were combined in a small volume 384-well plate and shaken at room temperature for 50 minutes. The reaction was terminated by adding a detection reagent prepared with a biotin:streptavidin ratio of 8:1 and shaken at room temperature for 60 minutes. All reactions were performed in triplicate and the signal was detected using Synergy NEO (Biotek).

[0609] Reagents, final assay concentration of IVKA in 2 mM ATP: Kit: HTRFKinEASE-STKS1, CisBio 62ST1PEB; 1x Kinase Buffer (KB); 5 mM MgCl2; 1 mM DTT; 2 mM ATP; 2.5 μM STK1; 50 pg / μL (Lats1, Carna 01-123); 1 mg / mL BSA; detection reagent: 8:1 biotin:streptavidin ratio; Streptavidin-XL665: 156 nM; final reaction volume: 10 μL; final assay volume: 20 μL. Assays were performed in 384-well plates. Each drug dose curve was performed in triplicate using 16 points of three-fold dilutions in 100% DMSO.

[0610] The Lats1 activities determined as described above are listed in the following table

[0611]

[0612]

[0613]

[0614] Compound T01 was extensively tested to explore its mechanism of action and its utility in otic hair cell regeneration. The compounds were tested on utricles isolated from 8- to 12-week-old mice. The inner ears were dissected from mice euthanized with halothane, placed in ice-cold Hank's balanced salt solution, and treated as described by Gnedeva, K. & Hudspeth [Proc. Natl. Acad. Sci. 112, 14066-14071 (2015)].

[0615] For proliferation assays, cells were incubated with 10 μM 5-ethynyl-2'deoxyuridine (EdU) detected by click chemistry.

[0616] Immunohistochemical analysis showed that T01 drove robust nuclear translocation of Yap in supporting cells after 24 hours of treatment at a concentration of 10 μM (quantified as the ratio to the constitutively expressed protein Sall2; control = 0.6; T01-treated = 1.0; p < 0.0001, unpaired two-tailed t-test, n = 570 control and 680 treated nuclei), which resulted in a significant reduction in Yap phosphorylation levels detected by Western blotting. After 5 days of treatment, T01 induced robust reentry into the cell cycle of adult utricle supporting cells, generating hundreds of EdU+ supporting cells (control = 20 EdU+ supporting cells; T01-treated = 250 EdU+ supporting cells; p = 0.021, unpaired one-tailed t-test, control n = 2, T01 n = 3).

Claims

1. A compound having the following structure: in: R 1 Selected from: (a) cyclohexyl; (b) phenyl or phenyl substituted by a substituent selected from halogen, (C1-C6)alkyl, hydroxy(C1-C3)alkyl, methoxycarbonyl, formamido and amino(C1-C3)alkyl; R 2 is selected from hydrogen, methyl, ethyl, propyl, cyclopropyl and hydroxymethyl; R 3 and R 4 selected from hydrogen; R 10 is independently selected in each instance from hydrogen and methyl; n is 0 or 1; X is S; The proviso is that when R 1 When it is phenyl and n is 1, R 2 、R 3 、R 4 and R 10 At least one of them is not hydrogen.

2. The compound according to claim 1, wherein n is 0.

3. The compound according to claim 1, wherein n is 1.

4. The compound according to claim 3, wherein R 10 It's hydrogen.

5. The compound according to claim 1, wherein R 1 is an ortho-substituted phenyl group and n is 0.

6. The compound according to claim 1, wherein R 2 It is hydroxymethyl.

7. A compound selected from Examples T14, T19, T20, T21, T24, T25, T27, T29, T32, T36, T39, T43, T49, T55, T57, T61, T62, T65, T67, T68, T70, T72, T73, T74, T75, T78, T79, T80, T81, and T82.

8. The compound of Example T72, whose structure is 9. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and the compound according to any one of claims 1 to 8.

10. Use of a compound of the following formula in the preparation of a medicament for treating hair cell damage: in: R 1 Selected from: (a) cyclohexyl; (b) phenyl or phenyl substituted by a substituent selected from halogen, (C1-C6)alkyl, hydroxy(C1-C3)alkyl, methoxycarbonyl, formamido and amino(C1-C3)alkyl; R 2 is selected from hydrogen, methyl, ethyl, propyl, cyclopropyl and hydroxymethyl; R 3 and R 4 selected from hydrogen; R 10 is independently selected in each instance from hydrogen and methyl; n is 0 or 1; and X is S.

11. Use of a compound of the following formula in the preparation of a medicament for treating hearing loss, tinnitus, vertigo, dizziness, imbalance, bilateral vestibulopathy, tremor or balance disorder caused by hair cell damage: in: R 1 Selected from: (a) cyclohexyl; (b) phenyl or phenyl substituted by a substituent independently selected from halogen, (C1-C6)alkyl, hydroxy(C1-C3)alkyl, methoxycarbonyl, formamido and amino(C1-C3)alkyl; R 2 is selected from hydrogen, methyl, ethyl, propyl, cyclopropyl and hydroxymethyl; R 3 and R 4 selected from hydrogen; R 10 is independently selected in each instance from hydrogen and methyl; n is 0 or 1; and X is S.

Images