Quinazolinone derivatives for HDAC inhibition

Abstract

The invention discloses a histone deacetylase 6 (HDAC6) inhibitor and a composition containing the histone deacetylase 6 (HDAC6) inhibitor. Compounds that are HDAC6 inhibitors are useful for treating, alleviating, ameliorating, and / or reducing the severity of HDAC6-related diseases or disorders in a subject in need thereof. HDAC6-related diseases or conditions include fibrosis, neurodegenerative diseases, kidney diseases, cancer, and increased cyst growth indicators.

Description

Technical Field

[0001] The present invention relates to compounds having histone deacetylase 6 (HDAC6) enzyme inhibitory activity and their uses. Background Art

[0002] Histone deacetylases (HDACs) catalyze the deacetylation of histones and non-histones and play important roles in epigenetic regulation. Autosomal dominant polycystic kidney disease (ADPKD) is characterized by slow progressive bilateral kidney enlargement caused by a large number of fluid-filled cysts. ADPKD is caused by mutations in the PKD1 or PKD2 genes, the disruption of whose normal functions leads to excessive proliferation of renal tubular epithelium, thus resulting in cyst formation. More than 12 million people worldwide suffer from ADPKD, making it one of the most common monogenic diseases. 50% of ADPKD patients eventually develop end-stage renal disease (ESRD) by the age of 60, accounting for 10% and 5% of the general ESRD patients in Europe and the United States, respectively. Unfortunately, the treatment options for this life-threatening disease remain limited and insufficient.

[0003] HDAC6 inhibitors have been proposed as a possible strategy for inhibiting cyst growth. It has also been found that HDAC6 inhibition downregulates cAMP levels, inhibits cell proliferation, and inhibits cAMP-activated CFTR chloride currents in MDCK cells. HDAC6 inhibition can also inhibit cyst growth in vitro. Based on previous studies showing that cystic cholangiocytes in polycystic liver disease have abnormal cell cycle profiles and dysfunctional cilia, Gradilone and co-workers investigated the role of HDAC6 in polycystic liver disease (PLD), which targets the epithelial layer of the biliary system. They found that the expression of HDAC6 protein was six-fold higher in cystic liver tissues and cultured cholangiocytes isolated from PCK rats (an animal model of PLD) and humans with PLD. As in our ADPKD study, inhibition of HDAC6 activity by HDAC6 inhibitors reduced the proliferation of cystic cholangiocytes in a dose- and time-dependent manner and inhibited cyst growth in three-dimensional (3D) culture.

[0004] Recent studies have reported that HDAC6 may play a role in cyst formation in ADPKD as well as PLD, and thus can be used as a potential therapeutic target. Summary of the Invention

[0005] On the one hand, the present invention relates to compounds of formula (I):

[0006]

[0007] or a pharmaceutically acceptable salt thereof,

[0008] wherein

[0009] R 1 is hydrogen, trifluoromethyl, (C 1-5 )alkyl, (C 3-5 )cycloalkyl, (C 1-6 )heterocycloalkyl, aralkyl, substituted phenyl or substituted heteroaryl;

[0010] R 2 is hydrogen, halogen, trifluoromethyl, cyano, (C 1-5 )alkyl, (C 1-6 )alkoxy, (C 1-6 )heterocycloalkyl, aralkyl, substituted phenyl or substituted heteroaryl;

[0011] R 3 is hydrogen, halogen, trifluoromethyl, cyano, (C 1-5 )alkyl, (C 1-6 )alkoxy, (C 1-6 )heterocycloalkyl, aralkyl, substituted phenyl or their substituted analogs;

[0012] R 4 is hydrogen, halogen, trifluoromethyl, cyano, (C 1-5 )alkyl, (C 1-6 )alkoxy, (C 1-6 )heterocycloalkyl, aralkyl, substituted phenyl or their substituted analogs;

[0013] R 5 is hydrogen, halogen, trifluoromethyl, cyano, (C 1-5 )alkyl, (C 1-6 )alkoxy, (C 1-6 )heterocycloalkyl, aralkyl, substituted phenyl or their substituted analogs;

[0014] R 6 is hydrogen, halogen or hydroxyalkoxy;

[0015] Partially selected from:

[0016] or a pharmaceutically acceptable salt, hydrate or prodrug thereof.

[0017] In one embodiment, R 1 is -CH3, -CH2CH3, -isopropyl, -cyclopropyl, -cyclopropylmethyl, -C6H5, -(4-Cl)C6H5, -(2-Cl)C6H5, -2,6-dimethylphenyl or -(4-OMe)C6H5.

[0018] In another embodiment, R 1 is -CH3 or -CH2CH3.

[0019] In another embodiment, R 3 is -Cl, -F or -CN.

[0020] In another embodiment, is part

[0021] In another embodiment, the compound is selected from:

[0022] 4-{[3-Methylquinazolin-4(3H)-one-2-yl]methyl}-N-hydroxybenzamide (6a);

[0023] 4-{[3-Cyclopropylquinazolin-4(3H)-one-2-yl]methyl}-N-hydroxybenzamide (6b);

[0024] 4-{[3-Cyclopropylmethylquinazolin-4(3H)-one-2-yl]methyl}-N-hydroxybenzamide (6c);

[0025] 4-{[3-Phenylquinazolin-4(3H)-one-2-yl]methyl}-N-hydroxybenzamide (6d);

[0026] 4-[(6-Chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N-hydroxybenzamide (6e);

[0027] 4-[(6-Chloro-3-cyclopropyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N-hydroxybenzamide (6f);

[0028] 4-{[3-(2-Chlorophenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]methyl}-N-hydroxybenzamide (6g);

[0029] 4-{[3-(4-Chlorophenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]methyl}-N-hydroxybenzamide (6h);

[0030] 4-{[3-(2,6-Dimethylphenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]methyl}-N-hydroxybenzamide (6i);

[0031] 4-[(6-Fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N-hydroxybenzamide (6j);

[0032] N-Hydroxy-4-((3-(4-methoxyphenyl)-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)benzamide (6k);

[0033] 4-[(3-Ethyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N-hydroxybenzamide (6l);

[0034] 4-[(6-Cyano-3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N-hydroxybenzamide (6m);

[0035] 4-((3,4-Dihydro-3-isopropyl-4-oxoquinazolin-2-yl)methyl)-N-hydroxybenzamide (6n);

[0036] 4-((3-Ethyl-6-fluoro-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-N-hydroxybenzamide (6o); and pharmaceutically acceptable salts thereof.

[0037] In another aspect of the present invention, the present invention relates to a pharmaceutical composition comprising:

[0038] (a) a therapeutically effective amount of a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof; and

[0039] (b) a pharmaceutically acceptable carrier or vehicle.

[0040] Furthermore, in another aspect of the present invention, the present invention relates to the use of a compound of the present invention or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present invention, in the preparation of a medicament for treating, alleviating, improving, and / or reducing the severity of an HDAC6-related disease or disorder in a subject in need thereof.

[0041] Alternatively, the present invention relates to a compound of the present invention or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present invention, for treating, alleviating, improving, and / or reducing the severity of an HDAC6-related disease or disorder in a subject in need thereof.

[0042] The present invention also relates to a method for treating, alleviating, improving, and / or reducing the severity of an HDAC6-related disease or disorder, which comprises: administering to a subject in need thereof a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of the present invention, thereby treating, alleviating, improving, and / or reducing the severity of an HDAC6-related disease or disorder in a subject in need thereof.

[0043] In one embodiment, the HDAC6-related disease or disorder is selected from fibrosis, neurodegenerative diseases, kidney diseases, cancer, and increased cyst growth markers.

[0044] In another embodiment, the fibrosis associated with HDAC6 activity is pulmonary fibrosis, idiopathic pulmonary fibrosis, liver fibrosis, renal fibrosis, or myelofibrosis.

[0045] In another embodiment, the neurodegenerative diseases associated with HDAC6 activity are Huntington's disease, Alzheimer's disease, Parkinson's disease, or amyotrophic lateral sclerosis.

[0046] In another embodiment, the kidney diseases associated with HDAC6 activity are polycystic kidney disease, or autosomal dominant polycystic kidney disease, or autosomal recessive polycystic kidney disease.

[0047] In another embodiment, the cancers associated with HDAC6 activity are lung cancer, breast cancer, kidney cancer, liver cancer, multiple myeloma, or glioma.

[0048] In another embodiment, the compound or its pharmaceutically acceptable salt, or the pharmaceutical composition is used for preparing a drug for reducing a cyst growth index associated with HDAC6 activity.

[0049] In another embodiment, the compound or its pharmaceutically acceptable salt, or the pharmaceutical composition is used for manufacturing a drug for treating, alleviating, improving, and / or reducing the severity of a condition with an increased cyst growth index, wherein the increased cyst growth index is associated with HDAC6 activity.

[0050] In another embodiment, the HDAC6-related diseases or conditions are selected from liver fibrosis, acute respiratory distress syndrome, acute pulmonary inflammation, pulmonary fibrosis, coronavirus-induced pulmonary inflammation, idiopathic pulmonary fibrosis, liver fibrosis, renal fibrosis, myelofibrosis, Huntington's disease, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, idiopathic pulmonary fibrosis (IPF), polycystic kidney disease, autosomal dominant polycystic kidney disease, lung cancer, breast cancer, liver cancer, and glioma. BRIEF DESCRIPTION OF THE DRAWINGS

[0051] Figure 1 It shows that pretreatment with compound 6a inhibits the growth of primary human ADPKD cysts in a concentration-dependent manner.

[0052] Figure 2A -D shows that compound 6a prevents cyst formation in human ADPKD cells.

[0053] Figure 3A -D shows that compound 6b prevents cyst formation in human ADPKD cells.

[0054] Figure 4A -D shows that compound 6a is superior to ACY1215 in terms of cytotoxicity and prevention of ADPKD cyst formation.

[0055] Figure 5A -D shows that compound 6a can rescue the cyst formation in human ADPKD cells.

[0056] Figure 6A -D shows that compound 6b can rescue the cyst index in human ADPKD cells.

[0057] Figure 7A -D shows that compound 6a confers efficacy in the PC1-KO mouse model.

[0058] Figure 8 It is shown that compound 6a reduced the kidney weight of transgenic Pkd1 knockdown mice.

[0059] Figure 9A -B shows that compound 6a reduced renal cysts in transgenic Pkd1 knockdown mice.

[0060] Figure 10 It is shown that compound 6a inhibits fibrosis and inflammatory biomarkers in the fibrosis assay panel. Detailed Description

[0061] Definitions

[0062] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present document, including definitions, will control.

[0063] As used herein, the term "alkyl" refers to a saturated, straight-chain or branched-chain hydrocarbon moiety. Exemplary alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, n-pentyl, and the like.

[0064] The term "C1-4 alkyl" refers to an alkyl group containing from 1 to 4 carbon atoms. When the term "alkyl" is used in combination with other substituents (e.g., "aralkyl"), the term "alkyl" is intended to encompass a divalent straight-chain or branched-chain hydrocarbon group. For example, "aralkyl" means a group -alkaryl, where its alkyl part is a divalent straight-chain or branched-chain carbon group and its aryl part is as defined herein and is represented by the bonding arrangement present in benzyl (-CH2-phenyl).

[0065] The term "cycloalkyl" refers to a non-aromatic saturated cyclic hydrocarbon ring. The term "C3-8 cycloalkyl" refers to a non-aromatic cyclic hydrocarbon ring having from three to eight ring carbon atoms. Exemplary "C3-8 cycloalkyl" groups useful in the present invention include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[0066] The term "alkoxy" refers to a group containing an alkyl group attached through an oxygen linking atom. The term "C1-4 alkoxy" refers to a straight-chain or branched-chain hydrocarbon group having from at least 1 and at most 4 carbon atoms attached through an oxygen linking atom. Exemplary "(C 1-4"Alkoxy" includes, but is not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, and sec-butoxy.

[0067] The term "aryl" denotes a group or moiety containing an aromatic monocyclic or bicyclic hydrocarbon group having 6 to 10 carbon ring atoms and may be fused to one or more cycloalkyl rings. Additionally, terms such as alkyl, aryl, cycloalkyl, heteroaryl, etc. can be used to define divalent substituents, e.g., a group bonded to two other groups. In such cases, these terms are intended to cover the divalent moieties. For example, "pentyl" is intended to denote pentylenediyl, where the pentyl moiety is any of the divalent straight-chain (e.g., -CH2CH2CH2CH2CH2-) or branched-chain (e.g., -CH2CH(CH3)CH2CH2-, -CH2CH2CH(CH2CH3)-, -CH2CH2C(CH3)2-) 5-carbon groups.

[0068] Generally, in the compounds of the present invention, heterocycloalkyl is 5- and / or 6-membered heterocycloalkyl, such as pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydrofuranyl, oxazolinyl, thiazolinyl, or pyrazolinyl, piperidinyl, piperazinyl, morpholinyl, tetrahydropyranyl, dihydropyranyl, 1,3-dioxolanyl, tetrahydro-2H-1,4-thiazinyl, 1,4-dioxanyl, 1,3-oxathianyl, and 1,3-dithianyl.

[0069] The term "heteroaryl" denotes a group or moiety containing an aromatic monocyclic ring having 5 to 10 ring atoms, including 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur. The term is also intended to cover heterocyclic groups containing nitrogen and / or sulfur, where the nitrogen or sulfur heteroatom is optionally oxidized. Exemplary instances of heteroaryl include, but are not limited to, thienyl, pyrrolyl, imidazolyl, pyrazolyl, furanyl, isothiazolyl, furazanyl, isoxazolyl, oxazolyl, oxadiazolyl, thiazolyl, pyridyl, pyridyl-N-oxide, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, tetrazinyl, triazolyl, and tetrazolyl.

[0070] Some heteroaryl groups of the compounds according to the present invention are 5-6 membered monocyclic heteroaryl. The selected 5-membered heteroaryl contains one nitrogen, oxygen, or sulfur ring heteroatom and optionally contains 1, 2, or 3 additional nitrogen ring atoms. The selected 6-membered heteroaryl contains 1, 2, 3, or 4 nitrogen ring heteroatoms. The selected 5- or 6-membered heteroaryl includes thienyl, pyrrolyl, imidazolyl, pyrazolyl, furanyl, isothiazolyl, furazanyl, isoxazolyl, oxazolyl, oxadiazolyl, thiazolyl, triazolyl, and tetrazolyl or pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, and thiadiazolyl.

[0071] The terms "halogen" and "halo" denote a chlorine, fluorine, bromine or iodine substituent.

[0072] When the disclosed compound or its salt is named or depicted by structure, it should be understood that the compound or its salt, including solvates (especially hydrates), may exist in crystalline form, amorphous form, or a mixture thereof. The compound or its salt or solvate (especially hydrate) may also exhibit polymorphism (i.e., the ability to occur in different crystalline forms). These different crystalline forms are generally referred to as "polymorphs". It should be understood that when named or depicted by structure, the disclosed compound or its solvate (especially hydrate) also includes all of its polymorphs. Polymorphs have the same chemical composition but differ in the packing mode, geometric arrangement, and other descriptive properties of the crystalline solid state. Thus, polymorphs can have different physical properties, such as shape, density, hardness, deformability, stability, and solubility properties. Polymorphs generally exhibit different melting points, IR spectra, and X-ray powder diffraction patterns, which can be used for identification. Those of ordinary skill in the art will understand that different polymorphs can be produced, for example, by varying or adjusting the conditions used to crystallize / recrystallize the compound. Due to their potential use in medicine, the salts of the compounds of formula (I) are preferably pharmaceutically acceptable salts. Suitable pharmaceutically acceptable salts include those described by Berge, Bighley and Monkhouse, J. Pharm. Sci (1977) 66, pp 1-19. The salts included in the term "pharmaceutically acceptable salts" refer to non-toxic salts of the compounds of the present invention. Generally, the salts can be readily prepared by appropriately using the desired acid or base. The salts can be precipitated from solution and collected by filtration, or can be recovered by evaporation of the solvent. When the compound of the present invention is a base (containing a basic moiety), the desired salt form can be prepared by any suitable method known in the art, including treatment of the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc., or with an organic acid, such as acetic acid, trifluoroacetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, etc., or with a pyranoside acid, such as glucuronic acid or galacturonic acid, or with an α-hydroxy acid, such as citric acid or tartaric acid, or with an amino acid, such as aspartic acid or glutamic acid, or with an aromatic acid, such as benzoic acid or cinnamic acid, or with a sulfonic acid, such as p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, etc.Suitable addition salts are formed from acids which form non-toxic salts and examples include acetate, p-aminobenzoate, ascorbate, aspartate, benzenesulfonate, benzoate, bicarbonate, bis(methylene)salicylate, bisulfate, bitartrate, borate, calcium edetate, camphorsulfonate, carbonate, clavulanate, citrate, cyclohexylsulfamate, edetate, edisylate, etidronate, esylate, ethanedisulfonate, ethanesulfonate, formate, fumarate, glucoheptonate, gluconate, glutamate, glycolate, glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, dihydrochloride, hydrofumarate, hydrogenphosphate, hydroiodide, hydrogenmaleate, hydrogensuccinate, hydroxynaphthoate, isethionate, itaconate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, monopotassium maleate, mucate, naphthalenesulfonate, nitrate, N-methylglucamine, oxalate, oxaloacetate, pamoate (embonate), palmate (palmitate), pantothenate, phosphate / diphosphate, pyruvate, polygalacturonate, propionate, saccharate, salicylate, stearate, subacetate, succinate, sulfate, tannate, tartrate, teoclate, toluenesulfonate, triethiodide, trifluoroacetate and valerate.

[0073] Other exemplary acid addition salts include pyrosulfate, sulfite, bisulfite, caprate, caprylate, acrylate, isobutyrate, caproate, heptanoate, propiolate, oxalate, malonate, suberate, sebacate, butyne-1,4-dioate, hexyne-1,6-dioate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, phenylacetate, phenylpropionate, phenylbutyrate, lactate, γ-hydroxybutyrate, mandelate and sulfonates such as xylenesulfonate, propanesulfonate, naphthalene-1-sulfonate and naphthalene-2-sulfonate. If the basic compound of the invention is isolated as a salt, the corresponding free base form of the compound can be prepared by any suitable method known in the art, including treatment of the salt with an inorganic or organic base, suitably an inorganic or organic base having a higher pKa than the free base form of the compound.

[0074] When the compounds of the present invention are acids (containing acidic moieties), the desired salts can be prepared by any suitable method known in the art, including treating the free acid with an inorganic or organic base, such as an amine (primary, secondary or tertiary amine), an alkali metal or alkaline earth metal hydroxide, etc. Exemplary examples of suitable salts include organic salts derived from amino acids (such as glycine and arginine), ammonia, primary amines, secondary amines and tertiary amines and cyclic amines, such as N-methyl-D-glucamine, diethylamine, isopropylamine, trimethylamine, ethylenediamine, dicyclohexylamine, ethanolamine, piperidine, morpholine and piperazine, and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium. Certain compounds of the present invention can form salts with one equivalent or more equivalents of an acid (if the compound contains a basic moiety) or a base (if the compound contains an acidic moiety). The present invention includes within its scope all possible stoichiometric and non-stoichiometric salt forms.

[0075] Compounds of the present invention having both basic and acidic moieties can be in the form of zwitterions, acid addition salts of the basic moiety or base salts of the acidic moiety. The present invention also provides for the conversion of one pharmaceutically acceptable salt of a compound of the present invention, such as a hydrochloride salt, to another pharmaceutically acceptable salt of the compound of the present invention, such as a sulfate salt. For crystalline forms of the compounds of formula (I) or solvates of their salts, those skilled in the art will understand that pharmaceutically acceptable solvates can be formed, in which solvent molecules are incorporated into the crystal lattice during crystallization. Solvates can involve non-aqueous solvents, such as ethanol, isopropanol, DMSO, acetic acid, ethanolamine and ethyl acetate, or they can involve water as the solvent incorporated into the crystal lattice. Solvates in which water is the solvent incorporated into the crystal lattice are generally referred to as "hydrates". Hydrates include stoichiometric hydrates as well as compositions containing variable amounts of water. The present invention includes all such solvates. Since the compounds of formula (I) are intended for use in pharmaceutical compositions, it is readily understood that they are each preferably provided in a substantially pure form, for example at least 60% pure, more suitably at least 75% pure, and preferably at least 85% pure, especially at least 98% pure (% by weight). Impure preparations of the compounds can be used to prepare the more pure forms used in pharmaceutical compositions.

[0076] The term “(C m -C n )” or “Cm-n”, where m, n are integers and n>m, refers to all integer unit amounts in the range from m to n specifically disclosed as part of the present invention. Thus, “(C m -C n )” means including C m , C m+1 , C m+2 ,..., C n-2 , C n-1 , C n , (C m-C m+1 )、(C m -C m+2 )、(C m -C m+3 ),...,(C m -C n-2 )、(C m -C n-1 ),(C m -C n );(C m+1 -C m+2 )、(C m+1 -C m+3 )、(C m+1 -C m+4 ),...,(C m+1 -C n-2 )、(C m+1 -C n-1 )、(C m+1 -C n ),...,(C n-2 -C n-1 ),(C n-2 -C n );and (C n-1 -C n ) are used as embodiments of the present invention.

[0077] “(C1-C6)” or “C1-6” means all integer unit amounts within the range of 1 to 6 are specifically disclosed as part of the present invention. Thus, it includes C1, C2, C3, C4, C5, C6; (C1-C2), (C1-C3), (C1-C4), (C1-C5), (C1-C6); (C2-C3), (C2-C4), (C2-C5), (C2-C6); (C3-C4), (C3-C5), (C3-C6); (C4-C5), (C4-C6); and (C5-C6) unit amounts are used as embodiments of the present invention.

[0078] The term “treating” or “treatment” means administering an effective amount of a therapeutic agent to a subject having a disease or a symptom or predisposition to such a disease, with the aim of curing, alleviating, slowing down, remedying, improving, reducing the severity, or preventing the disease, its symptoms, or the predisposition thereto.

[0079] The terms “HDAC6-related disease or disorder”, “HDAC6-mediated disease or disorder”, and “disease or disorder associated with HDAC6” are interchangeable.

[0080] The terms "HDAC6-related disease or disorder", "HDAC6-mediated disease or disorder", or "disease or disorder associated with HDAC6" refer to a disease or disorder that can be treated, alleviated, improved, or have its severity reduced by inhibiting HDAC6 activity and / or a disease or disorder in which inhibiting HDAC6 can provide a benefit.

[0081] The "Guidance for Industry and Reviewers Estimating the Safe Starting Dose in Clinical Trials for Therapeutics in Adult Healthy Volunteers" issued by the Food and Drug Administration of the United States Department of Health and Human Services discloses that the "human equivalent dose" can be calculated by the following formula:

[0082] HED = animal dose (mg / kg) × (animal weight (kg) / human weight (kg)) 0.33 .

[0083] The compounds of the present invention can be obtained by using the synthetic procedures described in the following schemes or by utilizing the knowledge of a skilled organic chemist. The syntheses provided in these schemes are applicable to preparing the compounds of the present invention having a variety of different substituents using appropriate precursors, which, if necessary, are appropriately protected to achieve compatibility with the reactions outlined herein. Subsequent deprotection, if required, provides compounds of the nature generally disclosed. Although only compounds of formula (I) are shown in the schemes, they are exemplary methods that can be used to prepare the compounds of the present invention. Intermediates (compounds used to prepare the compounds of the present invention) may also exist as salts. Thus, with respect to intermediates, the phrase "compound of formula (number)" refers to a compound having that structural formula or a pharmaceutically acceptable salt thereof.

[0084]

[0085] General synthetic procedure for compounds 6a - 6m. Reagents and conditions: (a) substituted amine, DIPEA, HOBt, EDC·HCl, DCM, 0 °C to rt for 16 h; (b) (i) oxalyl chloride, DMF, DCM, 0 °C to rt for 2 h; (ii) aniline, TEA, DCM, 0 °C to rt for 3 h; (c) iron powder, NH4Cl, EtOH:H2O (8:2), 90 °C for 16 h; (d) 2-[4-(methoxycarbonyl)phenyl]acetic acid, DIPEA, HOBt, EDC·HCl, DCM, 0 °C to rt for 16 h; (e) ZnCl2, hexamethyldisilazane, DMF, 120 °C for 24 h; (f) 2 M NH2OH in MeOH solution, NaOH, rt for 1 h; (g) (i) NaCN, NiBr2, NMP MW 120 W, 200 °C for 10 min; (ii) NH2OBn·HCl, EDCI, HOBt, DIPEA, DMF, rt for 1 h; (iii) DCM solution containing 1 M BBr3 dimethyl sulfide complex, DCM, 0 °C for 30 min.

[0086] This series of compounds has 2 - substituted quinazolin - 4 - one as the core. This is readily synthesized from the corresponding diamides (key intermediate II, 3a - 3l). The diamide 3 is prepared from commercially available o - nitrobenzoic acid 1 via two different routes (a four - step reaction sequence involving acyl chloride and acid - amide coupling reactions), as shown in Scheme 1.

[0087] Commercially available nitrobenzoic acid 1 is treated with a DCM solution of oxalyl chloride to give the corresponding acyl chloride, which is then treated with aniline to give nitroamide 2. Alternatively, nitrobenzoic acid 1 is coupled with the corresponding amine in the presence of the coupling agents EDC·HCl and HOBt to give nitroamide 2. The nitro group of 2 is reduced in the presence of Fe - AcOH to give substituted anthranilamide 3. These aminoamides 3a - 3l react with 2-[4-(methoxycarbonyl)phenyl]acetic acid in the presence of EDC·HCl and HOBt to give intermediate I, 4a - 4l. The intermediate 4 treated with hexamethyldisilazane and zinc chloride undergoes smooth cyclization to give 2 - substituted quinazolin - 4-(3H)-one 5a - 5l. These 2 - substituted quinazolin - 4-(3H)-one intermediates 5 react with freshly prepared NH2OH in methanol in the presence of NaOH to give the final products 6a - 6l. They are purified using a combination of column chromatography and reverse - phase HPLC to give the target isohydroxamic acid compounds 6a - 6l as solids.

[0088] For the synthesis of compound 6m, a nitrile group was inserted from the chloride intermediate 5e using sodium cyanide and nickel bromide by microwave irradiation to obtain a nitrile intermediate. Surprisingly, it was found that half of the nitrile intermediate carried a carboxylic acid instead of the original ester group. Therefore, the carboxylic acid group reacted with NH2OBn under the action of a coupling agent and was then treated with BBr3 to obtain the hydroxamic acid compound 6m.

[0089] Example

[0090] Pharmacological research

[0091] Compounds 6a - p inhibit HDAC6.

[0092] We first evaluated a series of quinazoline derivatives against HDACs and found several compounds with effective and selective inhibitory activity against HDAC6 in the single - digit nM range. Table 1 shows the HDAC inhibitory activity of compounds 6a - 6o. 50 In the digital nM range of the compounds with effective and selective inhibitory activity against HDAC6. Table 1 shows the HDAC inhibitory activity of compounds 6a - 6o.

[0093] Table 1

[0094]

[0095] Compound 6a inhibits the growth of primary human ADPKD cysts.

[0096] We next verified our initial screening results and performed in vitro 3D primary human ADPKD cell assays. The anti - ADPKD cyst growth efficacy of compound 6a and compound 6b was evaluated using prevention and reduction assays:

[0097] Prevention (pre - treatment) assay: Cells were seeded on day 0 and treated on day 1. Additional treatments were added for 3 - 4 days throughout the study. The study was terminated based on the cyst size of any + stimulation group. End - point whole - well images were captured on the last day of the study, and cell counting was performed after that day ( Figure 1 , 2 and 3).

[0098] Reduction (rescue) assay: Drug treatment and precise experimental timing were based on cyst growth, but cells were seeded on day 0. After cysts began to form (around days 4 - 7), the compound was added. The study was terminated based on the cyst size of any stimulation group. Whole - well images were captured before drug administration and again before the end of the experiment. The cell number was evaluated at the end of the experiment. (Figures 5 and 6)

[0099] In the Cyst Reduction Assay, using DMSO as a control, screening of two therapeutic drugs (Compound 6a and Compound 6b) was performed on ADPKD cells with PKD1 mutations from 1 donor stimulated with forskolin, and was carried out together with the control drug (Ricolinostat). Human ADPKD cells were pre-treated with 6 serial dilutions of Compound 6a and Compound 6b, n = 8. The control drug Ricolinostat was run at a single point (3 μM) ( Figure 4D ).

[0100] Endpoint:

[0101] The main measures of treatment efficacy were the measurement of the number of cysts per well, cyst size, cyst cell proliferation (measured by CellTiter Glo), and cytotoxicity (measured by LDH release). Cyst number and size measurements were determined by post-acquisition image analysis.

[0102] Results:

[0103] At the end of this DBM prevention assay, cyst cell proliferation (CTG), cyst number (CN), total cyst area (TA), and mean size / cyst were measured. Overall, pre-treatment with Compound 6a and Compound 6b could prevent the growth of cyst cell proliferation, cyst number, and cyst size in the primary human 3D ADPKD cell model ( Figure 1 ).

[0104] Both Compound 6a and Compound 6b showed cytotoxicity at 90 μM, while the control drug (ACY1215) showed cytotoxicity levels at 3 μM and 10 μM. Compound 6a tended to have cytotoxicity above 30 μM and could refer to its similar cytotoxicity profile in the reduction assay (see NTU SOW 5). Among all the compounds tested, Compound 6a was the most effective in reducing the cyst formation index.

[0105] Our data from individual human ADPKD donor cyst cultures also demonstrated a positive correlation between HDAC6 inhibition and reduced cyst growth in all treatments (Table 1; Figures 2, 3, and 4), suggesting that HDAC6 inhibition may be a potential target for treating ADPKD. In addition, this effect was specific to ADPKD cells, as we did not observe cytotoxicity in non-ADPKD renal cells treated with Compound 6a, while ACY1215 was significantly present (Figure 4).

[0106] Figure 2A-D shows the effect of pretreatment with compound 6a on cyst indices in primary human ADPKD cells. (A) Cyst cell proliferation (measured by CTG) and cytotoxicity (measured by LDH release) (B) Number of cysts (C) Total cyst area (D) Average size / cyst are plotted in a 6-point concentration-response curve, where one-way ANOVA was used to determine the significant difference between the DMSO control group and the drug-treated group, followed by Dunnet's multiple comparison test (*p<0.05).

[0107] Figure 3A -D shows the effect of pretreatment with compound 6b on cyst indices in primary human ADPKD cells. (A) Cyst cell proliferation (measured by CTG) and cytotoxicity (measured by LDH release) (B) Number of cysts (CN) (C) Total cyst area (TA) (D) Average size / cyst are plotted in a 6-point concentration-response curve, where one-way ANOVA was used to determine the significant difference between the DMSO control group and the drug-treated group, followed by Dunnet's multiple comparison test (*p<0.05).

[0108] Figure 4A -D shows that compound 6a is superior to ACY1215 in terms of cytotoxicity and prevention of ADPKD cyst formation.

[0109] Figure 5A -D shows the reduction assay: the effect of compound 6a on cyst indices. (A) Cyst cell proliferation (measured by CTG) and cytotoxicity (measured by LDH release) (B) Number of cysts (CN) (C) Total cyst area (TA) (D) Average size / cyst are plotted in a 6-point concentration-response curve, where one-way ANOVA was used to determine the significant difference between the DMSO control group and the drug-treated group, followed by Dunnet's multiple comparison test (*p<0.05).

[0110] Figure 6A -D shows the reduction assay: the effect of compound 6b on cyst indices. (A) Cyst cell proliferation (measured by CTG) and cytotoxicity (measured by LDH release) (B) Number of cysts (CN) (C) Total cyst area (TA) (D) Average size / cyst are plotted in a 6-point concentration-response curve, where one-way ANOVA was used to determine the significant difference between the DMSO control group and the drug-treated group, followed by Dunnet's multiple comparison test (*p<0.05).

[0111] At the end of the DBM reduction assay, CTG, LDH, the number of cysts, and cyst size were measured. (Figures 5 and 6) All four of these benchmark metrics together constitute the standard cyst formation metrics, which are used to measure and compare the effects of compound 6a (Figure 5) and compound 6b (Figure 6) in inhibiting cyst growth and reducing the cyst formation characteristics of human ADPKD cells in vitro. Among all the tested compounds, compound 6a was the most effective in reducing the cyst formation metrics.

[0112] Compound 6a confers efficacy in a PKD mouse model.

[0113] The efficacy of compound 6a was further evaluated in a Pkd1 knockdown mouse model. A transgenic mouse line with a ~60 - 70% reduction in Pkd1 expression develops severe renal cystic disease at a rate similar to that of human ADPKD. (Figure 7) These results further support the haploinsufficiency hypothesis and suggest that the occurrence and development of renal cystic disease are related to the Pkd1 expression level. Therefore, we attempted to conduct a randomized, blinded, and statistically significant efficacy study in Pkd1 knockdown mice to determine whether treatment with compound 6a produces similar beneficial effects. From postnatal day P14 to P28, Pkd1 knockdown mice were orally administered PBS, 60 mg·kg -1 , 120 mg·kg -1 , and 150 mg·kg -1 of compound 6a daily. The mice were sacrificed at P28 and the kidneys were harvested. Most importantly, treatment of the animals with compound 6a led to a decrease in the kidney-to-body weight ratio (KW / BW) (Figure 9) and the cyst index (total cyst area / total kidney area) ( Figure 8 ) compared to the vehicle control group.

[0114] Figure 7A -D shows that compound 6a confers efficacy in a PC1-KO mouse model (A) Animal No. 86, cyst index: 49.9%; 50.5% (vehicle group) (B) Animal No. 84, cyst index: 30.2%; 23.6% (60 mg / Kg) (C) Animal No. 87, cyst index: 21.5%; 18.8% (120 mg / Kg) (D) Animal No. 83, cyst index: 26.5%; 11.4% (150 mg / Kg).

[0115] Figure 8 It shows that compound 6a significantly reduced the cyst index in transgenic Pkd1 knockdown mice.

[0116] Figure 9A -B shows that compound 6a reduced renal cysts in transgenic Pkd1 knockdown mice.

[0117] Compound 6a preferentially distributes to the kidneys and liver.

[0118] We next evaluated the in vivo pharmacokinetics and biodistribution profiles of compound 6a in wild-type mice following a single 30 mg kg -1 oral administration. Compound 6a was rapidly absorbed into the plasma, showing a tmax ≤ 0.5 h and a Cmax of 264 μg mL -1 , with a half-life of < 4 h. The rapid plasma clearance of compound 6a reflected extensive distribution to tissues such as the kidney and liver. Uniquely, compound 6a showed preferential renal distribution, with a kidney-to-plasma (K / P) ratio of approximately 2 / 1 based on AUClast.

[0119] Compound 6a inhibited inflammatory and fibrotic biomarkers in the fibrosis assay panel.

[0120] Compound 6a was characterized in the Eurofins fibrosis assay panel in three human primary cell-based systems, including the MyoF system, the REMyoF system, and the SAEMyoF system. These systems were designed to mimic complex human tissue and disease biology, which drives abnormal inflammation involved in fibrosis and wound healing. Renal fibrosis disease associated with advanced renal failure was captured in the REMyoF system, which consists of a co-culture of renal proximal tubular epithelial cells and adult fibroblasts.

[0121] Figure 10 The profiles of compound 6a in the fibrosis assay panel were shown.

[0122] The results showed that at the concentrations tested in this study, compound 6a was active (with 15 annotated readings) and non-cytotoxic. Compound 6a affected inflammation-related activities (decreased I-TAC, sIL-6, and MCP-1), myofibroblast activation-related activities (increased α-SMA), fibrosis-related matrix activities (decreased type I collagen, type III collagen; increased type IV collagen; regulated MMP-1), and tissue remodeling / wound healing activities (decreased tPA, uPA, and sVEGF).

[0123] ​​Four common activities were annotated in the REMyoF system: type I collagen, I-TAC, sIL-6, and sVEGF. Four distinct activities (not shown) were present in the following systems: SAEMyoF(αSMA) and MyoF(IL-8, TIMP-1, MMP-1). In the study, compound 6a was characterized by analysis in a human primary cell-based fibrosis assay combination that mimicked the fibrotic and inflammatory processes driving fibrosis in a tissue-specific context. The fibrosis assay combination evaluated the biological impact of the test drug under conditions that retained the complex crosstalk and feedback mechanisms relevant to in vivo results.

[0124] Compound 6a was non-cytotoxic and active, with 15 annotated readings, 9 in the renal fibrosis (REMyoF) system model, 3 in the MyoF system, and 3 in the SAEMyoF system. The annotated biomarker activities affected inflammation, myofibroblast activation, fibrosis-related matrix, and tissue remodeling / wound healing activities. When 10 μM of compound 6a was superimposed with 1.1 μM of the selected reference benchmark nintedanib, four common activities and four distinct activities were identified.

[0125] The early fibrosis screening service (SAEMyoF) is A high-throughput screening format in the fibrosis system SAEMyoF, consisting of small airway epithelial cells and lung fibroblasts. This system was stimulated with profibrotic and proinflammatory factors to mimic the biology of fibrotic lung diseases such as idiopathic pulmonary fibrosis. Biomarker readings captured the effects on matrix remodeling, tissue repair, and inflammation-related responses relevant to translation in diseased lungs. This service allowed for the rapid testing of the anti-fibrotic development potential of 18 compounds at 4 concentrations in a human fibrosis model.

[0126] Chemical synthesis

[0127] Example 1: Preparation of 4-{[3-cyclopropylquinazolin-4(3H)-one-2-yl]methyl}-N-hydroxybenzamide (Compound 6b)

[0128] Step 1: Preparation of the intermediate N-cyclopropyl-2-nitrobenzamide

[0129]

[0130] At 0 °C, DIPEA (13 mL, 89.92 mmol, 3 equiv), EDC·HCl (8.58 g, 44.78 mmol, 1.5 equiv), HOBt (4.04 g, 29.94 mmol, 1 equiv) were added to a solution of 2-nitrobenzoic acid (5 g, 29.94 mmol, 1 equiv) in DCM (50 mL), and then cyclopropylamine (2.56 mL, 35.93 mmol, 1.2 equiv) was added. The reaction mixture was warmed to room temperature and stirred for 16 h. Then, the reaction mixture was poured into ice water and extracted with DCM (100 mL x 2). The organic layer was washed with water (100 mL) and brine (100 mL). After drying over Na2SO4, the solvent was filtered off and removed in vacuo to give a crude residue. The crude compound was purified by column chromatography (eluting with 0 to 20% EtOAc in petroleum ether), and the desired fraction (R f = 0.5, EtOAc / petroleum ether = 3:7) was collected to give the title compound as an off-white solid. (5 g, 81% yield)

[0131] Step 2: Preparation of Intermediate 2-Amino-N-cyclopropylbenzamide

[0132]

[0133] Iron powder (5.43 g, 96.96 mmol, 4 equiv) and NH4Cl (5.4 g, 96.96 mmol, 4 equiv) were added to a solution of N-cyclopropyl-2-nitrobenzamide (5 g, 24.27 mmol, 1 equiv) in EtOH:H2O (8:2), and the reaction mixture was stirred at 90 °C for 16 h. The solvent was removed in vacuo, extracted with EtOAc (100 mL x 2), and washed with water (100 mL). After drying over anhydrous Na2SO4, the solvent was filtered and concentrated under reduced pressure to give the title compound as an off-white solid. (3.7 g, 90% yield) R f = 0.4 (EtOAc / petroleum ether = 3:7);

[0134] Step 3: Preparation of Intermediate Methyl 4-(2-{[2-(Cyclopropylcarbamoyl)phenyl]amino}-2-oxoethyl)benzoate

[0135]

[0136] At 0 °C, DIPEA (9.06 mL, 54.12 mmol, 3 equiv), EDC·HCl (5.18 g, 27.06 mmol, 1.5 equiv), and HOBt (2.43 g, 18.04 mmol, 1 equiv) were added to a solution of 2-[4-(methoxycarbonyl)phenyl]acetic acid (3.5 g, 18.04 mmol, 1 equiv) in DCM (35 mL). Then, N-cyclopropyl-2-nitrobenzamide (3.17 g, 18.04 mmol, 1 equiv) was added, and the reaction mixture was warmed to room temperature and stirred for 16 h. Then, the reaction mixture was poured into ice water and extracted with DCM (50 mL x 2). The organic layer was washed with water (50 mL), dried over Na2SO4, and concentrated under reduced pressure to give a crude residue. The crude compound was purified by column chromatography (eluting with 0 to 35% EtOAc in petroleum ether), and the desired fraction (R f = 0.4, EtOAc / petroleum ether = 1:4) was collected to give the title compound as an off-white solid. (3.8 g, 60% yield)

[0137] Step 4: Preparation of Intermediate Methyl 4-[(3-cyclopropyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]benzoate

[0138]

[0139] To a solution of methyl 4-(2-{[2-(cyclopropylcarbamoyl)phenyl]amino}-2-oxoethyl)benzoate (3 g, 8.52 mmol, 1 equiv) in DMF (10 mL) was added ZnCl2 (0.58 g, 4.26 mmol, 0.5 equiv), followed by hexamethyldisilazane (5.34 mL, 25.56 mmol, 3 equiv). The reaction mixture was stirred at 120 °C for 24 h. Then, the reaction mixture was poured into ice water and extracted with EtOAc (50 mL x 2). The organic layer was washed with water (100 mL), dried over Na2SO4, and concentrated under reduced pressure to give a crude residue. The crude compound was purified by column chromatography (eluting with 0 to 50% EtOAc in petroleum ether), and the desired fraction (R f = 0.5, EtOAc / petroleum ether = 1:1) was collected to give the title compound as an off-white solid. (1.7 g, 60% yield)

[0140] Step 5: Preparation of 4-{[3-Cyclopropylquinazolin-4(3H)-one-2-yl]methyl}-N-hydroxybenzamide (Compound 6b)

[0141]

[0142] At room temperature, a solution of 2 M NH2OH in anhydrous MeOH (20 ml) and NaOH (0.26 g, 6.49 mmol) were added to the ester intermediate (1 g, 3.24 mmol), and the reaction mixture was stirred for 1 h. The reaction mixture was filtered through a Celite pad and washed with MeOH (10 ml). The filtrate was concentrated under reduced pressure to give a crude residue. The crude product was purified by column chromatography (silica gel 100 - 200 mesh, eluted with 5% - 20% MeOH in DCM), and the desired fraction (R f = 0.3, MeOH / DCM = 1 / 9) was collected to give the target compound as a brown solid. (0.5 g, 50% yield) mp = 200 - 202 °C; 1 1H NMR (600 MHz, DMSO - d6) δ 11.18 (brs, 1H), 9.05 (brs, 1H), 8.05 (d, J = 8.4 Hz, 1H), 7.74 (m, 1H), 7.70 (d, J = 7.8 Hz, 2H), 7.53 (d, J = 7.8 Hz, 1H), 7.47 - 7.44 (m, 1H), 7.38 (d, J = 7.8 Hz, 2H), 4.44 (s, 2H), 2.69 (m, 1H), 1.18 (m, 2H), 0.91 (m, 2H); 13 13C NMR (150 MHz, DMSO - d6) δ 164.5, 162.8, 158.4, 147.0, 140.2, 134.6, 131.7, 129.5, 127.6, 127.1, 127.0, 126.5, 121.3, 41.3, 27.7, 10.8; LC - MS: Mol Wt: 335.13, [M + H] + : 336.24; HR - ESI MS m / z [M + H] + Calculated 336.1343 Found 336.1332; HPLC purity = 96.1% (t R = 2.6 min, eluted by mobile phase: B: aqueous solution of 0.05% TFA, A: acetonitrile solution of 0.05% TFA; gradient (T / %A): 0 / 3, 8.5 / 100, 9.0 / 100, 9.5 / 3, 10 / 3; column temperature: 50 °C, flow rate: 0.55 ml / min; diluent: ACN:H2O, Acquity BEH C18 100 mm x 2.1 mm, 1.7 μm).

[0143] Example 2: Preparation of 4-{[3 - methylquinazolin - 4(3H) - one - 2 - yl]methyl}-N - hydroxybenzamide (Compound 6a)

[0144] Steps 1 to 4 (the same as Steps 1 to 4 of Example 1):Preparation of methyl 4-[(3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]benzoate

[0145]

[0146] The title compound was prepared by a method similar to Steps 1 to 4 of Example 1 above. Yellow solid (2.12 g, 56.7% yield). R f = 0.35 (EtOAc / heptane = 3 / 2); mp = 142 - 144 °C; 1 1H NMR (600 MHz, DMSO-d6) δ = 8.11 (dd, J = 7.8, 0.6 Hz, 1H), 7.92 (d, J = 7.8 Hz, 2H), 7.78 (m, 1H), 7.58 (d, J = 7.8 Hz, 1H), 7.49 (m, 1H), 7.43 (d, J = 8.4 Hz, 2H), 4.38 (s, 2H), 3.83 (s, 3H), 3.45 (s, 3H); 13 13C NMR (150 MHz, DMSO-d6) δ 166.0, 161.5, 155.7, 146.8, 141.4, 134.2, 129.5, 129.2, 128.2, 126.8, 126.6, 126.1, 119.8, 52.0, 41.2, 30.4; ESIMS(+) m / z 309.0 [M+H] + 。

[0147] Step 5: Preparation of 4-{[3-methylquinazolin-4(3H)-one-2-yl]methyl}-N-hydroxybenzamide (Compound 6a)

[0148]

[0149] The title compound was prepared by a method similar to Step 5 of Example 1 above. Brown solid (3.555 g, 65.4% yield). mp = 228 - 230 °C; 1 1H NMR (600 MHz, DMSO-d6) δ 11.19 (s, 1H), 9.02 (s, 1H), 8.11 (dd, J = 8.4, 1.2 Hz, 1H), 7.79 - 7.76 (m, 1H), 7.72 (d, J = 8.4 Hz, 2H), 7.59 (d, J = 7.8 Hz, 1H), 7.51 - 7.48 (m, 1H), 7.36 (d, J = 7.8 Hz, 2H), 4.33 (s, 2H), 3.45 (s, 3H); 1313C NMR (150 MHz, DMSO-d6) δ 164.0, 161.5, 155.9, 146.8, 139.0, 134.2, 131.4, 128.7, 127.3, 126.8, 126.6, 126.2, 119.8, 41.1, 30.5; LC-MS: Mol Wt: 309.11, [M+H] + : 310.11; HR-ESIMS m / z [M+H] + Calcd 310.1186 Found 310.1177; HPLC purity = 94.7% (t R = 2.3 min, eluted with mobile phase: B: aqueous solution of 0.05% TFA, A: acetonitrile solution of 0.05% TFA; gradient (T / %A): 0 / 3, 8.5 / 100, 9.0 / 100, 9.5 / 3, 10 / 3; column temperature: 50 °C, flow rate: 0.55 ml / min; diluent: ACN:H2O, Acquity BEH C18 100 mm x 2.1 mm, 1.7 μm).

[0150] Example 3: Preparation of 4-{[3-(cyclopropylmethyl)quinazolin-4(3H)-one-2-yl]methyl}-N-hydroxybenzamide (Compound 6c)

[0151]

[0152] The title compound was prepared by a method similar to Steps 1 to 5 of Example 1 above. Solid. 1 1H NMR (600 MHz, DMSO-d6) δ 11.20 (brs, 1H), 9.05 (brs, 1H), 8.12 (d, J = 7.8 Hz, 1H), 7.81 - 7.78 (m, 1H), 7.71 (d, J = 7.8 Hz, 2H), 7.59 (d, J = 8.4 Hz, 1H), 7.52 - 7.50 (m, 1H), 7.37 (d, J = 7.8 Hz, 2H), 4.37 (s, 2H), 3.95 (d, J = 7.2 Hz, 2H), 1.7 (m, 1H), 0.44 - 0.38 (m, 4H); 13 13C NMR (150 MHz, DMSO-d6) δ 161.7, 155.4, 146.8, 134.4, 128.7, 127.2, 126.9, 126.7, 126.3, 120.1, 47.0, 40.6, 10.5, 3.8; LC-MS: Mol Wt: 349.14, [M+H] + : 350.31; HR-ESIMS m / z [M+H] +Calculated value: 350.1499, Measured value: 350.1490; HPLC purity = 96% (t R = 3.2 min, eluted with mobile phase: B: aqueous solution of 0.05% TFA, A: acetonitrile solution of 0.05% TFA; gradient (T / %A): 0 / 3, 8.5 / 100, 9.0 / 100, 9.5 / 3, 10 / 3; column temperature: 50 °C, flow rate: 0.55 ml / min; diluent: ACN:H2O, Acquity BEH C18 100 mm x 2.1 mm, 1.7 μm).

[0153] Example 4: Preparation of 4-{[3-phenylquinazolin-4(3H)-one-2-yl]methyl}-N-hydroxybenzamide (Compound 6d)

[0154] Step 1: Preparation of 2-nitro-N-phenylbenzamide

[0155]

[0156] Cool a solution of 2-nitrobenzoic acid (12 g, 1 equivalent) in DCM (120 ml) to 0 - 5 °C, and add oxalyl chloride (16.4 g, 1.8 equivalents) and one drop of DMF dropwise thereto. Stir the reaction mixture and warm it to room temperature for 2 hours. After indicating the completion of the reaction by tlc, evaporate the solvent under vacuum and use it without further purification. Dissolve aniline (8.85 g, 1.2 equivalents) and triethylamine (9.65 g, 1.2 equivalents) in DCM (70 ml) and cool to 0 - 5 °C, and add a solution of the crude acyl chloride (14.70 g, 1 equivalent) in DCM (70 ml) to this solution. Warm the reaction mixture to room temperature and stir for 3 hours. Evaporate the reaction mixture in vacuo and redissolve it in DCM (50 ml). Wash the organic layer with saturated NaHCO 3(水溶液) (25 ml) and water (25 ml × 2). After drying over Na2SO4, concentrate the organic layer under reduced pressure and purify it by column chromatography. Collect the desired fraction (R f = 0.4, EtOAc / hexane = 3 / 7) to obtain the target compound. (15.65 g, 90% yield)

[0157] Steps 2 to 4: Same as Example 1.

[0158] Step 5: Preparation of 4-{[3-phenylquinazolin-4(3H)-one-2-yl]methyl}-N-hydroxybenzamide (Compound 6d)

[0159]

[0160] The title compound was prepared by a method similar to Steps 1 to 5 of Example 1 above. Pale pink solid. mp = 167 - 169 °C; 1 H NMR (600 MHz, DMSO-d6) δ 11.15 (s, 1H), 8.98 (s, 1H), 8.11 (dd, J = 7.8 Hz, 1H), 7.86 - 7.83 (m, 1H), 7.67 (d, J = 8.4 Hz, 1H), 7.58 (d, J = 7.8 Hz, 2H), 7.55 - 7.53 (m, 1H), 7.45 - 7.44 (m, 3H), 7.24 (d, J = 7.2 Hz, 2H), 7.00 (d, J = 7.8 Hz, 2H), 3.85 (s, 2H); 13 C NMR (150 MHz, DMSO-d6) δ 163.9, 161.5, 155.2, 147.1, 139.0, 137.0, 134.7, 131.0, 129.2, 128.9, 128.8, 128.7, 127.0, 126.9, 126.7, 126.3, 120.6, 41.6; LC-MS: Mol Wt: 371.39, [M+H] + : 372.14; HR-ESIMS m / z [M+H] + Calculated value 372.1343, Measured value 372.1336; LC-MS = 98.6%.

[0161] Example 5: Preparation of 4-[(6-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N-hydroxybenzamide (Compound 6e)

[0162] Steps 1 to 4: Preparation of methyl 4-[(6-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]benzoate

[0163]

[0164] The title compound was prepared by a method similar to Steps 1 to 4 of Example 1 above. White solid. (460 mg, 49% yield) (R f = 0.35, EtOAc / heptane = 1:1); mp = 173 - 175 °C; 11H NMR (600 MHz, DMSO-d6) δ 8.02 (d, J = 2.4 Hz, 1H), 7.92 (d, J = 8.4 Hz, 2H), 7.78 (dd, J = 9.0, 2.4 Hz, 1H), 7.58 (d, J = 9.0 Hz, 1H), 7.43 (d, J = 8.4 Hz, 2H), 4.37 (s, 2H), 3.83 (s, 3H), 3.45 (s, 3H); 13 13C NMR (150 MHz, DMSO-d6) δ 166.0, 160.5, 156.4, 145.5, 141.2, 134.3, 130.7, 129.5, 129.3, 129.1, 128.2, 125.1, 121.0, 52.1, 41.1, 30.6; ESIMS (+) m / z 343 [M+H] + 。

[0165] Step 5: Preparation of 4-[(6-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N-hydroxybenzamide (Compound 6e)

[0166]

[0167] The title compound was prepared by a method similar to Step 5 of Example 1 above. White solid. (184 mg, 44% yield) R f = 0.31 (MeOH / DCM = 1 / 9); mp = 214 - 216 °C; 1 1H NMR (600 MHz, DMSO-d6) δ 11.19 (s, 1H), 9.01 (s, 1H), 8.01 (d, J = 2.4 Hz, 1H), 7.78 (dd, J = 8.4, 2.4 Hz, 1H), 7.72 (d, J = 8.4 Hz, 2H), 7.60 (d, J = 9.0 Hz, 1H), 7.36 (d, J = 8.4 Hz, 2H), 4.33 (s, 2H), 3.45 (s, 3H); 13 13C NMR (150 MHz, DMSO-d6) δ 164.0, 160.6, 156.6, 145.5, 138.8, 134.3, 131.4, 130.7, 129.1, 128.8, 127.2, 125.1, 121.0, 41.0, 30.6; HR-ESIMS m / z [M+H] + Calculated value 344.0796 Measured value 344.0787; HPLC purity = 95.5% (t R = 8.2 min, eluted with ACN solution of 0.1% FA / H2O solution of 0.1% FA = 10 / 90 to 100 / 0, 3.5μm XB-C18 LC column (100x4.6mm).

[0168] Example 6: Preparation of 4-[(6-chloro-3-cyclopropyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N-hydroxybenzamide (Compound 6f)

[0169] Steps 1 to 4: Preparation of methyl 4-[(6-chloro-3-cyclopropyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]benzoate

[0170] The title compound was prepared by a method similar to Steps 1 to 4 of Example 1 above. White solid. (1.26 g, 61% yield) (R f = 0.4, EtOAc / heptane = 1:1); mp = 161 - 163 °C; 1 1H NMR (600 MHz, DMSO-d6) δ 7.99 (d, J = 2.4 Hz, 1H), 7.91 (d, J = 7.8 Hz, 2H), 7.76 (dd, J = 9.0, 2.4 Hz, 1H), 7.53 (d, J = 9.0 Hz, 1H), 7.46 (d, J = 7.8 Hz, 2H), 4.48 (s, 2H), 3.83 (s, 3H), 2.74 (m, 1H), 1.17 (m, 2H), 0.92 (m, 2H); 13 13C NMR (150 MHz, DMSO-d6) δ 166.1, 161.3, 158.4, 145.2, 142.0, 134.2, 130.6, 129.6, 129.3, 129.0, 128.0, 125.0, 122.0, 52.1, 40.9, 27.3, 10.2; ESIMS(+) m / z 369 [M+H] + .

[0171] Step 5: Preparation of 4-[(6-chloro-3-cyclopropyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N-hydroxybenzamide (Compound 6f)

[0172]

[0173] The title compound was prepared by a method similar to Step 5 of Example 1 above. White solid. (90 mg, 11% yield) (R f = 0.2, MeOH / DCM = 1 / 9); mp = 240 - 243 °C (dec.); 11H NMR (600 MHz, DMSO-d6) δ 11.18 (s, 1H), 9.01 (s, 1H), 7.98 (s, 1H), 7.76 (d, J = 8.4 Hz, 1H), 7.70 (d, J = 7.2 Hz, 2H), 7.55 (d, J = 7.8 Hz, 1H), 7.38 (d, J = 7.2 Hz, 2H), 4.44 (s, 2H), 2.71 (s, 1H), 1.19 (s, 1H), 0.92 (s, 1H); 13 13C NMR (150 MHz, DMSO-d6) δ 164.1, 161.3, 158.6, 145.2, 139.5, 134.2, 131.2, 130.6, 129.1, 128.9, 127.1, 125.0, 122.0, 40.8, 27.3, 10.2; HR-ESI MS m / z [M + H] + Calculated value 370.0953, found value 370.0942; HPLC purity = 95.6% (t R = 8.6 min, eluted with ACN solution of 0.1% FA / H2O solution of 0.1% FA = 10 / 90 to 100 / 0, 3.5 μm XB-C18 LC column 100 x 4.6 mm).

[0174] Example 7: Preparation of 4-{[3-(4-chlorophenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]methyl}-N-hydroxybenzamide (Compound 6h)

[0175] Steps 1 to 4: Preparation of methyl 4-{[3-(4-chlorophenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]methyl}benzoate

[0176]

[0177] The title compound was prepared by a method similar to Steps 1 to 4 of Example 1 above. Brown solid. (1.34 g, 63% yield) (R f = 0.37, EtOAc / heptane = 1:1); mp = 169 - 171 °C; 11H NMR (600 MHz, DMSO-d6) δ 8.11 (d, J = 7.8 Hz, 1H), 7.84 (m, 1H), 7.79 (d, J = 7.2 Hz, 2H), 7.67 (d, J = 8.4 Hz, 1H), 7.55 (m, 1H), 7.48 (d, J = 7.8 Hz, 2H), 7.28 (d, J = 8.4 Hz, 2H), 7.10 (d, J = 7.8 Hz, 2H), 3.91 (s, 2H), 3.82 (s, 3H); 13 13C NMR (150 MHz, DMSO-d6) δ 166.1, 161.5, 154.6, 147.0, 141.2, 135.8, 134.8, 133.7, 130.8, 129.2, 128.4, 128.0, 127.1, 127.0, 126.4, 120.6, 52.0, 41.8; ESIMS (+) m / z 405 [M+H] + 。

[0178] Step 5: Preparation of 4-{[3-(4-chlorophenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]methyl}-N-hydroxybenzamide (Compound 6h)

[0179]

[0180] The title compound was prepared by a method similar to Step 5 of Example 1 above. Orange solid. (200 mg, 25% yield) (R f = 0.14, MeOH / DCM = 5 / 95); mp = 152 - 155 °C; 1 1H NMR (600 MHz, DMSO-d6) δ 11.16 (s, 1H), 8.98 (s, 1H), 8.11 (d, J = 7.8 Hz, 1H), 7.85 (m, 1H), 7.67 (d, J = 7.8 Hz, 1H), 7.59 (d, J = 8.4 Hz, 2H), 7.54 (m, 1H), 7.50 (d, J = 8.4 Hz, 2H), 7.29 (d, J = 8.4 Hz, 2H), 7.03 (d, J = 7.8 Hz, 2H), 3.87 (s, 2H); 13 13C NMR (150 MHz, DMSO-d6) δ 163.9, 161.5, 154.9, 147.0, 138.9, 135.9, 134.7, 133.6, 131.0, 130.8, 129.2, 128.7, 127.1, 127.0, 126.8, 126.3, 120.5, 54.9, 41.6; HR-ESIMS m / z [M+H] +Calculated value 406.0953, measured value 406.0938; HPLC purity = 97.4% (t R = 8.8 min, eluted with ACN solution of 0.1% FA / H2O solution of 0.1% FA = 10 / 90 to 100 / 0, 3.5 μm XB-C18 LC column 100 x 4.6 mm).

[0181] Example 8: Preparation of 4-{[3-(2-chlorophenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]methyl}-N-hydroxybenzamide (Compound 6g)

[0182] Steps 1 to 4: Preparation of methyl 4-{[3-(2-chlorophenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]methyl}benzoate

[0183]

[0184] The title compound was prepared by a method similar to Steps 1 to 4 of Example 1 above. Yellow solid. (1.46 g, 58% yield) (R f = 0.22, EtOAc / heptane = 1:1); mp = 167 - 169 °C; 1 1H NMR (600 MHz, DMSO-d6) δ 8.13 (d, J = 7.8 Hz, 1H), 7.89 (m, 1H), 7.77 (d, J = 7.8 Hz, 2H), 7.72 (d, J = 7.8 Hz, 1H), 7.57 (m, 3H), 7.51 (m, 2H), 7.04 (d, J = 7.8 Hz, 2H), 3.88 (dd, J = 19.8, 15.6, 2H), 3.82 (s, 3H); 13 13C NMR (150 MHz, DMSO-d6) δ 166.0, 160.6, 154.3, 147.0, 140.4, 135.1, 134.2, 132.0, 131.2, 131.0, 130.0, 129.1, 129.0, 128.3, 128.1, 127.24, 127.22, 126.4, 120.2, 52.0, 41.6; ESIMS(+) m / z 405 [M+H] + .

[0185] Step 5: Preparation of 4-{[3-(2-chlorophenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]methyl}-N-hydroxybenzamide (Compound 6g)

[0186]

[0187] The title compound was prepared by a method similar to Step 5 of Example 1 above. Orange solid. (297 mg, 37% yield) (R f = 0.08, MeOH / DCM = 5 / 95); mp = 137 - 140 °C; 1 1H NMR (600 MHz, DMSO-d6) δ 11.17 (s, 1H), 8.99 (s, 1H), 8.13 (d, J = 7.2 Hz, 1H), 7.87 (m, 1H), 7.71 (m, 1H), 7.62 - 7.46 (m, 7H), 6.98 (d, J = 6.6 Hz, 2H), 3.82 (dd, J = 25.2, 15.0 Hz, 2H); 13 13C NMR (150 MHz, DMSO-d6) δ 163.9, 160.7, 154.6, 147.0, 138.1, 135.1, 134.3, 132.0, 131.2, 131.1, 130.1, 128.8, 128.4, 127.2, 126.8, 126.5, 120.2, 48.6, 41.3; HR-ESIMSm / z [M+H] + Calculated 406.0953 Found 406.0944; HPLC purity = 96.6% (t R = 8.7 min, eluted with ACN solution of 0.1% FA / H2O solution of 0.1% FA = 10 / 90 to 100 / 0, 3.5 μm XB-C18 LC column 100x4.6 mm).

[0188] Example 9: Preparation of 4-{[3-(2,6-dimethylphenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]methyl}-N-hydroxybenzamide (Compound 6i)

[0189] Steps 1 to 4: Preparation of methyl 4-{[3-(2,6-dimethylphenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]methyl}benzoate

[0190]

[0191] The title compound was prepared by a method similar to Steps 1 to 4 of Example 1 above. Yellow solid. (1.3 g, 68% yield) (R f = 0.53, EtOAc / heptane = 1:1); mp = 120 - 123 °C; 11H NMR (600 MHz, DMSO-d6) δ 8.14 (dd, J = 7.8, 1.2 Hz, 1H), 7.89 (m, 1H), 7.79 (d, J = 7.8 Hz, 2H), 7.75 (d, J = 7.8 Hz, 1H), 7.56 (m, 1H), 7.35 (m, 1H), 7.20 (d, J = 7.2 Hz, 2H), 7.01 (8.4 Hz, 2H), 3.81 (s, 3H), 3.75 (s, 2H), 1.71 (s, 6H); 13 13C NMR (150 MHz, DMSO-d6) δ 165.9, 160.3, 154.6, 147.1, 140.2, 135.4, 135.0, 134.9, 129.4, 129.3, 129.0, 128.7, 128.3, 127.3, 127.2, 126.5, 120.2, 52.0, 41.2, 17.0; ESIMS (+) m / z 399.0 [M+H] + 。

[0192] Step 5: Preparation of 4-{[3-(2,6-Dimethylphenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]methyl}-N-hydroxybenzamide (Compound 6i)

[0193]

[0194] The title compound was prepared by a method similar to Step 5 of Example 1 above. Yellow solid. (335 mg, 42% yield) (R f = 0.09, MeOH / DCM = 5 / 95); mp = 133 - 136 °C; 1 1H NMR (600 MHz, DMSO-d6) δ 11.16 (s, 1H), 8.97 (s, 1H), 8.14 (d, J = 7.8 Hz, 1H), 7.89 (m, 1H), 7.75 (d, J = 7.8 Hz, 1H), 7.59 (d, J = 8.4 Hz, 2H), 7.56 (m, 1H), 7.35 (m, 1H), 7.21 (7.8 Hz, 2H), 6.93 (d, J = 8.4 Hz, 2H), 3.71 (s, 2H), 1.71 (s, 6H); 13 13C NMR (150 MHz, DMSO-d6) δ 163.8, 160.3, 154.9, 147.2, 137.8, 135.5, 135.0, 131.4, 129.3, 129.0, 128.7, 127.2, 127.1, 126.7, 126.5, 120.1, 54.9, 41.0, 17.0; HR-ESIMS m / z [M+H]+ Calculated value: 400.1656, Measured value: 400.1645; HPLC purity = 99% (t R = 8.9 min, eluted with ACN solution of 0.1% FA / H2O solution of 0.1% FA from 10 / 90 to 100 / 0 3.5μm XB-C18 LC column 100x4.6mm).

[0195] Example 10: Preparation of 4-[(6-Fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N-hydroxybenzamide (Compound 6j)

[0196] Steps 1 to 4: Preparation of methyl 4-[(6-fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]benzoate

[0197]

[0198] The title compound was prepared by a method similar to Steps 1 to 4 of Example 1 above. Yellow solid. (1.55 g, 82% yield) (R f = 0.35, EtOAc / heptane = 1:1); mp = 155 - 157 °C; 1 1H NMR (600 MHz, DMSO-d6) δ 7.92 (d, J = 8.4 Hz, 2H), 7.77 (m, 1H), 7.66 (m, 2H), 7.43 (d, J = 8.4 Hz, 2H), 4.37 (s, 2H), 3.83 (s, 3H), 3.45 (s, 3H); 13 13C NMR (150 MHz, DMSO-d6) δ 166.0, 160.9, 160.7, 159.1, 155.3, 143.7, 141.3, 129.8, 129.7, 129.5, 129.2, 128.2, 122.9, 122.7, 121.0, 121.0, 110.7, 110.6, 52.1, 41.0, 30.6; ESIMS(+) m / z 327.1 [M+H] + .

[0199] Step 5: Preparation of 4-[(6-Fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N-hydroxybenzamide (Compound 6j)

[0200]

[0201] The title compound was prepared by a method similar to Step 5 of Example 1 above. White solid. (280 mg, 35% yield) (R f = 0.07, MeOH / DCM = 5 / 95); mp = 226 - 228 °C; 1 1H NMR (600 MHz, DMSO-d6) δ 11.18 (s, 1H), 9.01 (s, 1H), 7.77 (d, J = 8.4 Hz, 1H), 7.71 (d, J = 8.4 Hz, 2H), 7.67 (m, 2H), 7.36 (d, J = 8.4 Hz, 2H), 4.33 (s, 2H), 3.45 (s, 3H); 13 13C NMR (150 MHz, DMSO-d6) δ 164.1, 161.0, 160.8, 159.2, 155.5, 143.8, 139.0, 131.4, 129.8, 128.8, 127.3, 123.0, 122.8, 121.1, 121.0, 110.8, 110.7, 41.0, 30.7; HR-ESI MS m / z [M+H] + Calculated value 328.1092, found 328.1086; HPLC purity = 98% (t R = 7.3 min, eluted by ACN solution of 0.1% FA / H2O solution of 0.1% FA from 10 / 90 to 100 / 0, 3.5 μm XB-C18 LC column 100 x 4.6 mm).

[0202] Example 11: Preparation of N-Hydroxy-4-((3-(4-methoxyphenyl)-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)benzamide (Compound 6k)

[0203] Steps 1 to 4: Preparation of Methyl 4-{[3-(4-methoxyphenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]methyl}benzoate

[0204]

[0205] The title compound was prepared by a method similar to Steps 1 to 4 of Example 1 above. White solid. (1.26 g, 64% yield) (R f = 0.32, EtOAc / heptane = 1:1; mp = 160 - 162 °C; 11H NMR (600 MHz, DMSO-d6) δ 8.11 (d, J = 7.8 Hz, 1H), 7.83 (m, 1H), 7.79 (d, J = 7.8 Hz, 2H), 7.65 (d, J = 7.8 Hz, 1H), 7.53 (m, 1H), 7.12 (d, J = 8.4 Hz, 2H), 7.09 (d, J = 8.4 Hz, 2H), 7.82 (d, J = 8.4 Hz, 2H), 3.91 (s, 2H), 3.82 (s, 3H), 3.79 (s, 3H); 13 13C NMR (150 MHz, DMSO-d6) δ 166.1, 161.7, 159.3, 155.5, 147.0, 141.5, 134.6, 129.9, 129.4, 129.1, 127.9, 127.0, 126.8, 126.4, 120.6, 114.4, 55.4, 52.0, 41.8; ESIMS (+) m / z 401.0 [M+H] + 。

[0206] Step 5: Preparation of N-Hydroxy-4-{[3-(4-methoxyphenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]methyl}benzamide (Compound 6k)

[0207]

[0208] The title compound was prepared by a method similar to Step 5 of Example 1 above. Orange solid. (152 mg, 19% yield) (R f = 0.11, MeOH / DCM = 5 / 95); mp = 214 - 215 °C; 1 1H NMR (600 MHz, DMSO-d6) δ 11.15 (s, 1H), 8.97 (s, 1H), 8.10 (dd, J = 7.8, 1.2 Hz, 1H), 7.83 (m, 1H), 7.65 (d, J = 7.8 Hz, 1H), 7.59 (d, J = 8.4 Hz, 2H), 7.53 (m, 1H), 7.14 (d, J = 9.0 Hz, 2H), 7.03 (d, J = 8.4 Hz, 2H), 6.97 (d, J = 9.0 Hz, 2H), 3.86 (s, 2H), 3.80 (s, 3H); 1313C NMR (150 MHz, DMSO-d6) δ 164.0, 161.7, 159.3, 155.7, 147.0, 139.1, 134.6, 131.0, 129.9, 129.5, 128.68, 128.67, 127.0, 126.7, 126.4, 120.6, 114.4, 55.4, 41.5; HR-ESIMS m / z [M+H] + Calcd for 402.1448, found 402.1436; HPLC purity = 99% (t R = 8.2 min, eluted with ACN solution of 0.1% FA / H2O solution of 0.1% FA = 10 / 90 to 100 / 0 3.5 μm XB-C18 LC column 100 x 4.6 mm).

[0209] Example 12: Preparation of 4-[(6-cyano-3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N-hydroxybenzamide (Compound 6m)

[0210]

[0211] To a mixture of the chloro-substituted ester intermediate (703 mg, 2.05 mmol), NaCN (200 mg, 4.08 mmol) and NiBr2 (448 mg, 2.05 mmol) was added NMP (6 ml), and the reaction mixture was irradiated with microwave for 5 minutes. The mixture was partitioned between EtOAc / H2O (50 ml / 30 ml) and washed with water (30 ml × 2). The organic phase was dried over MgSO4 and concentrated in vacuo. The crude product was purified by column chromatography using EtOAc / heptane as the eluent, and the required fraction ((R f= 0.09, EtOAc / heptane = 1 / 1), an acid intermediate was obtained. The acid intermediate (210 mg, 0.658 mmol), EDCI (420 mg, 2.19 mmol) and HOBt (154 mg, 1.01 mmol) were dissolved in DMF (5 ml), and the mixture was stirred at ambient temperature. After 15 minutes, NH2OBn.HCl (327 mg, 2.05 mmol) and DIPEA (0.36 ml, 2.06 mmol) were added to the mixture, and the reaction mixture was stirred for another 2 h. Then the mixture was poured into water (100 ml), and the resulting precipitate was filtered to obtain an orange solid (214 mg, 77% yield). To a suspension of the solid in anhydrous DCM (10 ml) was added BBr3 dimethyl sulfide complex (1 M DCM solution, 1 ml) at 0 °C, and the reaction mixture was stirred for 30 minutes. The mixture was quenched with water (10 ml) and washed with water (20 ml × 2). The H2O phase was extracted with DCM (20 ml x 3). All the organic phases were combined, dried over MgSO4 and concentrated in vacuo. The crude product was purified by column chromatography using DCM / MeOH as the eluent, and the desired fraction (R f = 0.08, MeOH / DCM = 5 / 95) was collected to give the title compound as a pink solid (24 mg, 43%); 1 1H NMR (600 MHz, DMSO-d6) δ 11.19 (s, 1H), 9.02 (s, 1H), 8.50 (s, 1H), 8.12 (d, J = 8.4 Hz, 1H), 7.71 (m, 3H), 7.37 (d, J = 7.8 Hz, 2H), 4.36 (s, 2H), 3.47 (s, 3H); 13 13C NMR (150 MHz, DMSO-d6) δ 164.0, 160.5, 159.3, 149.4, 138.5, 136.4, 131.9, 131.4, 128.9, 128.3, 127.3, 120.3, 118.2, 108.7, 41.2, 30.8; HR-ESIMS m / z [M+H] + Calculated 335.1139 Found 335.1129; HPLC purity = 95% (t R = 7.1 min, eluted by ACN solution of 0.1% FA / H2O solution of 0.1% FA = 10 / 90 to 100 / 0, 3.5 μm XB-C18 LC column 100 x 4.6 mm).

[0212] Scheme 2 Synthesis of 6n

[0213]

[0214] Example 13: Preparation of 4-((3,4-dihydro-3-isopropyl-4-oxoquinazolin-2-yl)methyl)-N-hydroxybenzamide (6n)

[0215] A solution of anthranilic acid (3.0 g, 21.87 mmol), 2-(4-bromophenyl)acetic acid (4.7 g, 21.87 mmol) and triphenyl phosphite (6.9 mL, 26.24 mmol) was dissolved in pyridine (20 mL) and irradiated with microwave at 250 W for 20 minutes. Prop-2-amine hydrochloride (2.39 g, 30.62 mmol) was added and irradiated with microwave at 250 W for 15 minutes. The reaction mixture was concentrated in vacuo and Pd(OAc)2 (0.21 g, 0.9 mmol), Xantphos (0.54 g, 0.9 mmol), Mo(CO)6 (4.6 g, 17.42 mmol), DMAP (3.14 g, 25.7 mmol) and DIPEA (4 mL, 22.49 mmol) dissolved in DMAc / MeOH = 1:1 (20 mL) which had been degassed was added. The resulting 7 without separation was stirred at 100 °C for 16 hours. After completion of the reaction, it was diluted with EA and water, filtered through diatomaceous earth to obtain a solution. The organic layer was dried over MgSO4, filtered and concentrated in vacuo. The residue was purified by column (DCM solution of 1% EA to DCM solution of 10% EA) to obtain white solid 5n (2.4 g). A solution of 5n (2.4 g, 7.13 mmol) was dissolved in a solution of 2M NH2OH in MeOH (40 mL). The suspension was stirred for 16 hours and the reaction mixture became a clear solution. After completion of the reaction, it was concentrated in vacuo. The residue was purified by column (DCM solution of 10% MeOH), and R f = 0.3 was collected to obtain a pink solid 6n (1.15 g, 16% over four steps).

[0216] 1 H NMR (600 MHz, DMSO-d6) δ 11.21 (s, 1H), 9.01 (s, 1H), 8.09 (d, J = 7.92 Hz, 1H), 7.80 (t, J = 7.68 Hz, 1H), 7.74 (d, J = 7.86 Hz, 2H), 7.64 (d, J = 8.1 Hz, 1H), 7.5 (t, J = 7.56 Hz, 1H), 7.37 (d, J = 7.92 Hz, 2H), 4.51 (s, 1H), 4.39 (s, 2H), 1.30 (d, J = 6.6 Hz, 6H); 13CNMR (150 MHz, d6-dmso) δ 19.49, 42.48, 52.06, 122.04, 126.36, 127.05, 127.13, 127.8255, 128.79, 131.84, 134.67, 139.96, 147.04, 156.08, 162.27, 164.37; HPLC purity = 97.2% (t R = 8.07 min)

[0217] Synthesis of 6l in Scheme 3

[0218]

[0219] Methyl 4-((3-ethyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)benzoate (5l)

[0220]

[0221] Dissolve a solution of anthranilic acid (3.0 g, 21.87 mmol), 2-(4-bromophenyl)acetic acid (4.7 g, 21.87 mmol) and triphenyl phosphite (6.9 mL, 26.24 mmol) in pyridine (20 mL) and irradiate under 250 W microwave for 20 minutes. Add ethylamine hydrochloride (2.49 g, 30.62 mmol) and irradiate under 250 W microwave for 15 min. After the reaction is completed, dilute with DCM and quench with 3% HCl. Dry the organic layer over MgSO4, filter and concentrate in vacuo to obtain a colorless oil, then wash with EA / heptane to obtain white solid 8 (3.14 g, 53%); Add Pd(OAc)2 (79 mg, 0.35 mmol), Xantphos (0.41 g, 0.7 mmol), Mo(CO)6 (1.74 g, 6.57 mmol), DMAP (1.78 g, 9.64 mmol) and DIPEA (1.53 mL, 8.77 mmol) dissolved in DMAc / MeOH = 1:1 (15 mL) which has been degassed to the solution of 8 (1.5 g, 4.38 mmol). Stir the reaction mixture under reflux for 16 hours. After the reaction is completed, dilute with EA and water, filter through diatomaceous earth to obtain a solution. Dry the organic layer over MgSO4, filter and concentrate in vacuo. Purify the residue by column (DCM), collect R f = 0.3 to obtain white solid 5l (1.02 g, 71%).

[0222] 11H NMR (600 MHz, DMSO-d6) δ 8.28 (d, J = 8.1 Hz, H), 8.01 (d, J = 8.1 Hz, 2H), 7.76 (t, J = 7.92 Hz, 1H), 7.70 (d, J = 8.16 Hz, 1H), 7.48 (t, J = 7.8 Hz, 1H), 7.36 (d, J = 8.04 Hz, 2H), 4.31 (s, 2H), 4.04 (q, J = 7.02 Hz, 2H), 3.91 (s, 3H), 1.21 (t, J = 7.14 Hz, 3H)

[0223] Example 14: Preparation of 4-((3-Ethyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-N-hydroxybenzamide (6l)

[0224]

[0225] Dissolve a solution of 5l (1.6 g, 0.5 mmol) in MeOH (30 mL) of 2M NH2OH. Stir the suspension mixture for 16 h, and the mixture becomes a clear solution. After completion of the reaction, concentrate the mixture in vacuo. Purify the residue by column (DCM solution of 10% MeOH), collect R f = 0.32 to obtain a pink solid 61 (0.73 g, 45%).

[0226] 1 1H NMR (600 MHz, DMSO-d6) δ 11.20 (s, 1H), 9.02 (s, 1H), 8.12 (d, J = 7.98 Hz, 1H), 7.80 (t, J = 7.38 Hz, 1H), 7.73 (d, J = 8.22 Hz, 2H), 7.61 (d, J = 8.04 Hz, 1H), 7.51 (t, J = 7.86 Hz, 1H), 7.40 (d, J = 8.28 Hz, 2H), 4.34 (s, 2H), 4.02 (q, J = 7.02 Hz, 2H), 1.09 (d, J = 6.96 Hz, 3H); 13 13C NMR (150 MHz, d6-dmso) δ 13.99, 41.19, 120.61, 126.59, 127.11, 127.36, 127.75, 129.14, 131.89, 134.81, 139.89, 147.31, 155.84, 161.62, 164.46; HPLC purity = 97.7%

[0227] Scheme 4 Synthesis of 6o

[0228]

[0229] 2-(4-Bromobenzyl)-3-ethyl-6-fluoroquinazolin-4-one (9)

[0230]

[0231] A solution of 2-amino-5-fluorobenzoic acid (2.0 g, 12.89 mmol), 2-(4-bromophenyl)acetic acid (2.77 g, 12.89 mmol) and triphenyl phosphite (4 mL, 15.47 mmol) was dissolved in pyridine (20 mL) and refluxed for 15 h. Ethylamine hydrochloride (1.4 g, 18.05 mmol) was added and then refluxed for 15 h. After completion of the reaction, it was diluted with DCM and quenched with 3% HCl.

[0232] The organic layer was dried over MgSO4, filtered and concentrated in vacuo to give a colorless oil, which was then washed with EA / heptane to give white solid 9 (3.48 g, 75%).

[0233] 1 H NMR (600 MHz, DMSO-d6) δ 7.77 (t, J = 8.88 Hz, 1H), 7.68 (d, J = 5.7 Hz, 2H), 7.53 (d, J = 8.1 Hz, 2H), 7.29 (d, J = 7.92 Hz, 2H), 4.27 (s, 2H), 4.03 (m, 2H), 1.09 (t, J = 6.84 Hz, 3H)

[0234] Methyl 4-((3-ethyl-6-fluoro-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)benzoate (5o)

[0235]

[0236] To a solution of 9 (2.5 g, 6.94 mmol) was added Pd(OAc)2 (0.125 g, 0.56 mmol), Xantphos (0.64 g, 1.11 mmol), Mo(CO)6 (2.75 g, 10.42 mmol), DMAP (1.87 g, 15.28 mmol) and DIPEA (2.42 mL, 13.89 mmol) dissolved in DMAc / MeOH = 1:1 (20 mL) which had been degassed. The reaction mixture was stirred under reflux for 24 h. After completion of the reaction, it was diluted with EA and water, filtered through celite to give a solution. The organic layer was dried over MgSO4, filtered and concentrated in vacuo. The residue was purified by column (1% EA in DCM solution to 10% EA in DCM solution), and then the fraction with R f = 0.39 was collected to give white solid 5o (0.93 g, 40%).

[0237] 1 1H NMR (600 MHz, DMSO-d6) δ 7.92 (d, J = 7.98 Hz, 2H), 7.77 (d, J = 8.34 Hz, 2H), 7.66 (d, J = 6.6 Hz, 2H), 7.47 (d, J = 7.98 Hz, 2H), 4.43 (s, 2H), 4.01 (q, J = 6.9 Hz, 2H), 3.83 (s, 3H), 1.07 (t, J = 6.96 Hz, 3H)

[0238] Example 15: Preparation of 4-((3-Ethyl-6-fluoro-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-N-hydroxybenzamide (6o)

[0239]

[0240] A solution of 5o (0.45 g, 0.13 mmol) was dissolved in a solution of 2 M NH2OH in MeOH (10 mL). The suspension was stirred for 16 h and the reaction mixture became a clear solution. After completion of the reaction, the mixture was concentrated in vacuo. The residue was purified by column (from a solution of 10% MeOH in DCM to a solution of 20% MeOH in DCM), and then the fraction with R f = 0.3 was collected to give a pink solid 6o (0.25 g, 55%). 1 1H NMR (600 MHz, DMSO-d6) δ 11.19 (s, 1H), 9.01 (s, 1H), 7.79 (d, J = 7.92 Hz, 1H), 7.72 (d, J = 8.16 Hz, 2H), 7.39 (d, J = 8.16 Hz, 2H), 4.34 (s, 2H), 4.04 (q, J = 6.96 Hz, 2H), 1.09 (t, J = 7.02 Hz, 3H); HPLC purity = 97.6% (t R = 6.6 min).

Claims

1. Compound of formula (I): or a pharmaceutically acceptable salt thereof, wherein R 1 is hydrogen, trifluoromethyl, (C 1-5 )alkyl, (C 3-5 )cycloalkyl, (C 1-6 )heterocycloalkyl, aralkyl, substituted phenyl or substituted heteroaryl; R 2 is hydrogen, halogen, trifluoromethyl, cyano, (C 1-5 )alkyl, (C 1-6 )alkoxy, (C 1-6 )heterocycloalkyl, aralkyl, substituted phenyl or substituted heteroaryl; R 3 is hydrogen, halogen, trifluoromethyl, cyano, (C 1-5 )alkyl, (C 1-6 )alkoxy, (C 1-6 )heterocycloalkyl, aralkyl, substituted phenyl or their substituted analogues; R 4 is hydrogen, halogen, trifluoromethyl, cyano, (C 1-5 )alkyl, (C 1-6 )alkoxy, (C 1-6 )heterocycloalkyl, aralkyl, substituted phenyl or their substituted analogues; R 5 is hydrogen, halogen, trifluoromethyl, cyano, (C 1-5 )alkyl, (C 1-6 )alkoxy, (C 1-6 )heterocycloalkyl, aralkyl, substituted phenyl or their substituted analogs; R 6 is hydrogen, a halogen or a hydroxyalkoxy group; Partially selected from: or a pharmaceutically acceptable salt, hydrate or prodrug thereof.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein, R 1 is -CH3, -CH2CH3, -isopropyl, -cyclopropyl, -cyclopropylmethyl, -C6H5, -(4-Cl)C6H5, -(2-Cl)C6H5, -2,6-dimethylphenyl or -(4-OMe)C6H5.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein, R 1 is -CH3 or -CH2CH3.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein, R 3 is -Cl, -F or -CN.

5. The compound according to any one of claims 1-4 or a pharmaceutically acceptable salt thereof, wherein, Part is 6. The compound according to any one of claims 1 to 5, wherein the compound is selected from: and pharmaceutically acceptable salts thereof.

7. A pharmaceutical composition comprising: (a) a therapeutically effective amount of the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof; and (b) a pharmaceutically acceptable carrier or vehicle.

8. Use of the compound according to any one of claims 1 - 6 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 7 in the manufacture of a medicament for treating, alleviating, improving and / or reducing the severity of an HDAC6 - related disease or disorder in a subject in need thereof.

9. The use according to claim 8, wherein the HDAC6 - related disease or disorder is selected from fibrosis, neurodegenerative diseases, kidney diseases, cancer and increased cyst growth markers.

10. The use according to claim 9, wherein the fibrosis related to HDAC6 activity is pulmonary fibrosis, idiopathic pulmonary fibrosis, liver fibrosis, renal fibrosis or myelofibrosis.

11. The use according to claim 9, wherein the neurodegenerative disease related to HDAC6 activity is Huntington's disease, Alzheimer's disease, Parkinson's disease or amyotrophic lateral sclerosis.

12. The use according to claim 9, wherein the kidney disease related to HDAC6 activity is polycystic kidney disease, or autosomal dominant polycystic kidney disease, or autosomal recessive polycystic kidney disease.

13. The use according to claim 9, wherein the cancer related to HDAC6 activity is lung cancer, breast cancer, kidney cancer, liver cancer, multiple myeloma or glioma.

14. The use according to claim 8, wherein the compound or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition is for the manufacture of a medicament for reducing a cyst growth marker related to HDAC6 activity.

15. The use according to claim 9, wherein the compound or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition is for the manufacture of a medicament for treating, alleviating, improving and / or reducing the severity of a disorder with increased cyst growth markers related to HDAC6 activity.

16. The use according to claim 8, wherein the HDAC6 - related disease or disorder is selected from liver fibrosis, acute respiratory distress syndrome, acute pulmonary inflammation, pulmonary fibrosis, coronavirus - induced pulmonary inflammation, idiopathic pulmonary fibrosis, liver fibrosis, renal fibrosis, myelofibrosis, Huntington's disease, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, idiopathic pulmonary fibrosis (IPF), polycystic kidney disease, autosomal dominant polycystic kidney disease, lung cancer, breast cancer, liver cancer and glioma.

17. The compound according to claim 1, which is selected from: 4 - {[3 - methylquinazolin - 4(3H) - one - 2 - yl]methyl}-N - hydroxybenzamide (6a); 4-{[3-Cyclopropylquinazolin-4(3H)-one-2-yl]methyl}-N-hydroxybenzamide (6b); 4-{[3-(Cyclopropylmethyl)quinazolin-4(3H)-one-2-yl]methyl}-N-hydroxybenzamide (6c); 4-{[3-Phenylquinazolin-4(3H)-one-2-yl]methyl}-N-hydroxybenzamide (6d); 4-[(6-Chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N-hydroxybenzamide (6e); 4-[(6-Chloro-3-cyclopropyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N-hydroxybenzamide (6f); 4-{[3-(2-Chlorophenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]methyl}-N-hydroxybenzamide (6g); 4-{[3-(4-Chlorophenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]methyl}-N-hydroxybenzamide (6h); 4-{[3-(2,6-Dimethylphenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]methyl}-N-hydroxybenzamide (6i); 4-[(6-Fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N-hydroxybenzamide (6j); N-Hydroxy-4-((3-(4-methoxyphenyl)-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)benzamide (6k); 4-[(3-Ethyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N-hydroxybenzamide (6l); 4-[(6-Cyano-3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N-hydroxybenzamide (6m); 4-((3,4-Dihydro-3-isopropyl-4-oxoquinazolin-2-yl)methyl)-N-hydroxybenzamide (6n); 4-((3-Ethyl-6-fluoro-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-N-hydroxybenzamide (6o); and pharmaceutically acceptable salts thereof.

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