Quinazolinone derivatives for HDAC inhibition

Abstract

Histone deacetylases 6 (HDAC6) inhibitors and compositions comprising the same are disclosed. Compounds as HDAC6 inhibitors are useful for treating, alleviating, ameliorating, and / or decreasing severity of an HDAC6-associated disease or condition in a subject in need thereof. HDAC6-associated diseases or conditions include fibrosis, neurodegenerative diseases, kidney diseases, cancer, and increased growth of cyst metrics.

Description

[0001] QUINAZOLINONE DERIVATIVES FOR HDAC INHIBITION

[0002] FIELD OF THE IN VENTION

[0003] The present invention relates to compounds with histone deacetylase 6 (HDAC6) enzyme inhibition activity and use thereof.

[0004] BACKGROUND OF THE INVENTION

[0005] Histone deacetylases (HDACs) catalyze the deacetylation of histone and non-histone proteins and play important roles in epigenetic regulation. Autosomal dominant polycystic kidney disease (ADPKD) is characterized by slowly progressive, bilateral kidney enlargement due to numerous fluid-filled cysts. ADPKD is caused by mutations in either PKD1 or PKD2 genes, where disruption of their normal functions leads to excessive proliferation of the renal tubular epithelium causing cyst formation. Over 12 million people worldwide have ADPKD, making it amongst the most commonly known monogenetic disorders. Fifty percent of ADPKD patients eventually develop end-stage renal disease (ESRD) by the age of 60, accounting for 10% and 5% of prevalent patients with ESRD in Europe and the United States, respectively. Unfortunately, treatment options for this life-threatening disorder are still limited and inadequate.

[0006] HDAC6 inhibitors have been suggested as one possible strategy to inhibit cyst growth. It has been also found that HDAC6 inhibition down regulated cAMP levels, inhibited cell proliferation, and inhibited c AMP-activated CFTR chloride currents in MDCK cells. HDAC6 inhibition also can inhibit cyst growth in vitro. Based on previous studies showing that cystic cholangiocytes in polycystic liver disease have abnormal cell cycle profiles and malfunctioning cilia, Gradilone and collaborators studied the role of HDAC6 in polycystic liver disease (PLD), which targets the epithelial lining of the biliary tree. They found that expression of the HDAC6 protein is six times higher in cystic liver tissue and in cultured cholangiocytes isolated from both PCK rats (an animal model of PLD) and humans with PLD. As in our studies of ADPKD, inhibition of HDAC6 activity by HDAC6 inhibitors decreased the proliferation of cystic cholangiocytes in a dose- and timedependent manner and inhibited cyst growth in three-dimensional (3D) cultures.

[0007] Recent studies report that HDAC6 could play a role in cyst formation in ADPKD as well as PLD and could therefore serve as a potential therapeutic target.

[0008] SUMMARY OF THE INVENTION

[0009] In one aspect, the invention relates to a compound of Formula (I): or a pharmaceutically acceptable salt thereof,

[0010] Wherein

[0011] R1is hydrogen, trifluoromethyl, (C1-5) alkyl, (C3-5) cycloalkyl, (C1-6) heterocycloalkyl, arylalkyl, substituted phenyl, or substituted heteroaryl;

[0012] R2is hydrogen, halogen, trifluoromethyl, cyano, (C1-5) alkyl, (C1-6) alkoxy, (C1-6) heterocycloalkyl, arylakyl, substituted phenyl, or substituted heterocycloaryl;

[0013] R3is hydrogen, halogen, trifluoromethyl, cyano, (C1-5) alkyl, (C1-6) alkoxy, (C1-6,) heterocycloalkyl, arylalkyl, substituted phenyl, or their substituted analogues;

[0014] R4is hydrogen, halogen, trifluoromethyl, cyano, (C1-5) alkyl, (C1-6) alkoxy, (C1-6) heterocycloalkyd, arylalkyl, substituted phenyl, or their substituted analogues;

[0015] R5is hydrogen, halogen, trifluoromethyl, cyano, (C1-5) alkyl. (C1-6,) alkoxy, (C1-6,) heterocycloalkyl, arylalkyl, substituted phenyl, or their substituted analogues;

[0016] R6is hydrogen, halogen, or hydroxyl alkoxy; or a pharmaceutically acceptable salt, hydrate, or prodrug thereof.

[0017] In one embodiment, R1is -CH3, -CH2CH3, -isopropyl, -cyclopropyl, -cyclopropyhnethyl, - CeHj, -(4-C1 )C6H5-(2-C1)C6H5, -2,6-dimethylphenyl, or -(4-OMe)C6H5

[0018] In another embodiment, R1is -CH3or -CH2CH3

[0019] In another embodiment, R3is -C1, -F or -CN.

[0020] In another embodiment, the moiety

[0021] In another embodiment, the compound is selected from the group consisting of: -{[3-methylquinazolin-4(3H)-on-2-yl]methyl}-N-hydroxybenzamide (6a); 4-{[3-Cyclopropylquinazolin-4(3H)-on-2-yl]methyl}-N-hydroxybenzamide (6b); 4-{[3-cyclopropylmehtylquinazolin-4(3H)-on-2-yl]methyl}-N-hydroxybenzamide (6c); 4-{[3-phenylquinazoIin-4(3H)-on-2-yl]methyU-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);

[0022] 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-dihydroquinazoIin-2-yl)methyl]-N-hydroxybenzamide (6j);

[0023] 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 (61); 4-[(6-cyano-3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N-hydroxybenzamide (6m);

[0024] 4-((3,4-dihydro-3-isopropyl-4-oxoquinazolin-2-yl)methyl)-N-hydroxybenzamide (6n);

[0025] 4-((3-ethyl-6-fluoro-4-oxo-3,4-dihydroqiiinazolin-2-yl)methyl)-N-hydroxybenzamide (6o); and a pharmaceutically acceptable salt thereof.

[0026] In another aspect of the invention, the invention relates to a pharmaceutical composition comprising:

[0027] (a) a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 6; and

[0028] (b) a pharmaceutically acceptable carrier or vehicle.

[0029] Further in another aspect of the invention, the invention relates to a use of a compound or a pharmaceutically acceptable salt thereof of the invention, or use of a pharmaceutical composition of the invention in the manufacture of a medicament for treating, alleviating, ameliorating, and / or decreasing severity of an HDAC6-associated disease or condition in a subject in need thereof.

[0030] Alternatively, the invention relates to a compound or a pharmaceutically acceptable salt thereof of the invention, or a pharmaceutical composition of the invention for use in treating, alleviating, ameliorating, and / or decreasing severity of an HDAC6-associated disease or condition in a subject in need thereof.

[0031] The invention also relates to a method for treating, alleviating, ameliorating, and / or decreasing severity of an HDAC6-associated disease or condition, comprising: administering a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof of the invention, or a pharmaceutical composition of the invention to a subject in need thereof and thereby treating, G Sleviating, ameliorating, and / or decreasing severity of an HDAC6-associated disease or condition in the subject in need thereof.

[0032] In one embodiment, the HDAC6-associated disease or condition is selected from the group consisting of fibrosis, a neurodegenerative disease, a kidney disease, cancer, and an increase in cyst growth metrics.

[0033] In another embodiment, the fibrosis, associated with HDAC6 activity, is pulmonary’ fibrosis, idiopathic pulmonary' fibrosis, hepatic fibrosis, renal fibrosis, or myelofibrosis.

[0034] In another embodiment, the neurodegenerative disease, associated with HDAC6 activity', is Huntington’s disease, Alzheimer’s disease, Parkinson’s disease, or Amyotrophic lateral sclerosis.

[0035] In another embodiment, the kidney disease, associated with HDAC6 activity, is polycystic kidney disease, or autosomal dominant polycystic kidney disease, or autosomal recessive polycystic kidney disease.

[0036] In another embodiment, the cancer, associated with HDAC6 activity', is lung, breast, renal, liver cancer, multiple myeloma, or glioma.

[0037] In another embodiment, the compound or pharmaceutically acceptable salt thereof, or the pharmaceutical composition is used in the manufacture of a medicament for decreasing the growth of cyst metrics, said growth of cyst metrics associated with HDAC6 activity.

[0038] In another embodiment, the compound or pharmaceutically acceptable salt thereof, or the pharmaceutical composition is used in the manufacture of a medicament for treating, alleviating, ameliorating, and / or decreasing severity of the condition of the increase in cyst growth metrics, said increase in cyst growth metrics associated with HDAC6 activity.

[0039] In another embodiment, the HDAC6-associated disease or condition is selected from the group consisting of hepatic 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.

[0040] BRIEF DESCRIPTION OF THE DRAWIN GS FIG. 1 shows pretreatment with Compound 6a suppressed the growth of primary human ADPKD cysts in a concentration-dependent manner.

[0041] FIGs. 2A-D show compound 6a prevents cyst metrics in human ADPKD cells.

[0042] FIGs. 3A-D show compound 6b prevents cyst metrics in human ADPKD cells.

[0043] FIGs. 4A-D show compound 6a is superior to ACY12I5 in terms of cytotoxicity and prevention of ADPKD cysts formation.

[0044] FIGs. 5A-D show compound 6a rescues cyst metrics in human ADPKD cells.

[0045] FIGs. 6A-D show compound 6b rescues cyst metrics in human ADPKD cells.

[0046] FIGs. 7A-D show compound 6a confers efficacy in PC 1 -KO mouse models.

[0047] FIG. 8 shows compound 6a reduced kidney weight in transgenic Pkdl knockdown mice.

[0048] FIGs. 9A-B show compound 6a reduced renal cyst in transgenic Pkdl knockdown mice.

[0049] FIG. 10 shows compound 6a inhibits fibrosis and inflammation biomarkers in the BIOMAP® Fibrosis Panel.

[0050] DETAILED DESCRIPTION OF THE INVENTION

[0051] DEFINITIONS

[0052] 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 pertains. In the case of conflict, the present document, including definitions will control.

[0053] As used herein, the term "alkyl" represents a saturated, straight, or branched hydrocarbon moiety. Exemplary alkyls include, but are not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, n-pentyl, etc.

[0054] The term "C1 - 4alkyl" refers to an alkyl containing from 1 to 4 carbon atoms. When the term "alkyl” is used in combination with other substituent groups, such as "arylalkyl", the term "alkyl" is intended to encompass a divalent straight or branched-chain hydrocarbon radical. For example, "arylalkyl" is intended to mean the radical -alkylaryl, wherein the alkyl moiety thereof is a divalent straight or branched-chain carbon radical and the aryl moiety thereof is as defined herein, and is represented by the bonding arrangement present in a benzyl group (-CH2-phenyl).

[0055] The term "cycloalkyl" refers to a non-aromatic, saturated, cyclic hydrocarbon ring. The term "C3- 8cycloalkyl" refers to a non-aromatic cyclic hydrocarbon ring having from three to eight ring carbon atoms. Exemplary "C3-8cycloalkyl" groups useful in the present invention include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[0056] The term "Alkoxy" refers to a group containing an alkyl radical attached through an oxygen linking atom. The term "C1-4alkoxy" refers to a straight- or branched-chain hydrocarbon radical having at least 1 and up to 4 carbon atoms attached through an oxygen linking atom. Exemplary "(C1 -4)alkoxy" groups useful in the present invention include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, and s-butoxy.

[0057] The term "Aryl" represents a group or moiety comprising an aromatic, monocyclic or bicyclic hydrocarbon radical containing from 6 to 10 carbon ring atoms and to which may be fused one or more cycloalkyl rings. In addition, the terms alkyl, aryl, cycloalkyl, heteroaryl, etc. may be used to define a divalent substituent, such as a group bonded to two other groups. In this instance, such terms are intended to encompass divalent moieties. For example, "pentyl" is intended to represent a pentylene diradical -wherein the pentyl moiety is any one of a divalent straight (e.g. -CH2CH2CH2CH2CH2-) or branched (e.g. -CH2CH(CH3)CH2CH2- ,-CH2CH2CH(CH2CH3)-, -CH2CH2C(CH3)2-) chain 5-carbon radical.

[0058] Generally, in the compounds of this invention, heterocycloalkyl groups are 5-membered and / or 6- membered heterocycloalkyl groups, such as pyrrolidyl (or pyrrolidinyl), tetrahydrofuranyl, tetrahydrothienyl, dihydrofuranyl, oxazolinyl, thiazolinyl or pyrazolinyl, piperidyl (or piperidinyl), piperazinyl, morpholinyl, tetrahydropyranyl, dihydropyranyl, 1 ,3-dioxanyl, tetrahydro-2H- 1 ,4- thiazinyl, 1,4-dioxanyl, 1 ,3-oxathianyl, and 1 ,3-dithianyl.

[0059] The term "Heteroaryl" represents a group or moiety comprising an aromatic monocyclic containing 5 to 10 ring atoms, including 1 to 4 heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur. This term is also intended to encompass heterocyclic groups containing nitrogen and / or sulfur where the nitrogen or sulfur heteroatoms are optionally oxidized. Illustrative examples of heteroaryls include, but are not limited to, thienyl, pyrrolyl, imidazolyl, pyrazolyl, furanyl, isothiazolyl, fiirazanyl, isoxazolyl, oxazolyl, oxadiazolyl, thiazolyl, pyridinyl, pyridinyl-N-oxide, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, tetrazinyl, triazolyl. tetrazolyl.

[0060] Some of the heteroaryl groups according to the compounds of the invention are 5-6 membered monocyclic heteroaryl groups. Selected 5-membered heteroaryl groups contain one nitrogen, oxygen or sulfur ring heteroatom, and optionally contain 1 , 2 or 3 additional nitrogen ring atoms. Selected 6- membered heteroaryl groups contain 1 , 2, 3 or 4 nitrogen ring heteroatoms. Selected 5- or 6-membered heteroaryl groups include thienyl, pyrrolyl, imidazolyl, pyrazolyl, furanyl, isothiazolyl, fiirazanyl, isoxazolyl, oxazolyl, oxadiazolyl, thiazolyl, triazolyl, and tetrazolyl or pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl and thiadiazolyl.

[0061] The terms "halogen" and "halo" represent chloro, fluoro, bromo or iodo substituents.

[0062] When a disclosed compound or its salt is named or depicted by structure, it is to be understood that the compound or salt, including solvates (particularly, hydrates) thereof, may exist in crystalline forms, non-crystalline forms or a mixture thereof. The compound or salt, or solvates (particularly, hydrates) thereof, may also exhibit polymorphism (i.e. the capacity to occur in different crystalline forms). These different crystalline forms are typically known as "polymorphs." It is to be understood that when named or depicted by structure, the disclosed compound, or solvates (particularly, hydrates) thereof, also include all polymorphs thereof. Polymorphs have the same chemical composition but differ in packing, geometrical arrangement, and other descriptive properties of the crystalline solid state. Polymorphs, therefore, may have different physical properties such as shape, density, hardness, deformability, stability, and dissolution properties. Polymorphs typically exhibit different melting points, IR spectra, and X-ray powder diffraction patterns, which may be used for identification. One of ordinary skill in the art will appreciate that different polymorphs may be produced, for example, bychanging or adjusting the conditions used in crystallizing / recrystallizing the compound. Because of 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. Salts encompassed within the term "pharmaceutically acceptable salts" refer to non-toxic salts of the compounds of this invention. Typically, a salt may be readily prepared by using a desired acid or base as appropriate. The salt mayprecipitate from solution and be collected by filtration or may be recovered by evaporation of the solvent. When a compound of the invention is a base (contain a basic moiety ), a desired salt form may be prepared by any suitable method knowm 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, and the like, 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, and the like, or with a pyranosidyl acid, such as glucuronic acid or galacturonic acid, or with an alphahydroxy acid, such ascitric 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 or the like. Suitable addition salts are formed from acids which form non-toxic salts and examples include acetate, p-aminobenzoate, ascorbate, aspartate, benzenesulfonate, benzoate, bicarbonate, bismethylenesalicylate, bisulfate, bitartrate, borate, calcium edetate, camsylate, carbonate, clavulanate, citrate, cyclohexylsulfamate, edetate, edisylate, estolate, esylate, ethanedisulfonate, ethanesulfonate, formate, fumarate, gluceptate, gluconate, glutamate, glycollate, glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, dihydrochloride, hydrofumarate, hydrogen phosphate, hydroiodide, hydromaleate, hydrosuccinate, hydroxynaphthoate, isethionate, itaconate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, monopotassium maleate, mucate, napsylate, nitrate, / V -methylglucamine, oxalate, oxaloacetate, pamoate (embonate), palmate, palmitate, pantothenate, phosphate / diphosphate, pyruvate, polygalacturonate, propionate, saccharate, salicyiate, stearate, subacetate, succinate, sulfate, tannate, tartrate, teoclate, tosylate, triethiodide, trifluoroacetate and valerate.

[0063] Other exemplary acid addition salts include pyrosulfate, sulfite, bisulfite, decanoate, 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, phenylbutrate, lactate, g- hydroxybutyrate, mandelate, and sulfonates, such as xylenesulfonate, propanesulfonate, naphthalene- 1 -sulfonate and naphthalene-2-sulfonate. If an inventive basic compound is isolated as a salt, the corresponding free base form of that compound may be prepared by any suitable method known to 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.

[0064] When a compound of the invention is an acid (contains an acidic moiety), a desired salt may be prepared by any suitable method known to the art, including treatment of the free acid with an inorganic or organic base, such as an amine (primary, secondary; or tertiary'), an alkali metal or alkaline earth metal hydroxide, or the like. Illustrative examples of suitable salts include organic salts derived from amino acids such as glycine and arginine, ammonia, primary, secondary, and tertiary amines, and cyclic amines, such as N-methyl-D-glucamine, diethylamine, isopropylamine, trimethylamine, ethylene diamine, dicyclohexylamine, ethanolamine, piperidine, morpholine, and piperazine, as tvell as inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum, and lithium. Certain of the compounds of this invention may form salts with one 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.

[0065] Compounds of the invention having both a basic and acidic moiety may be in the form of zwitterions, acid-addition salt of the basic moiety or base salts of the acidic moiety'. This invention also provides for the conversion of one pharmaceutically acceptable salt of a compound of this invention, e.g., a hydrochloride salt, into another pharmaceutically acceptable salt of a compound of this invention, e.g., a sulfate salt. For solvates of the compounds of Formula (I), or salts thereof that are in crystalline form, the skilled artisan will appreciate that pharmaceutically-acceptable solvates may be formed wherein solvent molecules are incorporated into the crystalline lattice during crystallization. Solvates may involve nonaqueous solvents such as ethanol, isopropanol, DMSO, acetic acid, ethanolamine, and ethyl acetate, or they may involve water as the solvent that is incorporated into the crystalline lattice. Solvates w'herein water is the solvent that is incorporated into the crystalline lattice are typically referred to as "hydrates." Hydrates include stoichiometric hydrates as w'ell as compositions containing variable amounts of water. The invention includes all such solvates. Because the compounds of Formula (I) are intended for use in pharmaceutical compositions it will readily be understood that they are each preferably provided in substantially pure form, for example at least 60% pure, more suitably at least 75% pure and preferably at least 85%, especially at least 98% pure (% are on a weight for weight basis). Impure preparations of the compounds may be used for preparing the more pure forms used in the pharmaceutical compositions.

[0066] The term “(Cm-Cn)” or “Cm-n”, wherein m, n are integers, and n>m, means that all integer unit amounts within the range m to n are specifically disclosed as part of the invention. Thus, by “(Cm- Cn) , it means that Cm, Cm+1, Cm+2, . . . , Cn-2, Cn-1, Cn, (Cm“Cm+1), (Cm-Cm+2), (Cm-Cm+3), . . . , (Cm-Cn- 2), (Cm-Cn-1), (Cm-Cn); (Cm+1-Cm-2), (Cm+1-Cm+3), (Cm+1 Cm+4), . . . , (Cm+1-Cn-2), (Cm+1-Cn-1), (Cm+1-Cn), . . . , (Cn-2-Cn-1), (Cn-2-Cn); and (Cn-1-Cn) are included as embodiments of this invention.

[0067] By “(Ci-Ce)” or “Cl-6”, it means that all integer unit amounts within the range 1 to 6 are specifically disclosed as part of the invention. Thus, 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 (Cs-Ce) units amounts are included as embodiments of this invention.

[0068] The term “treating” or "treatment" refers to administration of an effective amount of a therapeutic agent to a subject, who has a disease, or a symptom or predisposition toward such a disease, with the purpose to cure, alleviate, relieve, remedy, ameliorate, decrease severity, or prevent the disease, the symptoms of it, or the predispositions towards it.

[0069] The terms “an HDAC6-associated disease or condition”, “an HDAC6-mediated disease or condition”, and “ a disease or a condition associated with HDAC6” are interchangeable.

[0070] The term “an HDAC6-associated disease or condition”, “an HDAC6-mediated disease or condition”, or “ a disease or a condition associated with HDAC6” refers to a disease or a condition that can be treated, alleviated, ameliorated, or decreased severity by inhibition of HDAC6 activity, and / or for which inhibition of HDAC6 can provides a benefit.

[0071] The “Guidance for Industry and Reviewers Estimating the Safe Starting Dose in Clinical Trials for Therapeutics in Adult Healthy Volunteers” published by the U.S. Department of Health and Human Services Food and Drug Administration discloses “a human equivalent dose” may be obtained by calculations from the following formula:

[0072] HED = animal dose in mg / kg x (animal weight in kg-'ltuman weight in kg) °33.

[0073] The compounds of the invention may be obtained by using synthetic procedures illustrated in the Schemes below or by drawing on the knowledge of a skilled organic chemist. The synthesis provided in these Schemes are applicable for producing compounds of the invention having a variety of different substituent groups employing appropriate precursors, which are suitably protected if needed, to achieve compatibility with the reactions outlined herein. Subsequent deprotection, where needed, affords compounds of the nature generally disclosed. While the Schemes are shown with compounds only of Formula (I), they are illustrative of processes that may be used to make the compounds of the invention. Intermediates (compounds used in the preparation of the compounds of the invention) may also be present as salts. Thus, in reference to intermediates, the phrase "compound(s) of formula (number)" means a compound having that structural formula or a pharmaceutically acceptable salt thereof.

[0074] Scheme 1 General synthesis procedures of compound 6a-6m. Reagents and conditions: (a) substituted amines, DIPEA, HOBt, EDC.HC1, DCM, 0°C to rt 16 h; (b) (i) oxalyl chloride, DMF, DCM, 0°C to rt 2 h (ii) aniline, TEA, DCM. 0°C to rt 3 h; (c) iron powder, NH4CI, EtOH:H2O (8: 2), 90°C 16 h; (d) 2-[4-(methoxycarbonyl)phenyl]acetic acid, DIPEA, HOBt, EDC.HC1, DCM, 0°C to rt 16 h; (e) ZnCh, hexamethyl disilazane, DMF, 120°C 24 h; (I) 2M NH2OH in MeOH, NaOH, rt I h; (g) (i) NaCN, NiBr2, NMP MW I20W 200°C 10 min (ii) NH2OBn.HCl, EDCI, HOBt, DIPEA, DMF, rt Ih (iii) IM BBr2, dimethyl sulfide complex in DCM, DCM, 0°C 30 min.

[0075] This series of compounds have 2-substituted quinazoline-4-one as a core. This w as readily synthesized from corresponding diamide as key intermediate II, 3a-31. This diamide 3 in turn was obtained from commercially available o-nitro benzoic acid 1 using two different routes a four-step reaction sequence involving acid chloride and acid amine coupling reaction as shown in Scheme 1 . Commercially available nitrobenzoic acid 1 on treatment with oxalyl chloride in DCM afforded the corresponding acid chloride which was then treated with aniline to afford nitro amide 2. Alternatively, the nitrobenzoic acid 1 on coupling reaction with corresponding amine in presence of coupling agent EDC.HC1 and HOBt afforded the nitro amide 2. Nitro group reduction of 2 in presence of Fe-AcOH afforded substituted anthraliamides 3. These amino amide 3a-31 on reaction with 2-[4-(methoxycarbonyl)phenyi]acetic acid in presence of EDC.HC1 and HOBt afforded the intermediate I, 4a-41. The intermediate 4 on treatment with hexamethyi disilazine and Zinc chloride underwent smooth cyclization to afford 2-substituted quinazo!in-4(3H)-one 5a-51. These 2- substituted quinazolin-4(3H)-one intermediates 5 on reaction with freshly prepared NH2OH in presence of NaOH in methanol afforded the final product 6a-61. The purification of which was carried out using combination of column chromatography, reverse phase HPLC to give the target hydroxamic acid compound 6a-61 as solids.

[0076] For the synthesis of compound 6m, the nitrile group was inserted from chloride intermediate 5e with sodium cyanide and nickel bromide via microwave irradiation to give nitrile intermediate. Surprisingly, half of the nitrile intermediates was found to bear a carboxylic acid instead of the original ester group. Hence, the carboxylic acid group was reacted with NH2OB11 with coupling agents and then treated with BBn to afford hydroxamic acid compound 6m.

[0077] EXAMPLES

[0078] Pharmacological studies

[0079] Compound 6a-p inhibits HDAC6.

[0080] We first evaluated a series of quinazolines derivatives against HDACs and found several compounds with potent and selective inhibiting activity toward HDAC6 with an IC50 in digital nM range. Table 1 shows HD AC inhibition activities of compound 6a-6o.

[0081] Compound 6a suppresses the growth of primary human ADPKD cysts.

[0082] We next validated our initial screening results and performed in vitro 3D primary human ADPKD cell assay. The anti-ADPKD cyst growth efficacy of compound 6a and compound 6b was evaluated with prevention and reduction assays:

[0083] Prevention (pretreatment) assay: Cells were seeded on Day 0 and treated on Day 1. Additional treatments were added 3-4 days throughout the study. The study was terminated based on the cyst size of any +stimulus groups. End point whole-well images were acquired on the last day of the study with cell number performed the day after (FIGs. 1, 2 and 3).

[0084] Reduction (rescue) assay: Drug treatments and precise experimental timing were based on cyst growth, but cells were seeded on Day 0. Compounds were added after cysts had begun to form, approximately on Day 4-7. The study was terminated based on the cyst size of any stimulus groups. Whole-well images were captured before dosing and captured again right before the conclusion of the experiment. Cell numbers were assessed at the conclusion of the experiment. (FIGs. 5 and 6)

[0085] Two therapeutic assets (compound 6a and compound 6b) were screened along the comparator (Ricolinostat) with DMSO control on ADPKD cells with PKD1 mutations from 1 Donor stimulated with Forskolin in a Cyst Reduction Assay. Human ADPKD cells were pretreated with 6 serial dilutions of compound 6a and compound 6b with n=8. The comparator Ricolinostat was run at a single point (3 pM) (FIG. 4D).

[0086] End Points:

[0087] The principal measurements of therapeutic effectiveness were measurements of cyst number per well, cyst size, cyst cell proliferation (measured via Cell Titer Gio) and cytotoxicity' (via LDH release). Cyst number and size measurements are determined via post-acquisition image analysis.

[0088] Results:

[0089] Cyst cell proliferation (CTG), Cyst Number (CN), Total Cyst Area (TA), and Average Size / Cyst were determined at the conclusion of this DBM Prevention Assay. Collectively, pretreatment 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 (FIG. 1).

[0090] Both compound 6a and compound 6b showed cytotoxicity at 90 pM, whereas the comparator (ACY1215) displayed levels of cytotoxicity at 3 and 10 pM. Compound 6a trends toward being cytotoxic past 30 pM and can be referenced having a similar cytotoxic profile in a reduction assay (see NTU SOW 5). Of all compounds tested, compound 6a was the most effective at reducing cystogenic metrics.

[0091] Our data from individual human ADPKD donor cyst cultures also demonstrate a positive correlation between HDAC6 inhibition and cyst growth reduction across all treatments (Table 1; FIGs. 2, 3 and 4), suggested HDAC6 inhibition might be a potential target for the treatment of ADPKD. Moreover, this effect is specific to ADPKD cells, as we observed no cytotoxicity in compound 6a-treated non-ADPKD kidney cells where ACY1215 was evident (FIG. 4).

[0092] FIGs. 2A-D show pretreatment with compound 6a in primary human ADPKD cells against cyst metrics. (A) Cyst cell proliferation (measured via CTG) and cytotoxicity (via LDH release) (B) Cyst Number (C) Total Cyst Area (D) Average Size / Cyst were plotted in a 6-point concentration-response curve where significance of DMSO control vs. drug-treated groups was determined using One-Way ANOVA, followed by Dunnet’s multiple comparisons test (*p<0.05).

[0093] FIGs. 3A-D show pretreatment with compound 6b in the primary human ADPKD cell against cyst metrics. (A) Cyst cell proliferation (measured via CTG) and cytotoxicity (via LDH release) (B) Cyst Number (CN) (C) Total Cyst Area (TA) (D) Average Size / Cyst w ere plotted in a 6-point concentration-response curve w'here significance of DMSO control vs. drug-treated groups was determined using One-Way ANOVA, followed by Dunnet’s multiple comparisons test (*p<0.05).

[0094] FIGs 4A-D show Compound 6a is superior to ACY1215 in terms of cytotoxicity and prevention of ADPKD cysts formation.

[0095] FIGs. 5A-D show reduction assay: Compound 6a against cyst metrics. (A) Cyst cell proliferation (measured via CTG) and cytotoxicity (via LDH release) (B) Cyst Number (CN) (C) Total Cyst Area (TA) (D) Average Size / Cyst were plotted in a 6-point concentration-response curve w'here significance of DMSO control vs drug-treated groups w'as determined using One-Way ANOVA, followed by Dunnet’s multiple comparisons test (*p<0.05).

[0096] FIGs. 6A-D show- reduction assay: Compound 6b against cyst metrics. (A) Cyst cell proliferation (measured via CTG) and cytotoxicity (via LDH release) (B) Cyst Number (CN) (C) Total Cyst Area (TA) (D) Average Size / Cyst were plotted in a 6-point concentration-response curve w'here significance of DMSO control vs drug-treated groups was determined using One-Way ANOVA, followed by Dunnet’s multiple comparisons test (*p<0.05). CTG, LDH, Cyst Number and Cyst Size were determined at the conclusion of this DBM Reduction Assay. (FIGs. 5 and 6) Collectively, all four benchmarks form standard cystogenic metrics that were used to measure and compare efficacy of compound 6a (FIG. 5) and compound 6b (FIG. 6) on inhibiting cyst growth and reducing the cystogenic profile of human ADPKD cells in vitro. Of all compounds tested, compound 6a was the most effective at reducing cystogenic metrics.

[0097] Compound 6a confers efficacy in PKD mouse model

[0098] The efficacy of compound 6a was further evaluated in the Pkd1 knockdown mice model. A transgenic line which had ~60-70% reduction of Pkdl expression was developed severe renal cystic disease at a rate similar to that of human ADPKD. (FIG. 7) These results further support the haploinsufficiency hypothesis, and suggest that the onset and progression of the renal cystic diseases are correlated with the level of Pkdl expression. Therefore, we sought to determine whether compound 6a treatment produces similar beneficial effects in a randomized, blinded and statistically powered efficacy study in Pkdl knockdown mice. Pkdl knockdown mice were daily oral administrated with PBS, 60 mg kg-1, 120 mg kg1and 150 mg kg1of compound 6a from post-natal day P14 to P28. Mice were sacrificed and kidneys were harvested on P28. Most importantly, animals treated with compound 6a led to reduction in kidney-to-body-weight ratio (KW / BW) (Fig. 9) and cyst index (total cystic area / total kidney area) (Fig. 8) when compared to the vehicle control group.

[0099] FIGs. 7A-D show compound 6a confers efficacy in PC1-KO mouse models (A) Animal No.86, Cystic index: 49.9%; 50.5% (Vehicle group) (B) Animal No.84, Cystic index: 30.2%; 23.6% (60 mg / Kg) (C) Animal No.87, Cystic index: 21.5%; 18.8% (120 mg / Kg) (D) Animal No.83, Cystic index: 26.5%; 1 1.4% (150 mg / Kg)

[0100] FIG. 8 shows Compound 6a significantly reduced cyst index in transgenic Pkdl knockdown mice.

[0101] FIGs. 9A-B show compound 6a reduced renal cystic in transgenic Pkdl knockdown mice.

[0102] Compound 6a preferentially distributes to kidney and liver.

[0103] We next evaluated the in vivo pharmacokinetic and biodistribution profile of compound 6a in wild-type mice following a single 30 mg kg1oral administration. Compound 6a is rapidly absorbed into plasma, showing tmax of <0.5 h, Cmax of 264 μg mL-1,and half life of <4 h. The rapid plasma clearance of compound 6a reflects the extensive distribution to tissues such as kidney and liver. Uniquely, compound 6a showed preferential kidney distribution, with kidney-to-plasma (K / P) ratio of approximately 2 / 1 by AUClast, respectively. Compound 6a inhibit inflammation and fibrosis biomarkers in the BIOMAP® Fibrosis Panel.

[0104] Compound 6a was subjected to characterize in the Eurofms BIOMAP® Fibrosis panel of 3 human primary cell-based systems including MyoF system, REMyoF system and SAEMyoF system. These systems are designed to model complex human tissue and disease biology that drives the aberrant inflammation involved in fibrosis and wound healing. Fibrotic disease of the kidney associated with end-stage renal failure is captured in the REMyoF system consisting of a co-culture of renal proximal tubule epithelial cells and adult fibroblasts.

[0105] FIG. 10 shows BIOMAP® profile of compound 6a in the Fibrosis Panel.

[0106] The result demonstrated that compound 6a is active with 15 annotated readouts and noncytotoxic at the concentrations tested in this study. Compound 6a impacts inflammation-related activities (decreased I-TAC, sIL-6, and MCP-1 ), myofibroblast activation-related activities (increaseda-SMA), fibrosis-related matrix activities (decreased collagen I, collagen III; increased collagen IV: modulated MMP-1), and tissue remodeling / wound healing activities (decreased tPA, uPA, and sVEGF).

[0107] There are four common activities that are annotated within the REMyoF system: Collagen I, I- TAC, sIL-6, and sVEGF. There are four differentiating activities (not shown) within the following systems: SAEMyoF (ctSMA) and MyoF (IL-8, TIMP-1, MMP-1). In study compound 6a was characterized by profiling in the Fibrosis Panel of human primary cell-based assays modeling the fibrotic and inflammatory processes that drive fibrosis in tissue-specific contexts. The Fibrosis panel evaluates tire biological impact of test agents in conditions that preserve the compl ex crosstalk and feedback mechanisms that are relevant to in vivo outcomes.

[0108] Compound 6a was noncytotoxic and active with 15 annotated readouts, nine in the Renol Fibrosis (REMyoF) system model, three in the MyoF system, and three in the SAEMyoF system. The annotated biomarker activities impact inflammation, myofibroblast activation, fibrosis-related matrix, and tissue remodelingAvound healing activities. When compound 6a at 10 pM was overlaid with the selected reference benchmark nintedanib at 1 .1 μM, there were four common and four differentiating activities identified.

[0109] The BIOMAP® Fibrosis Early Screening Service (SAEMyoF) is a high quantity screening format in the BIOMAP® fibrosis system SAEMyoF, comprised of small airway epithelial cells and lung fibroblasts. This system is stimulated with profibrotic and proinflammatory factors to model the biology of fibrotic lung diseases such as idiopathic pulmonary fibrosis. Biomarker readouts capture impacts on translationally relevant matrix remodeling, tissue repair, and inflammation related responses in the diseased lung. This service allows for rapid testing of 18 compounds at 4 concentrations for antifibrotic development potential in a human fibrosis model. Chemical Synthesis

[0110] Example 1: Preparation of 4-{[3-Cyclopropylquinazolin-4(3H)-on-2-yl]methyl}-N- hydroxybenzamide (compound 6b)

[0111] Step 1: Preparation of the intermediate N-cyclopropyl-2-nitrobenzamide

[0112] To a solution of 2 -nitrobenzoic acid (5 g, 29.94 mmol, 1 eq.) in DCM (50 ml) was added DIPEA (13 ml, 89.92 mmol, 3 eq.), EDC.HC1 (8.58 g, 44.78 mmol, 1.5 eq.), HOBt (4.04 g, 29.94 mmol, 1 eq) then add cyclopropyl amine (2.56 mL, 35.93 mmol, 1.2 eq) at 0°C, and the reaction mixture was allowed to warm to rt and stirred for 16h. Then, the reaction mixture was poured into ice water and extracted with DCM (100 ml x2). The organic layers were washed with water (100 ml) and brine (100 ml). After dried over Na2SC>4, the solvent was filtered and removed in vacuo to give crude residue. The crude compound was purified by column chromatography (eluted by 0 to 20% EtOAc in pet ether) and the desired fraction (R / = 0.5, EtOAc / pet ether = 3:7) was collected to afford the title compound as an off white solid. (5 g, 81% yield)

[0113] Step 2: Preparation of the intermediate 2-amino-N-cyclopropylbenzamide

[0114] To a solution of N-cyclopropyl-2-nitrobenzamide (5g, 24.27 mmol, 1 eq) in EtOH: H2O (8: 2) was added iron power (5.43g, 96.96 mmol, 4 eq) and NH4CI (5.4 g, 96.96 mmol, 4 eq), and the reaction mixture was stirred for 16h at 90°C. The solvent was removed in vacuo, extracted with EtOAc (100 ml x2), and washed with water (100 ml). After dried over anhydrous Na2SO4, the solvent was filtered and concentrated under reduced pressure to afford the title compound as an off white solid. (3.7 g, 90% yield) Rf= 0.4 (EtOAc / pet ether = 3:7);

[0115] Step 3: Preparation of the intermediate methyl 4-(2-{[2- (eyclopropylcarbamoyl)phenyl]amino}-2-oxoethyl)benzoate To a solution of 2-[4-(methoxycarbonyl)phenyl]acetic acid (3.5 g, 18.04 mmol, 1 eq) in DCM (35 ml) was added DIPEA (9.06 mL, 54.12 mmol, 3 eq), EDC.HC1 (5.18 g, 27.06 mmol, 1.5 eq), HOBt (2.43 g, 18.04 mmol, 1 eq) then added N-cyclopropyl-2-nilrobenzamide (3.17 gm, 18.04 mmol, 1 eq.) at 0°C, and the reaction mixture was allowed to warm to rt and stirred for 16h. Then, the reaction mixture was poured into ice water and extracted with DCM (50 ml x2). The organic layers were washed with water (50 ml), dried over Na2SO4, and concentrated under reduced pressure to give crude residue. The crude compound was purified by column chromatography (eluted by 0 to 35% EtOAc in pet ether) and the desired fraction (Rf= 0.4, EtOAc / pet ether = 1:4) was collected to afford the title compound as an off white solid. (3.8 g, 60% yield)

[0116] Step 4: Preparation of the intermediate methyl 4-[(3-cyclopropyl-4-oxo-3,4- dihydroquinazolin-2-yl)methyl]benzoate

[0117] To a solution of methyl 4-(2-{[2-(cyclopropylcarbamoyl)phenyl]amino}-2-oxoethyl)benzoate (3 gm, 8.52 mmol, 1 eq) in DMF (10 ml) was added ZnCh (0.58g, 4.26 mmol, 0.5 eq) followed by hexamethyl disilazane (5.34 mL, 25.56 mmol, 3 eq), and the reaction mixture was stirred at 120°C for 24 h. Then, the reaction mixture was poured into ice water and extracted wdth EtOAc (50 ml. x2). The organic layers w ere washed w ith winter (100 ml), dried over Na2SO4, and concentrated under reduced pressure to give crude residue. The crude compound was purified by column chromatography (eluted by 0 to 50% EtOAc in pet ether) and the desired fraction (Ry = 0.5, EtOAc / pet ether = 1 :1) was collected to afford the title compound as an off-white solid. (1 .7 g, 60% yield)

[0118] Step 5: Preparation of 4-{[3-Cyclopropylquinazolin-4(3H)-on-2-yl]methyl}-N- hydroxybenzamide (compound 6b)

[0119] To the ester intermediate ( 1g, 3.24 mmol) was added the solution of 2M NH2OH in anhydrous MeOH (20 ml) and NaOH (0.26 g, 6.49 mmol) at rt, and the reaction mixture was stirred for Ih. The reaction mixture was filtered through a pad of celite and washed with MeOH ( 10 ml). The filtrate was concentrated under reduced pressure to give crude residue. The crude product was purified by column chromatography (silica gel 100-200 mesh, eluted by 5%-20% MeOH in DCM) and the desired fraction (Rf= 0.3, MeOH / DCM = 1 / 9) was collected to afford target compound as a brown solid. (0.5 g, 50% yield) mp = 200-202 °C;1H NMR (600 MHz, DMSO-d6) δ 1 1.18 (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, <7= 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); 13C NMR (150 MHz, DMSO-rfe) 5 164.5, 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-ESIMS m / z [M + Hp calcd 336.1343 found 336.1332; HPLC Purity = 96.1% (IR = 2.6 min, eluted by Mobile Phase: B: 0.05%TFA in water. A: 0.05%TFA in Acetonitrile; Gradient (T / %A): 0 / 3, 8.5 / 100, 9.0 / 100, 9.5 / 3, 10 / 3; Column Temp; 50 °C, Flow rate: 0.55 ml / rnin; Diluent: ACN: H2O, Acquity BEH Cl 8 100mm x 2.1 mm. 1.7 pm).

[0120] Example 2: Preparation of 4-{[3-methylquinazolin-4(3H)-on-2-yl]methyl}-N-hydroxybenz.amide (compound 6a)

[0121] Step 1 to 4 (same as step 1 to 4 of Example 1): Preperation of the intermediate methyl 4- [(3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]benzoate

[0122] The title compound was prepared by using the similar procedure described above of the step 1 to 4 of example 1. Yellow solid (2.12 g, 56.7% yield). R / = 0.35 (EtOAc / n-Heptane = 3 / 2); mp = 142- 144 °C ; ‘H NMR (600 MHz, DMSO-d6) δ = 8. 11 (dd, J= 7.8, 0.6 Hz, 1H), 7.92 (d, <7= 7.8 Hz, 2H), 7.78 (m, 1H), 7.58 (d, J= 7.8 Hz, 1H), 7.49 (m, 1H), 7.43 (d, <7 = 8.4 Hz, 2H), 4.38 (s, 2H), 3.83 (s, 3H), 3.45 (s, 3H);I3C NMR (150 MHz, DMSCM,) 6 166.0, 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]+.

[0123] Step 5: Preparation of 4-{[3-methylquinazolin-4(3H)-on-2-yl]methyl}-N-hydroxybenzamide (compound 6a) The title compound was prepared by using the similar procedure described above of the step 5 of example 1. Brown solid (3.555 g, 65.4 %yield). mp = 228-230 °C;1H NMR (600 MHz. DM SO-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, IH), 7.51-7.48 (m, IH), 7.36 (d, J= 7.8 Hz, 2H), 4.33 (s, 2H), 3.45 (s, 3H); 13C NMR (150 MHz, DMSO-cAs) 8 164.0, 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+Hp : 310.1 1; HR-ESIMS m / z [M + H]+calcd 310.1186 found 310.1 177; HPLC Purity = 94.7% (tR= 2.3 min, eluted by Mobile Phase: B: 0.05%TFA in water. A: 0.05%TFA in Acetonitrile; Gradient(T / %A): 0 / 3, 8.5 / 100, 9.0 / 100, 9.5 / 3, 10 / 3 ; Column Temp: 50 °C, Flow rate: 0.55 ml / min; Diluent: ACN: H2O, Acquity BEH C18 100mm x 2.1 mm, 1.7 pm).

[0124] Example 3: Preparation of 4-{[3-cyclopropylmehtylquinazolin-4(3H)-on-2-yl]methyl}-N- hydroxybenzamide (compound 6c)

[0125] The title compound was prepared by using the similar procedure described above of the step 1 to 5 of example 1. solid.1H NMR (600 MHz, DMSO-cfc) 8 11.20 (brs, 1H), 9.05 (brs, 1H), 8.12 (d, J = 7.8 Hz, 1H), 7.81-7.78 (m. IH). 7.71 (d, J= 7.8 Hz. 2H). 7.59 (d, J = 8.4 Hz, 1H), 7.52-7.50 (m, IH). 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);I3C NMR (150 MHz, DMSO-cp) 5 161.7, 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 | calcd 350.1499 found 350.1490; HPLC Purity = 96% (tR= 3.2 min, eluted by Mobile Phase: B: 0.05%TFA in water, A: 0.05%TFA in Acetonitrile; Gradient(T / %A): 0 / 3, 8.5 / 100, 9.0 / 100, 9.5 / 3, 10 / 3; Column Temp: 50 °C, Flow rate: 0.55 ml / min; Diluent: ACN: H2O, Acquity BEH C18 100mm x 2.1 mm, 1.7 pm).

[0126] Example 4: Preparation of 4-{[3-phenylquinazolin-4(3H)-on-2-yl]methyl}-N-hydroxybenzamide (compound 6d)

[0127] Step 1; Preparation of 2-nitro-N-phenylbenzamide

[0128] To a solution of 2 -nitrobenzoic acid (12 g, 1 eq) in DCM (120 ml) was cooled to 0-5°C, and the solution was added oxalyl chloride ( 16.4 g, 1.8 eq) dropwise and a drop of DMF. The reaction mixture was allowed to stir and warm to rt for 2 h. Upon completion of reaction as indicated by tic, the solvent was evaporated in vacuo and used without further purification. To the aniline (8.85 g, 1.2 eq) and triethyl amine (9.65 g, 1.2 eq) were dissolved in DCM (70 ml) and cooled to 0-5°C, and the solution was added a solution of crude acyl chloride (14.70 g, 1 eq) in DCM (70 ml). The reaction mixture was allowed to warm to rt and stirred for 3 h. The reaction mixture was evaporated in vacuo and redissolved in DCM (50 ml). The organic layer was washed with sat. NaHCO.va.p (25 ml) and water (25 ml x 2). After dried over Na2SOa, the organic layer was concentrated under reduced pressure and purified by column chromatography. The desired fraction (Ry = 0.4, EtOAc / hexane = 3 / 7) was collected to afford target compound. (15.65 g, 90% yield)

[0129] Step 2 to 4: same as Example 1.

[0130] Step 5: Preparation of 4-{[3-phenylquinazolm-4(3H)-on-2-yl]methyl}-N-hydroxybenzamide (compound 6d)

[0131] The title compound was prepared by using the simi lar procedure described above of the step 1 to 5 of example 1. Light pink solid, mp = 167-169 °C;1H NMR (600 MHz, DMSO-d6) δ 1 1.15 (s, 1H), 8.98 (s, 1H), 8.11 (dd, J= 7.8 Hz, 1H), 7.86-7.83 (m, IH), 7.67 (d, .7= 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);I3C NMR (150 MHz, DMSO-d6) 5 163.9, 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+Hf : 372.14; HR-ES1MS m / z [M + H]+calcd 372.1343 found 372.1336; LC-MS = 98.6%. Example 5: Preparation of 4-[(6-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N- hydroxybenzamide (compound 6e)

[0132] Steps 1 to 4; Preparation of methyl 4-[(6-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-2- yl)methyl]benzpate

[0133] The title compound was prepared by using the similar procedure described above of the step 1 to 4 of example 1. White solid. (460 mg, 49% yield) (R<= 0.35, EtOAc / heptane = 1:1); mp = 173-175 °C; *H NMR (600 MHz, DMSO-) 6 8.02 (d, J= 2.4 Hz, IH), 7.92 (d, J= 8.4 Hz, 2H), 7.78 (dd, J = 9.0, 2.4 Hz, IH), 7.58 (d, J= 9.0 Hz, IH), 7.43 (d, J= 8.4 Hz, 2H), 4.37 (s, 2H), 3.83 (s, 3H), 3.45 (s, 3H);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]+.

[0134] Step 5: Preparation of 4-[( 6-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N- hydroxybenzflmide (compound 6e)

[0135] The title compound was prepared by using the similar procedure described above of the step 5 of example 1. White solid. (184 mg, 44% yield) R; = 0.31 (MeOH / DCM = 1 / 9); mp = 214-216 °C; *H NMR (600 MHz, DMSO-d6) δ 11.19 (s, IH), 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, . / = 9.0 Hz, 1H), 7.36 (d, J = 8.4 Hz, 2H), 4.33 (s, 2H), 3.45 (s, 3H);13C NMR (150 MHz, DMS0-<4) 8 164.0, 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]+calcd 344.0796 found 344.0787; HPLC purity = 95.5% (tR= 8.2 min, eluted by 0.1 %FA in ACN / 0.1%FA in H2O = 10 / 90 to 100 / 0, Kinetex® 3.5 pm XB-C18 100 A . LC Column 100 x 4.6 mm).

[0136] Example 6: Preparation of 4-[(6-chloro-3-cyclopropyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]- N-hydroxybenzamide (compound 6f)

[0137] Step 1 to 4: Preparaton of methyl 4-[(6-chloro-3-cyclopropyl-4-oxo-3,4-dihydroquinazolin-2- yl)methyl]ben~oate

[0138] The title compound was prepared by using the similar procedure described above of the step 1 to 4 of example 1. White solid. (1.26 g, 61 % yield) (Rf= 0.4, EtOAc / heptane = 1 : 1); mp = 161-163 °C; 1H NMR (600 MHz, DMSO-de) 3 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);13C NMR (150 MHz, DMSO-< / ..) 5 166.1, 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]+.

[0139] Step 5: Preparation of 4-[( 6-chloro-3-cyclopropyl-4-oxo-3,4-dihydro(iuinazolin-2-yl)methyl]- N-hydroxybenzamide (compound 6f)

[0140] The title compound was prepared by using the similar procedure described above of the step 5 of example 1. White solid. (90 mg, 11% yield) (R / = 0.2, MeOH / DCM = 1 / 9); mp = 240-243 °C (dec.); ‘H NMR (600 MHz, DMSO-Je) 8 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);13C NMR (150 MHz. DMSO-J6) 6 164.1, 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-ESIMS m / z [M + H]+calcd 370.0953 found 370.0942; HPLC purity = 95.6% (tR = 8.6 min, eluted by 0.1%FA in ACN / 0.1%FA in H2O = 10 / 90 to 100 / 0, Kinetex® 3.5 pm XB-C18 100 A , LC Column 100 x 4.6 mm).

[0141] Example 7: Preparation of 4-{[3-(4-chlorophenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]methyl}-N- hydroxybenzamide (compound 6h)

[0142] Step 1 to 4: Preparation of methyl 4-{[3-(4-chlorophenyl)-4-oxo-3,4-dihydroquinazolin-2- yl]methyl}benzoate

[0143]

[0144] The title compound was prepared by using the similar procedure described above of the step 1 to 4 of example 1. Brown solid. (1.34 g, 63% yield) (Rf = 0.37, EtOAc / heptane = 1 :1); mp = 169-171 °C;1H NMR (600 MHz, DMSO-ds) 6 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);13C NMR (150 MHz, DMSO-ck) 5 166.1, 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] +

[0145] Step 5: Preparation of 4-{[3-(4-cMorophenyl)-4-oxo-3,4-dihydroquinazoIin-2-yl]methyl}-N- hydroxybenzamide (compound 6h)

[0146] The title compound was prepared by using the similar procedure described above of the step 5 of Example 1. Orange solid. (200 mg, 25% yield) (R / = 0.14, MeOH / DCM = 5 / 95); mp = 152-155 °C; 1H NMR (600 MHz, DMSO-de) 8 11 .16 (s, 1H), 8.98 (s, 1H), 8. 11 (d, J = 7.8 Hz, 1H), 7.85 (m, 1H),

[0147] 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);13C NMR ( 150 MHz, DMSO-d6) δ 163.9, 161.5,

[0148] 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]+calcd 406.0953 found 406.0938; HPLC purity = 97.4% (tR = 8.8 min, eluted by 0.1 %FA in ACN / 0.1%FA in H2O = 10 / 90 to 100 / 0, Kinetex® 3.5 pm XB- C18 100 A , LC Column 100 x 4.6 mm). Example 8: Preparation of 4-{[3-(2-chlorophenyl)-4-oxo-3,4-dihydroqumazolin-2-yl]methyl}-N- hydroxybenzamide (compound 6g)

[0149] Step 1 to 4: Preparation of methyl 4-{[3-(2-chlorophenyl)-4-oxo-3,4-dihydroquinazolin-2- yl]methyl}benzoate

[0150] The title compound was prepared by using the similar procedure described above of the step 1 to 4 of example 1. Yellow solid. (1.46 g, 58% yield) (Rf= 0.22, EtOAc / heptane = 1:1); mp = 167-169 °C;1H NMR (600 MHz, DMSO-tfe) 5 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, 1 H), 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);3C 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]+.

[0151] Step 5: Preparation of 4-{[3-(2-chlorophenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]methyl}-N- hydroxybenzamide (compound 6g)

[0152] The title compound was prepared by using the similar procedure described above of the step 5 of example 1. Orange solid. (297 mg, 37% yield) (Rf = 0.08, MeOH / DCM = 5 / 95); mp = 137-140 °C; 'H NMR (600 MHz, DMSO-<4) 6 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);13C NMR (150 MHz, DMSO-tA) 8 163.9, 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-ESIMS m / z [M + H]+calcd 406.0953 found 406.0944; HPLC purity = 96.6% (tR = 8.7 min, eluted by 0.1 %FA in ACN / 0.1%FA in H2O = 10 / 90 to 100 / 0. Kinetex® 3.5 pm XB-C18 100 A , EC Column 100 x 4.6 mm). Example 9: Preparation of 4-{[3-(2,6-dimethylphenyl)-4-oxo-3,4-dihydroquinazoliii-2-yl]methyl}- N-hydroxybenzamide (compound 6i)

[0153] Step 1 to 4: Preparation of methyl 4-{[3-(2,6-dimethylphenyl)-4-oxo-3,4-dihydroquinazolin-

[0154] 2-yl]methyl}benzoate

[0155] The title compound was prepared by using the similar procedure described above of the step 1 to 4 of example 1. Yellow solid. (1.3 g, 68% yield) (R- = 0.53, EtOAc / heptane = 1:1 ); mp = 120-123 °C; ‘H 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);13C NMR (150 MHz, DMSO-A) 8 165.9, 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]+.

[0156] Step 5: Preparation of 4-{[3-(2, 6-dimethyIphenyl)-4-oxo-3,4-dihydroqiiinazolin-2-yl]methyl}- N-hydroxybenzamide (compound 6i)

[0157] The title compound was prepared by using the similar procedure described above of the step 5 of example 1. Yellow solid. (335 mg, 42% yield) (R / = 0.09, MeOH'DCM = 5 / 95); mp = 133-136 °C; 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, .7= 7,8 Hz, 1H), 7.59 (d. J= 8.4 Hz, 2H), 7.56 (m, IH), 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);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]+calcd 400.1656 found 400.1645; HPLC purity = 99% (tR= 8.9 min, eluted by 0.1 %F A in ACN / 0.1 %FA in H2O = 10 / 90 to 100 / 0, Kinetex® 3.5 um XB-C 18 100 A , LC Column 100 x 4.6 mm).

[0158] Example 10: Preparation of 4-[(6-fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methylJ-N- hydroxybenzamide (compound 6j)

[0159] Step 1 to 4: Preparation of methyl 4-[(6-fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-2- yl)methyl] benzoate

[0160] The title compound was prepared by using the similar procedure described above of the step 1 to 4 of example 1. Yellow solid. (1.55 g. 82% yield) (Rf = 0.35, EtOAc / heptane = 1: 1); mp = 155-157 °C; NMR (600 MHz, DMSO-<A) 5 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);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, 1 10.7, 110.6, 52.1, 41.0, 30.6; ESIMS(+) m / z 327.1 [M + H]+.

[0161] Step 5: Preparation of 4-[(6-fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl]-N- hydroxybenzamide (compound 6j)

[0162] The title compound was prepared by using the similar procedure described above of the step 5 of example 1. White solid. (280 mg, 35% yield) (Rf= 0.07, MeOH / DCM = 5 / 95); mp = 226-228 °C;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);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, 1 10.8, 1 10.7, 41.0, 30.7; HR-ESIMS m / z [M + H]+calcd 328.1092 found 328.1086; HPLC purity = 98% (tR= 7.3 min, eluted by 0.1%FA in ACN / 0.1%FA in H2O = 10 / 90 to 100 / 0, KINETEX® 3.5 pm XB-C18 100 Å , LC Column 100 x 4.6 mm).

[0163] Example 11: Preparation of N-hydroxy-4-((3-(4-methoxyphenyl)-4-oxo-3,4-dihydroquinazolin-2- yl)methyl)benzamide (compound 6k)

[0164] Step 1 to 4: Preparation of methyl 4-{[3-(4-methoxyphenyl)-4-oxo-3,4-dihydroquinazolin-2- yljmethyljbenzoate

[0165] The title compound was prepared by using the similar procedure described above of the step 1 to 4 of example 1. White solid. (1.26 g, 64% yield) (Rf = 0.32. EtOAc / heptane = 1:1); mp = 160-162 °C; NMR (600 MHz, DMSO-rfc) 5 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);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]+.

[0166] Step 5; Preparation otN-hydroxy-4-{[3-(4-methoxyphenyl)-4-oxo-3,4-dihydroquinazolin-2- yl]methyl}benzamide (compound 6k)

[0167] The title compound was prepared by using the similar procedure described above of the step 5 of example 1. Orange solid. (152 mg, 19% yield) (Rf = 0.11 , MeOH / DCM = 5 / 95); mp = 214-215 °C; 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, IH), 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);53C 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, 1 14.4, 55.4, 41.5; HR-ESIMS m / z [M + Hf calcd 402.1448 found 402.1436; HPLC purity = 99% (tR= 8.2 min, eluted by 0. 1%FA in ACN / 0.1%FA in H2O = 10 / 90 to 100 / 0, Kinetex® 3.5 pm XB-C18 100 A , LC Column 100 x 4.6 mm).

[0168] Example 12: Preparation of 4-[(6-cyano-3-methyl-4-oxo-3,4-dihydroqumazolin-2-yl)methyl]-N- hydroxybenzamide (compound 6m)

[0169] To the mixture of the chloro-substituted ester intermediate (703 mg, 2.05 mmol), NaCN (200 mg, 4.08 mmol), and NiBn (448 mg, 2.05 mmol) was added NMP (6 ml), and the reaction mixture was irradiated with microwave for 5 min. The mixture was partitioned between EtOAc / FhO (50 ml / 30 ml) and w-ashed with w ater (30 ml x2). The organic phase w as dried over MgSCL and concentrated in vacuo. The crude product was purified by column chromatography with EtOAc / heptane as the eluent and the desired fraction (R / = 0.09, EtOAc / heptane = 1 / 1) was collected to give acid intermediate. To the acid intermediate (210 mg, 0.658 mmol), EDCI (420 mg, 2.19 mmol), and HOBt (154 mg, 1.01 mmol) was dissolved in DMF (5 ml), and the mixture w'as stirred at ambient temperature. After 15 min, the mixture was added NEEOBn.ElCl (327 mg, 2.05 mmol) and DIPEA (0.36 ml, 2.06 mmol), and the reaction mixture was stirred for further 2 h. The mixture was then poured into water (100 ml) and the resulting precipitate was filtered to give orange solid (214 mg, 77% yield). To the suspension of the solid in dry DCM (10 ml) was added BBr3, dimethyl sulfide complex (IM in DCM, 1 ml) at 0°C, and the reaction mixture was stirred for 30 min. The mixture was quenched by water (10 ml) and washed by water (20 ml x2). The H2O phase was extracted with DCM (20 ml x3). All the organic phase was combined, dried over MgSCU. and concentrated in vacuo. The crude product was purified by column chromatography with DCM'MeOH as the eluent and the desired fraction (Rf= 0.08, MeOH / DCM = 5 / 95) was collected to give the titled compound as pink solid (24 mg, 43%);1H NMR (600 MHz, DMSO-d6) δ 11.19 (s, IH), 9.02 (s, 1H), 8.50 (s, 1H), 8.12 (d, 7= 8.4 Hz, 1H), 7.71 (m, 3H). 7.37 (d, <7 = 7.8 Hz, 2H), 4.36 (s, 2H), 3.47 (s, 3H);13C NMR (150 MHz, DMS0-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, 1 18.2, 108.7, 41.2, 30.8; HR-ESIMS m / z [M + H]+calcd 335.1139 found 335.1129; HPLC purity = 95% (tR= 7.1 min, eluted by 0.1%FA in ACN / 0.1%FA in H20 = 10 / 90 to 100 / 0, Kinetex® 3.5 gm XB-C18 100 A , LC Column 100 x 4.6 mm).

[0170] Scheme 2 Synthesis of 6n

[0171] Example 13: Preparation of 4-((3,4-dihydro-3-isopropyl-4-oxoquinazolin-2-yl)methyl)-N- hydroxy benzamide (6n)

[0172] The 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) were dissolved in pyridine (20 mL), and microwave irradiation 250W for 20 min. Propan-2 -amine hydrochloride (2.39 g, 30.62 mmol) was added and microwave irradiation 250W for 15min. The reaction mixture was concentrated in vacuo and added PdfOAch (0.21 g, 0.9 mmol), Xantphos (0.54 g, 0.9 mmol), Mo(CO)r (4.6g, 17.42 mmol), DMAP (3.14 g, 25.7 mmol) and DIPEA (4 mL, 22.49 mmol) were dissolved in DMAc / MeOH = 1 : 1 (20 mL) which already degassed. The the resulting 7 without isolation was stirred at 100 °C for 16 hours. After the reaction finished, diluted with EA and water, filtered by celite to get solution. The organic layer was dried over Mg Sth. filtered and concentrated in vacuo. The reside was purified by column ( 1% EA in DCM to 10% EA in DCM) to get the white solid 5n (2.4 g). The solution of 5n (2.4 g, 7.13 mmol) was dissolved in 2M NH2OH of MeOH (40 mL). The suspension solution was stirred for 16 hours and the reaction mixture was turned to clear solution. After the reaction finished, concentrated in vacuo. The reside was purified by column (10% MeOH in DCM) and the Rf= 0.3 was collected to get the pink solid 6n (1 . 15g, 16% of four steps).

[0173] 1H 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);13C NMR (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% (tR= 8.07 min)

[0174] Scheme 3 Synthesis of 61 The 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) were dissolved in pyridine (20 mL), and under microwave irradiation 250W for 20 min. Ethylamine hydrochloride (2.49 g, 30.62 mmol) was added and under microwave irradiation 250W for 15min. After the reaction finished, diluted with DCM and quenched with 3% HC1. The organic layer was dried over MgSCu, filtered and concentrated in vacuo to get colorless oil, then washed with EA / heplane to get white solid 8 (3. 14 g, 53%); The solution of 8 (1.5 g, 4.38 mmol) was added Pd(OAc)2 (79 mg, 0.35 mmol), Xantphos (0.41 g, 0.7 mmol), Mo(CO)c (1.74 g , 6.57 mmol), DMAP (1.78 g, 9.64 mmol) and DIPEA (1.53 mL, 8.77 mmol) were dissolved in DMAc / MeOH = 1 : 1 ( 15mL) which already degassed. The reaction mixture was stirred at reflux for 16 hours. After the reaction finished, diluted with EA and water, filtered by celite to get solution. The organic layer was dried over MgSCfi, filtered and concentrated in vacuo. The reside was purified by column (DCM) and the Rf= 0.3 was collected to get the white solid 51 (1.02 g, 71%).

[0175] 1H NMR (600 MHz, DMSO-d6) 58.28 (d, 7 = 8.1 Hz, H), 8.01 (d, J = 8.1 Hz, 2H), 7.76 (t. J = 7.92 Hz, 1H), 7.70 (d, <7 = 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)

[0176] Example 14: Preparation of 4-((3-ethyl-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-N- hydroxybenzamide (61)

[0177] The solution of 51 (1.6 g, 0.5 mmol) was dissolved in 2M NH2OH of MeOH (30 mL). The suspension mixture was stirred for 16 hours and the mixture was turned to clear solution. After the reaction finished, the mixture was concentrated in vacuo. The reside was purified by column ( 10% MeOH in DCM) and the Rf= 0.32 was collected to get the pink solid 61 (0.73 g, 45%).

[0178] 'H 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, 7.86 Hz, 1H), 7.40

[0179] (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);13C NMR (150 MHz, tfc-dmso) 513.99, 41.19, 120.61, 126.59, 127.11, 127.36, 127.75, 129.14, 131.89, 134.81,

[0180] 139.89, 147.31, 155.84, 161.62, 164.46; HPLC purity = 97.7%

[0181] The solution of 2-amino-5 -fluorobenzoic acid (2.0 g, 12.89 mmol), 2-(4-broinophenyl)acelic acid (2.77g, 12.89 mmol) and triphenyl phosphite (4 mL, 15.47 mmol) were dissolved in pyridine (20 mL), and refluxed for 15 hours. Ethylamine hydrochloride (1.4 g, 18.05 mmol) was added and then was refluxed for 15 hours. After the reaction finished, diluted with DCM and quenched with 3% HCI.

[0182] The organic layer was dried over MgSCfi. filtered and concentrated in vacuo to get colorless oil, then washed with EA / heptane to get white solid 9 (3.48 g, 75%)

[0183] 1H 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) methyl 44(3-ethyl-6-fluoro-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)benzoate (5o)

[0184] The 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 mmo1), DMAP (1.87 g, 15.28 mmol) and DIPEA (2.42 mL, 13.89 mmol) were dissolved in DMAc / MeOH = 1: 1 (20 mL) which already degassed. The reaction mixture was stirred at reflux for 24 hours. After the reaction finished, diluted with EA and water, filtered through celite to get solution. The organic layer was dried over MgSO4. filtered and concentrated in vacuo. The reside was purified by column (1% EA in DCM to 10% EA in DCM), then the Rf= 0.39 layer was collected to get the white solid 5o (0.93 g, 40%).

[0185] 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)

[0186] Example 15: Preparation of 4-((3-ethyI-6-fluoro-4-oxo-3,4-dihydroquinazolin-2-yl)methyl)-N- hydroxybenzamide (6o)

[0187] The solution of 5o (0.45 g, 0.13 mmol) was dissolved in 2M NH2OH of MeOH ( 10 mL). The suspension solution was stirred for 16 hours and the reaction mixture was turned to clear solution. After the reaction was finished, the mixture was concentrated in vacuo. The reside was purified by column (10% MeOH in DCM to 20% MeOH in DCM), then the Rf= 0.3 layer was collected to get the pink solid 60 (0.25 g, 55%).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% ( / R = 6.6 min)

Claims

CLAIMSWhat is claimed is:1 . A compound of F ormula (I) :or a pharmaceutically acceptable salt thereof, whereinR1is hydrogen, trifluoromethyl, (C1-5) alkyl, (C3-5) cycloalkyl, (C1-6) heterocycloalkyl, arylalkyl, substituted phenyl, or substituted heteroaryl;R2is hydrogen, halogen, trifluoromethyl, cyano, (C1-5) alkyl, (C1-6) alkoxy, (C1-6) heterocycloalkyl, arylakyl, substituted phenyl, or substituted heterocycloaryl;R3is hydrogen, halogen, trifluoromethyl, cyano, (C1-5) alkyl, (C1-6) alkoxy, (C1-6) heterocycloalkyl, arylalkyl, substituted phenyl, or their substituted analogues;R4is hydrogen, halogen, trifluoromethyl, cyano, (C1-5) alkyl, (C1-6) alkoxy, (C1-6) heterocycloalkyl, arylalkyl, substituted phenyl, or their substituted analogues:R5is hydrogen, halogen, trifluoromethyl, cyano, (C1-5) alkyl, (C1-6) alkoxy, (C1-6) heterocycloalkyl, arylalkyl, substituted phenyl, or their substituted analogues;R6is hydrogen, halogen, or hydroxyl alkoxy;The moietyis selected from the group consisting of or a pharmaceutically acceptable salt, hydrate,or prodrug thereof.

2. The compound or the pharmaceutically acceptable salt thereof of claims 1 , whereinR1is -CH3, -CH2CH3, -isopropyl, -cyclopropyl, -cyclopropylmethyl, -C6H5. -(4-C1)C6H5, -(2- C1)C6H5, -2,6-dimethylphenyl. or -(4-C)Me)CeH5.

3. The compound or the pharmaceutically acceptable salt thereof of claim 1, wherein R1is -CHs or - CH2CH3.

4. The compound or the pharmaceutically acceptable salt thereof of claims 1 , wherein R3is -CL -F or -CN.

5. The compound or the pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 4, wherein the moiety6. A compound as claimed in any one of claims 1 to 5, wherein the compound is selected from the group consisting of:

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

8. Use of a compound or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 6, or use of a pharmaceutical composition as claimed in claim 7 in the manufacture of a medicament for treating, alleviating, ameliorating, and / or decreasing severity of an HDAC6- associated disease or condition in a subject in need thereof.

9. The use as claimed in claim 8, wherein the HDAC6-associated disease or condition is selected from the group consisting of fibrosis, a neurodegenerative disease, a kidney disease, cancer, and an increase in cyst growth metrics.

10. The use as claimed in claim 9, wherein the fibrosis, associated with HDAC6 activity, is pulmonary fibrosis, idiopathic pulmonary fibrosis, hepatic fibrosis, renal fibrosis, or myelofibrosis.

11. The use as claimed in claim 9, wherein the neurodegenerative disease, associated with HDAC6 activity', is Huntington’s disease, Alzheimer’s disease, Parkinson’s disease, or Amyotrophic lateral sclerosis.

12. The use as claimed in claim 9, wherein the kidney disease, associated with HDAC6 activity, is polycystic kidney disease, or autosomal dominant polycystic kidney disease, or autosomal recessive polycystic kidney disease.

13. The use as claimed in claim 9, wherein the cancer, associated with HDAC6 activity, is lung, breast, renal, liver cancer, multiple myeloma, or glioma.

14. The use as claimed in claim 8, wherein the compound or pharmaceutically acceptable salt thereof, or the pharmaceutical composition is used in the manufacture of a medicament for decreasing the growth of cyst metrics, said growth of cyst metrics associated with HDAC6 activity.

15. The use as claimed in claim 9, wherein the compound or pharmaceutically acceptable salt thereof, or the pharmaceutical composition is used in the manufacture of a medicament for treating, alleviating, ameliorating, and / or decreasing severity of the condition of the increase in cyst growth metrics, said increase in cyst growth metrics associated with HDAC6 activity.

16. The use as claimed in claim 8, wherein the HDAC6-associated disease or condition is selected from the group consisting of hepatic 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. A compound as claimed in claims 1 , selected from the group consisting of:4-{[3-methylquinazolin-4(3H)-on-2-yl]methyl}-N-hydroxybenzamide (6a);4- {[3-Cyclopropylquinazolin-4(3H)-on-2-yl]methyl} -N-hydroxybenzamide (6b);4- { [3-cyclopropylmehtyIquinazolin-4(3H)-on-2-y I jmethyl } -N-hydroxybenzamide (6c) ;4- {[3-phenylquinazolin-4( 3 H)-on-2 -y I] methyl } -N -hydroxy benzamide ( 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-dihydroquinazoIin-2-yI]methyl} -N-hydroxybenzamide (6g);4- {[3-(4-chlorophenyl)-4-oxo-3,4-dihydroquinazolin-2-yl]melhyl} -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-dihydroqumazolin-2-yl)methyl)benzamide (6k); 4-[(3-ethyl-4-oxo-3,4-dihydroquinazoIin-2-yl)methyl]-N-hydroxybenzamide (61);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 a pharmaceutically acceptable salt thereof.

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