CN105439972A

CN105439972A - Heat shock protein inhibitor and preparation method and application thereof

Info

Publication number: CN105439972A
Application number: CN201510425553.6A
Authority: CN
Inventors: 张健存; 陈超南; 孙建
Original assignee: Guangzhou Institute of Biomedicine and Health of CAS
Current assignee: Guangzhou Institute of Biomedicine and Health of CAS
Priority date: 2015-07-17
Filing date: 2015-07-17
Publication date: 2016-03-30

Abstract

The present invention discloses a heat shock protein inhibitor and a preparation method and an application thereof, and belongs to the technical field of medicinal chemistry. The heat shock protein inhibitor has the structure features of a formula I. The compound can inhibit activity of the heat shock protein 90, and then can be used for preparing anti-tumor drugs.

Description

Heat shock protein inhibitors and its preparation method and application

Technical field

The present invention relates to field of pharmaceutical chemistry technology, particularly relate to a kind of heat shock protein inhibitors and its preparation method and application.

Background technology

Heat shock protein 90 (HSP90) is a kind of ATP dependence protein, and the HSP90 expressed in normal cell accounts for the 1-2% of total protein in cell, and wherein 3% plays a part to regulate nucleus function in nucleus.Under stress, as in tumour, the expression of HSP90 is increased to the 4-6% of whole cell protein group.Mankind HSP90 comprises three family: HSP90A (being present in the HSP90 α 1 in endochylema, HSP90 α 2, HSP90 β), HSP90B (be present in the glucose associated protein in endoplasmic reticulum---GRP94) and TPAR (be present in the tumor necrosis factor receptor proteins 1 in plastosome---TRAP1).75kDA-HSP90N has myristoylation signal, and link with RAF-1 in the mode that correlated series Ras in reversion rate virus relies on, it is construed to a newcomer of HSP90 family.Wherein be made up of 730 and 724 amino acid respectively the HSP90 α of two kinds of distinct genetic expressions and HSP90 β, the protein sequence of 86% has similarity.

HSP90 monomer comprises three structural domains: N-end structure, intermediate field, C-end structure.N-end and C-end all include the different binding sites to HSP90, and there is special functionally active each pole.The energy that Function protein matter can be provided to fold and transport when ATP is attached to N-end, simultaneously nucleotide bond is incorporated into C-end and can assists ATP allosteric thus make ATP be bonded to N-end.The dimerization of C-terminal regulatory HSP90.The interaction of region intermediate regulation and control client protein, can combine with other molecular chaperones and other albumen, and it also contains the γ-phosphoric acid binding site of an ATP simultaneously.Most HSP90 inhibitor is incorporated into N-end, and small part is incorporated into C-end.

Under stress, as in tumour, the expression of HSP90 is increased to the 4-6% of whole cell protein group.In kinds cancer, HSP90 and some co-chaperone are high expression level, and in normal cell, HSP90 is in silent status, and is in active state in cancer cells.HSP90 client protein known at present more than 200, as EGFR, HER2, AKT, CDK4, MET etc.These client protein relate to the transmission of signal, the transhipment of protein, the maturation of signal, congenital and adaptive immunity.And most oncogene belongs to HSP90 client protein, as the fusion protein BCR-ABL etc. in the EGFR in non-small cell lung cancer cell, the HER2 in mammary cancer, chronic myelognous white corpuscle.Oncogene ErBB2 is the substrate of HSP90 partner complexes, plays an important role to the generation of cancer, survival, metabolism and vasculogenesis.Therefore, suppress HSP90 that ErBB2 will be caused to dissociate from mixture, produce lower tonal signal thus the growth of inhibition tumor cell and vasculogenesis.Scr family (v-Scr and c-Scr) in TYR can gather with HSP90, and it can cause sarcoma of uterus virus.Intracellular signaling after the cell substrate of factors stimulated growth and mediated by integrin sticks is relevant with it.The activity of Scr regulates and controls by HSP90, to the deterioration of tumour with shift relevant.

HSP90 inhibitor can block multiple paths that cancer cells is formed simultaneously, thus avoids using single inhibitor to produce resistance for during a certain path tumour cell treatment.Inhibitor based on HSP90 also will not act on by kinases itself, but the associated molecule companion of maintenance kinase activity conformation is inhibited, thus a large amount of substrate client protein enzyme body is degraded.Compared with traditional kinase inhibitor, HSP90 inhibitor can be degraded multiple similar kinase whose level simultaneously, can play restraining effect widely.Experiment shows that the single or drug combination of HSP90 inhibitor all has antineoplastic action.Therefore, HSP90 is the potential target of one of cancer therapy, and exploitation target HSP90 inhibitor is rational choice.

Summary of the invention

Based on this, the invention provides a kind of new heat shock protein inhibitors, this compounds can suppress the activity of heat shock protein 90, and then may be used for preparing in anti-tumor drug.

There is heat shock protein inhibitors or its pharmacy acceptable salt of formula I constitutional features:

Wherein:

R ₁, R ₇, R ₈independently be selected from: H, C ₁-C ₆alkyl, C ₁-C ₆unsaturated alkyl, halogen, hydroxyl, C ₁-C ₆alkoxyl group, NHCOOR, SO ₂nHR, NHSO ₂r, CN, NHCOR, CONHR;

R is selected from: C ₁-C ₆alkyl, C ₁-C ₆unsaturated alkyl;

R ₂, R ₃independently be selected from: H, D, C ₁-C ₆alkyl, C ₁-C ₆unsaturated alkyl, C ₃-C ₈cycloalkyl, phenyl, the phenyl of replacement, heteroaryl, acyl group;

R ₄be selected from: H, D, C ₁-C ₆alkyl, C ₁-C ₆unsaturated alkyl, C ₃-C ₈cycloalkyl, substituted or non-substituted aryl, substituted or non-substituted heteroaryl, (CR ₆r ₉) _nr ₅;

N is selected from: 1,2 or 3;

R ₅be selected from: amino, the amido of replacement, substituted or non-substituted amide group.

R ₆, R ₉independently be selected from: H, D, C ₁-C ₆alkyl, C ₃-C ₈cycloalkyl;

X, Y, Z are independently selected from: CH, N, O.

The above-mentioned compound with formula I constitutional features, for the inhibitor of heat shock protein 90, and heat shock protein 90 is that ATP is according to resistant protein, after HSP90 inhibitor occupies ATP-binding site, cause client protein abnormal conformation, it can be removed by Ubiquitin-proteasome path fast by cell, thus plays antitumous effect.

Wherein in an embodiment, described X, Y, Z are selected from following group:

Wherein in an embodiment, described heat shock protein inhibitors is selected from the compound of following general formula I I:

Wherein:

R ₇, R ₈be selected from: H, C ₁-C ₆alkyl, C ₁-C ₆unsaturated alkyl, halogen, hydroxyl, C ₁-C ₆alkoxyl group, CN;

R ₁be selected from: C ₁-C ₆alkyl, C ₁-C ₆unsaturated alkyl, C ₁-C ₆alkoxyl group.

Wherein in an embodiment, described R ₄be selected from: substituted or non-substituted aryl, substituted or non-substituted heteroaryl, C ₃-C ₈cycloalkyl, (CR ₆r ₉) _nr ₅,

Wherein: R ₆, R ₉independently be selected from: H, C ₁-C ₆alkyl, and this R ₆, R ₉h can not be selected simultaneously;

R ₅be selected from: amino, C ₁-C ₆saturated alkyl amine, substituted or non-substituted saturated heterocyclic amine, substituted or non-substituted unsaturated heterocyclic amine, substituted or non-substituted C ₁-C ₆cyclic alkyl amides base, substituted or non-substituted C ₁-C ₆alkylamidoalkyl, substituted or non-substituted aryl amido group, substituted or non-substituted benzo aryl amido group, substituted or non-substituted heteroaryl amide base, substituted or non-substituted benzo heteroaryl amide base, substituted or non-substituted heterocyclyl amides base, substituted or non-substituted aralkyl amido, substituted or non-substituted sulfoamido, substituted or non-substituted saturated heterocyclyl amide group, substituted or non-substituted C ₁-C ₆alkyl urea;

N is selected from: 1.

Wherein in an embodiment, described R ₂, R ₃independently be selected from: H, C ₃-C ₈cycloalkyl, C ₁-C ₆alkyl, C ₁-C ₆unsaturated alkyl.

Wherein in an embodiment, described heat shock protein inhibitors is selected from following compound:

5-(2,4-dihydroxyl-5-isopropyl phenyl)-4-(3-morpholino third-1-alkynes-1-base) isoxzzole-3-methane amide;

N-cyclopropyl-5-(2,4-dihydroxyl-5-isopropyl phenyl)-4-(3-morpholino third-1-alkynes-1-base) isoxzzole-3-methane amide;

N-cyclopropyl-5-(2,4-dihydroxyl-5-isopropyl phenyl)-4-(phenylene-ethynylene) isoxzzole-3-methane amide;

4-((4-(tertiary butyl) phenyl) ethynyl)-5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(phenylene-ethynylene) isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-((4-fluorophenyl) ethynyl) isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(pyridine-3-ethynyl) isoxzzole-3-methane amide;

4-(cyclohexyl-acetylene base)-5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl isoxzzole-3-methane amide;

4-(cyclopentyl ethynyl)-5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl isoxzzole-3-methane amide;

4-(cyclopropyl acethlene base)-5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-morpholino third-1-alkynes-1-base) isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-(4-(methoxyl group imido grpup) piperidin-1-yl) third-1-alkynes-1-base) isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-4-(3-((2S, 6R)-2,6-thebaine) the third-1-alkynes-1-base)-N-ethyl isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-4-(3-(1,1-sulphur dioxide is for morpholino) third-1-alkynes-1-base)-N-ethyl isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-thiomorpholine generation the third-1-alkynes-1-base) isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-(4-(6-(trifluoromethyl) nicotinoyl amido) piperidin-1-yl) the third-1-alkynes-1-base) isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-(4-(6-(trifluoromethyl) nicotinoyl) piperazine-1-base) the third-1-alkynes-1-base) isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-(4-(trifluoromethyl) benzoylamino) third-1-alkynes-1-base) isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-(6-(trifluoromethyl) nicotinoyl amido) third-1-alkynes-1-base) isoxzzole-3-methane amide;

4-(3-(2-naphthoyl amido) third-1-alkynes-1-base)-5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-(quinoline-3-formamido-) third-1-alkynes-1-base) isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-methyl-3-(sulfonyloxy methyl amido) fourth-1-alkynes-1-base) isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-methyl-3-(4-Methyl benzenesulfonyl amido) fourth-1-alkynes-1-base) isoxzzole-3-methane amide;

4-(3-acetylaminohydroxyphenylarsonic acid 3-methyl fourth-1-alkynes-1-base)-5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl isoxzzole-3-methane amide;

4-(3-(cyclopropyl carboxamide base)-3-methyl fourth-1-alkynes-1-base)-5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl isoxzzole-3-methane amide;

4-(3-(cyclopentyl formamido-)-3-methyl fourth-1-alkynes-1-base)-5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl isoxzzole-3-methane amide;

4-(3-(cyclohexyl formamido-)-3-methyl fourth-1-alkynes-1-base)-5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-(3-isopropylureido)-3-methyl fourth-1-alkynes-1-base) isoxzzole-3-methane amide;

N-(4-(5-(2,4-dihydroxyl-5-isopropyl phenyl)-3-(ethamine formyl radical) isoxzzole-4-base)-2-methyl fourth-3-alkynes-2-base) morpholine-4-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-(furans-3-formamido-)-3-methyl fourth-1-alkynes-1-base) isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-methyl-3-((4-(trifluoromethyl) phenyl) amido) fourth-1-alkynes-1-base) isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-methyl-3-(4-(trifluoromethyl) benzamido) fourth-1-alkynes-1-base) isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-methyl-3-(picoline amide group) fourth-1-alkynes-1-base) isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-methyl-3-(6-methylnictotinyl amido) fourth-1-alkynes-1-base) isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-methyl-3-(quinoline-3-formamido-) fourth-1-alkynes-1-base) isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-(6-methoxypyridine amide group)-3-methyl fourth-1-alkynes-1-base) isoxzzole-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-methyl-3-(6-(trifluoromethyl) picolinamide base) fourth-1-alkynes-1 isoxzzole-Ji)-3-methane amide;

5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-(6-fluorine nicotinoyl amido)-3-methyl fourth-1-alkynes-1-base) isoxzzole-3-methane amide.

The invention also discloses above-mentioned heat shock protein inhibitors or the preparation method of its pharmacy acceptable salt, adopt following circuit combination:

Wherein: R ₁₀be selected from: benzyl, trimethylsilyl, three second are silica-based, t-Butyldimethylsilyl, and tert-butyl diphenyl is silica-based, di-tert-butyl methylsilyl, methyl, ethanoyl, to methoxy-benzyl, and methoxymethyl.

A is selected from: halogen.

The invention also discloses above-mentioned heat shock protein inhibitors or its pharmacy acceptable salt, prepare prevention and therapy have expression of heat shock protein 90 increase pathological characteristics disease medicine in application.

Wherein in an embodiment, described in the disease of pathological characteristics that there is expression of heat shock protein 90 increases be: cancer, myelodysplastic syndrome, systemic mastocytosis, Xi Peier-lindau's syndrome, multicenter type Castleman is sick, psoriatic.

Described cancer comprises bladder cancer, colorectal carcinoma, liver cancer, lung cancer, mammary cancer, carcinoma of vagina, ovarian cancer, carcinoma of the pancreas, kidney, cancer of the stomach, gastrointestinal cancer, prostate cancer, head and neck cancer, abdominal cavity cancer, thyroid carcinoma, osteocarcinoma, the cancer of the brain, central nervous system cancer, hematologic cancers.

The invention also discloses a kind of pharmaceutical composition, comprise above-mentioned heat shock protein inhibitors or its pharmacy acceptable salt, and the acceptable vehicle of pharmacy or carrier.

Compared with prior art, the present invention has following beneficial effect:

Novel heat shock protein inhibitors of the present invention, compared with inhibitor similar with routine, remain in original molecule with the interactional atom of HSP90 receptor protein amino-acid residue and group, introduce the phenyl ring in the alkynes replacement original molecule with bioisostere effect, thus retain the effect of parent compound, make it have following advantage:

1. suppress HSP90 α and HSP90 β two enzyme acceptors, and alkynyl chain more former phenyl ring chain in C-4 position is spatially closer to the amino-acid residue of receptor protein, is easier to be combined with receptor protein simultaneously.

2. the HSP90 α of two kinds of distinct genetic expressions and HSP90 β is made up of 730 and 724 amino acid respectively, and the protein sequence of 86% has similarity.Therefore such inhibitor is effective to most cancer patientss.

3., compared with single inhibitor, use two medicines acting on single target spot respectively to compare, the medicine patient acting on two target spots uses more convenient, can also avoid the interaction of medicine and medicine simultaneously.

The preparation method of heat shock protein inhibitors disclosed by the invention, have raw material and be easy to get, mild condition, productive rate is high, is separated simple, is suitable for industrialized advantage.

Embodiment

Below in conjunction with embodiment, the present invention is described further, but do not cause any restriction to the present invention.

Compound of the present invention and salt thereof also can by becoming known for the method preparation of preparative chemistry related compound, and the raw material related in an embodiment all obtains by the similar approach of prior art.

The straight line pointed to outward in phenyl ring by phenyl ring in general formula I of the present invention represents the unfixed replacement in position.

" alkyl " refers to saturated hydrocarbyl, comprises the alkyl of straight or branched, as C ₁-C ₆alkyl refers to have the saturated straight chain of 1 to 6 carbon atom or the alkyl of side chain, and wherein the example of straight chain saturated alkyl includes but not limited to ethyl, n-propyl etc., and the example of saturated branched-chain alkyl includes but not limited to sec.-propyl, the tertiary butyl etc.; " unsaturated alkyl " refers to the alkyl with alkenyl or alkynyl, comprise the unsaturated alkyl of straight or branched, wherein the example of unsaturated straight chained alkyl includes but not limited to vinyl, propenyl etc., and the example of unsaturated side chain alkyl includes but not limited to 2-methylpropenyl etc.; " cycloalkyl " refers to the alkyl with ring texture, as C ₃-C ₈cyclic alkyl refers to have the saturated of 3 to 8 carbon atoms or the undersaturated alkyl with ring texture, wherein the example of saturated cyclic alkyls includes but not limited to cyclopropyl, cyclopentyl, ethyl substituted cyclohexyl etc., the example of unsaturated cyclic alkyl includes but not limited to cyclopentenes etc., preferred C in the present invention ₃-C ₆ring-type alkane.

" replacement " refers to the hydrogen base with specifying in substituent group displacement ad hoc structure.When the substituting group that position can be selected from designated group through more than more than in any ad hoc structure replaces, described substituting group can be identical or different in each position.As used herein, term " replacement " expection includes all of organic compounds and allows substituting group.One extensively in, described allow substituting group to include organic compounds non-annularity and ring-type, branch and non-branch, carbocyclic ring and heterocycle, aromatic series and non-aromatic substituting group.The heteroatomss such as such as nitrogen can have any substituting group that allows of hydrogen substituting group and/or organic compound as herein described, and described substituting group meets heteroatomic valence mumber.

" heteroaryl " refers to the 4n+2 aromatic nucleus system containing 5-6 unit monocycle 6, and aromatic ring system has ring carbon atom and 1-4 ring hetero atom, and wherein each ring hetero atom is independently selected from nitrogen, oxygen and sulphur.Comprise the heteroaryl of one or more nitrogen-atoms, tie point can be carbon or nitrogen-atoms, allows if cross chemical valence.The multi-loop system of heteroaryl can comprise one or more heteroatoms." heteroaryl " also comprises described heteroaryl ring-member as defined above, and condensed one or more carbocyclic ring or heterocyclic group, wherein tie point is on heteroaryl ring, and in this case, the quantity of ring members only includes the number of member on heteroaryl ring system." heteroaryl " also comprises described heteroaryl ring-member as defined above, condense one or more aromatic yl group, wherein tie point can be on aryl or heteroaryl ring, and in this case, the quantity of ring members only is included in the ring members number in fused polycycle (aryl/hetaryl) member ring systems.Polyheteroaromatic, one of them is not containing heteroatomic ring (such as, indyl, quinolyl, carbazyl, and analogue), its tie point can be on any ring, that is, can be to comprise on heteroatomic ring (such as, 2-indyl) or do not containing on heteroatomic ring (such as, 5-indyl).Include but not limited to containing 2 heteroatomic exemplary 5 yuan of heteroaryls, imidazoles, pyrazoles, oxazole, isoxazolyl, thiazolyl, and isothiazole.Include but not limited to containing 1 heteroatomic exemplary 6 yuan of heteroaryl, pyridyl.Include but not limited to containing 2 heteroatomic exemplary 6 yuan of heteroaryls, pyridazinyl, pyrimidyl, and pyrazinyl.

" heterocyclic radical " refers to cyclic alkyl as herein defined, and wherein main chain also comprises one or more heteroatomss (as oxygen, sulphur, nitrogen, boron, silicon, phosphorus).

" amido " refer to the hydrogen atom of ammonia replaced by alkyl after organic compound, as heterocyclic amine, aralkyl amido etc.

" amide group " refers to that the hydrogen of ammonia (or amine) is by the derivative of acyl substituted, as cyclic alkyl amides base, alkylamidoalkyl, aryl amido group, benzo aryl amido group, heteroaryl amide base, benzo heteroaryl amide base, heterocyclyl amides base, sulfoamido, alkyl urea etc.

Following examples are prepared with reference to following reaction scheme,

Reaction conditions: (a) cylite, salt of wormwood, in acetonitrile, backflow is spent the night; (b) MePPh3I, potassium tert.-butoxide, tetrahydrofuran (THF), 0 DEG C-room temperature, overnight; (c) H ₂(10atm), 10% palladium carbon, acetic acid, ethanol, room temperature, 2 days; (d) acetic acid, 46% boron trifluoride ether solution; (e) cylite, salt of wormwood, in acetonitrile, backflow is spent the night; (f) oxalic acid diethyl ester, sodium Metal 99.5, ethanol, room temperature, 4h; (g) oxammonium hydrochloride, ethanol, room temperature; (h) hydronium(ion) Lithium Oxide 98min, tetrahydrofuran (THF)-methanol-water, room temperature, cyclopropylamine, EDCI, HOBt, methylene dichloride room temperature; I () carbinolamine solution, spends the night by 60 DEG C; (j) 70% ethylamine solution, alcohol reflux, 5h; (k) N-iodosuccinimide, ceric ammonium nitrate, acetonitrile backflow is spent the night; (l) terminalalkyne, two triphenylphosphine palladium, cuprous iodide, triethylamine or diisopropylamine, DMF, spends the night by 55 DEG C; The dichloromethane solution of (m) 1M boron tribromide, methylene dichloride, 0 DEG C of-rt, 1h.

Embodiment 1

The preparation of 5-(2,4-dihydroxyl-5-isopropyl phenyl)-4-(3-morpholino third-1-alkynes-1-base) isoxzzole-3-methane amide.

According to above-mentioned circuit combination, method steps is as follows:

Step a) prepares Compound I-2, i.e. 1-(two (benzyloxy) phenyl of 2,4-) ethyl ketone.

2 are added in the 500mL two-mouth bottle that reflux condensing tube is housed, 4-resacetophenone I-1 (9.12g, 60.00mmol), salt of wormwood (21.00g, 151.00mmol), acetonitrile 200mL in oil bath pan at 80 DEG C reflux 1h, then inject cylite (14.70mL, 144.00mmol) with syringe and continue backflow and spend the night.After TLC detection reaction to starting raw material reacts completely, be cooled to room temperature, Büchner funnel suction filtration, washed with dichloromethane resistates, collect filtrate, revolve and steam except desolventizing obtains pale tan oil.In oily matter, add ether, stir and produce white flock precipitate, sand core funnel suction filtration then washed with diethylether, repeat above operation and obtain white solid 15.80g for twice, be Compound I-2, yield is 79%.Mass spectrum monitoring ESI-MSm/z:333.0 (M+H) ⁺.

Step b) prepare Compound I-3, i.e. (((4-third-1-alkene-2-base)-1,3-(dioxygen base)) two (methylene radical)) hexichol.

Potassium tert.-butoxide (4.72g, 42.15mmol) is added, triphenyl phosphorus methyl iodide (17.88g, 42.15mmol), at 0 DEG C and N in the 250mL two-mouth bottle that dropping funnel is housed ₂inject anhydrous THF (tetrahydrofuran (THF)) under provide protection and stir 1h.Then by 1-(2, two (benzyloxy) phenyl of 4-) ethyl ketone I-2 (10.76g, 32,42mmol) be dissolved in completely and inject dropping funnel in anhydrous THF and be slowly added dropwise to reaction system through dropping funnel again, after dropping completely, stir 1h, then will room temperature for overnight be reacted on.After TLC monitoring starting raw material reacts completely, revolve and steam removing reaction solvent, acetic acid ethyl dissolution gained resistates, extracts three times with water, and saturated sodium-chloride water solution extraction once, collect ethyl acetate layer, use anhydrous sodium sulfate drying again, revolve and steam removing organic solvent, obtain white solid 8.45g finally by silica column purification, be Compound I-3, yield 79%.

The characterization data of this Compound I-3 is: ¹hNMR (400MHz, CDCl ₃, δ _ppm): 7.38-7.28 (m, 10H), 7.13 (d, J=8.4Hz, 1H), 6.59 (s, 1H), 6.52 (d, J=8.4Hz, 1H), 5.08 (s, 2H), 5.03 (s, 2H), 5.00 (s, 2H), 2.12 (s, 3H).

Step c) prepare Compound Compound I-4, i.e. 4-isopropyl benzene-1,3-glycol.

In the autoclave of 2L, add I-3 (350.00g, 1.06mol), the 10% palladium carbon of 35g, 1mL formic acid, 1L ethanol, pass into hydrogen and be heated to 78 DEG C of maintenances two days.Being cooled to room temperature after reacting completely, by reaction solution with diatomaceous Büchner funnel suction filtration being housed, washing with alcohol, collecting filtrate, revolve and steam removing organic solvent, finally obtain white solid 130g with silica column purification, be Compound I-4, yield 80%.

The characterization data of this Compound I-4 is: ¹hNMR (400MHz, CDCl ₃, δ _ppm): 7.01 (d, J=8.4Hz, 1H), 6.39 (d, J=8.4Hz, 1H), 6.32 (s, 1H), 5.50 (brs, 1H), 5.33 (brs, 1H), 3.12 (m, 1H), 1.21 (d, J=6.4Hz, 6H).

Steps d) prepare Compound I-5, i.e. 1-(2,4-dihydroxyl-5-isopropyl phenyl)-ethyl ketone.

First in the 250mL two-mouth bottle being provided with reflux condensing tube, 4-isopropyl benzene-1 is added, 3-glycol I-4 (5.36g, 35.25mmol), then under the provide protection of nitrogen, inject 100mL47% boron trifluoride ether solution stir after half an hour, finally inject 4.03mL acetic acid in oil bath pan heated overnight at reflux, TLC monitors after starting raw material reacts completely and is cooled to room temperature, the sodium acetate aqueous solution adding 70mL10% stirs 2h, be diluted with water to 250mL, add extraction into ethyl acetate again three times, collected organic layer, revolve steaming removing organic solvent after anhydrous sodium sulfate drying and obtain brown-red solid, add methylene dichloride making beating and produce faint yellow insolubles, sand core funnel suction filtration also obtains faint yellow solid 6.80g by washed with dichloromethane, be Compound I-5, yield 99.4%.

The characterization data of this Compound I-5 is: ¹hNMR (400MHz, CDCl ₃, δ _ppm): 12.56 (s, 1H), 7.50 (s, 1H), 6.31 (s, 1H), 5.89 (s, 1H), 3.14 (m, 1H), 2,57 (s, 3H), 1.25 (d, J=6.0Hz, 6H).

Step e) prepare Compound I-6, i.e. 1-(two (the benzyloxy)-5-sec.-propyl of 2,4-)-ethyl ketone.

1-(2 is added in the 250mL two-mouth bottle that reflux condensing tube is housed, two hydroxyl-5-the sec.-propyl of 4-) methyl phenyl ketone I-5 (6.80g, 35.05mmol), salt of wormwood (12.09g, 87.63mmol), 100mL acetonitrile, 80 DEG C of backflow 1h in oil bath pan, then inject cylite (10mL, 84.15mmol) backflow with syringe to spend the night.After TLC monitoring reaction starting raw material reacts completely, reaction system is cooled to room temperature, Büchner funnel suction filtration, and residue from dichloromethane washs, and collects filtrate, revolves steaming removing organic solvent and obtains red-brown oily matter.Sherwood oil is added stirring to pulp in oily matter, produce white precipitate, sand core funnel suction filtration, petroleum ether, obtains white solid 6.72g, is Compound I-6, yield 51.26%.

The characterization data of this Compound I-6 is: ¹hNMR (400MHz, CDCl ₃, δ _ppm): 7.76 (s, 1H), 7.32-7.42 (m, 10H), 6.51 (s, 1H), 5.10 (s, 2H), 5.09 (s, 2H), 3.29 (m, 1H), 2,56 (s, 3H), 1.22 (d, J=6.8Hz, 6H).

Step f) prepare Compound I-7, i.e. (Z)-ethyl-4-(two (the benzyloxy)-5-isopropyl phenyl of 2,4-)-2-hydroxyl-4-oxo but-2-ene acid methyl esters.

Sodium Metal 99.5 (0.90g is added in the 250mL two-mouth bottle being provided with reflux condensing tube, 39.00mmol), 100mL ethanol stirs in 0 DEG C of ice bath until after sodium Metal 99.5 reacts completely, at room temperature add 1-(2, two (the benzyloxy)-5-isopropyl phenyl of 4-) ethyl ketone I-6 (6.72g, 17.98mmol) stir 30min, then inject oxalic acid diethyl ester (3.94mL, 29.11mmol) in oil bath pan 80 DEG C backflow 4h.Room temperature is cooled to after TLC monitoring is complete to substrate reactions, the aqueous hydrochloric acid adding 2M regulates pH value to being less than 7 generation light yellow precipitate, vacuum-drying removing organic solvent, then add methylene dichloride to dissolve, water extracts three times, and saturated nacl aqueous solution extraction once, is revolved after anhydrous sodium sulfate drying and steamed removing methylene dichloride, obtain yellow-brown solid 8.18g, be Compound I-7 yield 96%.

The characterization data of this Compound I-7 is: ¹hNMR (400MHz, CDCl3, δ ppm): 7.87 (s, 1H), 7.45-7.35 (m, 11H), 6.53 (s, 1H), 5.13 (s, 2H), 5.11 (s, 2H), 4.28 (q, J=14.0Hz, 2H), 3.30 (m, 1H), 1.28 (t, J=6.8Hz, 3H), 1.23 (d, J=6.8Hz, 6H).

Step g) prepare Compound I-8, i.e. 5-(two (benzyloxy-5-isopropyl phenyl) isoxzzole-3-carboxylic acid, ethyl ester of 2,4-.

(Z)-ethyl-4-(2 is added in the 250mL two-mouth bottle being provided with reflux condensing tube, two (the benzyloxy)-5-isopropyl phenyl of 4-)-2-hydroxyl-4-oxo but-2-ene acid methyl esters I-7 (8.18g, 17.26mmol), oxammonium hydrochloride (1.56g, 22.48mmol) and 100mL ethanol is 80 DEG C of backflow 5h in oil bath pan.Be cooled to room temperature after TLC monitoring reacts completely, vacuum-drying obtains white solid.Acetic acid ethyl dissolution solid, water extracts three times, and once, anhydrous sodium sulfate drying organic layer, then revolves steaming removing organic solvent and obtain white solid 6.74g, be Compound I-8, yield 83% in saturated sodium-chloride water solution extraction.

The characterization data of this Compound I-8 is: 1HNMR (400MHz, CDCl3, δ ppm): 7.83 (s, 1H), 7.41-7.33 (m, 10H), 7.00 (s, 1H), 6.58 (s, 1H), 5.16 (s, 2H), 5.07 (s, 2H), 4.42 (q, J=7.2Hz, 2H), 3.34 (m, 1H), 1.41 (t, J=7.2Hz, 3H), 1.23 (d, J=6.8Hz, 6H).

Step h) prepare compound 5-(two (the benzyloxy)-5-isopropyl phenyl of 2,4-) isoxzzole-3-methane amide I-9a, method is as follows:

5-(2 is added in the 100mL two-mouth bottle being provided with reflux condensing tube, two (the benzyloxy)-5-isopropyl phenyl of 4-) isoxzzole-3-carboxylic acid, ethyl ester I-8 (1.88g, 4mmol), the methanolic ammonia solution that 50mL is saturated heated and stirred in 60 DEG C of oil bath pans is spent the night.Be cooled to room temperature after TLC monitoring reacts completely, revolve after steaming removing reaction solvent and obtain white solid 1.57g through silica column purification, be target product I-9a, yield 89%.

The characterization data of this target product I-9a is: ¹hNMR (400MHz, CDCl3, δ ppm): 7.80 (s, 1H), 7.41-7.31 (m, 10H), 7.09 (s, 1H), 6.75 (brs, 1H), 6.55 (s, 1H), 5.18 (s, 2H), 5.03 (s, 2H), 3.33 (m, 1H), 1.25 (d, J=6.8Hz, 6H).

The structural formula of this target product I-9a is:

Step I) prepare compound 5-(two (the benzyloxy)-5-isopropyl phenyl of 2,4-)-N-cyclopropyl isoxzzole-3-methane amide I-9b, method is as follows:

5-(2 is added in 100mL single port bottle, two (the benzyloxy)-5-isopropyl phenyl of 4-) isoxzzole-3-carboxylic acid, ethyl ester I-8 (0.265g, 0.55mmol), one hydronium(ion) Lithium Oxide 98min (0.116g, 2.77mmol), and 7.76mL tetrahydrofuran (THF), 3.88mL methyl alcohol and 0.78mL water, in room temperature for overnight.After TLC monitoring reacts completely, revolve after steaming removing organic solvent and add acetic acid ethyl dissolution, water extracts three times, and once, anhydrous sodium sulfate drying, collected organic layer final vacuum is dry obtains intermediate in saturated sodium-chloride water solution extraction.Intermediate is added at the dry two-mouth bottle of 100mL; cyclopropylamine (0.05mL; 0.72mmol), EDCI (0.172g, 0.90mmol); HOBt (0.121g; 0.90mmol), the anhydrous N of DMAP (0.73mg, 0.006mmol) and 20mL; dinethylformamide, room temperature for overnight stir 30min in 0 DEG C of ice bath under nitrogen protection effect after.After TLC monitoring reacts completely, revolve after steaming removing organic solvent and add acetic acid ethyl dissolution, water extracts three times, saturated sodium-chloride water solution extraction once, anhydrous sodium sulfate drying, collected organic layer final vacuum is dry, finally by silica column purification to white solid 213mg, be Compound I-9b two step yield 64%.

The characterization data of this Compound I-9b is: ¹hNMR (400MHz, CDCl3, δ ppm): 7.74 (s, 1H), 7.34-7.29 (m, 10H), 6.87 (s, 1H), 6.58 (s, 1H), 5.10 (s, 2H), 5.06 (s, 2H), 3.31 (m, 1H), 2.95-2.88 (m, 1H), 1.22 (d, J=6.8Hz, 6H), 0.90-0.85 (m, 2H), 0.68-0.64 (m, 2H).

The structural formula of this compound is:

Step j) prepare compound 5-(two (the benzyloxy)-5-isopropyl phenyl of 2,4-)-N-ethyl isoxzzole-3-methane amide I-9c, method is as follows:

5-(2 is added in the 100mL two-mouth bottle being provided with backflow cold energy pipe, two (the benzyloxy)-5-isopropyl phenyl of 4-) isoxzzole-3-carboxylic acid, ethyl ester I-8 (1.88g, 4mmol) and 50mL ethanolic soln, 5mL70% ethylamine solution is return stirring 5h in 80 DEG C of oil bath pans.Be cooled to room temperature after TLC monitoring reacts completely, revolve after steaming removing reaction solvent and cool precipitation solid after ethanol heat of solution, sand core funnel suction filtration, cold washing with alcohol solid, obtains white solid 1.69g, is Compound I-9c, yield 90%.

The characterization data of this Compound I-9c is: ¹hNMR (400MHz, CDCl3, δ ppm): 7.79 (s, 1H), 7.33-7.39 (m, 10H), 7.07 (s, 1H), 6.81 (brs, 1H), 6.55 (s, 1H), 5.17 (s, 2H), 5.14 (s, 2H), 3.48 (m, 2H), 3.33 (m, 1H), 1.27 (t, J=6.8Hz, 3H), 1.24 (d, J=6.8Hz, 6H).

The structural formula of this Compound I-9c is:

Step k) prepare compound 5-(two (the benzyloxy)-5-isopropyl phenyl of 2,4-)-4-iodo isoxzzole-3-methane amide I-10a, method is as follows:

5-(2 is added in the 100mL two-mouth bottle being provided with reflux condensing tube, two (the benzyloxy)-5-isopropyl phenyl of 4-) isoxzzole-3-methane amide I-9a (1.68g, 3.80mmol), 50mL acetonitrile heated and stirred, N-iodosuccinimide (1.03g is added after solid is dissolved completely, 4.56mmol) and ceric ammonium nitrate (0.208g, 0.38mmol) 85 DEG C of backflows in oil bath pan spend the night.Be cooled to room temperature after TLC monitoring reacts completely, vacuum-drying obtains brown-red solid.Acetic acid ethyl dissolution solid, water extracts three times, and saturated sodium-chloride water solution extraction is once, anhydrous sodium sulfate drying organic layer, then revolves and steams removing organic solvent, obtain brown white solid 1.58g finally by silica column purification, be target product I-10a, yield 73%.

The characterization data of this target product I-10a is: ¹hNMR (400MHz, CDCl3, δ ppm): 7.41-7.34 (m, 11H), 6.78 (brs, 1H), 6.60 (s, 1H), 5.93 (brs, 1H), 5.10 (s, 2H), 5.06 (s, 2H), 3.35 (m, 1H), 1.24 (d, J=6.8Hz, 6H).

The structural formula of this target product I-10a is:

Prepare compound 5-(two (the benzyloxy)-5-isopropyl phenyl of 2,4-)-N-cyclopropyl isoxzzole-3-methane amide I-10b, the same I-10a of method, replaces to I-9b unlike by raw material I-9a.

The characterization data of this Compound I-10b is: ¹hNMR (400MHz, CDCl3, δ ppm): 7.34-7.29 (m, 10H), 6.87 (s, 1H), 6.58 (s, 1H), 5.10 (s, 2H), 5.06 (s, 2H), 3.31 (m, 1H), 2.95-2.88 (m, 1H), 1.22 (d, J=6.8Hz, 6H), 0.90-0.85 (m, 2H), 0.68-0.64 (m, 2H).

The structural formula of this Compound I-10b is:

Prepare compound 5-(two (the benzyloxy)-5-isopropyl phenyl of 2,4-)-N-ethyl isoxzzole-3-methane amide I-10c, the same I-10a of method, replaces to I-9c unlike by raw material I-9a.

The characterization data of this Compound I-10c is: ¹hNMR (400MHz, CDCl3, δ ppm): 7.40-7.29 (m, 11H), 6.81 (brs, 1H), 6.59 (s, 1H), 5.08 (s, 2H), 5.05 (s, 2H), 3.48 (m, 2H), 3.34 (m, 1H), (1.27 t, J=6.8Hz, 3H), 1.23 (d, J=6.8Hz, 6H).

The structural formula of this Compound I-10c is:

Prepare compound 1:5-(2,4-dihydroxyl-5-isopropyl phenyl)-4-(3-morpholino third-1-alkynes-1-base) isoxzzole-3-methane amide.

Step l) in 25mL two-mouth bottle, add 5-(2; two (the benzyloxy)-5-isopropyl phenyl of 4-)-4-iodo isoxzzole-3-methane amide (0.285g; 0.50mmol); 4-(the third-2-alkynes-1-base) morpholine (0.094g; 0.75mmol); two triphenyl palladium chloride (0.036g; 0.05mmol); cuprous iodide (0.020g; 0.10mmol) and 1.75mL diisopropylamine and the anhydrous N of 1.50mL; dinethylformamide is under the protection of argon gas, and in 55 DEG C of oil bath pans, heated and stirred is spent the night.After TLC monitoring reacts completely, revolve and steam removing reaction solvent, add acetic acid ethyl dissolution, water extracts three times, and saturated sodium-chloride water solution extracts once, anhydrous sodium sulfate drying, white solid 159mg is obtained through silica column purification, yield 56% after collected organic layer vacuum-drying.

Step m) in 50mL two-mouth bottle, add 5-(2; two (the benzyloxy)-5-isopropyl phenyl of 4-) preparation (0.16g of-4-(3-morpholino third-1-alkynes-1-base) isoxzzole-3-methane amide; 0.28mmol); 10mL anhydrous methylene chloride; under-5 DEG C of ice baths, the boron tribromide dichloromethane solution (0.84mL of 1M is added again under argon shield effect; 0.84mmol) stir 20min; in stirred at ambient temperature 50min; and after reacting completely with LC-MS monitoring, add 16mL saturated aqueous sodium carbonate cancellation reaction.Revolve after steaming removing organic solvent and add 16mL water, 160mL extraction into ethyl acetate, then water extracting twice, each 24.5mL, then extract once with 42mL saturated sodium-chloride water solution, anhydrous sodium sulfate drying, obtains 24mg white powder compound 1 through silica column purification after collected organic layer vacuum-drying, yield 23%.

The characterization data of this compound 1 is: 1HNMR (400MHz, CD ₃oDandCDCl3, δ ppm): 7.72 (s, 1H), 6.55 (s, 1H), 3.85 (t, J=4.4Hz, 4H), 3.65 (s, 2H), 3.45 (m, 2H), 2.77 (t, J=4.4Hz, 4H), 1.33 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oDandCDCl ₃, δ ppm): 173.7,162.1,159.4,158.2,155.3,128.1,128.0,106.0,104.0,98.0,90.8,75.4,67.2,52.9,48.5,27.1,23.1.ESI-MSm/z:386.2 (M+H)+, 384.3 (M-H)-.

The structural formula of this compound 1 is:

Embodiment 2

The preparation of compound 2:N-cyclopropyl-5-(2,4-dihydroxyl-5-isopropyl phenyl)-4-(3-morpholino third-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 2 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.54 (s, 1H), 6.43 (s, 1H), 3.70 (t, J=4.4Hz, 4H), 3.52 (s, 2H), 3.20 (m, 1H), 2.90-2.85 (m, 1H), 2.63 (t, J=4.4Hz, 4H), 1.20 (d, J=6.8Hz, 6H), 0.86-0.79 (m, 2H), 0.71-0.67 (m, 2H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.8,162.6,160.0,159.4,156.1,128.4,128.3,106.2,104.0,98.4,91.0,75.6,67.6,53.4,48.7,27.7,23.7,23.2,6.6.ESI-MSm/z:426.2 (M+H)+, 424.3 (M-H)-.

The structural formula of this compound 2 is:

Embodiment 3

The preparation of compound 3:N-cyclopropyl-5-(2,4-dihydroxyl-5-isopropyl phenyl)-4-(phenylene-ethynylene) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 3 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.73 (s, 1H), 7.53-7.50 (m, 2H), 7.36-7.35 (m, 3H), 6.46 (s, 1H), 3.19 (m, 1H), 2.91 (m, 1H), 1.17 (d, J=6.8Hz, 6H), 0.87-0.82 (m, 2H), 0.71-0.67 (m, 2H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.5,162.7,160.1,160.1,159.2,156.3,132.5,129.7,129.5,128.5,128.4,124.3,106.4,104.0,98.3,96.0,79.3,27.6,23.6,23.2,6.6.ESI-MSm/z:403.1 (M+H)+, 401.2 (M-H)-.

The structural formula of this compound 3 is:

Embodiment 4

The preparation of compound 4:4-((4-(tertiary butyl) phenyl) ethynyl)-5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 4 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.75 (s, 1H), 7.44 (d, J=8.0Hz, 2H), 7.38 (d, J=8.0Hz, 2H), 6.47 (s, 1H), 3.45 (q, J=7.2Hz, 2H), 3.20 (m, 1H), 1.31 (s, 9H), 1.25 (t, J=7.2Hz, 3H), 1.18 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.3,161.1,160.1,159.3,156.2,153.2,132.3,128.5,128.4,126.4,121.3,106.4,104.1,98.4,96.3,78.6,35.6,35.5,31.5,27.6,23.2,14.8.ESI-MSm/z:447.1 (M+H)+, 445.1 (M-H)-.

The structural formula of this compound 4 is:

Embodiment 5

The preparation of compound 5:5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(phenylene-ethynylene) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 5 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.74 (s, 1H), 7.51-7.49 (m, 2H), 7.34-7.32 (m, 2H), 6.48 (s, 1H), 3.46 (q, J=7.2Hz, 2H), 3.20 (m, 1H), 1.25 (t, J=7.2Hz, 3H), 1.17 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.5,161.0,160.1,159.2,156.2,132.5,129.6,129.4,128.5,128.4,124.2,106.3,104.0,98.3,96.0,79.3,35.5,27.6,23.1,14.7.ESI-MSm/z:391.0 (M+H)+, 389.1 (M-H)-.

The structural formula of this compound 5 is:

Embodiment 6

The preparation of compound 6:5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-((4-fluorophenyl) ethynyl) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 6 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.70 (s, 1H), 7.54 (dd, J=8.8Hz, J=5.6Hz, 2H), 7.12 (t, J=8.8Hz, 2H), 6.46 (s, 1H), 3.44 (q, J=7.2Hz, 2H), 3.19 (m, 1H), 1.25 (t, J=7.2Hz, 3H), 1.16 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.6,163.1,161.1,160.1,159.3,156.3,134.7,134.6,128.5,128.4,120.6,120.6,116.7,116.5,106.3,104.0,98.3,94.8,79.1,35.5,27.6,23.2,14.8.ESI-MSm/z:409.0 (M+H)+, 407.1 (M-H)-.

The structural formula of this compound 6 is:

Embodiment 7

The preparation of compound 7:5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(pyridine-3-ethynyl) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 7 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 8.68 (s, 1H), 8.50 (d, J=4.4Hz, 1H), 7.95 (d, J=8.0Hz, 1H), 7.65 (s, 1H), 7.45 (dd, J=8.0Hz, J=4.8Hz, 1H), 6.47 (s, 1H), 3.45 (q, J=7.2Hz, 2H), 3.20 (m, 1H), 1.26 (t, J=7.2Hz, 3H), 1.18 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 174.3,160.9,160.4,159.3,156.4,152.3,149.2,140.5,128.6,128.4,125.0,122.3,106.2,104.0,97.9,91.9,83.5,35.5,27.6,23.2,14.8.ESI-MSm/z:392.1 (M+H)+, 390.1 (M-H)-.

The structural formula of this compound 7 is:

Embodiment 8

The preparation of compound 8:4-(cyclohexyl-acetylene base)-5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 8 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.63 (s, 1H), 6.43 (s, 1H), 3.42 (d, J=7.2Hz, 2H), 3.20 (m, 1H), 2.67-2.62 (m, 1H), 1.89-1.86 (m, 2H), 1.78-1.74 (m, 2H), 1.57-1.52 (m, 3H), 1.40-1.30 (m, 3H), 1.25 (t, J=7.2Hz, 3H), 1.21 (d, J=6.4Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 172.9,161.3,159.8,159.7,156.0,128.5,128.3,106.5,104.1,101.5,98.7,70.2,35.5,33.7,31.2,27.9,27.0,25.8,23.2,14.8.ESI-MSm/z:397.1 (M+H)+, 395.1 (M-H)-.

The structural formula of this compound 8 is:

Embodiment 9

The preparation of compound 9:4-(cyclopentyl ethynyl)-5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 9 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.65 (s, 1H), 6.43 (s, 1H), 3.42 (d, J=7.2Hz, 2H), 3.20 (m, 1H), 2.92-2.85 (m, 1H), 2.00-1.95 (m, 2H), 1.77-1.65 (m, 4H), 1.63-1.60 (m, 2H), 1.25 (t, J=7.2Hz, 3H), 1.21 (d, J=6.4Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 172.9,161.3,159.8,159.6,156.0,128.4,128.3,106.5,104.0,101.7,98.7,69.7,35.5,34.8,32.2,27.8,26.0,23.2,14.8.ESI-MSm/z:383.2 (M+H)+, 381.1 (M-H)-.

The structural formula of this compound 9 is:

Embodiment 10

The preparation of compound 10:4-(cyclopropyl acethlene base)-5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 10 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.69 (s, 1H), 6.43 (s, 1H), 3.42 (d, J=7.2Hz, 2H), 3.20 (m, 1H), 1.53-1.46 (m, 1H), 1.27-1.21 (m, 9H), 0.90-0.87 (m, 2H), 0.79-0.75 (m, 2H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.3,161.2,159.8,159.6,156.0,128.3,106.5,104.0,100.8,98.6,65.1,35.5,27.6,23.2,14.7,9.0.ESI-MSm/z:355.1 (M+H)+, 353.0 (M-H)-.

The structural formula of this compound 10 is:

Embodiment 11

The preparation of compound 11:5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-morpholino third-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 11 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.54 (s, 1H), 6.43 (s, 1H), 3.70 (t, J=4.0Hz, 4H), 3.53 (s, 2H), 3.44 (q, J=7.2Hz, 2H), 3.20 (m, 1H), (2.63 t, J=4.0Hz, 4H), 1.26-1.20 (m, 9H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.9,161.0,160.0,159.6,156.2,128.4,128.3,106.2,104.0,98.4,91.0,75.6,67.6,53.4,35.5,27.7,23.2,14.8.ESI-MSm/z:414.2 (M+H)+, 412.1 (M-H)-.

The structural formula of this compound 11 is:

Embodiment 12

Compound 12:5-(2,4-dihydroxyl-5-isopropyl phenyl) preparation of-N-ethyl-4-(3-(4-(methoxyl group imido grpup) piperidin-1-yl) third-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 12 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.54 (s, 1H), 6.43 (s, 1H), 3.78 (s, 3H), 3.59 (s, 2H), 3.42 (q, J=7.2Hz, 2H), 3.20 (m, 1H), 2.74 (t, J=6.0Hz, 2H), 2.69-2.65 (m, 2H), 2.63-2.58 (m, 2H), 2.34 (t, J=6.0Hz, 2H), 1.25 (t, J=7.2Hz, 3H), 1.21 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.8,161.0,160.0,159.6,157.4,156.1,128.4,128.3,106.3,104.0,98.4,91.0,75.5,61.4,53.6,52.3,48.2,35.5,31.8,27.7,25.6,23.2,14.8.ESI-MSm/z:455.0 (M+H)+, 453.1 (M-H)-.

The structural formula of this compound 12 is:

Embodiment 13

Compound 13:5-(2,4-dihydroxyl-5-isopropyl phenyl)-4-(3-((2S, 6R)-2,6-thebaine) the third-1-alkynes-1-base) preparation of-N-ethyl isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 13 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.53 (s, 1H), 6.43 (s, 1H), 3.69-3.61 (m, 2H), 3.50 (s, 2H), (3.42 q, J=7.2Hz, 2H), 3.20 (m, 1H), 2.79 (d, J=10.8Hz, 2H), 1.98 (t, J=10.8Hz, 2H), 1.25 (t, J=7.2Hz, 3H), 1.21 (d, J=6.8Hz, 6H), 1.12 (d, J=6.4Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.9,161.0,160.0,159.6,156.1,128.4,128.3,106.2,104.0,98.5,91.1,72.8,59.0,48.3,35.5,27.6,23.3,19.3,14.8.ESI-MSm/z:442.2 (M+H)+, 440.3 (M-H)-.

The structural formula of this compound 13 is:

Embodiment 14

The preparation of compound 14:5-(2,4-dihydroxyl-5-isopropyl phenyl)-4-(3-(1,1-sulphur dioxide is for morpholino) third-1-alkynes-1-base)-N-ethyl isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 14 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.54 (s, 1H), 6.44 (s, 1H), 3.67 (s, 2H), 3.42 (q, J=7.2Hz, 2H), 3.20 (m, 1H), 3.13 (s, 8H), 1.25 (t, J=7.2Hz, 3H), 1.21 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.8,161.0,160.0,159.6,156.1,128.4,106.3,104.0,98.4,91.1,75.7,52.2,51.3,48.0,35.5,27.6,23.2,14.8.ESI-MSm/z:462.1 (M+H)+, 460.1 (M-H)-.

The structural formula of this compound 14 is:

Embodiment 15

The preparation of compound 15:5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-thiomorpholine generation the third-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 15 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.45 (s, 1H), 6.45 (s, 1H), 3.62 (s, 2H), 3.42 (q, J=7.2Hz, 2H), 3.25-3.16 (m, 3H), 2.99-2.96 (m, 4H), 2.88-2.84 (m, 2H), 1.25 (t, J=7.2Hz, 3H), 1.21 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.8,161.0,160.0,159.6,156.0,128.4,128.3,106.3,104.0,98.8,91.0,75.8,48.9,46.8,44.0,35.5,27.7,23.2,14.8.ESI-MSm/z:430.2 (M+H)+, 428.1 (M-H)-.

The structural formula of this compound 15 is:

Embodiment 16

Compound 16:5-(2,4-dihydroxyl-5-isopropyl phenyl) preparation of-N-ethyl-4-(3-(4-(6-(trifluoromethyl) nicotinoyl amido) piperidin-1-yl) the third-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 16 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 9.91 (d, J=2.0Hz, 1H), 8.42 (dd, J=8.0Hz, J=2.0Hz, 1H), 7.91 (d, J=8.4Hz, 1H), 7.48 (s, 1H), 6.47 (s, 1H), 4.02 (s, 2H), 3.49-3.23 (m, 4H), 3.19 (m, 1H), 3.06 (t, J=11.6Hz, 2H), 2.21-2.17 (m, 2H), 1.96-1.87 (m, 2H), 1.23 (t, J=7.2Hz, 3H), 1.15 (d, J=6.4Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 174.4,166.5,160.7,160.3,159.4,156.1,155.0,151.2,150.9,150.7,150.4,150.1,138.5,134.4,128.6,128.4,126.0,123.8,121.6,121.6,121.6,121.6,121.5,120.4,119.4,106.1,104.0,103.4,98.0,87.1,78.8,52.1,46.5,35.5,27.7,23.2,14.8.ESI-MSm/z:600.1 (M+H)+, 598.1 (M-H)-.

The structural formula of this compound 16 is:

Embodiment 17

Compound 17:5-(2; 4-dihydroxyl-5-isopropyl phenyl) preparation of-N-ethyl-4-(3-(4-(6-(trifluoromethyl) nicotinoyl) piperazine-1-base) the third-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 17 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 8.82 (d, J=1.6Hz, 1H), 8.10 (dd, J=8.0Hz, J=2.0Hz, 1H), 7.89 (d, J=8.0Hz, 1H), 7.56 (s, 1H), 6.43 (s, 1H), 3.83 (s, 2H), 3.63, (s, 2H), 3.45-3.39 (m, 4H), 3.19 (m, 1H), 2.72 (d, J=28.8Hz, 4H), 1.22 (t, J=7.2Hz, 3H), 1.15 (d, J=6.4Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.7,168.2,160.9,160.0,159.5,156.1,149.8,149.7,149.4,138.1,136.0,128.4,128.3,123.8,121.7,121.7,121.6,106.3,104.0,98.5,90.8,75.9,48.2,35.4,27.6,23.2,14.8.ESI-MSm/z:586.2 (M+H)+, 584.1 (M-H)-.

The structural formula of this compound 17 is:

Embodiment 18

Compound 18:5-(2,4-dihydroxyl-5-isopropyl phenyl) preparation of-N-ethyl-4-(3-(4-(trifluoromethyl) benzoylamino) third-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 18 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 8.02 (d, J=8.0Hz, 2H), 7.78 (d, J=8.0Hz, 2H), 7.63 (s, 1H), 6.43 (s, 1H), 4.45 (s, 2H), 3.43 (q, J=7.2Hz, 2H), 3.15 (m, 1H), 1.24 (t, J=7.2Hz, 3H), 1.16 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 174.1,168.2,160.9,160.1,159.3,156.1,138.8,129.3,129.1,128.5,128.4,126.5,126.5,126.4,124.2,106.2,104.0,97.8,92.2,73.0,35.5,31.2,27.6,23.2,14.7.ESI-MSm/z:516.2 (M+H)+, 514.3 (M-H)-.

The structural formula of this compound 18 is:

Embodiment 19

Compound 19:5-(2,4-dihydroxyl-5-isopropyl phenyl) preparation of-N-ethyl-4-(3-(6-(trifluoromethyl) nicotinoyl amido) third-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 19 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 9.13 (d, J=2.0Hz, 1H), 8.44 (dd, J=8.4Hz, J=2.0Hz, 1H), 7.93 (d, J=8.0Hz, 1H), 7.62 (s, 1H), 6.42 (s, 1H), 4.46 (s, 2H), 3.43 (q, J=7.2Hz, 2H), 3.16 (m, 1H), 1.23 (t, J=7.2Hz, 3H), 1.16 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 174.2,166.2,160.9,160.1,159.4,156.2,150.1,138.6,134.0,128.5,128.4,121.6,106.2,104.0,97.8,91.8,73.2,49.3,35.5,31.3,27.6,23.2,14.7.ESI-MSm/z:517.2 (M+H)+, 515.1 (M-H)-.

The structural formula of this compound 19 is:

Embodiment 20

The preparation of compound 20:4-(3-(2-naphthoyl amido) third-1-alkynes-1-base)-5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 20 is: ¹hNMR (400MHz, CD ₃oDandCDCl3, δ ppm): 8.41 (s, 1H), 7.96-7.90 (m, 4H), 7.66 (s, 1H), 7.59-7.55 (m, 2H), 6.43 (s, 1H), 4.49 (s, 2H), 3.44 (q, J=7.2Hz, 2H), 3.15 (m, 1H), 1.24 (t, J=7.2Hz, 3H), 1.16 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oDandCDCl3, δ ppm): 174.0,169.6,160.8,159.9,159.2,156.0,136.2,133.9,132.2,130.0,129.2,129.0,128.8,128.7,128.4,128.3,127.7,124.8,106.1,104.0,97.7,92.4,73.0,35.4,31.2,27.4,23.2,14.7.ESI-MSm/z:498.2 (M+H)+, 496.3 (M-H)-.

The structural formula of this compound 20 is:

Embodiment 21

The preparation of compound 21:5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-(quinoline-3-formamido-) third-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 21 is: ¹hNMR (400MHz, CD ₃oDandCDCl3, δ ppm): 9.27 (s, 1H), 8.78 (s, 1H), 8.08 (d, J=8.0Hz, 1H), 7.99 (d, J=8.0Hz, 1H), 7.84 (t, J=8.0Hz, 1H), 7.67 (s, 1H), 7.57 (s, 1H), 6.41 (s, 1H), 4.49 (s, 2H), 3.45 (q, J=7.2Hz, 2H), 3.16 (m, 1H), 1.24 (t, J=7.2Hz, 3H), 1.17 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oDandCDCl3, δ ppm): 173.7,166.6,160.1,159.4,158.3,155.3,149.2,137.3,132.4,130.0,128.8,128.4,128.2,128.0,127.8,127.4,106.0,104.0,97.3,91.4,73.2,35.1,31.2,26.9,23.0,14.6.ESI-MSm/z:499.1 (M+H)+, 497.1 (M-H)-.

The structural formula of this compound 21 is:

Embodiment 22

The preparation of compound 22:5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-methyl-3-(sulfonyloxy methyl amido) fourth-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 22 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.40 (s, 1H), 6.44 (s, 1H), 3.44 (q, J=7.2Hz, 2H), 3.20 (m, 1H), 3.05 (s, 3H), 1.64 (s, 6H), 1.25 (t, J=7.2Hz, 3H), 1.22 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 174.1,160.8,160.2,159.4,156.3,128.6,128.5,106.1,103.9,98.7,98.0,73.2,51.3,42.6,35.5,31.5,28.1,23.1,14.8.ESI-MSm/z:450.2 (M+H)+, 448.3 (M-H)-.

The structural formula of this compound 22 is:

Embodiment 23

The preparation of compound 23:5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-methyl-3-(4-Methyl benzenesulfonyl amido) fourth-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 23 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.61 (d, J=8.0Hz, 2H), 7.30 (s, 1H), 7.01 (d, J=8.0Hz, 2H), 6.45 (s, 1H), 3.46 (q, J=7.2Hz, 2H), 3.16 (m, 1H), 2.18 (s, 3H), 1.60 (s, 6H), 1.29 (t, J=7.2Hz, 3H), 1.22 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.4,160.6,160.2,159.2,156.2,144.2,140.2,130.1,128.5,128.3,106.2,104.0,98.1,97.3,73.6,51.2,35.5,31.3,28.3,23.2,21.4,14.9.ESI-MSm/z:526.2 (M+Na)+, 524.3 (M-H)-.

The structural formula of this compound 23 is:

Embodiment 24

The preparation of compound 24:4-(3-acetylaminohydroxyphenylarsonic acid 3-methyl fourth-1-alkynes-1-base)-5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 24 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.51 (s, 1H), 6.44 (s, 1H), 3.50 (q, J=7.2Hz, 2H), 3.20 (m, 1H), 1.93 (s, 3H), 1.64 (s, 6H), 1.27 (t, J=7.2Hz, 3H), 1.23 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.6,172.4,160.7,160.1,159.3,156.2,128.5,128.4,106.3,104.1,100.7,97.4,70.9,35.7,29.3,28.0,23.3,23.2,14.9.ESI-MSm/z:414.2 (M+H)+, 412.1 (M-H)-.

The structural formula of this compound 24 is:

Embodiment 25

The preparation of compound 25:4-(3-(cyclopropyl carboxamide base)-3-methyl fourth-1-alkynes-1-base)-5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 25 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.63 (s, 1H), 6.43 (s, 1H), 3.43 (q, J=7.2Hz, 2H), 3.20 (m, 1H), 2.65 (m, 1H), 1.89-1.86 (m, 2H), 1.78-1.75 (m, 2H), 1.57-1.52 (m, 3H), 1.36 (s, 1H), 1.26 (t, J=7.2Hz, 3H), 1.22 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 172.9,161.3,159.8,159.7,156.0,128.5,128.3,106.5,104.1,101.5,98.7,70.2,35.5,33.7,31.2,27.9,27.0,25.8,23.2,14.8.ESI-MSm/z:440.1 (M+H)+, 438.1 (M-H)-.

The structural formula of this compound 25 is:

Embodiment 26

The preparation of compound 26:4-(3-(cyclopentyl formamido-)-3-methyl fourth-1-alkynes-1-base)-5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 26 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.52 (s, 1H), 6.44 (s, 1H), 3.52 (q, J=7.2Hz, 2H), 3.20 (m, 1H), 2.61 (m, 1H), 1.83-1.80 (m, 2H), 1.79-1.72 (m, 4H), 1.71 (s, 6H), 1.70-1.50 (m, 2H), 1.27 (t, J=7.2Hz, 3H), 1.23 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 179.1,174.5,161.6160.9,160.3,157.0,129.4,129.3,107.5,105.4,102.0,98.5,71.7,47.4,36.5,32.2,30.2,28.9,27.9,24.0,15.7.ESI-MSm/z:468.2 (M+H)+, 466.3 (M-H)-.

The structural formula of this compound 26 is:

Embodiment 27

The preparation of compound 27:4-(3-(cyclohexyl formamido-)-3-methyl fourth-1-alkynes-1-base)-5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 27 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.52 (s, 1H), 6.44 (s, 1H), 3.52 (q, J=7.2Hz, 2H), 3.20 (m, 1H), 2.17-2.15 (m, 2H), 1.75 (d, J=6,0Hz, 4H), 1.68-1.60 (m, 1H), 1.63 (s, 6H), 1.47-1.28 (m, 4H), 1.26 (t, J=7.2Hz, 3H), 1.22 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 179.1,174.4,161.6,160.9,160.3,157.0,129.6,129.4,107.6,105.5,102.0,98.7,71.8,47.4,36.5,31.5,30.2,29.0,27.8,27.6,24.0,15.7.ESI-MSm/z:482.2 (M+H)+, 480.2 (M-H)-.

The structural formula of this compound 27 is:

Embodiment 28

The preparation of compound 28:5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-(3-isopropylureido)-3-methyl fourth-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 28 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.51 (s, 1H), 6.44 (s, 1H), 3.82 (m, 1H), 3.50 (q, J=7.2Hz, 2H), 3.20 (m, 1H), 1.62 (s, 6H), 1.26 (t, J=7.2Hz, 3H), 1.22 (d, J=6.8Hz, 6H), 1.09 (d, J=6.4Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.5,160.7,160.1,159.3,159.2,156.2,128.4,106.4,104.2,101.6,97.6,70.8,56.1,42.7,35.7,30.5,29.5,23.5,23.2,14.9.ESI-MSm/z:457.2 (M+H)+, 455.2 (M-H)-.

The structural formula of this compound 28 is:

Embodiment 29

Compound 29:N-(4-(5-(2; 4-dihydroxyl-5-isopropyl phenyl)-3-(ethamine formyl radical) isoxzzole-4-base)-2-methyl fourth-3-alkynes-2-base) preparation of morpholine-4-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 29 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.51 (s, 1H), 6.44 (s, 1H), 3.64 (t, J=4.4Hz, 4H), 3.52 (q, J=7.2Hz, 2H), 3.23 (s, 4H), 3.20 (m, 1H), 1.65 (s, 6H), 1.26 (t, J=7.2Hz, 3H), 1.22 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 174.2,161.6,160.8,159.9,156.9,129.7,129.4,107.7,105.6,103.1,98.7,71.6,68.5,46.3,36.5,31.5,30.9,29.0,24.0,15.7.ESI-MSm/z:485.2 (M+H)+, 483.3 (M-H)-.

The structural formula of this compound 29 is:

Embodiment 30

The preparation of compound 30:5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-(furans-3-formamido-)-3-methyl fourth-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 30 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 8.09 (s, 1H), 7.53 (s, 2H), 6.83 (s, 1H), 6.45 (s, 1H), 3.50 (q, J=7.2Hz, 2H), 3.17 (m, 1H), 1.74 (s, 6H), 1.24 (t, J=7.2Hz, 3H), 1.20 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.6,164.5,160.6,160.1,159.1,156.2,146.6,145.0,128.4,128.3,124.0,109.9,106.2,104.0,100.0,97.3,71.2,49.3,35.7,29.5,28.0,23.2,15.0.ESI-MSm/z:466.1 (M+H)+, 464.1 (M-H)-.

The structural formula of this compound 30 is:

Embodiment 31

Compound 31:5-(2,4-dihydroxyl-5-isopropyl phenyl) preparation of-N-ethyl-4-(3-methyl-3-((4-(trifluoromethyl) phenyl) amido) fourth-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 31 is: ¹hNMR (400MHz, CDCl3, δ ppm): 7.70 (s, 1H), 7.47-7.46 (m, 4H), 7.00-6.96 (m, 1H), 6.49 (s, 1H), 4.02 (s, 2H), 3.49 (q, J=7.2Hz, 2H), 3.15 (m, 1H), 1.52 (s, 6H), 1.24 (t, J=7.2Hz, 3H), 1.19 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CDCl3, δ ppm): 172.0, 158.9, 158.7, 158.0, 154.2, 144.4, 129.7, 129.4, 129.2, 128.7, 127.2, 125.6, 125.5, 125.5, 123.5, 105.7, 104.7, 102.3, 97.0, 71.6, 51.7, 48.8, 35.1, 29.5, 27.1, 23.0, 14.7.ESI-MSm/z:530.2 (M+H)+, 528.1 (M-H)-.

The structural formula of this compound 31 is:

Embodiment 32

Compound 32:5-(2,4-dihydroxyl-5-isopropyl phenyl) preparation of-N-ethyl-4-(3-methyl-3-(4-(trifluoromethyl) benzamido) fourth-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 32 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 7.64 (d, J=8.4Hz, 2H), 7.57 (d, J=8.4Hz, 2H), 7.26 (s, 1H), 6.33 (s, 1H), 6.02 (s, 1H), 3.37 (q, J=7.2Hz, 2H), 3.18 (m, 1H), 1.53 (s, 6H), 1.21 (t, J=7.2Hz, 3H), 1.18 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 168.1,162.6,162.0,160.5,159.3,158.0,155.8,134.6,130.9,129.8,128.3,128.0,126.4,126.3,110.1,108.7,103.5,89.3,71.5,35.4,29.9,27.7,23.2,14.7.ESI-MSm/z:544.1 (M+H)+, 542.1 (M-H)-.

The structural formula of this compound 32 is:

Embodiment 33

The preparation of compound 33:5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-methyl-3-(picoline amide group) fourth-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 33 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 8.57 (d, J=2.2Hz, 1H), 8.07 (d, J=6.8Hz, 1H), 7.51 (t, J=6.0Hz, 2H), 6.44 (s, 1H), 3.51 (q, J=7.2Hz, 2H), 3.14 (m, 1H), 1.80 (s, 6H), 1.24 (t, J=7.2Hz, 3H), 1.14 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.7,165.4,160.6,160.1,159.3,156.2,151.0,149.5,138.8,128.4,128.3,127.8,122.9,106.2,104.1,100.1,97.3,71.6,49.1,35.7,29.4,27.9.23.1,15.0.ESI-MSm/z:477.2 (M+H)+, 475.1 (M-H)-.

The structural formula of this compound 33 is:

Embodiment 34

The preparation of compound 34:5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-methyl-3-(6-methylnictotinyl amido) fourth-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 34 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 8.86 (s, 1H), (8.14 d, J=8.4Hz, 1H), 7.52 (s, 1H), 7.39 (d, J=8.4Hz, 1H), 6.43 (s, 1H), 3.52 (q, J=7.2Hz, 2H), 3.17 (m, 1H), 2.59 (s, 3H), 1.78 (s, 6H), 1.25 (t, J=7.2Hz, 3H), 1.20 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.7,167.3,162.5,160.7,160.1,159.3,156.2,148.8,137.6,129.7,128.5,128.4,124.6,106.3,104.0,100.5,97.4,71.4,49.7,35.7,29.4,28.1,23.7.23.2,15.0.ESI-MSm/z:491.2 (M+H)+, 489.1 (M-H)-.

The structural formula of this compound 34 is:

Embodiment 35

The preparation of compound 35:5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-methyl-3-(quinoline-3-formamido-) fourth-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 35 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 9.21 (s, 1H), 8.77 (s, 1H), 8.05 (d, J=8.0Hz, 1H), 8.00 (d, J=8.0Hz, 1H), 7.83 (t, J=8.0Hz, 1H), 7.65 (t, J=0.8Hz, 1H), 7.53 (s, 1H), 6.45 (s, 1H), 3.52 (q, J=7.2Hz, 2H), 3.16 (m, 1H), 1.83 (s, 6H), 1.24 (t, J=7.2Hz, 3H), 1.19 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.7,167.3,160.7,160.1,159.3,156.2,149.9,149.6,137.8,132.8,130.3,129.1,129.0,128.9,128.5,128.4,128.3,106.3,104.1,100.5,97.4,71.5,49.8,35.7,29.4,28.1,23.2,15.0.ESI-MSm/z:527.1 (M+H)+, 525.0 (M-H)-.

The structural formula of this compound 35 is:

Embodiment 36

The preparation of compound 36:5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-(6-methoxypyridine amide group)-3-methyl fourth-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 36 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 8.66 (s, 1H), 8.08 (dd, J=8.8Hz, J=2.4Hz, 1H), 7.52 (s, 1H), (6.80 d, J=8.8Hz, 1H), 6.44 (s, 1H), 3.94 (s, 3H), 3.52 (q, J=7.2Hz, 2H), 3.16 (m, 1H), 1.77 (s, 6H), 1.25 (t, J=7.2Hz, 3H), 1.20 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 174.4,168.4,161.6,160.9,160.2,157.0,149.2,140.2,129.5,129.4,126.3,112.1,107.4,105.3,101.7,98.4,72.3,55.3,36.5,30.3,28.9,23.9,15.8.ESI-MSm/z:507.2 (M+H)+, 505.1 (M-H)-.

The structural formula of this compound 36 is:

Embodiment 37

Compound 37:5-(2,4-dihydroxyl-5-isopropyl phenyl) preparation of-N-ethyl-4-(3-methyl-3-(6-(trifluoromethyl) picolinamide base) fourth-1-alkynes-1 isoxzzole-Ji)-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 37 is: ¹hNMR (400MHz, CD ₃oD, δ ppm): 9.10 (d, J=2.0Hz, 1H), 8.41 (dd, J=8.4Hz, J=2.0Hz, 1H), 7.90 (d, J=8.0Hz, 1H) 7.52 (s, 1H), 6.44 (s, 1H), 3.52 (q, J=7.2Hz, 2H), 3.18 (m, 1H), 1.80 (s, 6H), 1.26 (t, J=7.2Hz, 3H), 1.20 (d, J=6.8Hz, 6H) .13CNMR (125MHz, CD ₃oD, δ ppm): 173.7,166.2,160.7,160.2,159.3,156.2,150.2,138.6,135.0,128.5,128.4,121.5,106.3,104.1,100.1,97.4,71.6,49.9,35.7,29.3,28.1,23.1,14.9.ESI-MSm/z:545.2 (M+H)+, 543.1 (M-H)-.

The structural formula of this compound 37 is:

Embodiment 38

The preparation of compound 38:5-(2,4-dihydroxyl-5-isopropyl phenyl)-N-ethyl-4-(3-(6-fluorine nicotinoyl amido)-3-methyl fourth-1-alkynes-1-base) isoxzzole-3-methane amide, with reference to the method for embodiment 1.

The characterization data of this compound 38 is: ¹hNMR (400MHz, CD ₃oD, δ _ppm): 8.66 (s, 1H), 8.34 (dt, J=7.6Hz, J=2.4Hz, 1H), 7.51 (s, 1H), 7.13 (dd, J=7.2Hz, J=2.4Hz, 1H), 6.44 (s, 1H), 3.52 (q, J=7.2Hz, 2H), 3.16 (m, 1H), 1.78 (s, 6H), 1.25 (t, J=7.2Hz, 3H), 1.20 (d, J=6.8Hz, 6H). ¹³cNMR (125MHz, CD ₃oD, δ _ppm): 173.7,167.2,166.3,165.3,160.7,160.1,159.3,156.2,148.7,148.6,142.7,142.6,130.5,130.5,128.5,128.4,110.6,110.3,106.3,104.1,100.4,97.4,71.5,49.7,35.7,29.3,28.0,23.2,15.0.ESI-MSm/z:495.2 (M+H) ⁺, 493.1 (M-H) ^-.

The structural formula of this compound 38 is:

Experimental example

Get the compound that above-described embodiment prepares, each compound all arranges multiple various dose group, and high dose group is configured to mother liquor, and three doubling dilutions are to lowest dose level group successively for doses remaining group, and whole sample to dissolve in DMSO and saves backup at-20 DEG C.

Experiment buffer includes 20mmol/lHEPES (K), 50mmol/lKCl, 5mmol/lMgCl ₂, 20mmol/lNa ₂moO ₄with 0.01%NP40, pH are 7.3.Add the reaction buffer of 5 μ l containing 40mMDTT and 2mg/mlBSA before each experiment, then add the fluorescently-labeled geldanamycin of 2.5 μ l (reaction density 5nM).Then utilize compound transferring apparatus LiquidHandlerEcho520 to join in reaction solution by the compound that 10nl1000 × gradient concentration has diluted, finally add 2.5 μ lHsp90 β orHsp90 α enzyme solution (reaction density 35ng/ μ l).Room temperature slight oscillatory reacts 2 hours, and finally read plate instrument with microwell plate and measure reading, exciting light is 485nm, and utilizing emitted light is 530nm, data GraphpadPrism5 software processes.

Each cancer cells is suspended in corresponding nutrient solution and is mixed with suitable concentration, and 50 μ l/ pore volumes are planted in 384 orifice plates, are incubated at 37 DEG C, 5%CO ₂24h in constant incubator.Compound DMSO is dissolved into 10mM (mmol/L) mother liquor, and become 1000 × series of compounds concentration storage liquid of 10 concentration gradients by 1:3 gradient dilution with DMSO, compound transferring apparatus LiquidHandlerEcho520 is utilized to be transferred in the respective aperture of cell 384 orifice plate by 1000 × series concentration compound storage liquid, every hole 50nL, adds equal-volume solvent DMSO in blank control wells.Soft mixing, 37 DEG C are continued to cultivate.Again change corresponding substratum after 72h, and add the CCK8 cell proliferation-toxicity detection reagent of 3 μ l to every hole.Continue culture plate to hatch 2h at 37 DEG C of incubators, be then determined at the absorbancy at 450nm place by microplate reader.

This experiment adopts aforesaid method to determine the inhibit activities of synthesized target compound to two of HSP90 albumen kinds of hypotype HSP90 α and HSP90 β respectively; To the human lung cancer cell line A549 of HSP90 high expression level, human breast carcinoma cell lines MCF-7, chronic myeloid leukemia cells system K562, prostate cancer cell line DU145 and the test inhibit activities with proliferating epidermal cancerous cell line Hela.Result is as shown in table 1.

Table 1 target compound is to the inhibit activities IC of HSP90 albumen and tumor cell line ₅₀(unit be μM)

From the above results, compared with positive control NVP/AUY-922 (CAS:747412-49-3), heat shock protein 90 provided by the present invention (HSP90) inhibitor is to inhibited while of HSP90 α and HSP90 β two kinds of enzymes, also inhibited to 5 quasi-cancer cells, particularly compound (2,12,13,40 and 43) all shows good inhibit activities to HSP90 molecular chaperones and five quasi-cancer cell systems.And compound (3,5,8,9,42 and 51) reaches the level of sub-nmole to HSP90 α and five quasi-cancer cell system inhibit activities, more weak to the inhibit activities of HSP90 β.To in synthesized compound, as compound (16,26,41 and 49) shows good inhibit activities to HSP90 molecular chaperones, but more weak to the anti-proliferative capacity of five quasi-cancer cell systems.

Each technical characteristic of the above embodiment can combine arbitrarily, for making description succinct, the all possible combination of each technical characteristic in above-described embodiment is not all described, but, as long as the combination of these technical characteristics does not exist contradiction, be all considered to be the scope that this specification sheets is recorded.

The above embodiment only have expressed several embodiment of the present invention, and it describes comparatively concrete and detailed, but can not therefore be construed as limiting the scope of the patent.It should be pointed out that for the person of ordinary skill of the art, without departing from the inventive concept of the premise, can also make some distortion and improvement, these all belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.

Claims

1. there is heat shock protein inhibitors or its pharmacy acceptable salt of formula I constitutional features:

Wherein:

R is selected from: C ₁-C ₆alkyl, C ₁-C ₆unsaturated alkyl;

N is selected from: 1,2 or 3;

X, Y, Z are independently selected from: CH, N, O.

2. heat shock protein inhibitors according to claim 1 or its pharmacy acceptable salt, is characterized in that, described X, Y, Z are selected from following group:

3. heat shock protein inhibitors according to claim 1 or its pharmacy acceptable salt, is characterized in that, described heat shock protein inhibitors is selected from the compound of following general formula I I:

Wherein:

4. heat shock protein inhibitors according to claim 1 or its pharmacy acceptable salt, is characterized in that, described R ₄be selected from: substituted or non-substituted aryl, substituted or non-substituted heteroaryl, C ₃-C ₈cycloalkyl, (CR ₆r ₉) _nr ₅,

N is selected from: 1.

5. heat shock protein inhibitors according to claim 1 or its pharmacy acceptable salt, is characterized in that, described R ₂, R ₃independently be selected from: H, C ₃-C ₈cycloalkyl, C ₁-C ₆alkyl, C ₁-C ₆unsaturated alkyl.

6. heat shock protein inhibitors according to claim 1 or its pharmacy acceptable salt, is characterized in that, is selected from following compound:

7. the heat shock protein inhibitors described in claim 1-6 or the preparation method of its pharmacy acceptable salt, is characterized in that, adopts following circuit combination:

Wherein: R ₁₀be selected from: benzyl, trimethylsilyl, three second are silica-based, t-Butyldimethylsilyl, and tert-butyl diphenyl is silica-based, di-tert-butyl methylsilyl, methyl, ethanoyl, to methoxy-benzyl, and methoxymethyl;

A is selected from: halogen.

8. the heat shock protein inhibitors described in any one of claim 1-6 or its pharmacy acceptable salt, prepare prevention and therapy have expression of heat shock protein 90 increase pathological characteristics disease medicine in application.

9. application according to claim 8, is characterized in that, described in the disease of pathological characteristics that there is expression of heat shock protein 90 increases be: cancer, myelodysplastic syndrome, systemic mastocytosis, Xi Peier-lindau's syndrome, multicenter type Castleman is sick, psoriatic.

10. a pharmaceutical composition, is characterized in that, comprises the heat shock protein inhibitors described in any one of claim 1 to 6 or its pharmacy acceptable salt, and the acceptable vehicle of pharmacy or carrier.