CN109232541B

CN109232541B - Prolyl hydroxylase and histone deacetylase dual inhibitor and preparation method and application thereof

Info

Publication number: CN109232541B
Application number: CN201811163270.9A
Authority: CN
Inventors: 李祎亮; 魏会强; 高骏; 毕常芬; 宁洪鑫; 于江; 勾文峰; 段玉清
Original assignee: Institute of Radiation Medicine of CAMMS
Current assignee: Institute of Radiation Medicine of CAMMS
Priority date: 2018-09-30
Filing date: 2018-09-30
Publication date: 2020-04-24
Anticipated expiration: 2038-09-30
Also published as: CN109232541A

Abstract

The invention discloses a novel prolyl hydroxylase and histone deacetylase dual inhibitor shown in a formula I, a preparation method and application thereof, and the inhibitor can selectively improve the level of hypoxia inducible factor-2 α (HIF-2 α) by inhibiting the activities of Prolyl Hydroxylase (PHD) and Histone Deacetylase (HDAC) at the same time, and is expected to become a novel radioprotectant.

Description

Prolyl hydroxylase and histone deacetylase dual inhibitor and preparation method and application thereof

Technical Field

The invention relates to a novel small-molecule prolyl hydroxylase and histone deacetylase dual inhibitor, a preparation method of the compound, and application of the compound as the prolyl hydroxylase and histone deacetylase dual inhibitor in preparation of radioprotective medicaments.

Background

The following discussion is presented in the context of a description of the invention to aid in understanding the invention, but is not to be construed as prior art to the invention. All cited publications are incorporated herein by reference in their entirety.

The hypoxia-inducible factor (HIF) is a nuclear transcription factor induced during hypoxia, and is a heterodimer consisting of a functional α subunit and a structural β subunit, the HIF mainly comprises 3 subtypes, namely HIF-l, HIF-2 and HIF-3, wherein the HIF mainly participates in adjusting adaptive hypoxia of cells, and is mainly HIF-l and HIF-2.

The proline residues at specific sites of the oxygen-dependent degradation domains (ODDs) of HIF-l α and HIF-2 α can be hydroxylated by Prolyl Hydroxylases (PHDs) and then rapidly degraded by the ubiquitin-like pathway Prolyl hydroxylase inhibitors can inhibit this process, leading to increased stability of HIF-1 α 0/2 α in vivo. studies of the Giacacia subject group of Stanford university, USA, show that blocking HIF degradation in mice, can significantly reduce mortality upon exposure to lethal doses, the oral administration of the nonspecific Prolyl hydroxylase inhibitor oxalglycine dimethyl ester (DMOG), and selectively stabilize HIF-1 α and HIF-2 α, respectively, show that HIF-2 α radioprotection is significant, HIF-1 α is not significant (Science, translational, 2014.6(236 ra 7-236 ra64), the existing HIF-2 inhibitors are associated with increased HIF 1- α at the same time, and thus, the issue of elevated HIF-21-7-148, 685-148, HIF-18, HIF-7-18, HIF-19, HIF-18, HIF-7-18, HIF-9, HIF-7-9, HIF-7-9, HIF.

Degradation of the α subunit of HIF occurs mainly in the oxygen dependent degradation domain (ODD), HIF-1 α and HIF-2 α differ significantly in this domain in that the K532 lysine residue of the ODD domain of HIF-1 α can be acetylated (cell.2002,111: 709. cndot. 720.), acetylated HIF-1 α is more easily degraded, whereas the absence of a similar acetylation modifying regulatory mechanism in the corresponding region of HIF-2 α (Nature Reviews cancer.2012,12:9-22) Histone deacetylases of type I and IIa (Histone deacylase, HDAC) can inhibit the acetylation modification process of the ODD domain of HIF-1 α, thereby increasing the HDAC-1 α level (Nature mechanical considerations. 2016,7: 10539). HDAC 27 (type IIa) and HDAC6 (HIF II HIF) can inhibit the formation of the heat-resistant transcriptional shock-activity complex of the HDAC5, HSP-5, HIF-5, and HIF-5, respectively, thereby increasing the effect of inhibiting the formation of the heat resistant transcriptional shock-resistant protein from the HSP-5, and HIF-5, and HSP-5, respectively, and HSP-5, respectively, and HSP-5, and HIF-5, and a corresponding, respectively, and a corresponding transcriptional regulatory mechanisms.

Disclosure of Invention

The invention aims to provide a preparation method and application of a novel prolyl hydroxylase and histone deacetylase dual inhibitor compound aiming at the defects of the prior art, so as to solve the technical problem that a similar compound is lacked in the prior art;

another technical problem to be solved by the present invention is that the scope of use of the compounds described in the background of the invention is relatively limited;

yet another technical problem to be solved by the present invention is that the existing compounds do not select for a novel stable HIF-2 α protein;

the invention also provides an application of the compound serving as a dual inhibitor of prolyl hydroxylase and histone deacetylase in preparation of radioprotectants.

In order to achieve the technical purpose, the invention adopts the following technical scheme:

a compound represented by formula I:

wherein:

a) x1, X2 and X3 are respectively and independently selected from a C atom, a N atom, a S atom and an O atom;

b) r is independently selected from C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, -CO (C1-C4 alkyl), -CO (aryl), -CO (heteroaryl), -CO (C3-C6 cycloalkyl), -CO (C3-C6 heterocycloalkyl), -SO2(C1-C4 alkyl), C3-C8 cycloalkyl, C3-C8 heterocycloalkyl, aryl, C1-C10 alkyl-aryl, heteroaryl and C1-C10 alkyl-heteroaryl;

c) n1 is 1-2;

preferably, X is₁、X₂、X₃Each independently selected from a C atom or an N atom, and X₁，X₂、X₃At least one of which is a nitrogen atom.

Preferably, R is independently selected from the following chemical structures:

preferably, n is₁Is 1.

Preferably, the compound or a pharmaceutically acceptable salt thereof is selected from:

the invention provides a pharmaceutical composition, which comprises the compound or the pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, including a diluent.

Meanwhile, the invention provides the compound or the pharmaceutically acceptable salt thereof and the application of the pharmaceutical composition in preparing prolyl hydroxylase inhibitor drugs.

Meanwhile, the invention provides the compound or pharmaceutically acceptable salt thereof and application of the pharmaceutical composition in preparing histone deacetylase inhibitor medicines.

Preferably, the compound or a pharmaceutically acceptable salt thereof and the pharmaceutical composition are drugs against radioprotection.

Meanwhile, the invention provides a preparation method of the compound, which comprises the following steps:

1) taking the compound A, and reacting the compound A with diethyl ethoxymethylidene malonate and organic base in ethanol to obtain a compound B;

2) reacting the product B obtained in the step 1) with p-methoxy benzyl chloride, potassium carbonate and potassium iodide in a polar aprotic solvent to obtain a compound C;

3) reacting the product C obtained in the step 2) with sodium hydroxide in a mixed solution of water and tetrahydrofuran to obtain a compound D;

4) taking a compound E, reacting with methyl p-formylcinnamate, acetic acid and sodium triacetoxyborohydride in 1, 2-dichloroethane, or reacting with methyl 2-chloropyrimidine-5-carboxylate and organic base in N, N-dimethylformamide, or taking a compound F, reacting with methyl p-hydroxycinnamate or methyl p-hydroxybenzoate and organic base in N, N-dimethylformamide, and obtaining a compound G;

5) reacting the product G obtained in the step 4) in a dichloromethane solution of trifluoroacetic acid to obtain a compound H;

6) reacting the product H or the amino aliphatic carboxylic acid ester obtained in the step 5) with a compound D, 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride, 1-hydroxybenzotriazole and an organic base in a polar aprotic solvent to obtain a compound I;

7) reacting the product I obtained in the step 6) in a dichloromethane solution of trifluoroacetic acid to obtain a compound J;

8) reacting the product J obtained in the step 7) with hydroxylamine hydrochloride and sodium hydroxide aqueous solution in 1, 4-dioxane to obtain the compound with the structure shown in the formula (I) in the claims 1-6;

wherein the structure of each compound is shown as follows:

preferably, the reactions of step 1) and step 2) are carried out under heating.

Preferably, the polar aprotic solvent of step 2) and step 6) is N, N-dimethylformamide, dimethyl sulfoxide, acetonitrile, acetone, methyl ethyl ketone; more preferably, the solvent is N, N-dimethylformamide.

Preferably, the organic base in step 1), step 4) and step 6) is triethylamine or N, N-diisopropylethylamine.

Preferably, the concentration of the dichloromethane solution of trifluoroacetic acid in the step 5) and the step 7) is 10-50%, and more preferably, the solution solubility is 30%.

In addition to the preparation methods provided above, the compounds of the present invention can be prepared according to the following procedures. It should be understood that the following methods are only illustrative of the preparation process and the claims of the present invention are not limited thereto. A typical synthesis method of formula (I) can be summarized in the basic steps: (1) preparing PMB protected carboxylic acid, namely reacting a corresponding heterocyclic amidine compound with diethyl ethoxymethylidene malonate, and carrying out ester hydrolysis after PMB protection to obtain the carboxylic acid; (2) preparing an amino connecting group, namely carrying out reductive amination on a corresponding N-protective connecting group and methyl p-formylcinnamate, or reacting with 2-chloropyrimidine-5-methyl carboxylate, methyl p-hydroxycinnamate and methyl p-hydroxybenzoate, and then removing an N-Boc protective group to obtain the amino connecting group; (3) synthesizing the final product, namely, carrying out amide condensation on a carboxylic acid part and an amino connecting group part, and then condensing the corresponding methyl carboxylate with hydroxylamine hydrochloride to obtain the final product.

The aromatic heterocyclic amidine is that the compound A reacts with diethyl ethoxymethylidene malonate in ethanol to obtain a compound B, and the compound 2 is protected by PMB and hydrolyzed into acid to obtain carboxylic acid D. And reacting the compound E with methyl p-formyl cinnamate or 2-chloropyrimidine-5-carboxylate, reacting the compound F with methyl p-hydroxycinnamate or methyl p-hydroxybenzoate to obtain an N-protected methyl carboxylate intermediate G, and carrying out N-deprotection reaction on the G to obtain an amino connecting group H. And (3) condensing the carboxylic acid D and the amino connecting group H under the action of EDCI and HOBT to obtain an intermediate I, removing PMB protection from the intermediate I to obtain an intermediate J, and finally condensing the intermediate J with hydroxylamine hydrochloride to obtain a final product. The reaction flow of the preparation method is shown in the reaction formula.

Technical terms related to the above technical solutions, unless specifically explained, will follow the following definitions.

Term "Alkyl "refers to a straight or branched chain hydrocarbon group having the indicated number of carbon atoms, thus, for example, the term" C "as used herein₁-C₄Alkyl "and" C₁-C₁₀Alkyl "refers to alkyl groups having at least 1 and at most 4 or 10 carbon atoms, respectively. Examples of such branched or straight chain alkyl groups for use in the present invention include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, isobutyl, n-butyl, tert-butyl, n-pentyl, isopentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, and n-decyl. And branched analogs of the latter five n-alkanes.

When the term "alkenyl" (or "alkenylene") is used, it refers to a straight or branched hydrocarbon chain containing the specified number of carbon atoms and at least 1 and up to 5 carbon-carbon double bonds. Examples include ethenyl (or ethenylene) and propenyl (or propenylene).

When the term "alkynyl" (or "alkynylene") is used, it refers to a straight or branched hydrocarbon chain containing the specified number of carbon atoms and at least 1 and up to 5 carbon-carbon triple bonds. Examples include ethynyl (or ethynylene) and propynyl (or propynyl).

When the term "cycloalkyl" is used, it refers to a non-aromatic, saturated, cyclic hydrocarbon ring containing the specified number of carbon atoms. Thus, for example, the term "C₃-C₈Cycloalkyl "refers to a non-aromatic cyclic hydrocarbon ring having 3 to 8 carbon atoms. Exemplary "C" as used in the present invention₃-C₈Cycloalkyl "groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

When the term "C" is used₅-C₈Cycloalkenyl "refers to a non-aromatic monocyclic carbocyclic ring having the indicated number of carbon atoms and up to 3 carbon-carbon double bonds. Exemplary "cycloalkenyl" groups include cyclopentenyl and cyclohexenyl.

When the term "C" is used₃-C₈Heterocycloalkyl "means a non-aromatic heterocyclic ring containing the specified number of ring atoms, which is saturated or has one or more degrees of unsaturation, and contains one or more heteroatoms selected from O, S or N. This is achieved byThe quasi-ring may be optionally fused to one or more other "heterocyclic" or cycloalkyl groups. Examples of "heterocyclic" groups include, but are not limited to, aziridine, thietane, oxetane, azetidine, thietane, oxetane, tetrahydrofuran, pyran, 1, 4-dioxane, 1, 4-dithiane, 1, 3-dioxane, 1, 3-dioxolane, piperidine, piperazine, 2, 4-piperazinedione, pyrrolidine, 2-imidazoline, imidazolidine, pyrazolidine, pyrazoline, morpholine, thiomorpholine, tetrahydrofuran, tetrahydrothiophene, and the like.

The term "aryl" refers to an aromatic group containing 5 to 14 ring atoms, at least one of which possesses a conjugated pi-electron system, including aromatic, heteroaromatic and fused aromatic or biaryl rings having all carbon atoms, and may bear substituents. The aryl group may carry 1 to 6 substituents. Heteroaromatic or heteroaromatic rings are groups containing 5 to 14 ring atoms, of which 1 to 4 heteroatoms are aromatic ring atoms and the remaining ring atoms are carbon atoms. Suitable heteroatoms are oxygen, sulfur, nitrogen, and selenium atoms. Suitable aromatic heterocycles are furan, thiophene, pyridine, pyrrolidine with a lower alkyl substituent on the nitrogen, pyridine nitroxide, pyrimidine, pyrazine, imidazole and other similar heterocycles, all of which may bear substituents.

The term "optionally substituted" or "substituted" refers to groups bearing 1-4 different substituents and may be respectively lower alkyl, lower aryl, lower aralkyl, lower cycloalkyl, lower heterocycloalkyl, hydroxy, lower alkoxy, lower aryloxy, polyhaloalkoxy, aralkoxy, lower heteroaryl, lower heteroaryloxy, lower heteroaryralkyl, lower heteroaralkoxy, azido, amino, halogen, lower alkylthio, oxy, lower acylalkyl, lower carboxylate, carboxylic acid, amide, nitro, lower acyloxy, lower aminoalkyl, lower alkylaminoaryl, lower alkylaryl, lower alkylaminoalkyl, lower alkoxyaryl, lower arylamino, lower aralkylamino, sulfonyl, lower acylaminoalkylaryl, lower amidinoaryl, lower acylaminoaryl, lower aralkylamino, lower acylaminoaryl, lower heteroarylalkyl, sulfonyl, lower amidi, Lower hydroxyalkyl, lower haloalkyl, lower alkylaminoalkyl, lower ureidoalkyl, cyano, lower alkoxyalkyl, lower polyhaloalkyl, lower aralkoxyalkyl.

"substituted aryl" and "substituted heteroaryl" refer to aromatic or heteroaromatic groups having 1 to 6 substituents on the aromatic or heteroaromatic ring. These substituents may be lower alkyl, lower alkoxy, lower polyhaloalkyl, halogen, hydroxy and amino.

Drawings

FIG. 1: results of the HIF-1 and HIF-2 overexpression assay.

Detailed Description

The compounds and preparations of the present invention are better illustrated by the following examples. These examples should not be construed as limitations of the present invention, and variations of these compounds, now known or later developed, should also be considered within the scope of the present invention and claimed.

Hereinafter, specific embodiments of the present invention will be described in detail. Well-known structures or functions may not be shown in detail in a following embodiment in order not to obscure the unnecessary detail.

Approximating language, as used herein in the following examples, may be applied to identify quantitative representations that could permissibly vary in number without resulting in a change in the basic function. Accordingly, a numerical value modified by a language such as "about", "left or right" is not limited to the precise numerical value itself. In some embodiments, "about" indicates that the value allowed for correction varies within plus or minus ten percent (+ -10%), for example, "about 100" indicates that any value between 90 and 110 is possible. Further, in the expression "about a first value to a second value", both the first and second values are corrected at about the same time. In some cases, the approximating language may be related to the precision of a measuring instrument.

Unless defined otherwise, technical and scientific terms used in the following examples have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

Example 14-hydroxy-2-pyrazol-1-ylpyrimidine-5-carboxylic acid ethyl ester (2)

Pyrazole-1-carboxamide hydrochloride (1eq) and N, N-diisopropylethylamine (2.2eq) were added to 5 times the volume of ethanol, stirred at room temperature for 20min, diethyl ethoxymethylidene malonate (1eq) was added, the mixture was heated to reflux, and the reaction was allowed to proceed overnight. After the reaction was completed, about half volume of ethanol was distilled off under reduced pressure, water and ethyl acetate were added to extract twice, the organic layer was washed once with water, and the aqueous layers were combined. Adjusting pH of water layer to 3-4, precipitating a large amount of white insoluble substance, filtering, and drying to obtain white powder of 2 with yield of about 90.25%. m.p.162.8-163.8 ℃; ESI-MS (M/z) 235.0822[ M + H]⁺；¹H-NMR(300MHz,DMSO-d₆)δ13.28(s,1H),8.63–8.56(m,2H),7.96(dd,J＝1.6,0.7Hz,1H),6.67(dd,J＝2.8,1.6Hz,1H),4.26(q,J＝7.1Hz,2H),1.29(t,J＝7.1Hz,3H)

Example 24- (4-Methoxybenzyloxy) -2-pyrazol-1-ylpyrimidine-5-carboxylic acid ethyl ester (3)

Adding 2 and anhydrous potassium carbonate (1.4eq) into N, N-dimethylformamide with the volume of 5 times, stirring for 30min at room temperature, adding a catalytic amount of potassium iodide (0.1eq), dissolving p-methoxybenzyl chloride into N, N-dimethylformamide with the volume of 5 times, dropwise adding the mixture into the system, heating to 80 ℃, and reacting overnight. TLC monitoring, after the reaction is completed, 10 times volume of water is added into the system, then ethyl acetate is used for extraction twice, organic layers are combined, and saturated NaHCO is used for extraction₃The solution was washed 2 times, and the organic layer was dried over anhydrous sodium sulfate. After the solvent was removed under reduced pressure, the residue was slurried with methanol and the filter cake was washed with mother liquor. The filter cake was dried to give 3 as a white solid powder in about 53.35% yield. m.p.96.8-97.8 deg.C; ESI-MS (M/z) 355.2[ M + H]⁺；¹H-NMR(300MHz,DMSO-d₆)δ8.97(d,J＝0.6Hz,1H),8.75(dd,J＝2.8,0.7Hz,1H),7.94–7.93(m,1H),7.53–7.47(m,2H),6.98–6.91(m,2H),6.66(dd,J＝2.7,1.5Hz,1H),5.58(s,2H),4.28(q,J＝7.1Hz,2H),3.75(s,3H),1.29(t,J＝7.1Hz,3H).

Example 34- (4-Methoxybenzyloxy) -2-pyrazol-1-ylpyrimidine-5-carboxylic acid ethyl ester (4)

3 and sodium hydroxide (5eq) were dissolved in a 10-fold volume amount of a tetrahydrofuran/water mixed solvent (volume ratio 1:1), stirred at room temperature, and the reaction was monitored by TLC. After the reaction is completed, partial tetrahydrofuran is evaporated under reduced pressure, water and ethyl acetate are added for extraction for 1 time, the pH value of a water layer is adjusted to 3-4, white solid is separated out, filtration is carried out, and drying is carried out to obtain 4 white powder, wherein the yield is about 87.97%. m.p.126.3-126.5 ℃; ESI-MS (M/z):327.3[ M + H]⁺；¹H-NMR(300MHz,DMSO-d₆)δ13.17(s,1H),8.95(s,1H),8.74(d,J＝2.7Hz,1H),7.93(d,J＝1.4Hz,1H),7.50(d,J＝8.6Hz,2H),6.93(d,J＝8.6Hz,2H),6.64(dd,J＝2.8,1.6Hz,1H),5.57(s,2H),3.75(s,3H).

EXAMPLE 43- [4- (4-tert-Butoxycarbonylaminopiperidin-1-ylmethyl) -phenyl ] -acrylic acid methyl ester (5)

Methyl p-formylcinnamate (1eq), 4-tert-butoxycarbonylaminopiperidine (1.1eq), acetic acid (2eq) and sodium triacetoxyborohydride (1.5eq) were added to 10-fold volume of 1, 2-dichloroethane and reacted at room temperature overnight. After the reaction was completed, the organic layer was washed twice with saturated aqueous sodium bicarbonate solution, the organic layer was dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a crude product of 5, which was used in the next step without purification. m.p.146.0-146.4 ℃; ESI-MS (M/z) 375.27[ M + H]⁺；¹H-NMR(300MHz,CDCl₃)δ7.67(d,J＝16.0Hz,1H),7.47(d,J＝8.2Hz,2H),7.33(d,J＝8.0Hz,2H),6.42(d,J＝16.0Hz,1H),4.47(d,J＝8.2Hz,1H),3.80(s,3H),3.51(s,2H),2.81(d,J＝11.5Hz,2H),2.12(t,J＝10.8Hz,2H),1.92(d,J＝12.5Hz,2H),1.44(s,9H),1.27(d,J＝9.6Hz,2H).

Example 53- [4- (4-Aminopiperidin-1-ylmethyl) -phenyl ] -acrylic acid methyl ester (6)

The crude product of 5 was dissolved in a 5-fold volume of 30% trifluoroacetic acid in dichloromethane, reacted at room temperature for about 30min, after completion of the reaction, the solvent was evaporated under reduced pressure, and the residue was separated by silica gel column (dichloromethane: methanol 10:1 → 5:1 → 1:1) to give 6 as a white powder in about 75% yield. m.p.219.7-220.1 deg.C; ESI-MS (M/z) 275.17[ M + H]⁺；¹H-NMR(300MHz,Methanol-d4)δ7.76–7.66(m,3H),7.55(d,J＝7.9Hz,2H),6.62(d,J＝16.0Hz,1H),4.36(s,2H),3.79(s,3H),3.59(d,J＝12.7Hz,2H),3.52–3.38(m,1H),3.16(t,J＝12.8Hz,2H),2.26(d,J＝14.0Hz,2H),1.95(d,J＝13.4Hz,2H).

Example 62- (4-tert-Butoxycarbonylaminopiperidin-1-yl) -pyrimidine-5-carboxylic acid methyl ester (7)

9.3g of 4- (N-Boc-amino) piperidine, 100mL of N, N-dimethylacetamide, 7.5g of N, N-diisopropylethylamine and 4.0g of 2-chloropyrimidine-5-carboxylic acid methyl ester are added in sequence into a 250mL three-necked flask, and the mixture is stirred at room temperature for 3 hours under the protection of nitrogen. The reaction was monitored by TLC (petroleum ether: ethyl acetate ═ 2:1), 200mL of water was added, dichloromethane was extracted (100mL × 3), and the organic phases were combined. The organic phase was washed successively with 5% dilute hydrochloric acid (100 mL. times.3) and saturated sodium chloride (100 mL. times.3). The organic phase was dried over anhydrous sodium sulfate for 2h, filtered under suction, the solvent was evaporated under reduced pressure and the crude product was purified by flash column chromatography (petroleum ether: ethyl acetate: 5:1) to give 6.8g of a white solid with a yield of 87.1%. m.p.180.7-181.3 ℃; ESI-MS (M/z) 337.18[ M + H]⁺；¹H NMR(400MHz,Chloroform-d)δ8.81(s,2H),4.76(dd,J＝13.7,3.9Hz,2H),4.50(d,J＝7.9Hz,1H),3.86(s,3H),3.75(s,1H),3.11(ddd,J＝14.2,11.7,2.8Hz,2H),2.05(dt,J＝13.7,3.4Hz,2H),1.44(s,9H),1.41–1.29(m,2H).¹³C NMR(101MHz,CDCl₃)δ165.27,161.96,159.80,155.11,112.16,79.51,51.71,48.07,42.93,32.35,28.40.

Example 72- (4-Aminopiperidin-1-yl) -pyrimidine-5-carboxylic acid methyl ester (8)

The intermediate 7 obtained in example 6 was subjected to the same procedure as in example 5, step 5 to obtain an intermediate 8. The product was purified by flash column chromatography (dichloromethane: methanol ═ 20:1) to afford a white solid in 91.2% yield. m.p.237.5-237.8 ℃; ESI-MS (M/z):237.1[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ8.79(s,2H),8.14(s,2H),4.76(dt,J＝13.6,3.3Hz,2H),3.79(s,3H),3.38(tt,J＝11.3,4.1Hz,1H),3.13–3.03(m,2H),2.01(dd,J＝13.2,3.9Hz,2H),1.44(qd,J＝12.2,4.2Hz,2H).

EXAMPLE 8N-tert-Butoxycarbonyl-2-bromoethylamine (9)

10.0g of BOC anhydride, 100mL of dichloromethane and 9.4g of 2-bromoethylamine hydrobromide were added successively to a 100mL eggplant-shaped bottle, and stirred at 0 ℃ to add 7.0g of triethylamine dropwise thereto. Gradually warming to room temperature and stirring for 13 h. 100mL of methylene chloride was added thereto, and the mixture was washed with saturated ammonium chloride (100 mL. times.2), saturated sodium bicarbonate (100 mL. times.2) and saturated sodium chloride (100 mL. times.2) in this order. The organic phase was dried overnight with anhydrous sodium sulfate and concentrated to dryness under reduced pressure to give 9.2g of a pale yellow oil, 89.6% yield. . ESI-MS (M/z) 247.98[ M + Na ]]⁺；¹H-NMR(400MHz,Chloroform-d)δ4.98(s,1H),3.53(q,J＝5.9Hz,2H),3.45(d,J＝5.8Hz,2H),1.44(s,9H).

Example 93- [4- (2-tert-Butoxycarbonylaminoethoxy) -phenyl ] -acrylic acid methyl ester (10)

In a 250mL three-necked flask, 10.0g of methyl 4-hydroxycinnamate, 100mL of dried DMF, and 15.5g of potassium carbonate were added, and the mixture was stirred at room temperature for 0.5 h. 01A, 12.5g are added, the temperature is raised to 90 ℃ and stirred for 5 h. TLC monitoring (chlorine)Carrying out simulation: acetone ═ 30:1) after the reaction was complete, cool to room temperature. To the reaction solution, 100mL of water was added, extracted with ethyl acetate (100 mL. times.3), and the organic phases were combined. The organic phase was washed with saturated sodium bicarbonate (100 mL. times.2) and saturated sodium chloride (100 mL. times.2) in this order. The organic phase was dried over anhydrous sodium sulfate overnight. After suction filtration and concentration under reduced pressure, the crude product was purified by flash column chromatography (petroleum ether: ethyl acetate: 2:1) to obtain 15.6g of a white solid with a yield of 86.4%. ESI-MS (M/z) 344.15[ M + Na ]]⁺；¹H NMR(400MHz,Chloroform-d)δ7.63(d,J＝16.0Hz,1H),7.48–7.43(m,2H),6.92–6.85(m,2H),6.30(d,J＝15.9Hz,1H),5.01(s,1H),4.04(t,J＝5.1Hz,2H),3.78(s,3H),3.54(q,J＝5.4Hz,2H),1.44(s,9H).¹³C NMR(101MHz,CDCl₃)δ167.70,160.34,155.86,144.37,129.74,127.44,115.49,114.78,79.63,67.28,51.60,40.01,28.38.

Example 103- [4- (2-Aminoethoxy) -phenyl ] -acrylic acid methyl ester (11)

The intermediate 10 obtained in example 9 was subjected to the same procedure as in example 5, step 5 to obtain an intermediate 11. The crude product was purified by flash column chromatography (dichloromethane: methanol ═ 20:1) to give 10.1g of a white solid in 94.0% yield. m.p.201.4-202.1 deg.C; ESI-MS (M/z) 222.04[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ8.23(s,2H),7.70(d,J＝8.8Hz,2H),7.62(d,J＝16.0Hz,1H),7.02(d,J＝8.8Hz,2H),6.51(d,J＝16.0Hz,1H),4.22(t,J＝5.1Hz,2H),3.70(s,3H),3.25(t,J＝5.0Hz,2H).¹³C NMR(101MHz,DMSO)δ166.84,159.64,144.12,130.11,127.16,115.43,114.94,64.46,51.27,38.19.

EXAMPLE 11N-tert-Butoxycarbonyl-2-bromoethylamine (12)

Compound 12 was obtained in the same manner as in example 8 using 3-bromopropylamine hydrobromide as the starting material. The product is light yellow oilThe yield was 91.1%.¹H NMR(400MHz,Chloroform-d)δ4.73(s,1H),3.41(td,J＝6.7,2.3Hz,2H),3.23(td,J＝6.8,2.3Hz,2H),2.06–1.96(m,2H),1.41(d,J＝2.4Hz,9H).

Example 123- [4- (3-tert-Butoxycarbonylaminopropoxy) -phenyl ] -acrylic acid methyl ester (13)

Intermediate 12 obtained in example 11 was subjected to the same procedure as in example 9, step 4 to give intermediate 13. The product was a white solid with a yield of 89.5%. m.p.87.3-87.9 ℃; ESI-MS (M/z) 358.24[ M + Na ]]⁺；¹H NMR(400MHz,DMSO-d₆)δ7.66(d,J＝8.5Hz,2H),7.61(d,J＝16.2Hz,1H),6.95(d,J＝9.0Hz,2H),6.90(d,J＝5.9Hz,1H),6.48(d,J＝15.9Hz,1H),4.01(t,J＝6.1Hz,2H),3.70(s,3H),3.08(q,J＝6.3Hz,2H),1.83(p,J＝6.7Hz,2H),1.37(s,9H).¹³C NMR(101MHz,CDCl₃)δ167.73,160.59,156.02,144.47,129.71,127.20,115.31,114.81,79.28,65.83,51.55,37.90,29.54,28.40.

Example 133- [4- (3-Aminopropoxy) -phenyl ] -acrylic acid methyl ester (14)

The intermediate 13 obtained in example 12 was subjected to the same procedure as in example 5, step 5 to obtain an intermediate 20. The product was purified by flash column chromatography (dichloromethane: methanol ═ 20:1) to afford a white solid in 91.2% yield. m.p.80.5-81.8 deg.C; ESI-MS (M/z) 236.08[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ7.96(s,2H),7.68(d,J＝8.6Hz,2H),7.61(d,J＝16.0Hz,1H),6.97(d,J＝8.4Hz,2H),6.49(d,J＝16.0Hz,1H),4.10(t,J＝6.1Hz,2H),3.70(s,3H),2.96(d,J＝8.3Hz,2H),2.01(p,J＝6.6Hz,2H).¹³C NMR(101MHz,DMSO)δ167.36,160.65,144.72,130.58,127.24,115.66,115.33,65.23,51.75,36.68,27.21.

Example 144- (2-tert-Butoxycarbonylaminoethoxy) -benzoic acid methyl ester (15)

Intermediate 15 was obtained in the same manner as in example 9, step 4, starting from 9 and methyl paraben. The product was a pale yellow oil, yield 87.7%. The crude product was used in the next reaction without further treatment.¹H-NMR(400MHz,Chloroform-d)δ7.97(dd,J＝8.8,2.7Hz,2H),6.89(dd,J＝8.9,2.7Hz,2H),5.02(s,1H),4.06(t,J＝4.4Hz,2H),3.87(d,J＝2.7Hz,3H),3.54(d,J＝5.8Hz,2H),1.44(d,J＝2.7Hz,9H).

Example 154- (2-Aminoethoxy) -benzoic acid methyl ester (16)

Intermediate 15 obtained in example 14 was subjected to the same procedure as in example 5, step 5 to give intermediate 16. The product was purified by flash column chromatography (dichloromethane: methanol ═ 20:1) to afford a pale yellow white solid in 85.8% yield. m.p.129.0-129.9 ℃; ESI-MS (M/z) 210.43[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ7.98(s,2H),7.91(d,J＝8.5Hz,2H),7.04(d,J＝8.4Hz,2H),4.14(t,J＝6.3Hz,2H),3.81(s,3H),2.98(t,J＝7.4Hz,2H),2.03(p,J＝6.8Hz,2H).

Example 164- (3-tert-Butoxycarbonylaminopropoxy) -benzoic acid methyl ester (17)

Intermediate 17 was obtained in the same manner as in example 9, step 4, starting from 12 and methyl paraben. The product was a pale yellow oil, 91.3% yield. ESI-MS (M/z) 332.17[ M + Na ]]⁺；¹H NMR(400MHz,Chloroform-d)δ7.99–7.94(m,2H),6.91–6.86(m,2H),4.76(s,1H),4.05(t,J＝6.0Hz,2H),3.87(s,3H),3.31(t,J＝6.7Hz,2H),1.99(p,J＝6.4Hz,2H),1.42(s,9H).

Example 174- (2-Aminoethoxy) -benzoic acid methyl ester (18)

Intermediate 17 obtained in example 16 was subjected to the same procedure as in example 5, step 5 to give intermediate 18. The product was purified by flash column chromatography (dichloromethane: methanol ═ 20:1) to afford a pale yellow white solid in 85.8% yield. m.p.129.0-129.9 ℃; ESI-MS (M/z) 210.43[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ7.98(s,2H),7.91(d,J＝8.5Hz,2H),7.04(d,J＝8.4Hz,2H),4.14(t,J＝6.3Hz,2H),3.81(s,3H),2.98(t,J＝7.4Hz,2H),2.03(p,J＝6.8Hz,2H).

Example 183- [4- (4- { [4- (4-methoxybenzyloxy) -2-pyrazol-1-yl-pyrimidine-5-carbonyl ] -amino } -piperidin-1-ylmethyl) -phenyl ] -acrylic acid methyl ester (19)

A100 mL bottle of eggplant was charged with 65.0 g of intermediate, 47.4 g of intermediate, 4.8g of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride, 3.4g of 1-hydroxybenzotriazole, 25mL of N, N-dimethylformamide, and 8.8g of N, N-diisopropylethylamine in that order, and stirred at room temperature for 2.0 hours. The reaction was terminated by TLC (dichloromethane: methanol ═ 20:1), 100mL of dichloromethane was added, and 5% dilute hydrochloric acid (100mL × 2), saturated sodium bicarbonate extraction (100mL × 2), and saturated sodium chloride (100mL × 2) were added in this order to wash the mixture. The organic phase was dried over anhydrous sodium sulfate for 2.0h, filtered, evaporated under reduced pressure to remove the solvent, and the product was purified by flash column chromatography (petroleum ether: ethyl acetate: 2:1) to give 10.5g of a white solid with a yield of 87.5%. m.p.133.5-133.9 deg.C; ESI-MS (M/z) 530.11[ M + H]⁺；¹H NMR(400MHz,Chloroform-d)δ9.30(s,1H),8.58(d,J＝2.8Hz,1H),7.91–7.87(m,1H),7.86(d,J＝1.5Hz,1H),7.63(d,J＝15.9Hz,1H),7.46–7.40(m,2H),7.37(d,J＝8.5Hz,2H),6.81–6.76(m,2H),6.75–6.71(m,2H),6.51(dd,J＝2.7,1.5Hz,1H),6.30(d,J＝16.0Hz,1H),5.61(s,2H),4.06(t,J＝5.1Hz,2H),3.83–3.79(m,2H),3.77(s,3H),3.72(s,3H).¹³C NMR(101MHz,CDCl₃)δ167.64,166.97,163.37,162.04,160.13,160.11,155.97,144.58,144.26,130.32,129.83,129.70,127.54,126.38,115.60,114.79,114.22,110.71,109.31,70.23,66.57,55.23,51.64,39.11.

Example 193- (4- {4- [ (4-hydroxy-2-pyrazol-1-yl-pyrimidine-5-carbonyl) -amino ] -piperidin-1-ylmethyl } -phenyl) -acrylic acid methyl ester (20)

199.0 g of the intermediate and 150mL of methylene chloride were added to a 250mL eggplant-shaped bottle, and 5.8g of trifluoroacetic acid was added dropwise at 0 ℃ and stirred at 0 ℃ for 1.0 hour. The completion of the reaction was monitored by TLC (petroleum ether: ethyl acetate: 1:3), and the solvent was distilled off under reduced pressure, 100mL of dichloromethane was added thereto, and the solvent was distilled off again. 20mL of methanol is added for pulping for 0.5h, and after suction filtration and drying, 6.7g of white solid is obtained with the yield of 96.3%. m.p.264.0-264.7 ℃; ESI-MS (M/z) 410.1453[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ13.51(s,1H),9.61(s,1H),8.62(d,J＝2.8Hz,1H),8.45(s,1H),8.06(d,J＝1.6Hz,1H),7.68(d,J＝8.8Hz,2H),7.61(d,J＝16.0Hz,1H),7.02(d,J＝8.8Hz,2H),6.72(dd,J＝2.8,1.6Hz,1H),6.49(d,J＝16.0Hz,1H),4.16(t,J＝5.6Hz,2H),3.70(d,J＝3.6Hz,5H).¹³C NMR(101MHz,TFA-D)δ174.91,171.12,166.84,162.86,155.66,151.30,149.92,149.49,133.28,132.62,130.23,117.49,116.10,114.92,113.99,68.35,54.75,42.20.

Example 203- (4- {4- [ (4-hydroxy-2-pyrazol-1-yl-pyrimidine-5-carbonyl) -amino ] -piperidin-1-ylmethyl } -phenyl) -propen-e hydroxamic acid (21)

202.0 g of intermediate, 3.4g of hydroxylamine hydrochloride and 20mL of 1, 4-dioxane are added into a 100mL eggplant-shaped bottle in sequence under a water bath at 15 ℃. 3.9g NaOH was dissolved in 20mL water and added dropwise to the reaction flask, one by oneThe temperature was gradually raised to room temperature and stirred overnight. The reaction was monitored by TLC the next day (dichloromethane: methanol ═ 15:1), 40mL of water was added, ethyl acetate was added for extraction (20mL × 2), the organic phase was discarded, 1N of dilute hydrochloric acid was added dropwise to the aqueous phase to adjust the pH to 3-4, a large amount of white solid was precipitated, stirred for 0.5h, filtered under suction, and dried to give 1.8g of pale purple solid with a yield of 89.3%. m.p.196.5-197.6 ℃; ESI-MS (M/z) 411.1404[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ13.56(s,1H),10.68(s,1H),9.60(s,1H),9.00(s,1H),8.62(d,J＝2.8Hz,1H),8.46(s,1H),8.06(s,1H),7.50(d,J＝8.2Hz,2H),7.40(d,J＝15.8Hz,1H),7.01(d,J＝8.3Hz,2H),6.72(t,J＝2.1Hz,1H),6.32(d,J＝15.7Hz,1H),4.14(t,J＝5.6Hz,2H),3.70(q,J＝5.7Hz,2H).¹³C NMR(101MHz,D₂O)δ173.15,167.55,162.21,158.46,158.14,156.11,143.22,134.47,129.63,128.55,128.45,116.95,114.92,110.47,108.57,66.39,38.42.

Example 212- (4- (4- (4-methoxybenzyloxy) -2-pyrazol-1-yl-pyrimidine-5-carbonyl) -aminopiperidin-1-yl) -pyrimidine-5-carboxylic acid methyl ester (22)

Intermediate 22 was obtained in the same manner as in example 18, step 6, starting from intermediate 4 and intermediate 8. The product was purified by flash column chromatography (petroleum ether: ethyl acetate 2:1) to give a white solid in 86.9% yield. m.p.155.2-156.1 ℃; ESI-MS (M/z) 583.34[ M + H]⁺；¹H NMR(400MHz,Chloroform-d)δ9.27(s,1H),8.59(d,J＝2.8Hz,1H),7.86(s,1H),7.67(d,J＝16.0Hz,1H),7.44(dd,J＝17.7,7.9Hz,5H),7.31(d,J＝7.7Hz,2H),6.95(d,J＝8.1Hz,2H),6.51(s,1H),6.41(d,J＝16.1Hz,1H),5.58(s,2H),3.97(s,1H),3.79(d,J＝2.6Hz,6H),3.40(s,2H),2.49(s,2H),2.18(t,J＝10.5Hz,2H),1.93–1.82(m,2H),1.39(d,J＝11.3Hz,2H).¹³C NMR(101MHz,CDCl₃)δ167.47,166.90,163.24,160.96,160.34,155.89,144.59,144.51,141.16,133.25,130.63,129.83,129.43,128.03,126.40,117.40,114.41,111.01,109.23,70.43,62.60,55.34,51.69,51.57,46.04,31.60.

Example 222- (4- (4-hydroxy-2-pyrazol-1-yl-pyrimidine-5-carbonyl) -aminopiperidin-1-yl) -pyrimidine-5-carboxylic acid methyl ester (23)

Intermediate 22 obtained in example 21 was subjected to the same procedure as in example 19, step 7 to give intermediate 23. Pulping with methanol for 0.5h, filtering, and drying to obtain white solid with yield of 97.4%. m.p.242.9-243.4; ESI-MS (M/z) 463.22[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ10.05(s,1H),9.44(s,1H),8.62(d,J＝2.8Hz,1H),8.45(s,1H),8.05(s,1H),7.84(d,J＝7.8Hz,2H),7.69(d,J＝16.1Hz,1H),7.57(d,J＝7.8Hz,2H),6.73(d,J＝16.2Hz,2H),4.39(s,2H),4.01(s,1H),3.73(s,3H),3.41(s,2H),3.14(s,2H),2.11(s,2H),1.73(s,2H).¹³C NMR(101MHz,TFA-D)δ173.76,171.00,166.09,155.66,151.41,149.40,147.96,139.31,133.71,133.35,131.77,131.68,120.98,114.89,113.98,64.05,54.94,54.78,47.87,31.05.

Example 232- (4- (4-hydroxy-2-pyrazol-1-yl-pyrimidine-5-carbonyl) -aminopiperidin-1-yl) -pyrimidine-5-hydroxamic acid (24)

Intermediate 23 obtained in example 22 was subjected to the same procedure as in example 20, step 8 to give intermediate 24. The product was a white solid in 83.8% yield. m.p.223.8-224.3 ℃; ESI-MS (M/z) 426.2436[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ13.53(s,1H),11.08(s,1H),9.41(s,1H),9.02(s,1H),8.68(s,2H),8.62(d,J＝2.8Hz,1H),8.45(s,1H),8.06(d,J＝1.5Hz,1H),6.72(dd,J＝2.7,1.6Hz,1H),4.51(dt,J＝13.7,4.2Hz,2H),4.16–4.05(m,1H),3.29(ddd,J＝13.6,10.8,2.9Hz,2H),1.95(dt,J＝12.8,3.9Hz,2H),1.45(td,J＝10.5,6.9Hz,2H).¹³C NMR(101MHz,D₂O)δ173.23,166.59,161.26,160.25,158.27,156.15,156.06,143.22,129.57,116.62,110.53,108.56,46.15,42.67,30.51.

Example 243- [4- (2- { [4- (4-methoxybenzyloxy) -2-pyrazol-1-yl-pyrimidin-5-yl ] -amino } -ethoxy) -phenyl ] -acrylic acid methyl ester (25)

Intermediate 25 was obtained in the same manner as in example 18, step 6, starting from intermediate 4 and intermediate 11. The product was purified by flash column chromatography (petroleum ether: ethyl acetate 2:1) to give 10.5g of a white solid in 87.5% yield. m.p.133.5-133.9 deg.C; ESI-MS (M/z) 530.11[ M + H]⁺；¹H NMR(400MHz,Chloroform-d)δ9.30(s,1H),8.58(d,J＝2.8Hz,1H),7.91–7.87(m,1H),7.86(d,J＝1.5Hz,1H),7.63(d,J＝15.9Hz,1H),7.46–7.40(m,2H),7.37(d,J＝8.5Hz,2H),6.81–6.76(m,2H),6.75–6.71(m,2H),6.51(dd,J＝2.7,1.5Hz,1H),6.30(d,J＝16.0Hz,1H),5.61(s,2H),4.06(t,J＝5.1Hz,2H),3.83–3.79(m,2H),3.77(s,3H),3.72(s,3H).¹³C NMR(101MHz,CDCl₃)δ167.64,166.97,163.37,162.04,160.13,160.11,155.97,144.58,144.26,130.32,129.83,129.70,127.54,126.38,115.60,114.79,114.22,110.71,109.31,70.23,66.57,55.23,51.64,39.11.

Example 253- (4- {2- [ (4-hydroxy-2-pyrazol-1-yl-pyrimidine-5-carbonyl) -amino ] -ethoxy } -phenyl) -acrylic acid methyl ester (26)

Intermediate 25 obtained in example 24 was subjected to the same procedure as in example 19, step 7 to give intermediate 26. 6.7g of a white solid was obtained, the yield was 96.3%. m.p.264.0-264.7 ℃; ESI-MS (M/z) 410.1453[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ13.51(s,1H),9.61(s,1H),8.62(d,J＝2.8Hz,1H),8.45(s,1H),8.06(d,J＝1.6Hz,1H),7.68(d,J＝8.8Hz,2H),7.61(d,J＝16.0Hz,1H),7.02(d,J＝8.8Hz,2H),6.72(dd,J＝2.8,1.6Hz,1H),6.49(d,J＝16.0Hz,1H),4.16(t,J＝5.6Hz,2H),3.70(d,J＝3.6Hz,5H).¹³C NMR(101MHz,TFA-D)δ174.91,171.12,166.84,162.86,155.66,151.30,149.92,149.49,133.28,132.62,130.23,117.49,116.10,114.92,113.99,68.35,54.75,42.20.

Example 263- (4- {2- [ (4-hydroxy-2-pyrazol-1-yl-pyrimidin-5-yl) -amino ] -ethoxy } -phenyl) -propene hydroxamic acid (27)

Compound 27 was obtained according to the same method as that of example 20, step 8, and intermediate 26 obtained in example 25. 1.8g of light purple solid is obtained, and the yield is 89.3%. m.p.196.5-197.6 ℃; ESI-MS (M/z) 411.1404[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ13.56(s,1H),10.68(s,1H),9.60(s,1H),9.00(s,1H),8.62(d,J＝2.8Hz,1H),8.46(s,1H),8.06(s,1H),7.50(d,J＝8.2Hz,2H),7.40(d,J＝15.8Hz,1H),7.01(d,J＝8.3Hz,2H),6.72(t,J＝2.1Hz,1H),6.32(d,J＝15.7Hz,1H),4.14(t,J＝5.6Hz,2H),3.70(q,J＝5.7Hz,2H).¹³C NMR(101MHz,D₂O)δ173.15,167.55,162.21,158.46,158.14,156.11,143.22,134.47,129.63,128.55,128.45,116.95,114.92,110.47,108.57,66.39,38.42.

Example 273- [4- (3- { [4- (4-methoxybenzyloxy) -2-pyrazol-1-yl-pyrimidin-5-yl ] -amino } -propoxy) -phenyl ] -acrylic acid methyl ester (28)

Intermediate 28 was obtained in the same manner as in example 18, step 6, starting from intermediate 4 and intermediate 14. The product was purified by flash column chromatography (petroleum ether: ethyl acetate 2:1) to give a white solid in 91.5% yield. m.p.139.5-140.0 deg.C; ESI-MS (M/z) 544.33[ M + H]⁺；¹H NMR(400MHz,Chloroform-d)δ9.30(s,1H),8.60(d,J＝2.7Hz,1H),7.88(d,J＝1.5Hz,1H),7.64(d,J＝16.0Hz,1H),7.59(t,J＝6.4Hz,1H),7.48–7.42(m,2H),7.39–7.34(m,2H),6.88–6.84(m,2H),6.84–6.79(m,2H),6.55–6.51(m,1H),6.31(d,J＝15.9Hz,1H),5.59(s,2H),3.94(t,J＝6.0Hz,2H),3.79(s,3H),3.77(s,3H),3.58(q,J＝6.4Hz,2H),1.99(p,J＝6.3Hz,2H).¹³C NMR(101MHz,CDCl₃)δ167.70,166.90,163.25,161.97,160.45,160.19,155.90,144.56,144.38,130.36,129.83,129.71,127.24,126.42,115.37,114.76,114.27,110.91,109.29,70.26,65.46,55.29,51.62,36.83,28.75.

Example 283- (4- {3- [ (4-hydroxy-2-pyrazol-1-yl-pyrimidine-5-carbonyl) -amino ] -propoxy } -phenyl) -acrylic acid methyl ester (29)

Intermediate 28 obtained in example 27 was subjected to the same procedure as in example 19, step 7 to give intermediate 29. Pulping with methanol for 0.5h, vacuum filtering, and drying to obtain white solid with yield of 95.0%. m.p.; ESI-MS (M/z) 422.2908[ M-H]^-；¹H NMR(400MHz,DMSO-d₆)δ13.37(s,1H),9.47(s,1H),8.63(d,J＝2.8Hz,1H),8.43(s,1H),8.06(d,J＝1.6Hz,1H),7.68–7.64(m,2H),7.61(d,J＝16.0Hz,1H),7.04–6.99(m,2H),6.72(dd,J＝2.8,1.6Hz,1H),6.48(d,J＝16.0Hz,1H),4.09(t,J＝6.0Hz,2H),3.70(s,3H),3.48(q,J＝6.5Hz,2H),1.98(p,J＝6.5Hz,2H).¹³C NMR(101MHz,TFA-D)δ174.85,171.21,166.49,162.87,155.03,151.09,149.89,149.35,133.13,132.41,129.87,117.39,115.75,114.77,113.67,68.67,54.57,40.72,29.96.

Example 293- (4- {3- [ (4-hydroxy-2-pyrazol-1-yl-pyrimidin-5-yl) -amino ] -propoxy } -phenyl) -propen-e hydroximic acid (30)

Compound 30 was obtained according to the same method as that of example 20, step 8, and intermediate 29 obtained in example 28. The product was a white solid with a yield of 60.1%. m.p.208.8-209.0 ℃; ESI-MS (M/z) 423.3214[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ13.54(s,1H),10.67(s,1H),9.48(s,1H),8.99(s,1H),8.63(d,J＝2.8Hz,1H),8.44(s,1H),8.06(d,J＝1.5Hz,1H),7.50(d,J＝8.4Hz,2H),7.40(d,J＝15.8Hz,1H),7.03–6.98(m,2H),6.72(dd,J＝2.8,1.6Hz,1H),6.31(d,J＝15.8Hz,1H),4.07(t,J＝6.0Hz,2H),3.48(q,J＝6.4Hz,2H),1.98(p,J＝6.4Hz,2H).¹³C NMR(101MHz,DMSO)δ163.63,162.82,160.09,148.87,145.27,138.51,130.36,130.20,129.48,127.86,116.93,115.31,113.36,111.07,99.97,66.40,36.53,29.23.

Example 304- (2- { [4- (4-methoxybenzyloxy) -2-pyrazol-1-yl-pyrimidine-5-carbonyl ] -amino } -ethoxy) -benzoic acid methyl ester (31)

Intermediate 31 was obtained in the same manner as in example 18, step 6, starting from intermediate 4 and intermediate 16. The product was purified by flash column chromatography (petroleum ether: ethyl acetate 2:1) to give a white solid in 87.9% yield. m.p.141.9-142.4 ℃; ESI-MS (M/z) 504.1920[ M + H]⁺；¹H NMR(400MHz,Chloroform-d)δ9.32(d,J＝2.0Hz,1H),8.61(d,J＝2.4Hz,1H),8.00–7.94(m,2H),7.91(s,1H),7.88(s,1H),7.42–7.35(m,2H),6.83–6.79(m,2H),6.78–6.74(m,2H),6.53(t,J＝2.2Hz,1H),5.63(d,J＝2.0Hz,2H),4.13–4.07(m,2H),3.89(d,J＝2.0Hz,3H),3.85(t,J＝5.5Hz,2H),3.76(d,J＝2.0Hz,3H).¹³C NMR(101MHz,CDCl₃)δ167.01,166.69,163.40,162.08,160.16,156.01,144.59,131.59,130.33,130.31,129.85,126.36,123.07,114.25,114.01,110.70,109.31,70.28,66.63,55.23,51.94,39.08.

Example 314- {2- [ (4-hydroxy-2-pyrazol-1-yl-pyrimidine-5-carbonyl) -amino ] -ethoxy } -benzoic acid methyl ester (32)

Intermediate 31 obtained in example 30 was subjected to the same procedure as in example 18, step 6 to give intermediate 32. Pulping with methanol for 0.5h, vacuum filtering, and drying to obtain white solid with yield of 97.2%. And m is selected.p.263.0-263.9℃；ESI-MS(m/z):384.1329[M+H]⁺；¹H NMR(400MHz,DMSO-d₆)δ9.61(s,1H),8.62(s,1H),8.45(s,1H),8.06(s,1H),7.91(d,J＝8.3Hz,2H),7.09(d,J＝8.5Hz,2H),6.72(s,1H),4.20(t,J＝5.6Hz,2H),3.81(s,3H),3.72(d,J＝7.5Hz,2H).¹³C NMR(101MHz,D₂O)δ173.45,171.13,166.89,165.45,155.78,151.35,149.48,134.56,133.30,124.52,116.80,114.92,114.09,68.27,54.93,42.15.

Example 324- {2- [ (4-hydroxy-2-pyrazol-1-yl-pyrimidine-5-carbonyl) -amino ] -ethoxy } -benzohydroxamic acid (33)

Compound 33 was obtained according to the same method as that of example 20, step 8, intermediate 32 obtained in example 31. The product was a white solid in 74.2% yield. m.p.224.3-225.0 deg.C; ESI-MS (M/z) 385.1272[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ13.50(s,1H),11.09(s,1H),9.59(s,1H),8.92(s,1H),8.62(d,J＝2.8Hz,1H),8.46(s,1H),8.05(d,J＝1.5Hz,1H),7.77–7.68(m,2H),7.06–7.00(m,2H),6.72(dd,J＝2.8,1.6Hz,1H),4.16(t,J＝5.6Hz,2H),3.70(q,J＝5.7Hz,2H).¹³C NMR(101MHz,DMSO)δ163.90,162.55,160.51,148.34,144.85,131.34,129.72,128.63,125.14,114.32,114.17,112.71,110.65,66.49,37.96.

Example 334- (3- { [4- (4-methoxybenzyloxy) -2-pyrazol-1-yl-pyrimidine-5-carbonyl ] -amino } -propoxy) -benzoic acid methyl ester (34)

Intermediate 34 was obtained in the same manner as in example 18, step 6, starting from intermediate 4 and intermediate 18. The product was purified by flash column chromatography (petroleum ether: ethyl acetate 2:1) to give a white solid in 84.8% yield. m.p.113.8-114.8 ℃; ESI-MS (M/z) 518.36[ M + H]⁺；¹H NMR(400MHz,Chloroform-d)δ9.31(s,1H),8.61(d,J＝2.7Hz,1H),8.01–7.95(m,2H),7.89(s,1H),7.60(s,1H),7.37(d,J＝8.2Hz,2H),6.85(t,J＝9.2Hz,4H),6.54(d,J＝2.5Hz,1H),5.60(s,2H),3.95(t,J＝6.0Hz,2H),3.89(d,J＝1.2Hz,3H),3.78(d,J＝1.2Hz,3H),3.59(q,J＝6.4Hz,2H),2.00(p,J＝6.3Hz,2H).¹³C NMR(101MHz,CDCl₃)δ166.94,166.77,163.30,162.41,162.01,160.24,155.95,144.57,131.60,130.39,129.85,126.42,122.78,114.31,114.02,110.90,109.28,70.31,65.49,55.31,51.90,36.79,28.74.

Example 344- {3- [ (4-hydroxy-2-pyrazol-1-yl-pyrimidine-5-carbonyl) -amino ] -propoxy } -benzoic acid methyl ester (35)

Intermediate 34 obtained in example 33 was subjected to the same procedure as in example 18, step 6 to give intermediate 35. Pulping with methanol for 0.5h, vacuum filtering, and drying to obtain white solid with yield of 95.1%. m.p.221.1-221.8; ESI-MS (M/z) 398.17[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ13.48(s,1H),9.52(s,1H),8.63(d,J＝2.6Hz,1H),8.42(s,1H),8.06(s,1H),7.90(dd,J＝8.8,2.3Hz,2H),7.08(dd,J＝8.8,2.3Hz,2H),6.73(t,J＝2.4Hz,1H),4.12(t,J＝5.5Hz,2H),3.81(d,J＝2.2Hz,3H),3.49(q,J＝6.6Hz,2H),2.00(p,J＝7.1Hz,2H).¹³C NMR(101MHz,D₂O)δ173.47,171.33,166.52,165.69,155.13,151.24,149.49,134.45,133.27,124.05,116.73,114.97,113.79,68.54,54.87,40.85,30.09.

Example 354- {3- [ (4-hydroxy-2-pyrazol-1-yl-pyrimidine-5-carbonyl) -amino ] -propoxy } -benzohydroxamic acid (36)

Compound 36 was obtained according to the same method as that of example 20, step 8, and intermediate 35 obtained in example 34. The product was a white solid in 73.9% yield. m.p.215.0-215.4 ℃; ESI-MS (M/z) 399.19[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ13.49(s,1H),11.07(s,1H),9.48(s,1H),8.91(s,1H),8.63(d,J＝2.7Hz,1H),8.44(s,1H),8.05(d,J＝1.5Hz,1H),7.75–7.68(m,2H),7.04–7.00(m,2H),6.72(dd,J＝2.8,1.6Hz,1H),4.08(t,J＝6.0Hz,2H),3.48(q,J＝6.4Hz,2H),1.98(p,J＝6.3Hz,2H).¹³C NMR(101MHz,DMSO)δ164.49,162.83,161.27,148.88,145.27,131.76,130.20,129.05,125.37,114.71,114.55,113.37,111.06,66.47,36.52,29.20.

Example 364- ({ [4- (4-Methoxybenzyloxy) -2-pyrazol-1-yl-pyrimidin-5-yl ] -amino } -methyl) -benzoic acid methyl ester (37)

Intermediate 37 was obtained in the same manner as in example 18, step 6, starting from intermediate 4 and methyl 4-aminomethylbenzoate. The product was purified by flash column chromatography (petroleum ether: ethyl acetate 2:1) to give a white solid in 93.0% yield. m.p.161.3-162.0 deg.C; ESI-MS (M/z) 496.12[ M + Na ]]⁺；¹H NMR(400MHz,Chloroform-d)δ9.33(s,1H),8.62(dd,J＝2.8,0.7Hz,1H),7.94–7.90(m,2H),7.88(dd,J＝1.6,0.7Hz,1H),7.78(t,J＝5.5Hz,1H),7.31–7.27(m,2H),7.23–7.17(m,2H),6.85–6.80(m,2H),6.54(dd,J＝2.7,1.6Hz,1H),5.57(s,2H),4.60(d,J＝5.4Hz,2H),3.93(s,3H),3.82(s,3H).¹³C NMR(101MHz,CDCl₃)δ166.98,166.70,163.42,161.83,160.27,156.08,144.65,142.62,130.49,130.02,129.90,129.33,127.47,126.05,114.29,110.63,109.34,70.55,55.28,52.16,43.58.

Example 374- { [ (4-hydroxy-2-pyrazol-1-yl-pyrimidine-5-carbonyl) -amino ] -methyl } -benzoic acid methyl ester (38)

Intermediate 37 obtained in example 36 was subjected to the same procedure as in example 18, step 6 to give intermediate 38. Pulping with methanol for 0.5h, filtering, and drying to obtain white solid with yield of 97.3%. m.p.280.0-281.4；ESI-MS(m/z):352.20[M-H]^-；¹H NMR(400MHz,DMSO-d₆)δ13.54(s,1H),9.84(s,1H),8.64(d,J＝2.8Hz,1H),8.47(s,1H),8.07(d,J＝1.5Hz,1H),7.96–7.91(m,2H),7.48–7.43(m,2H),6.73(dd,J＝2.8,1.6Hz,1H),4.61(d,J＝6.1Hz,2H),3.84(s,3H).¹³C NMR(101MHz,D₂O)δ173.17,171.32,166.87,155.71,151.38,149.46,144.80,133.32,132.83,131.08,129.79,114.93,114.08,55.14,46.35.

Example 384- { [ (4-hydroxy-2-pyrazol-1-yl-pyrimidine-5-carbonyl) -amino ] -methyl } -benzohydroxamic acid (39)

Compound 39 was obtained according to the same method as that of example 20, step 8, and intermediate 38 obtained in example 37. The product was a white solid with a yield of 71.6%. m.p.258.7-259.8 ℃; ESI-MS (M/z) 355.1172[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ11.23(s,1H),10.67(t,J＝6.0Hz,1H),9.11(s,1H),8.64–8.48(m,2H),7.80–7.76(m,1H),7.72(d,J＝8.0Hz,2H),7.37(d,J＝7.9Hz,2H),6.57–6.48(m,1H),4.55(d,J＝6.0Hz,2H).¹³C NMR(101MHz,DMSO)δ171.05,165.08,164.11,156.25,154.78,143.09,142.46,131.23,129.09,127.10,126.97,111.40,108.15,41.57.

Example 394- [ (4-methoxybenzyl) oxy-2- (1H-pyrazol-1-yl) pyrimidine-5-carboxamido ] butyric acid methyl ester (40)

Intermediate 40 was obtained in the same manner as in example 18, step 6, starting from intermediate 4 and methyl 4-aminobutyrate. The product was purified by flash column chromatography (petroleum ether: ethyl acetate 2:1) to give a white solid in 84.8% yield. m.p.115.8-116.5 ℃; ESI-MS (M/z) 426.22[ M + H]⁺；¹H NMR(400MHz,Chloroform-d)δ9.27(s,1H),8.59(d,J＝2.8Hz,1H),7.89–7.83(m,1H),7.51(t,J＝5.6Hz,1H),7.46–7.41(m,2H),6.96–6.90(m,2H),6.51(dd,J＝2.8,1.6Hz,1H),5.61(s,2H),3.81(s,3H),3.64(s,3H),3.40(td,J＝6.8,5.5Hz,2H),2.22(t,J＝7.4Hz,2H),1.78(p,J＝7.1Hz,2H).¹³C NMR(101MHz,CDCl₃)δ173.32,166.91,163.27,161.90,160.28,155.92,144.53,130.54,129.83,126.46,114.35,110.91,109.26,70.33,55.31,51.67,38.95,31.28,24.52.

Example 404- [ 4-hydroxy-2- (1H-pyrazol-1-yl) pyrimidine-5-carboxamido ] butyric acid methyl ester (41)

The intermediate 40 obtained in example 39 was subjected to the same procedures as in example 18, step 6 to give an intermediate 41. Pulping with methanol for 0.5h, vacuum filtering, and drying to obtain white solid with yield of 90.1%. m.p.212.1-214.3; ESI-MS (M/z) 306.13[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ13.39(s,1H),9.33(s,1H),8.63(d,J＝2.9Hz,1H),8.43(s,1H),8.06(s,1H),6.72(t,J＝2.2Hz,1H),3.58(s,3H),3.32(q,J＝6.6Hz,2H),2.35(t,J＝7.4Hz,2H),1.77(p,J＝7.1Hz,2H).¹³C NMR(101MHz,TFA-D)δ180.69,171.48,166.91,155.48,151.47,149.54,133.41,115.06,113.71,55.12,42.22,33.57,26.17.

EXAMPLE 414- [ 4-hydroxy-2- (1H-pyrazol-1-yl) pyrimidine-5-carboxamido ] butylhydroxamic acid (42)

Compound 42 was obtained according to the same method as that of example 20, step 8, intermediate 41 obtained in example 40. The product was a white solid in 61.2% yield. m.p.216.8-217.1 ℃; ESI-MS (M/z) 307.1171[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ13.53(s,1H),10.54–10.26(m,1H),9.32(s,1H),8.62(d,J＝2.7Hz,1H),8.44(s,1H),8.06(d,J＝1.5Hz,1H),6.72(dd,J＝2.8,1.6Hz,1H),3.29(q,J＝6.7Hz,2H),2.00(dd,J＝8.3,6.7Hz,2H),1.72(p,J＝7.3Hz,2H).¹³C NMR(101MHz,D₂O)δ173.25,167.58,167.32,158.26,156.18,143.21,129.61,110.66,108.47,38.29,30.32,25.40.

Example 425- [ (4-methoxybenzyl) oxy-2- (1H-pyrazol-1-yl) pyrimidine-5-carboxamido ] pentanoic acid ethyl ester (43)

Intermediate 40 was obtained in the same manner as in example 18, step 6, starting from intermediate 4 and ethyl 5-aminopentanoate. The product was purified by flash column chromatography (petroleum ether: ethyl acetate 2:1) to give a white solid in 83.2% yield. m.p.122.3-123.8 ℃; ESI-MS (M/z) 476.19[ M + Na ]]⁺；¹H NMR(400MHz,Chloroform-d)δ9.29(s,1H),8.61(dd,J＝2.7,0.7Hz,1H),7.88(dd,J＝1.7,0.7Hz,1H),7.52–7.46(m,1H),7.46–7.42(m,2H),6.97–6.92(m,2H),6.53(dd,J＝2.8,1.6Hz,1H),5.62(s,2H),4.12(q,J＝7.1Hz,2H),3.83(s,3H),3.37(q,J＝6.3Hz,2H),2.24(t,J＝6.9Hz,2H),1.59–1.44(m,4H),1.25(t,J＝7.1Hz,3H).¹³C NMR(101MHz,CDCl₃)δ173.23,166.92,163.24,161.77,160.26,155.88,144.50,130.60,129.82,126.49,114.31,110.99,109.21,70.32,60.34,55.32,39.20,33.67,28.60,22.04,14.25.

Example 435- [ 4-hydroxy-2- (1H-pyrazol-1-yl) pyrimidine-5-carboxamido ] pentanoic acid ethyl ester (44)

Intermediate 43 obtained in example 42 was subjected to the same procedure as in example 18, step 6 to give intermediate 44. Pulping with methanol for 0.5h, filtering, and drying to obtain white solid with yield of 97.3%. m.p.178.7-179.8 ℃; ESI-MS (M/z) 334.15[ M + H]⁺；¹H NMR(400MHz,DMSO-d6)δ13.50(s,1H),9.32(s,1H),8.62(d,J＝2.9Hz,1H),8.43(s,1H),8.06(s,1H),6.72(s,1H),4.04(q,J＝7.1Hz,2H),3.30(q,J＝6.3Hz,2H),2.32(t,J＝6.9Hz,2H),1.53(h,J＝7.1Hz,4H),1.17(t,J＝7.1Hz,3H).¹³CNMR(101MHz,TFA-D)δ180.98,171.46,166.72,155.26,151.34,149.48,133.32,114.96,113.61,65.29,42.74,36.07,30.22,24.19,14.58.

EXAMPLE 445- [ 4-hydroxy-2- (1H-pyrazol-1-yl) pyrimidine-5-carboxamido ] pentahydroxamic acid (45)

Compound 45 was obtained according to the same method as that of example 20, step 8, intermediate 44 obtained in example 43. The product was a white solid in 69.7% yield. m.p.203.5-203.9 ℃; ESI-MS (M/z) 321.1275[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ13.48(s,1H),10.37(s,1H),9.32(s,1H),8.62(d,J＝2.8Hz,1H),8.43(s,1H),8.05(d,J＝1.5Hz,1H),6.71(dd,J＝2.7,1.7Hz,1H),3.28(q,J＝6.3Hz,2H),1.98(t,J＝6.8Hz,2H),1.59–1.41(m,4H).¹³C NMR(101MHz,D₂O)δ173.24,167.50,167.16,158.18,156.15,143.19,129.59,110.70,108.45,38.62,32.43,27.95,23.05.

Example 456- [ (4-methoxybenzyl) oxy-2- (1H-pyrazol-1-yl) pyrimidine-5-carboxamido ] hexanoic acid methyl ester (46)

Intermediate 46 was obtained in the same manner as in example 18, step 6, starting from intermediate 4 and methyl 6-aminocaproate. The product was purified by flash column chromatography (petroleum ether: ethyl acetate 2:1) to give a white solid in 86.5% yield. m.p.135.5-136.7 ℃; ESI-MS (M/z) 476.15[ M + Na ]]⁺；¹H NMR(400MHz,Chloroform-d)δ9.27(s,1H),8.59(dd,J＝2.8,0.7Hz,1H),7.85(dd,J＝1.6,0.7Hz,1H),7.46–7.38(m,3H),6.97–6.91(m,2H),6.50(dd,J＝2.7,1.6Hz,1H),5.59(s,2H),3.81(s,3H),3.64(s,3H),3.34(td,J＝6.9,5.5Hz,2H),2.22(t,J＝7.5Hz,2H),1.54(p,J＝7.6Hz,2H),1.42(p,J＝7.0Hz,2H),1.25–1.13(m,2H).¹³C NMR(101MHz,CDCl₃)δ173.86,166.90,163.22,161.70,160.29,155.87,144.49,130.55,129.81,126.43,114.31,110.98,109.21,70.36,55.31,51.50,39.44,33.73,28.81,26.34,24.46.

Example 466- [ 4-hydroxy-2- (1H-pyrazol-1-yl) pyrimidine-5-carboxamido ] hexanoic acid methyl ester (47)

Intermediate 46 obtained in example 45 was subjected to the same procedure as in example 18, step 6 to give intermediate 47. Pulping with methanol for 0.5h, filtering, and drying to obtain white solid with yield of 94.3%. m.p.174.6-176.0 ℃; ESI-MS (M/z) 334.15[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ13.51(s,1H),9.31(s,1H),8.61(d,J＝2.8Hz,1H),8.53–8.32(m,1H),8.05(d,J＝1.6Hz,1H),6.71(dd,J＝2.8,1.6Hz,1H),3.57(s,3H),3.27(q,J＝6.7Hz,2H),2.29(t,J＝7.4Hz,2H),1.52(dp,J＝16.8,7.3Hz,4H),1.35–1.24(m,2H).¹³C NMR(101MHz,TFA-D)δ181.99,171.54,166.73,155.28,151.41,149.53,133.38,115.03,113.60,54.84,43.15,36.17,30.48,28.37,26.53.

Example 476- [ 4-hydroxy-2- (1H-pyrazol-1-yl) pyrimidine-5-carboxamido ] hydroxamic acid (48)

Compound 48 was obtained from intermediate 47 obtained in example 46 according to the same method as in example 20, step 8. The product was a white solid in 73.9% yield. m.p.189.4-189.7 ℃; ESI-MS (M/z) 335.1535[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ13.49(s,1H),10.35(s,1H),9.31(s,1H),8.61(d,J＝2.8Hz,1H),8.43(s,1H),8.05(d,J＝1.5Hz,1H),6.74–6.68(m,1H),3.27(q,J＝6.6Hz,2H),1.95(t,J＝7.3Hz,2H),1.50(h,J＝7.5Hz,4H),1.28(td,J＝8.5,4.1Hz,2H).¹³CNMR(101MHz,D₂O)δ173.19,167.78,167.02,158.08,156.07,143.12,129.51,110.65,108.41,38.82,32.68,28.09,25.69,25.10.

Example 487- [ (4-methoxybenzyl) oxy-2- (1H-pyrazol-1-yl) pyrimidine-5-carboxamido ] heptanoic acid ethyl ester (49)

Intermediate 49 was obtained in the same manner as in example 18, step 6, starting from intermediate 4 and ethyl 7-aminoheptanoate. The product was purified by flash column chromatography (petroleum ether: ethyl acetate 2:1) to give a white solid in 87.2% yield. m.p.82.6-83.5 deg.C; ESI-MS (M/z) 482.2435[ M + H]⁺；¹H NMR(400MHz,Chloroform-d)δ9.27(s,1H),8.58(dd,J＝2.7,0.7Hz,1H),7.84(dd,J＝1.6,0.7Hz,1H),7.44–7.38(m,3H),6.95–6.90(m,2H),6.50(dd,J＝2.8,1.6Hz,1H),5.58(s,2H),4.09(q,J＝7.2Hz,2H),3.80(s,3H),3.32(td,J＝6.9,5.4Hz,2H),2.23(t,J＝7.5Hz,2H),1.57–1.48(m,2H),1.40(p,J＝7.1Hz,2H),1.28–1.11(m,7H).¹³C NMR(101MHz,CDCl₃)δ173.62,166.89,163.21,161.66,160.27,155.84,144.48,130.53,129.80,126.42,114.31,111.00,109.20,70.35,60.21,55.31,39.55,34.19,28.89,28.69,26.49,24.72,14.26.

Example 497- [ 4-hydroxy-2- (1H-pyrazol-1-yl) pyrimidine-5-carboxamido ] heptanoic acid ethyl ester (50)

Intermediate 49 obtained in example 48 was subjected to the same procedure as in example 18, step 6 to give intermediate 50. Pulping with methanol for 0.5h, filtering, and drying to obtain white solid with yield of 93.2%. m.p.169.6-170.8 ℃; ESI-MS (M/z) 362.13[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ13.53(s,1H),9.32(s,1H),8.62(d,J＝2.8Hz,1H),8.43(s,1H),8.06(d,J＝1.7Hz,1H),6.72(dd,J＝2.8,1.6Hz,1H),4.03(q,J＝7.1Hz,2H),3.28(q,J＝6.6Hz,2H),2.26(t,J＝7.3Hz,2H),1.54–1.46(m,4H),1.29(p,J＝3.6Hz,4H),1.16(t,J＝7.1Hz,3H).¹³C NMR(101MHz,TFA-D)δ182.06,171.55,166.82,155.39,151.46,149.55,133.40,115.03,113.64,65.22,43.43,36.68,30.72,30.70,28.57,26.96,14.78.

Example 507- [ 4-hydroxy-2- (1H-pyrazol-1-yl) pyrimidine-5-carboxamido ] heptahydroxamic acid (51)

Compound 51 was obtained from intermediate 50 obtained in example 49 according to the same method as in example 20, step 8. The product was a white solid with a yield of 87.7%. m.p.195.8-196.4 ℃; ESI-MS (M/z) 349.16[ M + H]⁺；¹H NMR(400MHz,DMSO-d₆)δ13.50(s,1H),10.33(s,1H),9.32(s,1H),8.62(d,J＝2.7Hz,1H),8.43(s,1H),8.06(d,J＝1.5Hz,1H),6.72(dd,J＝2.8,1.6Hz,1H),3.28(q,J＝6.6Hz,2H),1.94(t,J＝7.4Hz,2H),1.54–1.43(m,4H),1.28(dh,J＝9.7,5.1,3.9Hz,4H).¹³CNMR(101MHz,D₂O)δ173.37,168.12,167.18,158.25,156.30,143.27,129.71,110.91,108.53,39.00,32.74,28.33,27.82,25.88,25.37.

Example 51 Histone Deacetylase (HDAC) inhibitory Activity test experiment

The drug to be tested is dissolved in a suitable solvent. With double distilled water (dd H)₂O) dilute the drug to 2-fold the working concentration. Adding 50 μ L of the sample to be tested into an EP tube, and adding 50 μ L of ddH into the positive control₂O, negative control plus 48. mu.L ddH₂O and 2. mu.Ltrichotain A. Add 50. mu.L Mix per tube and Mix 37. mu.L for 30 min.

After completion of incubation, 10. mu.L of Lysin Developer was added to each tube and mixed to 37. mu.l for 30min to terminate the reaction. Cells were washed 3 times for 10min each with PBS.

The measuring method comprises the following steps: ex 350-

The calculation method comprises the following steps: the inhibition rate (%) (1- (sample value/positive value)) × 100%.

Table 1 results of the HDAC inhibitory activity test of the compound

As can be seen from table 1, the compounds of the present invention showed significant Histone Deacetylase (HDAC) inhibitory activity.

Example 52 Prolyl Hydroxylase (PHD) inhibitory Activity test experiment

Seven identical culture mediums are respectively added with 5 muM of blank control DMSO, 5 muM of PHD single-target inhibitor T55 muM, 5 muM of HDAC single-target inhibitor SAHA, 5 muM of double-target inhibitor GJ09G, GJ12G, GJ13G and GJ14G, and are co-cultured under the same conditions for 48 hours, then the cells are collected, the total protein of the cells is extracted after cell lysis, 40 mug of total protein is taken from each group, Western Blot is used for analyzing the content of HIF-1 and HIF-2 in the protein, mouse anti-human HIF-1 α and HIF-2 α monoclonal antibodies are used as primary antibodies, and β -actin is used as internal reference.

And quantifying according to the concentration of the protein band scanned by the electrophoresis result, and correcting by using the concentration of the relative internal reference β -actin to obtain the contents of HIF-1 α and HIF-2 α expressed by HT1080 cells under the action of various inhibitors, and further comparing with a blank control group and a single-target control group to determine the influence of the PHD/HDAC double-target inhibitor on the expression level of HIF-1 α and HIF-2 α proteins.

Results of compound activity test:

the results of the expression level test of the compounds HIF-1 α and HIF-2 α are shown in figure 1 and Table 2 of the specification.

TABLE 2 test results of HIF-1 node and HIF-2 node expression levels and relative expression levels of the compounds

	β-actin	HIF-1α	HIF-1α/β-actin	HIF-2α	HIF-2a/β-actin
						DMSO	456000	855000	1.88	1000000	2.19
T5	478000	1570000	3.28	3130000	6.54
						SAHA	495000	822000	1.66	2930000	5.91
GJ09G	491000	1000000	2.04	1760000	3.58
						GJ13G	448000	1210000	2.70	3660000	8.16
GJ15G	451000	154000	0.34	3650000	8.09
						GJ14G	484000	219000	0.45	3240000	6.69

As can be seen from Table 2, the compounds of the present invention exhibit significant Prolyl Hydroxylase (PHD) inhibitory activity and are capable of selectively stabilizing the HIF-2I protein.

Claims

1. A compound of the formula I:

wherein:

a)X₁is a N atom, X₂、X₃Is a C atom;

b) r independently selects C₁-C₁₀Alkyl radical, C₂-C₁₀An alkenyl group;

c)n₁are 1 and 2.

2. A compound of the formula I:

wherein:

a)X₁is a N atom, X₂、X₃Is a C atom;

b)n₁is 1;

c) r independently selects the following chemical structure:

3. the following compounds, or pharmaceutically acceptable salts thereof, are selected from:

4. a process for the preparation of a compound according to any one of claims 1 to 3, characterized in that it comprises the following steps:

2) reacting the compound B obtained in the step 1) with p-methoxy benzyl chloride, potassium carbonate and potassium iodide in a polar aprotic solvent to obtain a compound C;

3) reacting the compound C obtained in the step 2) with sodium hydroxide in a mixed solution of water and tetrahydrofuran to obtain a compound D;

5) reacting the compound G obtained in the step 4) in a dichloromethane solution of trifluoroacetic acid to obtain a compound H;

6) reacting the compound H or the amino aliphatic carboxylic acid ester obtained in the step 5) with a compound D, 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride, 1-hydroxybenzotriazole and an organic base in a polar aprotic solvent to obtain a compound I;

7) reacting the compound I obtained in the step 6) in a dichloromethane solution of trifluoroacetic acid to obtain a compound J;

8) reacting the compound J obtained in the step 7) with hydroxylamine hydrochloride and sodium hydroxide aqueous solution in 1, 4-dioxane to obtain a compound with a structure shown in a formula I;

wherein the compound has the following structure:

wherein n is 1,2, n₁＝1、2。

5. A composition comprising a compound of formula I as described in any one of claims 1-3, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

6. Use of a compound according to any one of claims 1 to 3 and pharmaceutically acceptable salts thereof for the manufacture of a prolyl hydroxylase inhibitor drug, a histone deacetylase inhibitor drug.

7. Use of a compound according to any one of claims 1 to 3, and pharmaceutically acceptable salts thereof, for the manufacture of a radioprotective agent.