CN109810071A

CN109810071A - A kind of miRNA biosynthesis inhibitor

Info

Publication number: CN109810071A
Application number: CN201910245647.3A
Authority: CN
Inventors: 王飞; 卢晓霞; 张国林; 王涛; 彭婷
Original assignee: Chengdu Institute of Biology of CAS
Current assignee: Chengdu Institute of Biology of CAS
Priority date: 2019-03-28
Filing date: 2019-03-28
Publication date: 2019-05-28
Anticipated expiration: 2039-03-28
Also published as: CN109810071B

Abstract

The present invention provides a kind of Formulas I compound represented or its conformers or its optical isomer or its pharmaceutically acceptable salt.It can closely be combined with the Binding proteins in miRNA biosynthetic process, and can effectively inhibit the synthesis of miRNA-21.Reactive compound prepared by the present invention can be used as miRNA-21 inhibitor, be further used as the potential drug for the treatment of malignant tumour.

Description

A kind of miRNA biosynthesis inhibitor

Technical field

The invention belongs to chemical medicines, and in particular to a kind of miRNA biosynthesis inhibitor.

Background technique

Cancer has become the first cause of the death in Chinese city, is the second cause of the death in rural area.Although China is anticancer drug Research and development are developed as preference, but are primarily present following two predicament to the research of anticancer drug at present: first is that medicine Object target spot is very few: the target spot of more than 2,000 a drugs of clinical listing is only 300 or so at present, is distributed mainly on hormone receptor With the seven genoid families such as GPCR.Second is that generation to disease and protein signaling networks understand unclear, lead to drug Clinical development failure, cost is excessively high.

MiRNA has the function of important in vivo, and the abnormal expression of miRNA will lead to the generation of many diseases, therefore, MiRNA becomes the drug tar-get and diagnosis marker of growing interest in cancer treatment in recent years.And TRBP has due to it Biological function diversity, and it is related with the proliferation of Several Kinds of Malignancy cell, differentiation and migration, TRBP makees in zooblast For Dicer companion, TRBP can be mature by influencing the miRNA that Dicer and Ago2 is mediated, and then influences pernicious turn of cancer cell It moves.

But existing miRNA inhibitor, inhibitory effect far can not also reach clinical requirement at present.Therefore, into one Step further investigation compound develops the new suppression to the generation of miRNA with more preferably inhibitory effect to the inhibiting mechanism of miRNA Preparation, it is very necessary in oncotherapy.

Summary of the invention

The purpose of the present invention is to provide a kind of compounds for effectively inhibiting miRNA-21 biosynthesis.

The present invention provides Formulas I compound represented or its conformer or its optical isomer or its pharmaceutically Acceptable salt:

Wherein:

X is selected from O, S；Y is selected from N；

R₁、R₂、R₃、R₄、R₅It is each independently selected from hydrogen, substituted or unsubstituted C1-2 alkyl, substituted or unsubstituted C1- 2 alkoxies, halogen, cyano, nitro, substituted or unsubstituted phenyl ,-COOR₉；

Or, R₁、R₂、R₅It is each independently selected from hydrogen, substituted or unsubstituted C1-2 alkyl, substituted or unsubstituted C1-2 Alkoxy, halogen, cyano, nitro, substituted or unsubstituted phenyl ,-COOR₉, R₃、R₄Two carbon atoms connected to it are together Form phenyl ring；The substituent group is selected from halogen, C1-4 alkyl, hydroxyl, carboxyl, nitro, amino, carboxyl, C₂-C₄Alkenyl, C₁-C₄ Alkyl, C₂-C₄Alkynyl, carbocylic radical, heterocycle；

R₆、R₈Be each independently selected from hydrogen, hydroxyl, halogen, C1-4 alkyl, C3-6 cycloalkyl group, phenyl,--COOR₉、-CONHR₁₀, wherein R₉、R₁0 be each independently selected from C1-2 alkyl, Wherein L₀、L₁、L₂、L₃It is each independently selected from 1-4 methylene, R₁₁Selected from halogen or hydroxyl；

And when X is O, Y N, R₁、R₂、R₄、R₅For hydrogen, R₃For methoxyl group, R₆For methyl, R₈For-COOR₉When, R₉It is not second Base.

Further,

X is selected from O, S；Y is selected from N；

R₁、R₂、R₃、R₄、R₅It is each independently selected from hydrogen, substituted or unsubstituted C1-2 alkyl, substituted or unsubstituted C1- 2 alkoxies, halogen, substituted or unsubstituted phenyl；Or, R₁、R₂、R₅It is each independently selected from hydrogen, substituted or unsubstituted C1-2 Alkyl, substituted or unsubstituted C1-2 alkoxy, halogen, substituted or unsubstituted phenyl, R₃、R₄Two carbon originals connected to it Son is formed together phenyl ring；The substituent group is selected from halogen；

R₆、R₈It is each independently selected from C1-2 alkyl ,-COOR₉, wherein R₉Selected from C1-2 alkyl,Wherein L₀Selected from 1-4 methylene.

Further,

X is selected from O；Y is selected from N；

R₁、R₂、R₃、R₄、R₅It is each independently selected from hydrogen, substituted or unsubstituted C1-2 alkyl, substituted or unsubstituted C1- 2 alkoxies, halogen, phenyl substituted or unsubstituted；The substituent group is selected from halogen；

R₆、R₈It is each independently selected from C1-2 alkyl ,-COOR₉, wherein R₉Selected from C1-2 alkyl,Wherein L₀Selected from 2 methylene.

Further,

The structure of the compound is as shown in Formulas I -1:

Wherein:

R₆Selected from C1-2 alkyl；

R₇Selected from C1-2 alkyl,Wherein L₀Selected from 2 methylene；

R₃Selected from methyl, methoxyl group, phenyl, halogen, preferably methyl, methoxyl group, phenyl, chlorine, bromine.

Further,

X is selected from S；Y is selected from N；

Further,

X is selected from S；Y is selected from N；

R₁、R₂、R₄、R₅For hydrogen, R₃Selected from substituted or unsubstituted C1-2 alkyl, substituted or unsubstituted C1-2 alkoxy, Halogen, substituted or unsubstituted phenyl, the substituent group are selected from halogen；Or, R₁、R₂、R₅For hydrogen, R₃、R₄Two connected to it Carbon atom is formed together phenyl ring；

R₆、R₈It is each independently selected from C1-2 alkyl ,-COOR₉, wherein R₉Selected from C1-2 alkyl,L₀For 2 methylene.

Further,

Its structure such as Formulas I -2:

Wherein:

R₆、R₈In, one is C1-2 alkyl, another is-COOR₉, wherein R9 be selected from C1-2 alkyl, L₀For 2 methylene；

R₃Selected from halogenated or not halogenated C1-2 alkyl, C1-2 alkoxy, halogen, phenyl, preferably trifluoromethyl, methoxy Base, ethyoxyl, methyl；R₄Selected from hydrogen, methyl；Or, R₃、R₄Two carbon atoms connected to it are formed together phenyl ring.

Further,

The compound are as follows:

The present invention also provides above compounds or its pharmaceutically acceptable salt to inhibit as miRNA-21 biosynthesis The purposes of agent.

Further, the inhibitor is the drug for treating tumour, preferably treats the drug of malignant tumour.

Prove that the present invention provides a kind of I compound represented of formula or its pharmaceutically acceptable salt, energy by test It is enough closely to be combined with the Binding proteins in miRNA biosynthetic process, and can effectively inhibit the conjunction of miRNA-21 At.Reactive compound prepared by the present invention can be used as miRNA-21 inhibitor, be further used as the potential medicine for the treatment of malignant tumour Object.

" substitution " refers to that the hydrogen atom in molecule is replaced by other different atoms or molecule.

The minimum value and maximum value of carbon content are indicated by prefix in hydrocarbon group, for example, C1-4 alkyl shows to appoint What alkyl containing 1-4 carbon atom.

Obviously, above content according to the present invention is not being departed from according to the ordinary technical knowledge and customary means of this field Under the premise of the above-mentioned basic fundamental thought of the present invention, the modification, replacement or change of other diversified forms can also be made.

The specific embodiment of form by the following examples remakes further specifically above content of the invention It is bright.But the range that this should not be interpreted as to the above-mentioned theme of the present invention is only limitted to example below.It is all to be based on above content of the present invention The technology realized all belongs to the scope of the present invention.

Detailed description of the invention

Fig. 1 is the relative intensity of fluorescence of compound prepared by the present invention, control compound 3b in screening active ingredients experiment.

Fig. 2 is the relative intensity of fluorescence of the amount of activated compound of the present invention, control compound 3b.

Fig. 3 is the concentration dependency curves that the amount of activated compound on luciferase reporter gene of the present invention influences.

Fig. 4 is the 2D (left side) and 3D (right side) figure of the compounds of this invention and Dicer-TRBP molecular docking, the color in 2D figure Represent: amino acid residue colours haloing in addition, indicates the Solvent accessible surface of interaction residue；Wherein, each number corresponds to Color respectively indicate: (1): with compound alkyl formed alkyl-π effect amino acid residue；(2): being formed and divided with compound The amino acid residue of van der Waals interaction between son；(3): the amino acid residue of intermolecular hydrogen bonding effect is formed with compound； (4): the amino acid residue of intermolecular C-H hydrogen bond action is formed with compound；(5): forming Cation-π effect with compound Amino acid residue；(6): the amino acid residue of intermolecular Halogen-H (Cl, Br, I) hydrogen bond action is formed with compound；(7): The amino acid residue of intermolecular π-πconjugation is formed with compound.

Fig. 5 is the 2D (left side) and 3D (right side) figure of the compounds of this invention and TRBP (dsRBD2) molecular docking, the face in 2D figure Color represents: amino acid residue colours haloing in addition, indicates the Solvent accessible surface of interaction residue；Wherein, each number pair The color answered respectively indicates: (1): the amino acid residue of alkyl-π effect is formed with compound alkyl；(2): being formed with compound The amino acid residue of intermolecular van der Waals interaction；(3): the amino acid residue of intermolecular hydrogen bonding effect is formed with compound； (4): the amino acid residue of intermolecular C-H hydrogen bond action is formed with compound；(5): forming Cation-π effect with compound Amino acid residue；(6): the amino acid residue of intermolecular Halogen-H (Cl, Br, I) hydrogen bond action is formed with compound；(7): The amino acid residue of intermolecular π-πconjugation is formed with compound.

Fig. 6 is cell screening illustraton of model.

Specific embodiment

Raw material, equipment used in the specific embodiment of the invention are known product, are obtained by purchase commercial product.

The synthesis of embodiment 1, the compounds of this invention

Ethyl 2- (4-methoxyphenyl) -4-methylthiazole-5-carboxylate (T2): 25mL's 4- methoxy benzamide (152.2mg, 1mmol) is added in round-bottomed flask, lawesson reagent (485.4mg, 1.2mmol) and THF (20mL).Lawesson reagent and solvent are removed after reacting 4h under normal temperature conditions, it is thio to obtain intermediate through column chromatographic isolation and purification Then benzamide is used as solvent, 70 DEG C of reactions at ethyl alcohol (10mL) with the bromo- 2-Oxobutyric acid ethyl ester (209mg, 1mmol) of 3- Under the conditions of flow back 4h, end of reaction post-processing removes solvent, obtains compound T2 through column chromatographic isolation and purification.Yield 92% is white Color solid is unfolded system PE/EA 10:1 (Rf=0.5, PE/EA=3:1),¹H NMR(400MHz,CDCl₃) 7.91 (d, J= 8.8Hz, 2H), 6.96 (d, J=8.8Hz, 2H), 4.37 (q, J=7.1Hz, 2H), 3.86 (s, 3H), 2.76 (s, 3H), 1.68 (s, 1H), 1.40 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ169.9,162.4,161.9,161.0, 128.4,125.9,120.9,114.4,61.1,55.5,17.6,14.4；HR-ESI-MS(positive mode)m/z: 278.0851[M+H]⁺(Calcd for C₁₄H₁₆NO₃S:278.0851),m/z:300.0668[M+Na]⁺(Calcd for C₁₄H₁₅NO₃SNa:300.0670).

Ethyl 2- (4-methoxyphenyl) -5-methyl-1H-imidazole-4-carboxylate (T3): Ethoxycarbonyl methylene triphenyl phosphine (CEMTPP) (1044.2mg, 3mmol) is added in the round-bottomed flask of 25mL, chloroacetic chloride (474.5mg, 6mmol), n,N-diisopropylethylamine (DIPEA) (516.9mg, 4mmol), methylene chloride (DCM) (20mL) are anti- It answers 1h to obtain 3- triphenyl phosphorus ethyl acetoacetate, Oxone (614.7mg, 1mmol) oxidation is then added in THF, reacts 4h Afterwards intermediate 2,3- dioxobutyric acid ethyl ester, then with 4-methoxybenzaldehyde (272.3mg, 2mmol) in acetic acid solvent (10mL), 12h is reacted at a temperature of 100 DEG C, and end of reaction post-processing removes solvent, obtains compound through column chromatographic isolation and purification T3.Yield 69%, white solid are unfolded system PE/EA 10:1 (Rf=0.5, PE/EA=5:1),¹H NMR(400MHz, CDCl₃) 7.84 (d, J=8.8Hz, 2H), 6.94 (d, J=8.8Hz, 2H), 4.38 (q, J=7.1Hz, 2H), 3.84 (s, 3H), 2.54 (s, 3H), 1.39 (t, J=7.1Hz)；¹³C NMR(100MHz,CDCl₃)δ160.8,127.3,121.8,114.3, 60.6,55.4,14.5；HR-ESI-MS(positive mode)m/z:261.1236[M+H]⁺(Calcd for C₁₄H₁₇N₂O₃: 261.1239),m/z:283.1052[M+Na]⁺(Calcd for C₁₄H₁₆N₂O₃Na:283.1059).

2-hydroxyethyl 2-(4-methoxyphenyl)-5-methyloxazole-4-carboxylate(T4): In the round-bottomed flask of 25mL be added compound 3b (5- methyl -2- (4- methoxyphenyl) -4- oxazole Ethyl formate, 261.3mg, 1mmol), EtOH/10%NaOH=1:1 (20mL) reacts 1 hour at 70 DEG C, then adjusts pH to 1 with concentrated hydrochloric acid Between~2, dichloromethane extraction, concentration organic phase obtains intermediate 3,3 and does not have to be further purified directly progress next step reaction.Xiang Zhong Methylene chloride 20mL, 1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride (EDCI) are added in mesosome 3 (383.4mg, 2mmol), 4-dimethylaminopyridine (DMAP) (24.4mg, 0.2mmol), ethylene glycol (124.1mg, 2mmol), instead System is answered to be heated to 80 DEG C, flow back 6h.20mL water quenching reaction is added after reaction, methylene chloride extraction merges organic phase And it is concentrated, then through column chromatographic isolation and purification (petrol ether/ethyl acetate/methanol=10/1/0.01), obtain compound T4.Yield 77%, white solid is unfolded system PE/EA/MeOH 1:1:0.01 (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃) 7.97 (d, J=8.9Hz, 2H), 6.97 (d, J=8.9Hz, 2H), 4.46 (t, J=9.2Hz, 2H), 3.97 (t, J=9.2Hz, 2H), 3.86 (s, 3H), 2.67 (s, 3H), 2.21 (s, 1H)；¹³C NMR(100MHz,CDCl₃)δ 162.6,161.8,159.9,156.3,128.3,128.0,119.0,114.3,66.8,61.0,55.4,12.1；HR-ESI-MS (positive mode)m/z:278.1023[M+H]⁺(Calcd for C₁₄H₁₆NO₅:278.1028),m/z:300.0839[M+ Na]⁺(Calcd for C₁₄H₁₅NO₅Na:300.0848).

2-hydroxyethyl 2-(4-methoxyphenyl)-4-methylthiazole-5-carboxylate (T5): compound T5 be using T2 as raw material, and with reference to T4 synthetic method obtain compound T5.Yield 80%, white solid, exhibition System PE/EA/MeOH 1:1:0.01 (Rf=0.5, PE/EA/MeOH=1:1:0.01) is opened,¹H NMR(400MHz,CDCl₃) 7.91 (d, J=8.8Hz, 2H), 6.96 (d, J=8.9Hz, 2H), 4.44 (t, J=4.6Hz, 2H), 3.94 (q, J=4.1Hz, 2H),3.86(s,3H),2.76(s,3H),1.67(s,1H)；¹³C NMR(100MHz,CDCl₃)δ170.3,162.6,162.1, 161.7,128.5,125.7,120.1,114.4,66.7,61.3,55.5,17.6；HR-ESI-MS(positive mode)m/ z:294.0796[M+H]⁺(Calcd for C₁₄H₁₆NO₄S:294.0800),m/z:316.0612[M+Na]⁺(Calcd for C₁₄H₁₅NO₄SNa:316.0619).

Ethyl 2- (4-methoxyphenyl) -4-methyloxazole-5-carboxylate (T6): 25mL's 4- methoxy benzamide (152.2mg, 1mmol) is added in round-bottomed flask, 2- chloro ethyl acetoacetate (329.2mg, 2mmol) and etoh solvent (20mL), it is reacted in 100 DEG C of tube sealing and obtains compound T6 for 24 hours.Yield 87%, compound T6, white solid are unfolded system PE/EA 10:1 (Rf=0.5, PE/EA=3:1),¹H NMR(400MHz,CDCl₃)8.06(d, J=9.0Hz, 2H), 6.97 (d, J=9.0Hz, 2H), 4.42 (q, J=7.2Hz, 2H), 3.86 (s, 3H), 2.51 (s, 3H), 1.42 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ162.4,162.3,159.0,147.1,137.0,129.0, 119.1,114.3,61.0,55.4,14.4,13.5；HR-ESI-MS(positive mode)m/z:262.1072[M+H]⁺ (Calcd for C₁₄H₁₆NO₄:262.1079),m/z:284.0895[M+Na]⁺(Calcd for C₁₄H₁₅NO₄Na: 284.0899).

Ethyl 2- (4-methoxyphenyl) -4-methylthiazole-5-carboxylate (T7): 25mL's 4- methoxy benzamide (152.2mg, 1mmol) is added in round-bottomed flask, lawesson reagent (485.4mg, 1.2mmol) and THF (20mL) removes lawesson reagent and solvent after reacting 4h under normal temperature conditions, obtains intermediate thio phenyl through column chromatographic isolation and purification Formamide, then 70 DEG C of reflux 6 are small in etoh solvent (20mL) with 2- chloro ethyl acetoacetate (329.2mg, 2mmol) When, end of reaction post-processing removes solvent, obtains compound T7 through column chromatographic isolation and purification.Yield 92%, white solid, expansion System PE/EA 10:1 (Rf=0.5, PE/EA=3:1),¹H NMR(400MHz,CDCl₃) 7.88 (d, J=8.8Hz, 2H), 6.95 (d, J=8.8Hz, 2H), 4.46 (q, J=7.1Hz, 2H), 3.85 (s, 3H), 2.78 (s, 3H), 1.46 (t, J= 7.1Hz,3H)；¹³C NMR(100MHz,CDCl₃)δ163.7,162.7,161.3,143.7,142.0,128.2,125.9, 114.2,61.1,55.4,14.4,13.3；HR-ESI-MS(positive mode)m/z:278.0850[M+H]⁺(Calcd for C₁₄H₁₆NO₃S:278.0851),m/z:300.0670[M+Na]⁺(Calcd for C₁₄H₁₅NO₃SNa:300.0670).

Ethyl 2- (4-methoxyphenyl) -4-methyl-1H-imidazole-5-carboxylate (T8): DL-Alanine (540mg, 6mmol) is added in the round-bottomed flask of 25mL, 10% NaOH solvent (10mL, pH=11~12), 4- Methoxy benzoyl chloride (1026mg, 6mmol) reacts under room temperature, by TLC monitoring to end of reaction, remove reaction raw materials and Solvent obtains crude product, adds EDCI (223mg, 1mmol), is added three after stirring 1 hour under DCM solvent (20mL) room temperature Butyl phosphine (202.3mg, 0.5mmol), cyanoformic ester (79mg, 0.8mmol), TLC monitoring reaction, to end of reaction, after Processing removes solvent and raw material, obtains compound T8 through column chromatographic isolation and purification.System PE/ is unfolded in yield 64%, white solid EA 10:1 (Rf=0.5, PE/EA=5:1),¹H NMR(400MHz,CD₃OD) 7.88 (d, J=8.8Hz, 2H), 7.03 (d, J= 8.8Hz, 2H), 4.39 (q, J=7.1Hz, 2H), 3.86 (s, 3H), 2.56 (s, 3H), 1.43 (t, J=7.0Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ160.8,127.3,121.8,114.3,60.6,55.4,29.7,29.4,14.5；HR-ESI-MS (positive mode)m/z:261.1246[M+H]⁺(Calcd for C₁₄H₁₇N₂O₃:261.1239),m/z:283.1063[M +Na]⁺(Calcd for C₁₄H₁₆N₂O₃Na:283.1059).

2-hydroxyethyl 2-(4-methoxyphenyl)-4-methyloxazole-5-carboxylate(T9): The synthetic method of compound T9 synthesized reference T4.System PE/EA/MeOH 1:1:0.01 (Rf=0.5, PE/ is unfolded in yield 81% ), EA/MeOH=1:1:0.01¹H NMR(400MHz,CDCl₃) 8.06 (d, J=9.0Hz, 2H), 6.98 (d, J=9.0Hz, 2H), 4.49 (t, J=4.6Hz, 2H), 3.97 (t, J=4.6Hz, 2H), 3.88 (s, 3H), 2.53 (s, 3H), 1.69 (s, 1H) ；¹³C NMR(100MHz,CDCl₃)δ162.7,162.4,159.1,148.0,136.5,129.1,118.9,114.3,66.5, 61.3,55.5,13.6；HR-ESI-MS(positive mode)m/z:278.1017[M+H]⁺(Calcd for C₁₄H₁₆NO₅: 278.1028),m/z:300.0832[M+Na]⁺(Calcd for C₁₆H₁₉NO₄Na:300.0848).

2-hydroxyethyl 2-(4-methoxyphenyl)-4-methylthiazole-5-carboxylate (T10): compound T10 obtains compound T10 with reference to the synthetic method of T4 using T7 as raw material.Yield 84%, white solid, Expansion system PE/EA/MeOH 1:1:0.01 (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃) 7.81 (d, J=8.8Hz, 2H), 6.94 (d, J=8.8Hz, 2H), 4.45 (t, J=4.4Hz, 2H), 3.96 (t, J=4.4Hz, 2H),3.85(s,3H),2.76(s,3H)；¹³C NMR(100MHz,CDCl₃)δ164.1,162.6,161.5,144.7,141.2, 128.2,125.5,114.3,67.0,60.9,55.4,13.3；HR-ESI-MS(positive mode)m/z:294.0800[M+ H]⁺(Calcd for C₁₄H₁₆NO₄S:294.0800),m/z:316.0618[M+Na]⁺(Calcd for C₁₄H₁₅NO₄SNa: 316.0619).

Ethyl 5-methyl-2- (pyridin-4-yl) oxazole-4-carboxylate (T11): compound T11's The synthetic method of synthesized reference 3b.Yield 61%, white solid, expansion system PE/EA 10:1 (Rf=0.5, PE/EA=5: 1),¹H NMR(400MHz,CDCl₃) 8.74 (d, J=6.1Hz, 2H), 7.92 (d, J=6.2Hz, 2H), 4.46 (q, J= 7.1Hz, 2H), 2.73 (s, 3H), 1.44 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ162.0,157.4, 157.3,150.6,133.5,129.5,120.1,61.3,14.4,12.3；HR-ESI-MS(positive mode)m/z: 233.0920[M+H]⁺(Calcd for C₁₁H₁₂NO₅:233.0926),m/z:255.0740[M+Na]⁺(Calcd for C₁₁H₁₁NO₅Na:255.0746).

Ethyl 5-methyl-2- (pyridin-2-yl) oxazole-4-carboxylate (T12): compound T12's The synthetic method of synthesized reference 3b.Yield 67%, white solid, expansion system PE/EA 10:1 (Rf=0.5, PE/EA=5: 1),¹H NMR(400MHz,CDCl₃) 8.71 (d, J=4.2Hz, 1H), 8.25 (d, J=8.0Hz, 1H), 7.83-7.79 (m, 1H), 7.39-7.36 (m, 1H), 4.44 (q, J=7.2Hz, 2H), 2.74 (s, 3H), 1.42 (t, J=7.1Hz, 3H)；¹³C NMR (100MHz,CDCl₃)δ162.2,158.3,157.5,149.9,145.4,137.0,129.1,125.1,122.5,61.2, 14.4,12.4；HR-ESI-MS(positive mode)m/z:233.0925[M+H]⁺(Calcd for C₁₁H₁₂NO₅: 233.0926),m/z:255.0749[M+Na]⁺(Calcd for C₁₁H₁₁NO₅Na:255.0746).

2-hydroxyethyl 5-methyl-2- (pyridin-2-yl) oxazole-4-carboxylate (T13): change Object T13 is closed using T12 as raw material, and obtain with reference to T4 synthetic method.System PE/EA/MeOH is unfolded in yield 60%, white solid (1:1:0.01 Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃) 7.70 (dd, J=1.1Hz, 1H), 7.46 (dd, J=1.1Hz, 1H), 7.12 (q, J=3.7Hz, 1H), 4.46 (t, J=4.6Hz, 2H), 3.97 (t, J= 4.6Hz,2H),3.21(s,1H),2.66(s,3H)；¹³C NMR(100MHz,CDCl₃)δ162.3,156.3,155.9,129.1, 128.7,128.6,128.2,128.0,66.8,60.9,12.1；HR-ESI-MS(positive mode)m/z:247.1772 [M-H]^-(Calcd for C₁₂H₁₁N₂O₄:247.0719),m/z:271.0686[M+Na]⁺(Calcd for C₁₂H₁₂N₂O₄Na: 271.0695).

Ethyl 2- (furan-2-yl) -5-methyloxazole-4-carboxylate (T14): the conjunction of compound T14 At the synthetic method of reference 3b.Yield 66%, colourless liquid are unfolded system PE/EA 10:1 (Rf=0.5, PE/EA=5:1),¹H NMR(400MHz,CDCl₃) 7.54 (dd, J=0.6Hz, 1H), 7.08 (dd, J=0.5Hz, 1H), 6.53 (q, J=1.8Hz, 1H), 4.42 (q, J=7.1Hz, 2H), 2.68 (s, 3H), 1.41 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ 162.2,155.7,152.3,144.7,142.1,128.6,112.3,111.9,61.1,14.4,12.1；HR-ESI-MS (positive mode)m/z:222.0763[M+H]⁺(Calcd for C₁₁H₁₂NO₄:222.0766),m/z:244.0590[M+ Na]⁺(Calcd for C₁₁H₁₁NO₄Na:244.0586).

2-hydroxyethyl 2- (furan-2-yl) -5-methyloxazole-4-carboxylate (T15): chemical combination Object T15 synthetic line be using T14 as raw material, and with reference to T4 synthetic method obtain.System PE/ is unfolded in yield 60%, colourless liquid EA/MeOH1:1:0.01 (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃) 7.57 (dd, J= 0.6Hz, 1H), 7.06 (dd, J=0.5Hz, 1H), 6.54 (q, J=1.8Hz, 1H), 4.45 (t, J=4.6Hz, 2H), 3.97 (t, J=4.7Hz, 2H), 3.59 (s, 1H), 2.66 (s, 3H)；¹³C NMR(100MHz,CDCl₃)δ162.2,156.2,152.3, 144.9,141.8,128.1,112.6,112.0,66.8,60.7,12.0；HR-ESI-MS(positive mode)m/z: 238.0718[M+H]⁺(Calcd for C₁₁H₁₂NO₅:238.0715),m/z:260.0538[M+Na]⁺(Calcd for C₁₁H₁₁NO₅Na:260.0535).

Ethyl 5-methyl-2- (thiophen-2-yl) oxazole-4-carboxylate (T16): compound T16 Synthesized reference 3b synthetic method.Yield 70%, white solid, expansion system PE/EA 10:1 (Rf=0.5, PE/EA=5: 1),¹H NMR(400MHz,CDCl₃) 7.72 (dd, J=1.2Hz, 1H), 7.44 (dd, J=1.2Hz, 1H), 7.11 (q, J= 3.7Hz, 1H), 4.43 (q, J=7.1Hz, 2H), 2.67 (s, 3H), 1.42 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz, CDCl₃)δ162.3,155.9,155.7,129.0,128.8,128.7,128.5,127.9,61.1,14.4,12.2；HR-ESI- MS(positive mode)m/z:238.0545[M-H]^-(Calcd for C₁₁H₁₂NO₃S:238.0538),m/z:260.0359 [M+Na]⁺(Calcd for C₁₁H₁₁NO₃SNa:260.0357).

2-hydroxyethyl 5-methyl-2-(thiophen-2-yl)oxazole-4-carboxylate(T17): Compound T17 synthetic line be using T16 as raw material, and with reference to T4 synthetic method obtain.Yield 74%, white solid, expanding body It is PE/EA/MeOH 1:1:0.01 (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃)7.70 (dd, J=1.2Hz, 1H), 7.46 (dd, J=1.2Hz, 1H), 7.12 (q, J=3.7Hz, 1H), 4.45 (t, J=4.6Hz, 2H), 3.97 (t, J=4.6Hz, 2H), 3.11 (s, 1H), 2.66 (s, 3H)；¹³C NMR(100MHz,CDCl₃)δ162.3, 156.3,155.9,129.1,128.7,128.6,128.2,128.0,66.8,60.9,12.1；HR-ESI-MS(positive mode)m/z:254.0465[M-H]^-(Calcd for C₁₁H₁₂NO₄S:254.0487),m/z:276.0288[M+Na]⁺ (Calcd for C₁₁H₁₁NO₄SNa:276.0306).

Ethyl 5-methyl-2-phenyloxazole-4-carboxylate (T18): the synthesized reference of compound T18 The synthetic method of 3b.Yield 64%, white solid are unfolded system PE/EA 10:1 (Rf=0.6, PE/EA=3:1),¹H NMR (400MHz,CDCl₃) 8.06-8.03 (m, 3H), 7.74 (d, J=2.6Hz, 2H), 4.43 (q, J=7.1Hz, 2H), 2.68 (s, 3H), 1.41 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ162.5,159.7,156.2,130.7,128.8, 128.7,126.6,61.1,14.4,12.2；HR-ESI-MS(positive mode)m/z:232.0965[M+H]⁺(Calcd for C₁₃H₁₄NO₃:232.0974),m/z:254.0787[M+Na]⁺(Calcd for C₁₃H₁₃NO₃Na:254.0793).

2-hydroxyethyl 5-methyl-2-phenyloxazole-4-carboxylate (T19): compound T19 Synthesized reference T4 synthetic method.Yield 61%, white solid, expansion system PE/EA/MeOH 1:1:0.01 (Rf=0.5, ), PE/EA/MeOH=1:1:0.01¹H NMR(400MHz,CDCl₃)8.02-8.00(m,2H),7.46-7.45(m,3H),4.45 (t, J=4.5,2H), 3.97 (t, J=4.6,2H), 2.69 (s, 3H)；¹³C NMR(100MHz,CDCl₃)δ162.4,159.8, 156.8,131.0,128.9,128.3,126.5,126.3,66.8,60.9,12.1；HR-ESI-MS(positive mode)m/ z:248.0917[M+H]⁺(Calcd for C₁₃H₁₄NO₄:248.0923),m/z:270.0723[M+Na]⁺(Calcd for C₁₃H₁₃NO₄Na:270.0742).

Ethyl 2- (2-methoxyphenyl) -5-methyloxazole-4-carboxylate (T20): compound The synthetic method of the synthesized reference 3b of T20.System PE/EA 10:1 (Rf=0.5, PE/EA is unfolded in yield 69%, colourless liquid =5:1),¹H NMR(400MHz,CDCl₃) 7.93 (dd, J=1.7Hz, 1H), 7.42-7.38 (m, 1H), 7.01-6.97 (m, 2H), 4.41 (q, J=7.1Hz, 2H), 3.91 (s, 3H), 2.68 (s, 3H), 1.40 (t, J=7.1Hz, 3H)；¹³C NMR (100MHz,CDCl₃)δ162.6,158.3,157.7,156.0,132.1,130.7,128.5,120.5,115.8,111.7, 69.9,55.9,14.4,12.2；HR-ESI-MS(positive mode)m/z:262.1082[M+H]⁺(Calcd for C₁₄H₁₆NO₄:262.1079),m/z:284.0920[M+Na]⁺(Calcd for C₁₄H₁₅NO₄Na:284.0899).

2-hydroxyethyl 2-(2-methoxyphenyl)-5-methyloxazole-4-carboxylate (T21): the synthetic method of the synthesized reference T4 of compound T21.System PE/EA/MeOH 1 is unfolded in yield 66%, colourless liquid: 1:0.01 (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃) 7.90 (dd, J=1.7Hz, 1H), 7.46-7.41 (m, 1H), 7.04-7.00 (m, 2H), 4.43 (t, J=4.5Hz, 2H), 3.93-3.91 (m, 5H), 3.33 (s, 1H),2.68(s,3H)；¹³C NMR(100MHz,CDCl₃)δ162.6,158.3,157.7,156.6,132.3,130.5, 128.0,120.6,115.5,111.8,66.7,60.8,55.9,12.1；HR-ESI-MS(positive mode)m/z: 278.1027[M+H]⁺(Calcd for C₁₄H₁₆NO₅:278.1028),m/z:300.0848[M+Na]⁺(Calcd for C₁₄H₁₅NO₅Na:300.0848).

Ethyl 2- (3-methoxyphenyl) -5-methyloxazole-4-carboxylate (T22): compound The synthetic method of the synthesized reference 3b of T22.System PE/EA 10:1 (Rf=0.5, PE/EA is unfolded in yield 70%, white solid =5:1),¹H NMR(400MHz,CDCl₃) 7.63 (d, J=7.2Hz, 1H), 7.58 (t, J=2.4Hz, 1H), 7.34 (t, J= 8.0Hz, 1H), 6.99 (dd, J=2.5Hz, 1H), 4.43 (q, J=7.1Hz, 2H), 3.85 (s, 3H), 2.68 (s, 3H), 1.42 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ162.5,159.8,159.6,156.2,129.8,128.8, 127.8,119.0,117.5,111.0,61.1,55.5,14.4,12.3；HR-ESI-MS(positive mode)m/z: 262.1073[M+H]⁺(Calcd for C₁₄H₁₆NO₄:262.1079),m/z:284.0899[M+Na]⁺(Calcd for C₁₄H₁₅NO₄Na:284.0899).

2-hydroxyethyl 2-(3-methoxyphenyl)-5-methyloxazole-4-carboxylate (T23): the synthetic method of the synthesized reference T4 of compound T23.System PE/EA/MeOH 1 is unfolded in yield 73%, white solid: 1:0.01 (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃) 7.61 (d, J=7.2Hz, 1H), 7.55 (t, J=2.4Hz, 1H), 7.37 (t, J=8.0Hz, 1H), 7.02 (dd, J=2.5Hz, 1H), 4.47 (t, J=4.6Hz, 2H), 3.98 (t, J=4.6Hz, 2H), 3.86 (s, 3H), 3.27 (s, 1H), 2.66 (s, 3H)；¹³C NMR(100MHz,CDCl₃) δ162.5,159.9,159.6,156.7,130.0,128.3,127.5,119.0,117.5,111.1,66.8,60.9,55.5, 12.2；HR-ESI-MS(positive mode)m/z:278.1017[M+H]⁺(Calcd for C₁₄H₁₆NO₅:278.1028), m/z:300.0841[M+Na]⁺(Calcd for C₁₄H₁₅NO₅Na:300.0848).

Ethyl 5-methyl-2- (p-tolyl) oxazole-4-carboxylate (T24): the synthesis of compound T24 With reference to the synthetic method of 3b.Yield 76%, colourless liquid are unfolded system PE/EA 10:1 (Rf=0.5, PE/EA=5:1),¹H NMR(400MHz,CDCl₃) 7.99 (d, J=8.2Hz, 2H), 7.28 (d, J=8.0Hz, 2H), 4.47 (q, J=7.1Hz, 2H), 2.71 (s, 3H), 2.41 (s, 3H), 1.45 (t, J=7.2Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ162.6,159.9, 155.9,141.1,129.4,128.7,126.6,123.9,61.0,21.5,14.4,12.2；HR-ESI-MS(positive mode)m/z:246.1122[M+H]⁺(Calcd for C₁₄H₁₆NO₃:246.1130),m/z:268.0949[M+Na]⁺(Calcd for C₁₄H₁₅NO₃Na:268.0950).

2-hydroxyethyl 5-methyl-2- (p-tolyl) oxazole-4-carboxylate (T25): compound The synthetic method of the synthesized reference T4 of T25.System PE/EA/MeOH 1:1:0.01 (Rf=is unfolded in yield 71%, white solid 0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃) 7.92 (d, J=8.2Hz, 2H), 7.28 (d, J= 8.0Hz, 2H), 4.48 (t, J=4.6Hz, 2H), 3.99 (t, J=4.7Hz, 2H), 3.89 (s, 1H), 2.66 (s, 3H), 2.41 (s,3H)；¹³C NMR(100MHz,CDCl₃)δ162.5,159.9,156.4,141.4,129.5,128.1,126.5,123,6, 66.7,60.8,21.6,12.1；HR-ESI-MS(positive mode)m/z:262.1082[M+H]⁺(Calcd for C₁₄H₁₆NO₄:261.1079),m/z:284.0905[M+Na]⁺(Calcd for C₁₄H₁₅NO₄Na:284.0899).

Ethyl 2- (4- (fluorooxy) phenyl) -5-methyloxazole-4-carboxylate (T26): chemical combination The synthetic line of object T26 refers to the synthetic method of 3b.Yield 80%, white solid, expansion system PE/EA 10:1 (Rf=0.5, ), PE/EA=5:1¹H NMR(400MHz,CDCl₃) 8.13 (d, J=8.8Hz, 2H), 7.31 (d, J=9.2Hz, 2H), 4.47 (q, J=7.2Hz, 2H), 2.72 (s, 3H), 1.45 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ162.3, (158.4,156.5,150.9 d, J=1.9Hz), 129.0,128.3,125.2,124.2 121.6,121.0,119.1,61.1, 14.4,12.2；HR-ESI-MS(positive mode)m/z:316.0792[M+H]⁺(Calcd for C₁₄H₁₃F₃NO₄: 316.0797),m/z:338.0612[M+Na]⁺(Calcd for C₁₄H₁₂F₃NO₄Na:338.0616).

2-hydroxyethyl 5-methyl-2-(4-(trifluoromethoxy)phenyl)oxazole-4- The synthetic method of carboxylate (T27): compound T27 synthesized reference T4.System is unfolded in yield 84%, white solid PE/EA/MeOH 1:1:0.01 (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃)8.08(d,J =8.9Hz, 2H), 7.32 (d, J=8.1Hz, 2H), 4.49 (t, J=4.6Hz, 2H), 4.00 (t, J=4.7Hz, 2H), 3.47 (s,1H),2.70(s,3H)；¹³C NMR(100MHz,CDCl₃) δ 162.3,158.5,157.0,151.0 (d, J=1.9Hz), 128.5,128.2,124.9,121.6,121.1,119.0,66.7,60.9,12.1；HR-ESI-MS(positive mode)m/ z:332.0740[M+H]⁺(Calcd for C₁₄H₁₃F₃NO₅:332.0746),m/z:354.0563[M+Na]⁺(Calcd for C₁₄H₁₂F₃NO₅Na:354.0565).

Ethyl 2- (4-fluorophenyl) -5-methyloxazole-4-carboxylate (T28): compound T28 Synthesized reference 3b synthetic method.Yield 83%, white solid, expansion system PE/EA 10:1 (Rf=0.5, PE/EA=5: 1),¹H NMR(400MHz,CDCl₃) 8.07-8.04 (m, 2H), 7.15-7.11 (m, 2H), 4.44 (q, J=7.1Hz, 2H), 2.69 (s, 3H), 1.43 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ165.5,163.0,162.4,158.8, 156.2,128.8 (d, J=8.8Hz), 123.0 (d, J=3.4Hz), 116.1 (d, J=22.1Hz), 61.1,14.4,12.2； HR-ESI-MS(positive mode)m/z:250.0877[M+H]⁺(Calcd for C₁₃H₁₃FNO₃:250.0879),m/z: 272.0698[M+Na]⁺(Calcd for C₁₃H₁₂FNO₃Na:272.0699).

2-hydroxyethyl 2-(4-fluorophenyl)-5-methyloxazole-4-carboxylate(T29): The synthetic method of the synthesized reference T4 of compound T29.System PE/EA/MeOH 1:1:0.01 is unfolded in yield 86%, white solid (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃)8.02-7.97(m,2H),7.16-7.10 (m, 2H), 4.45 (t, J=4.6Hz, 2H), 3.96 (t, J=4.7Hz, 2H), 3.71 (s, 1H), 2.64 (s, 3H)；¹³C NMR (100MHz,CDCl₃) δ 165.6,163.1,162.3,158.9,156.7,128.8 (d, J=8.8Hz), 128.3,122.7 (d, ), J=3.4Hz 116.2 (d, J=22.3Hz), 66.7,60.8,12.1；HR-ESI-MS(positive mode)m/z: 266.0829[M+H]⁺(Calcd for C₁₃H₁₃FNO₄:266.0829),m/z:288.0651[M+Na]⁺(Calcd for C₁₃H₁₂FNO₄Na:288.0648).

Ethyl 2- (4-chlorophenyl) -5-methyloxazole-4-carboxylate (T30): compound T30 Synthesized reference 3b synthetic method.Yield 80%, white solid, expansion system PE/EA 10:1 (Rf=0.5, PE/EA=5: 1),¹H NMR(400MHz,CDCl₃) 8.01-7.97 (m, 2H), 7.43-7.40 (m, 2H), 4.44 (q, J=7.1Hz, 2H), 2.69 (s, 3H), 1.42 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ162.3,158.7,156.4,136.9, 129.1,127.9,125.1,61.1,14.4,12.2；HR-ESI-MS(positive mode)m/z:266.0580[M+H]⁺ (Calcd for C₁₃H₁₃ClNO₃:266.0584),m/z:288.0409[M+Na]⁺(Calcd for C₁₃H₁₂ClNO₃Na: 288.0403).

2-hydroxyethyl 2-(4-chlorophenyl)-5-methyloxazole-4-carboxylate(T31): The synthetic method of the synthesized reference T4 of compound T31.System PE/EA/MeOH 1:1:0.01 is unfolded in yield 87%, white solid (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃) 7.95 (d, J=8.6Hz, 2H), 7.43 (d, J=8.6Hz, 2H), 4.47 (t, J=4.5Hz, 2H), 3.97 (t, J=4.6Hz, 2H), 3.20 (s, 1H), 2.67 (s, 3H)；¹³C NMR(100MHz,CDCl₃)δ162.3,158.8,156.9,137.2,129.2,128.5,127.8,124.8,66.8,60.9, 12.2；HR-ESI-MS(positive mode)m/z:282.0532[M+H]⁺(Calcd for C₁₃H₁₃ClNO₄: 282.0533),m/z:304.0348[M+Na]⁺(Calcd for C₁₃H₁₂ClNO₄Na:304.0353).

Ethyl 2- (4-bromophenyl) -5-methyloxazole-4-carboxylate (T32): compound T32 Synthesized reference 3b synthetic method.Yield 90%, white solid, expansion system PE/EA 10:1 (Rf=0.5, PE/EA=5: 1),¹H NMR(400MHz,CDCl₃) 7.92 (d, J=8.6Hz, 2H), 7.58 (d, J=8.6Hz, 2H), 4.43 (q, J= 7.2Hz, 2H), 2.68 (s, 3H), 1.42 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ162.3,158.8, 156.4,132.0,130.0,128.0,125.5,125.3,61.1,14.4,12.3；HR-ESI-MS(positive mode)m/ z:310.0087[M+H]⁺(Calcd for C₁₃H_13brNO₃:310.0079),m/z:331.9902[M+Na]⁺(Calcd for C₁₃H₁₂BrNO₃Na:331.9898).

2-hydroxyethyl 2-(4-bromophenyl)-5-methyloxazole-4-carboxylate(T33): The synthetic method of the synthesized reference T4 of compound T33.System PE/EA/MeOH 1:1:0.01 is unfolded in yield 87%, white solid (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃) 7.90 (d, J=8.6Hz, 2H), 7.60 (d, J=8.5Hz, 2H), 4.48 (t, J=4.5Hz, 2H), 3.97 (t, J=4.7Hz, 2H), 3.03 (s, 1H), 2.68 (s, 3H)；¹³C NMR(100MHz,CDCl₃)δ162.4,158.9,157.0,132.2,128.5,128.0,125.5,125.3,66.8,61.0, 12.2；HR-ESI-MS(positive mode)m/z:326.0027[M+H]⁺(Calcd for C₁₃H_13brNO₃: 326.0028),m/z:347.9847[M+Na]⁺(Calcd for C₁₃H₁₂BrNO₃Na:347.9847).

Ethyl 2- ([1,1'-biphenyl] -4-yl) -5-methyloxazole-4-carboxylate (T34): change Close the synthetic method of the synthesized reference 3b of object T34.Yield 87%, white solid, expansion system PE/EA 10:1 (Rf=0.5, ), PE/EA=5:1¹H NMR(400MHz,CDCl₃) 8.15 (d, J=8.3Hz, 2H), 7.69 (d, J=8.3Hz, 2H), 7.64 (d, J=7.3Hz, 2H), 7.48 (t, J=7.2Hz, 2H), 7.40 (t, J=7.3Hz, 1H), 4.46 (q, J=7.1Hz, 2H), 2.72 (s, 3H), 1.45 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ162.5,159.5,156.2,143.4, 140.0,128.9,128.0,127.4,127.1,127.1,125.5,61.1,14.4,12.3；HR-ESI-MS(positive mode)m/z:308.1284[M+H]⁺(Calcd for C₁₉H₁₈NO₃:308.1287),m/z:330.1097[M+Na]⁺(Calcd for C₁₉H₁₇NO₃Na:330.1106).

2-hydroxyethyl 2-([1,1'-biphenyl]-4-yl)-5-methyloxazole-4-carboxylate (T35): the synthetic method of the synthesized reference T4 of compound T35.System PE/EA/MeOH 1 is unfolded in yield 87%, white solid: 1:0.01 (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃) 8.09 (d, J=8.4Hz, 2H), 7.69 (d, J=8.7Hz, 2H), 7.64 (d, J=7.2Hz, 2H), 7.48 (t, J=7.2Hz, 2H), 7.40 (t, J=7.3Hz, 1H), 4.49 (t, J=4.5Hz, 2H), 4.00 (t, J=4.6Hz, 2H), 3.20 (s, 1H), 2.69 (s, 3H)；¹³C NMR (100MHz,CDCl₃)δ162.5,159.6,156.8,143.6,139.9,129.0,128.4,128.0,127.5,127.1, 125.2,66.8,61.0,12.2；HR-ESI-MS(positive mode)m/z:324.1242[M+H]⁺(Calcd for C₁₉H₁₈NO₄:324.1236),m/z:346.1058[M+Na]⁺(Calcd for C₁₉H₁₇NO₄Na:346.1055).

Ethyl 2- (3,5-difluorophenyl) -5-methyloxazole-4-carboxylate (T36): chemical combination The synthetic method of the synthesized reference 3b of object T36.System PE/EA 10:1 (Rf=0.5, PE/ is unfolded in yield 87%, white solid ), EA=5:1¹H NMR(400MHz,CDCl₃) 7.61-7.59 (m, 2H), 6.93-6.88 (m, 1H), 4.46 (q, J=7.1Hz, 2H), 2.71 (s, 3H), 1.44 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz,CDCl₃) δ 164.5 (d, J=12.6Hz), 162.1,162.0 (d, J=12.4Hz), 155.9,129.4 (t, J=13.8Hz), 109.8 (d, J=11.8Hz), 109.7 (d, ), J=11.8Hz 106.4 (t, J=25.4Hz), 61.3,14.4,12.3；HR-ESI-MS(positive mode)m/z: 268.0773[M+H]⁺(Calcd for C₁₃H₁₂F₂NO₃:268.0785),m/z:290.0592[M+Na]⁺(Calcd for C₁₃H₁₁F₂NO₃Na:290.0605).

2-hydroxyethyl 2-(3,5-difluorophenyl)-5-methyloxazole-4-carboxylate (T37): the synthetic method of the synthesized reference T4 of compound T37.System PE/EA/MeOH 1 is unfolded in yield 86%, white solid: 1:0.01 (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃)7.58(m,2H),6.94(m,H), 4.49 (t, J=4.6Hz, 2H), 3.98 (t, J=4.7Hz, 2H), 2.71 (s, 3H), 2.55 (s, 1H)；¹³C NMR(100MHz, CDCl₃) δ 164.5 (d, J=12.6Hz), 162.3,162.0 (d, J=12.4Hz), 157.4,129.2 (t, J=13.8Hz), 109.8 (d, J=11.8Hz), 109.6 (d, J=11.8Hz), 106.3 (t, J=25.4Hz), 66.8,61.1,12.2；HR- ESI-MS(positive mode)m/z:284.0727[M+H]⁺(Calcd for C₁₃H₁₂F₂NO₄:284.0734),m/z: 306.0549[M+Na]⁺(Calcd for C₁₃H₁₁F₂NO₄Na:306.0554).

Ethyl 2-(3,5-bis(trifluoromethyl)phenyl)-5-methyloxazole-4- The synthetic method of carboxylate (T38): compound T38 synthesized reference 3b.System is unfolded in yield 89%, white solid PE/EA 10:1 (Rf=0.5, PE/EA=5:1),¹H NMR(400MHz,CDCl₃)8.53(s,2H),7.95(s,1H),4.47 (q, J=7.1Hz, 2H), 2.75 (s, 3H), 1.45 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ161.9, (157.4,156.8,133.0 q, J=34.0Hz), 129.5,128.6,127.0,126.6 (d, J=3.1Hz), 124.3, 124.0 (q, J=7.4Hz), 121.5,118.8,61.4,14.4,12.3；HR-ESI-MS(positive mode)m/z: 368.0703[M+H]⁺(Calcd for C₁₅H₁₂F₆NO₃:368.0721),m/z:390.0532[M+Na]⁺(Calcd for C₁₅H₁₁F₆NO₃Na:390.0541).

2-hydroxyethyl 2-(3,5-bis(trifluoromethyl)phenyl)-5-methyloxazole-4- The synthetic method of carboxylate (T39): compound T39 synthesized reference T4.System is unfolded in yield 83%, white solid PE/EA/MeOH1:1:0.01 (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃)8.49(s, 2H), 7.96 (s, 1H), 4.50 (t, J=4.5Hz, 2H), 3.99 (t, J=4.6Hz, 2H), 2.75 (s, 3H), 2.52 (s, 1H) ；¹³C NMR(100MHz,CDCl₃) δ 162.0,158.0,156.9,133.1 (q, J=34.0Hz), 129.1,128.4,126.9, 126.5 (d, J=3.2Hz), 124.2 (q, J=7.6Hz), 121.5,118.8,66.9,60.9,12.3；HR-ESI-MS (positive mode)m/z:384.0667[M+H]⁺(Calcd for C₁₅H₁₂F₆NO₄:384.0671),m/z:406.0487 [M+Na]⁺(Calcd for C₁₅H₁₁F₆NO₄Na:406.0490).

Ethyl 5-ethyl-2- (4-methoxyphenyl) oxazole-4-carboxylate (T40): compound T40 Synthesized reference 3b synthetic method.Yield 94%, white solid, expansion system PE/EA 10:1 (Rf=0.5, PE/EA=5: 1),¹H NMR(400MHz,CDCl₃) 8.01 (d, J=8.9Hz, 2H), 6.96 (d, J=8.9Hz, 2H), 4.43 (q, J= 7.1Hz, 2H), 3.84 (s, 3H), 3.13 (q, J=7.7Hz, 2H), 1.42 (t, J=7.1Hz, 3H), 1.34 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ162.6,161.6,160.5,159.7,128.3,127.7,119.5,114.1, 61.0,55.4,19.8,14.4,12.2；HR-ESI-MS(positive mode)m/z:276.1221[M+H]⁺(Calcd for C₁₅H₁₈NO₄:276.1236),m/z:298.1038[M+Na]⁺(Calcd for C₁₅H₁₇NO₄Na:298.1055).

2-hydroxyethyl5-ethyl-2-(4-methoxyphenyl)oxazole-4-carboxylate(T41): The synthetic method of the synthesized reference T4 of compound T41.System PE/EA/MeOH 1:1:0.01 is unfolded in yield 91%, white solid (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃) 7.96 (d, J=8.7Hz, 2H), 6.95 (d, J=8.7Hz, 2H), 4.45 (t, J=4.3Hz, 2H), 3.96 (t, J=4.5Hz, 2H), 3.84 (s, 3H), 3.10 (q, J= 7.6Hz, 2H), 1.32 (t, J=7.6Hz, 3H), 1.24 (s, 1H)；¹³C NMR(100MHz,CDCl₃)δ162.5,161.7, 161.0,159.8,128.3,127.2,119.2,114.2,66.6,60.8,55.4,19.7,12.1；HR-ESI-MS (positive mode)m/z:292.1174[M+H]⁺(Calcd for C₁₅H₁₈NO₅:292.1185),m/z:314.1002[M+ Na]⁺(Calcd for C₁₅H₁₇NO₅Na:314.1004).

Ethyl 2- (4-methoxyphenyl) -5-propyloxazole-4-carboxylate (T42): compound The synthetic method of the synthesized reference 3b of T42.System PE/EA 10:1 (Rf=0.5, PE/EA is unfolded in yield 89%, white solid =5:1),¹H NMR(400MHz,CDCl₃) 8.01 (d, J=8.8Hz, 2H), 6.95 (d, J=8.9Hz, 2H), 4.43 (q, J= 7.1Hz, 2H), 3.84 (t, J=7.4Hz, 2H), 1.81-1.72 (m, 2H), 1.42 (t, J=7.1Hz, 3H), 1.02 (t, J= 7.4Hz,3H)；¹³C NMR(100MHz,CDCl₃)δ162.6,161.6,159.8,159.4,128.3,119.5,114.1, 60.9,55.4,28.0,21.4,14.4,13.7；HR-ESI-MS(positive mode)m/z:290.1382[M+H]⁺ (Calcd for C₁₆H₂₀NO₄:290.1392),m/z:312.1202[M+Na]⁺(Calcd for C₁₆H₁₉NO₄Na: 312.1212).

2-hydroxyethyl 2-(4-methoxyphenyl)-5-propyloxazole-4-carboxylate (T43): the synthetic method of the synthesized reference T4 of compound T43.System PE/EA/MeOH 1 is unfolded in yield 87%, white solid: 1:0.01 (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃) 7.98 (d, J=8.8Hz, 2H), 6.97 (d, J=8.8Hz, 2H), 4.46 (t, J=4.5Hz, 2H), 3.97 (t, J=4.7Hz, 2H), 3.85 (s, 3H), 3.10 (s, 1H), 3.07 (t, J=7.4Hz, 2H), 1.81-1.72 (m, 2H), 1.03 (t, J=7.4Hz, 3H)；¹³C NMR(100MHz, CDCl₃)δ162.7,161.7,160.0,159.9,128.3,127.8,119.2,114.2,66.7,61.0,55.4,27.9, 21.3,13.7；HR-ESI-MS(positive mode)m/z:306.1341[M+H]⁺(Calcd for C₁₆H₂₀NO₅: 306.1347),m/z:328.1153[M+Na]⁺(Calcd for C₁₆H₁₉NO₅Na:328.1161).

Ethyl 5-isopropyl-2- (4-methoxyphenyl) oxazole-4-carboxylate (T44): chemical combination The synthetic method of the synthesized reference 3b of object T44.System PE/EA 10:1 (Rf=0.5, PE/ is unfolded in yield 89%, white solid ), EA=5:1¹H NMR(400MHz,CDCl₃) 8.02 (d, J=8.8Hz, 2H), 6.96 (d, J=8.8Hz, 2H), 4.44 (q, J =7.1Hz, 2H), 3.85 (s, 3H), 3.84 (q, 7.0Hz, 1H), 1.43 (t, J=7.1Hz, 3H), 1.36 (d, J=7.0Hz, 6H)；¹³C NMR(100MHz,CDCl₃)δ163.7,162.6,161.6,159.5,128.3,126.7,119.5,114.1, 60.9,55.4,26.2,20.8,14.4；HR-ESI-MS(positive mode)m/z:290.1380[M+H]⁺(Calcd for C₁₆H₂₀NO₄:290.1392),m/z:312.1197[M+Na]⁺(Calcd for C₁₆H₁₉NO₄Na:312.1212).

2-hydroxyethyl 5-isopropyl-2-(4-methoxyphenyl)oxazole-4-carboxylate (T45): the synthetic method of the synthesized reference T4 of compound T45.System PE/EA/MeOH 1 is unfolded in yield 87%, white solid: 1:0.01 (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃) 7.97 (d, J=8.8Hz, 2H), 6.96 (d, J=8.8Hz, 2H), 4.45 (t, J=4.5Hz, 2H), 3.96 (t, J=4.7Hz, 2H), 3.84 (s, 3H), 3.83 (q, J=7Hz, 1H), 3.67 (s, 1H), 1.34 (d, J=7.0Hz, 6H)；¹³C NMR(100MHz,CDCl₃)δ164.3, 162.6,161.7,159.6,128.3,126.1,119.2,114.2,66.7,60.8,55.4,26.2,20.7；HR-ESI-MS (positive mode)m/z:306.1337[M+H]⁺(Calcd for C₁₆H₂₀NO₅:306.1341),m/z:328.1153[M+ Na]⁺(Calcd for C₁₆H₁₉NO₅Na:328.1161).

Ethyl 5- (tert-butyl) -2- (4-methoxyphenyl) oxazole-4-carboxylate (T46): change Close the synthetic method of the synthesized reference 3b of object T46.System Compound wt-3-7was is unfolded in yield 85%, white solid Obtained as a white solid in 62%yield；isolated by column chromatography PE/ EA 10:1 (Rf=0.5, PE/EA=5:1),¹H NMR(400MHz,CDCl₃) 8.00 (d, J=8.8Hz, 2H), 6.96 (d, J= 8.8Hz, 2H), 4.45 (q, J=7.1Hz, 2H), 3.86 (s, 3H), 1.50 (s, 9H), 1.44 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ165.1,162.6,161.5,157.9,128.3,127.3,119.5,114.1,61.2,55.4, 33.5,28.3,14.4；HR-ESI-MS(positive mode)m/z:304.1550[M+H]⁺(Calcd for C₁₇H₂₂NO₄: 306.1341),m/z:326.1374[M+Na]⁺(Calcd for C₁₇H₂₁NO₄Na:326.1368).

2-hydroxyethyl 5-(tert-butyl)-2-(4-methoxyphenyl)oxazole-4- The synthetic method of carboxylate (T47): compound T47 synthesized reference T4.System is unfolded in yield 88%, white solid PE/EA/MeOH 1:1:0.01 (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃)7.97(d,J =8.8Hz, 2H), 6.98 (d, J=8.8Hz, 2H), 4.46 (t, J=4.4Hz, 2H), 3.97 (t, J=4.6Hz, 2H), 3.86 (s,3H),2.36(s,1H),1.50(s,9H)；¹³C NMR(100MHz,CDCl₃)δ166.0,162.5,161.7,158.0, 128.2,126.7,119.2,114.3,67.1,61.0,55.4,33.5,28.1；HR-ESI-MS(positive mode)m/z: 320.1501[M+H]⁺(Calcd for C₁₇H₂₂NO₅:320.1498),m/z:342.1322[M+Na]⁺(Calcd for C₁₇H₂₁NO₅Na:342.1317).

Ethyl 5-cyclopropyl-2- (4-methoxyphenyl) oxazole-4-carboxylate (T48): change Close the synthetic method of the synthesized reference 3b of object T48.Yield 88%, white solid, expansion system PE/EA 10:1 (Rf=0.5, ), PE/EA=5:1¹H NMR(400MHz,CDCl₃) 7.93 (d, J=8.9Hz, 2H), 6.93 (d, J=d, J=8.9Hz, 2H), 4.45 (q, J=7.1Hz, 2H), 3.83 (s, 3H), 2.83-2.76 (m, 1H), 1.43 (t, J=7.1Hz, 3H), 1.18-1.15 (m,4H)；¹³C NMR(100MHz,CDCl₃)δ162.9,161.5,160.3,158.3,128.2,128.0,119.4,114.1, 60.9,55.4,14.5,9.2,8.1；HR-ESI-MS(positive mode)m/z:288.1244[M+H]⁺(Calcd for C₁₆H₁₈NO₄:288.1236),m/z:310.1059[M+Na]⁺(Calcd for C₁₆H₁₇NO₄Na:310.1055).

2-hydroxyethyl 5-cyclopropyl-2-(4-methoxyphenyl)oxazole-4-carboxylate (T49): the synthetic method of the synthesized reference T4 of compound T49.System PE/EA/MeOH 1 is unfolded in yield 86%, white solid: 1:0.01 (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃) 7.84 (d, J=8.6Hz, 2H), 6.91 (d, J=8.6Hz, 2H), 4.45 (t, J=4.5Hz, 2H), 4.27 (s, 1H), 3.96 (t, J=4.5Hz, 2H), 3.81 (s,3H),2.75-2.71(m,1H),1.13-1.09(m,4H)；¹³C NMR(100MHz,CDCl₃)δ162.8,161.6, 160.8,158.3,128.1,127.5,119.0,114.2,66.6,60.8,55.4,9.4,8.0；HR-ESI-MS(positive mode)m/z:304.1179[M+H]⁺(Calcd for C₁₆H₁₈NO₅:304.1185),m/z:326.0999[M+Na]⁺(Calcd for C₁₆H₁₇NO₅Na:326.1004).

Ethyl 2- (4-methoxyphenyl) -5-phenyloxazole-4-carboxylate (T50): compound The synthetic method of the synthesized reference 3b of T50.System PE/EA 10:1 (Rf=0.5, PE/EA is unfolded in yield 92%, white solid =5:1),¹H NMR(400MHz,CDCl₃) 8.10-8.06 (m, 4H), 7.50-7.42 (m, 3H), 6.98 (d, J=8.9Hz, 2H), 4.47 (q, J=7.1Hz, 2H), 3.84 (s, 3H), 1.43 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ 162.4,161.9,159.9,154.6,130.1,128.6,128.5,128.4,128.1,127.3,119.1,114.2,61.4, 55.4,14.3；HR-ESI-MS(positive mode)m/z:324.1237[M+H]⁺(Calcd for C₁₉H₁₈NO₄: 324.1236),m/z:346.1060[M+Na]⁺(Calcd for C₁₉H₁₇NO₄Na:346.1055).

2-hydroxyethyl 2-(4-methoxyphenyl)-5-phenyloxazole-4-carboxylate (T51): the synthetic method of the synthesized reference T4 of compound T51.System PE/EA/MeOH 1 is unfolded in yield 91%, white solid: 1:0.01 (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃) 8.04 (dd, J=4.0Hz, 2.2Hz, 2H), 7.97 (d, J=8.8Hz, 2H), 7.46 (dd, J=0.9Hz, 2.3Hz, 3H), 6.92 (d, J=8.9Hz, 2H), 4.47 (t, J=4.4Hz, 2H), 3.98 (t, J=4.6Hz, 3H), 3.81 (s, 3H)；¹³C NMR(100MHz,CDCl₃)δ 162.2,162.0,159.9,154.8,130.3,128.5,128.4,128.4,127.5,126.9,118.7,114.3,67.2, 60.7,55.4；HR-ESI-MS(positive mode)m/z:340.1175[M+H]⁺(Calcd for C₁₉H₁₈NO₅: 340.1185),m/z:362.0995[M+Na]⁺(Calcd for C₁₉H₁₇NO₅Na:362.1004).

4-hydroxybutyl 2-(4-methoxyphenyl)-5-methyloxazole-4-carboxylate (T52): compound 3b (261.3mg, 1mmol) is added in the round-bottomed flask of 25mL, EtOH/10%NaOH=1:1 (20mL), 1h is reacted at 70 DEG C, then is adjusted between pH to 1~2 with concentrated hydrochloric acid, and dichloromethane extraction, concentration organic phase obtains midbody compound 3 do not have to be further purified；The addition methylene chloride 20mL into intermediate 3, dicyclohexylcarbodiimide (DCC) (412.6mg, 2mmol), 4-dimethylaminopyridine (DMAP) (24.4mg, 0.2mmol) and alcohol (2mmol), reaction system are heated to 80 DEG C, It flows back 6 hours.20mL water quenching reaction is added after reaction, methylene chloride extraction merges organic phase and is concentrated, then through column Chromatography purifies (petrol ether/ethyl acetate/methanol=10/1/0.01), obtains compound T52.Yield 82%, white solid, Expansion system PE/EA/MeOH 1:1:0.01 (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃) 8.00 (d, J=9.0Hz, 2H), 6.96 (d, J=9.0Hz, 2H), 4.41 (t, J=6.5Hz, 2H), 3.85 (s, 3H), 3.74 (t, J=6.3Hz, 2H) 2.67 (s, 3H), 1.93-1.86 (m, 3H), 1.75-1.69 (m, 2H)；¹³C NMR(100MHz, CDCl₃)δ162.5,161.7,159.8,155.6,128.4,128.3,119.3,114.2,64.8,62.2,55.4,29.4, 25.2,12.2；HR-ESI-MS(positive mode)m/z:306.1341[M+H]⁺(Calcd for C₁₆H₂₀NO₄: 306.1333),m/z:328.1155[M+Na]⁺(Calcd for C₁₆H₁₉NO₄Na:328.1161).

N-(2-hydroxyethyl)-2-(4-methoxyphenyl)-5-methyloxazole-4-carboxamide (T53): compound T53 synthetic method refers to T52.System PE/EA/MeOH 1:1:0.01 is unfolded in yield 71%, white solid (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃) 7.91 (d, J=8.9Hz, 2H), 7.47 (s, 1H), 6.96 (d, J=8.9Hz, 2H), 3.85 (s, 3H), 3.83 (t, J=4.8Hz, 2H), 3.61 (q, J=5.6Hz, 2H), 2.67(s,3H)；¹³C NMR(100MHz,CDCl₃)δ163.4,161.6,158.8,152.6,129.8,128.0,119.5, 114.3,62.6,55.4,42.2,11.8；HR-ESI-MS(positive mode)m/z:277.1189[M+H]⁺(Calcd for C₁₄H₁₇N₂O₄:277.1188),m/z:299.1011[M+Na]⁺(Calcd for C₁₄H₁₆N₂O₄Na:299.1008).

2-(2-hydroxyethoxy)ethyl 2-(4-methoxyphenyl)-5-methyloxazole-4- The synthetic method of carboxylate (T54): compound T54 synthesized reference T52.System is unfolded in yield 80%, white solid PE/EA/MeOH1:1:0.01 (Rf=0.5, PE/EA/MeOH=1:1:0.01),¹H NMR(400MHz,CDCl₃)7.97(d,J =8.9Hz, 2H), 6.93 (d, J=8.9Hz, 2H), 4.47 (t, J=4.7Hz, 2H), 3.83-3.81 (m, 5H), 3.75 (t, J =4.2Hz, 2H), 3.64 (t, J=4.9Hz, 2H), 3.05 (s, 1H), 2.64 (s, 3H)；¹³C NMR(100MHz,CDCl₃)δ 162.4,161.7,159.8,155.9,128.3,128.2,119.2,114.2,72.6,68.8,63.7,61.6,55.4, 12.1；HR-ESI-MS(positive mode)m/z:322.1295[M+H]⁺(Calcd for C₁₆H₂₀NO₆:322.1291), m/z:344.1114[M+Na]⁺(Calcd for C₁₆H₁₉NO₆Na:344.1110).

5-chloro-2- (4-methoxyphenyl) -6-methylpyrimidin-4-ol (T55): in nitrogen protection Under, 4- methoxybenzene carbonamidine (330.4mg, 2mmol) is added in the round-bottomed flask of 25mL, NaOH (374mg, 2mmol), methanol (20mL), 60 DEG C of reaction 2h, removes solvent after completion of the reaction, obtains compound T55 through column chromatographic isolation and purification.Yield 54%, White solid is unfolded system PE/EA 4:1 (Rf=0.5, PE/EA=2:1),¹H NMR(400MHz,DMSO-d₆)13.01(s, 1H), 8.11 (d, J=8.8Hz, 2H), 7.09 (d, J=8.9Hz, 2H), 3.85 (s, 3H), 2.42 (s, 3H)；¹³C NMR (100MHz,DMSO-d₆)δ162.7,130.2,114.6,56.0,48.0,33.8,25.8,24.9,22.6；HR-ESI-MS (positive mode)m/z:251.0588[M+H]⁺(Calcd for C₁₂H₁₂ClN₂O₂:251.0587),m/z:273.0403 [M+Na]⁺(Calcd for C₁₄H₁₆N₂O₃Na:273.0407).

2- (chloromethyl) -5-phenyl-1,3,4-oxadiazole (T56): add in the round-bottomed flask of 25mL Enter benzoyl hydrazine (272.3mg, 2mmol), phosphorus oxychloride (306.7mg, 2mmol) and monoxone (20mL) reflux preparation T57. Yield 96%, white solid are unfolded system PE/EA 1:5 (Rf=0.5, PE/EA=1:2),¹H NMR(400MHz,CDCl₃) 8.05 (d, J=6.9Hz, 2H), 7.56-7.47 (m, 3H), 4.77 (s, 2H)；¹³C NMR(150MHz,CDCl₃)δ166.0, 162.2,132.2,129.1,127.1,123.3,33.0；HR-ESI-MS(positive mode)m/z:195.0315[M+H]⁺ (Calcd for C₉H₈N₂OCl:195.0325),m/z:217.0139[M+Na]⁺(Calcd for C₉H₇N₂OClNa: 217.0145).

2- (2-chloroethyl) -5-phenyl-1,3,4-oxadiazole (T57): the synthesized reference of compound T57 The synthetic method of T56.Yield 95%, white solid are unfolded system PE/EA 1:5 (Rf=0.5, PE/EA=1:2),¹H NMR (400MHz,CDCl₃) 8.04 (d, J=7.3Hz, 2H), 7.55-7.48 (m, 3H), 3.97 (t, J=6.9Hz, 2H), 3.43 (t, J=6.9Hz, 2H)；¹³C NMR(150MHz,CDCl₃)δ165.1,163.5,131.8,129.1,126.9,123.7,39.6, 29.2；HR-ESI-MS(positive mode)m/z:209.0470[M+H]⁺(Calcd for C₁₀H₁₀N₂OCl: 209.0482),m/z:231.0290[M+Na]⁺(Calcd for C₁₀H₉N₂OClNa:231.0301).

2-phenylthiazole (S1): compound S1 synthesized reference T7.System is unfolded in yield 97%, colourless liquid PE/EA20:1 (Rf=0.5, PE/EA=10:1),¹H NMR(400MHz,CDCl₃) 7.99-7.96 (m, 2H), 7.88 (d, J= 3.2Hz,1H),7.46-7.41(m,3H),7.33-7.31(m,1H)；¹³C NMR(100MHz,CDCl₃)δ168.5,143.6, 133.5,130.1,129.0,126.6,118.9；HR-ESI-MS(positive mode)m/z:162.0365[M+H]⁺ (Calcd for C₉H₈NS:162.0377),m/z:184.0199[M+Na]⁺(Calcd for C₉H₇NSNa:184.0197).

The synthesis of ethyl 2- (4-methoxyphenyl) thiazole-5-carboxylate (S2): compound S2 is joined Examine T7.Yield 91%, white solid are unfolded system PE/EA 20:1 (Rf=0.5, PE/EA=10:1),¹H NMR(400MHz, CDCl₃) 8.06 (s, 1H), 7.93 (d, J=8.8Hz, 2H), 6.93 (d, J=8.8Hz, 2H), 4.44 (q, J=7.1Hz, 2H), 3.82 (s, 3H), 1.41 (t, J=7.1Hz, 3H)；¹³C NMR(150MHz,CDCl₃)δ168.7,161.6,161.5,147.8, 128.5,126.2,125.8,114.3,61.4,55.4,14.3；HR-ESI-MS(positive mode)m/z:264.0689[M +H]⁺(Calcd for C₁₃H₁₄NO₃S:264.0694),m/z:286.0509[M+Na]⁺(Calcd for C₁₃H₁₃NO₃SNa: 286.0514).

2-hydroxyethyl2- (4-methoxyphenyl) thiazole-5-carboxylate (S3): compound S3 Synthesized reference T10.Yield 88%.White solid is unfolded system PE/EA 1:1 (Rf=0.5, PE/EA=1:1),¹H NMR (400MHz,CDCl₃) 8.12 (s, 1H), 7.91 (d, J=8.8Hz, 2H), 6.96 (d, J=8.8Hz, 2H), 4.47 (t, J= 4.5Hz, 2H), 3.95 (t, J=4.6Hz, 2H), 3.85 (s, 3H), 3.38 (s, 1H)；¹³C NMR(150MHz,CDCl₃)δ 169.1,161.8,161.6,147.2,128.6,127.0,126.5,125.5,114.4,114.3,67.1,60.9,55.4, 52.4,29.7；HR-ESI-MS(positive mode)m/z:280.0634[M+H]⁺(Calcd for C₁₃H₁₄NO₄S: 280.0644),m/z:302.0451[M+Na]⁺(Calcd for C₁₃H₁₃NO₄SNa:302.0463).

(2- (4-methoxyphenyl) -4-methylthiazol-5-yl) methanol (S4): being that starting is former with T7 Material uses nitrogen protection in the THF solvent of lithium aluminium hydride reduction, reacts 4~6h at 0 DEG C and obtain compound S4.Yield 85% is white Color solid is unfolded system PE/EA 1:1 (Rf=0.5, PE/EA=1:1),¹H NMR(400MHz,CDCl₃) 7.82 (d, J= 8.8Hz, 2H), 6.93 (d, J=8.8Hz, 2H), 4.78 (s, 2H), 3.83 (s, 3H), 2.39 (s, 3H), 2.24 (s, 1H)；¹³C NMR(100MHz,CDCl₃)δ166.2,161.1,150.1,130.1,127.9,126.5,114.2,55.8,55.4,15.1； HR-ESI-MS(positive mode)m/z:236.0751[M+H]⁺(Calcd for C₁₂H₁₄NO₂S:236.0745),m/z: 258.0569[M+Na]⁺(Calcd for C₁₂H₁₃NO₂SNa:258.0565).

Diethyl 2- (4-methoxyphenyl) thiazole-4,5-dicarboxylate (S5): compound S5's Synthesis be using chloro methyl-oxalacetic ester as starting material, and with reference to T7 synthetic method.Yield 78%, white solid, exhibition System PE/EA20:1 (Rf=0.5, PE/EA=10:1) is opened,¹H NMR(400MHz,CDCl₃) 7.94 (d, J=8.9Hz, 2H), 6.96 (d, J=8.9Hz, 2H), 4.50 (q, J=7.2Hz, 2H), 4.39 (q, J=7.2Hz, 2H), 3.87 (s, 3H), 1.44 (t, J=7.2Hz, 3H), 1.40 (t, J=7.2Hz, 3H)；¹³C NMR(150MHz,CDCl₃)δ171.4,163.4,162.4, 160.3,150.9,128.8,127.0,125.1,114.5,62.3,62.1,55.5,14.1,14.0；HR-ESI-MS (positive mode)m/z:336.0916[M+H]⁺(Calcd for C₁₆H₁₈NO₅S:336.0906),m/z:358.0734[M +Na]⁺(Calcd for C₁₆H₁₇NO₅SNa:358.0725).

Ethyl 2- (4-chlorophenyl) -4-methylthiazole-5-carboxylate (S6): compound s 6 Synthesized reference T7.Yield 87%, white solid are unfolded system PE/EA 20:1 (Rf=0.5, PE/EA=10:1),¹H NMR (400MHz,CDCl₃) 7.78 (d, J=8.5Hz, 2H), 7.32 (d, J=8.5Hz, 2H), 4.32 (q, J=7.1Hz, 2H), 2.70 (s, 3H), 1.36 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ168.1,161.9,161.0,136.9, 131.3,129.1,127.8,122.0,61.2,17.5,14.3；HR-ESI-MS(positive mode)m/z:282.0360[M +H]⁺(Calcd for C₁₃H₁₃ClNO₂S:282.0356),m/z:304.0182[M+Na]⁺(Calcd for C₁₃H₁₂ClNO₂SNa:304.0175).

2-hydroxyethyl 2-(4-chlorophenyl)-4-methylthiazole-5-carboxylate(S7): The synthesized reference T10 of compound S7.Yield 88%, white solid, expansion system PE/EA 1:1 (Rf=0.5, PE/EA=1: 1),¹H NMR(400MHz,CDCl₃) 7.81 (d, J=8.5Hz, 2H), 7.37 (d, J=8.5Hz, 2H), 4.40 (t, J= 4.6Hz, 2H), 3.92 (t, J=4.6Hz, 2H), 2.79 (S, 1H), 2.72 (s, 3H)；¹³C NMR(100MHz,CDCl₃)δ 168.9,162.2,161.7,137.2,131.1,129.3,128.0,121.4,66.9,61.0,17.5；HR-ESI-MS (positive mode)m/z:298.0308[M+H]⁺(Calcd for C₁₃H₁₃ClNO₃S:298.0305),m/z:320.0132 [M+Na]⁺(Calcd for C₁₃H₁₂ClNO₃SNa:320.0124).

Ethyl 2- (4-bromophenyl) -4-methylthiazole-5-carboxylate (S8): compound S8's Synthesized reference T7.Yield 98%, white solid are unfolded system PE/EA 20:1 (Rf=0.5, PE/EA=10:1),¹H NMR (400MHz,CDCl₃) 7.79 (d, J=8.5Hz, 2H), 7.55 (d, J=8.5Hz, 2H), 4.37 (q, J=7.1Hz, 2H), 2.76 (s, 3H), 1.40 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ168.3,162.1,161.1,132.2, 131.8,128.1,125.4,122.1,61.3,17.5,14.4；HR-ESI-MS(positive mode)m/z:325.9831[M +H]⁺(Calcd for C₁₃H_13brNO₂S:325.9850),m/z:347.9676[M+Na]⁺(Calcd for C₁₃H₁₂BrNO₂SNa:347.9670).

2-hydroxyethyl 2-(4-bromophenyl)-4-methylthiazole-5-carboxylate(S9): The synthesized reference T10 of compound S9.Yield 94%, white solid, expansion system PE/EA 1:1 (Rf=0.5, PE/EA=1: 1),¹H NMR(400MHz,CDCl₃) 7.82 (d, J=8.5Hz, 2H), 7.59 (d, J=8.5Hz, 2H), 4.44 (t, J= 4.6Hz, 2H), 3.95 (t, J=4.7Hz, 2H), 2.77 (s, 3H), 2.01 (s, 1H)；¹³C NMR(100MHz,CDCl₃)δ 168.9,162.3,161.8,132.3,131.7,128.2,125.6,121.4,66.9,61.2,17.6；HR-ESI-MS (positive mode)m/z:341.9807[M+H]⁺(Calcd for C₁₃H_13brNO₃S:341.9800),m/z:363.9624 [M+Na]⁺(Calcd for C₁₃H₁₂BrNO₃SNa:363.9619).

Ethyl 4-methyl-2-(4-(trifluoromethyl)phenyl)thiazole-5-carboxylate (S10): the synthesized reference T7 of compound S10.System PE/EA 20:1 (Rf=0.5, PE/ is unfolded in yield 87%, white solid ), EA=10:1¹H NMR(400MHz,CDCl₃) 8.03 (d, J=8.1Hz, 2H), 7.67 (d, J=8.2Hz, 2H), 4.36 (q, J =7.1Hz, 2H), 2.76 (s, 3H), 1.39 (t, J=7.2Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ167.6,161.9 (d, J=74.1Hz), 136.0,132.9 (q, J=32.7Hz), 127.0,126.0 (d, J=3.7Hz), 125.1,122.9, 122.4,61.4,17.5,14.3；HR-ESI-MS(positive mode)m/z:316.0600[M+H]⁺(Calcd for C₁₄H₁₃F₃NO₂S:316.0619),m/z:338.0425[M+Na]⁺(Calcd for C₁₄H₁₂F₃NO₂SNa:338.0439).

2-hydroxyethyl 4-methyl-2-(4-(trifluoromethyl)phenyl)thiazole-5- Carboxylate (S11): compound S11 synthesized reference T10.System PE/EA 1:1 is unfolded in yield 86%, white solid (Rf=0.5, PE/EA=1:1),¹H NMR(400MHz,CDCl₃) 7.91 (d, J=8.2Hz, 2H), 7.59 (d, J=8.3Hz, 2H), 4.38 (t, J=4.6Hz, 2H), 3.90 (t, J=4.8Hz, 2H), 3.38 (s, 1H), 2.68 (s, 3H)；¹³C NMR (100MHz,CDCl₃) δ 168.1,162.0 (d, J=38.1Hz), 135.6,133.0 (q, J=32.8Hz), 130.0,126.9, 126.0 (d, J=3.8Hz), 125.0,122.3,122.2,66.9,60.7,17.4；HR-ESI-MS(positive mode)m/ z:332.0560[M+H]⁺(Calcd for C₁₄H₁₃F₃NO₃S:332.0568),m/z:354.0386[M+Na]⁺(Calcd for C₁₄H₁₂F₃NO₃SNa:354.0388).

Ethyl 4-methyl-2- (4-nitrophenyl) thiazole-5-carboxylate (S12): compound S12 Synthesized reference T7.Yield 83%, white solid are unfolded system PE/EA 20:1 (Rf=0.5, PE/EA=10:1),¹H NMR (400MHz,CDCl₃) 8.31 (d, J=8.9Hz, 2H), 8.14 (d, J=8.9Hz, 2H), 4.40 (q, J=7.1Hz, 2H), 2.79 (s, 3H), 1.41 (t, J=7.1Hz, 3H)；¹³C NMR(150MHz,CDCl₃)δ166.4,161.8,161.5,149.0, 138.4,127.5,124.3,124.0,61.6,17.5,14.3；HR-ESI-MS(positive mode)m/z:293.0586[M +H]⁺(Calcd for C₁₃H₁₃N₂O₄S:293.0596),m/z:315.0418[M+Na]⁺(Calcd for C₁₃H₁₂N₂O₄SNa: 315.0415).

2-hydroxyethyl 4-methyl-2-(4-nitrophenyl)thiazole-5-carboxylate(S13): The synthesized reference T10 of compound S13.Yield 79%, white solid, expansion system PE/EA 1:1 (Rf=0.5, PE/EA=1: 1),¹H NMR(400MHz,CDCl₃) 8.32 (d, J=8.9Hz, 2H), 8.14 (d, J=8.9Hz, 2H), 4.48 (t, J= 4.6Hz, 2H), 3.98 (t, J=4.7Hz, 2H), 2.81 (s, 3H), 1.85 (s, 1H)；¹³C NMR(150MHz,CDCl₃)δ 166.8,162.2,162.0,149.1,138.2,127.5,124.4,123.2,67.0,61.2,17.6；HR-ESI-MS (positive mode)m/z:309.0531[M+H]⁺(Calcd for C₁₃H₁₃N₂O₅S:309.0545),m/z:331.0352 [M+Na]⁺(Calcd for C₁₃H₁₂N₂O₅SNa:331.0365).

Ethyl 2- (4-isocyanophenyl) -4-methylthiazole-5-carboxylate (S14): compound The synthesized reference T7 of S14.Yield 88%, white solid are unfolded system PE/EA 20:1 (Rf=0.5, PE/EA=10:1),¹H NMR(400MHz,CDCl₃) 8.07 (d, J=8.4Hz, 2H), 7.74 (d, J=8.4Hz, 2H), 4.39 (q, J=7.1Hz, 2H), 2.78 (s, 3H), 1.40 (t, J=7.2Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ166.9,161.9,161.4,136.7, 132.8,127.2,123.6,118.2,114.1,61.6,17.5,14.3；HR-ESI-MS(positive mode)m/z: 273.0692[M+H]⁺(Calcd for C₁₄H₁₃N₂O₂S:273.0698),m/z:295.0505[M+Na]⁺(Calcd for C₁₄H₁₂N₂O₂SNa:295.0517).

2-hydroxyethyl 2-(4-((2-hydroxyethoxy)carbonyl)phenyl)-4- Methylthiazole-5-carboxylate (S15): 4- cyano substitutive derivative S14 70 DEG C of heating in 10%NaOH solvent 1h, TLC detection, adjust Ph to 1~2 using dense HCl after the reaction was completed, take chloromethanes by force and extract and be concentrated；Then dichloromethane is added Alkane 20mL, 1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride (EDCI) (383.4mg, 2mmol), 4- diformazan ammonia Yl pyridines (DMAP) (24.4mg, 0.2mmol), ethylene glycol (124.1mg, 2mmol), reaction system are heated to 80 DEG C, flow back 6 Hour.To which 20mL water quenching reaction is added after reaction, methylene chloride extraction merges organic phase and is concentrated, then chromatographs through column It isolates and purifies, to obtain compound S15.System PE/EA 1:1 (Rf=0.5, PE/EA is unfolded in yield 77%, white solid =1:1),¹H NMR(400MHz,CDCl₃) 8.14 (d, J=8.4Hz, 2H), 8.05 (d, J=8.4Hz, 2H), 4.51 (t, J= 4.6Hz, 2H), 4.47 (t, J=4.6Hz, 2H), 4.00 (t, J=4.6Hz, 2H), 3.97 (t, J=4.6Hz, 2H), 2.81 (s, 3H),1.86(s,2H)；¹³C NMR(150MHz,CDCl₃)δ168.6,166.1,162.2,162.0,136.8,131.9, 130.4,129.7,126.8,67.0,66.9,61.4,61.2,17.6；HR-ESI-MS(positive mode)m/z: 352.0847[M+H]⁺(Calcd for C₁₆H₁₈NO₆S:352.0855),m/z:374.0657[M+Na]⁺(Calcd for C₁₆H₁₇NO₆SNa:374.0674).

Ethyl 4-methyl-2- (naphthalen-2-yl) thiazole-5-carboxylate (S16): compound The synthesized reference T7 of S16.Yield 80%, white solid are unfolded system PE/EA 20:1 (Rf=0.5, PE/EA=10:1),¹H NMR(400MHz,CDCl₃) 8.44 (s, 1H) 8.01 (dd, J=1.8Hz, 1.8Hz, 1H), 7.90-7.81 (m, 3H), 7.53- 7.48 (m, 2H), 4.38 (q, J=7.2Hz, 2H), 2.81 (s, 3H), 1.41 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz, CDCl₃)δ169.9,162.3,161.1,134.5,133.1,130.2,128.9,128.8,127.9,127.5,126.9, 126.6,123.8,121.9,61.3,17.6,14.4；HR-ESI-MS(positive mode)m/z:298.0894[M+H]⁺ (Calcd for C₁₇H₁₆NO₂S:298.0902),m/z:320.0706[M+Na]⁺(Calcd for C₁₇H₁₅NO₂SNa: 320.0721).

2-hydroxyethyl 4-methyl-2-(naphthalen-2-yl)thiazole-5-carboxylate (S17): the synthesized reference T10 of compound S17.System PE/EA 1:1 (Rf=0.5, PE/ is unfolded in yield 83%, white solid ), EA=1:1¹H NMR(400MHz,CDCl₃) 8.41 (s, 1H), 7.96 (dd, J=1.6Hz, 1.6Hz, 1H), 7.89-7.80 (m, 3H), 7.54-7.48 (m, 2H), 4.43 (t, J=4.5Hz, 2H), 3.95 (t, J=4.7Hz, 2H), 2.79 (s, 3H), 2.55(s,1H)；¹³C NMR(100MHz,CDCl₃)δ170.4,162.4,161.8,134.6,133.1,130.0,129.0, 128.9,127.9,127.6,127.0,126.7,123.7,121.1,66.9,61.1,17.6；HR-ESI-MS(positive mode)m/z:314.0846[M+H]⁺(Calcd for C₁₇H₁₆NO₃S:314.0851),m/z:336.0664[M+Na]⁺ (Calcd for C₁₇H₁₅NO₃SNa:336.0670).

Ethyl 2- (3,5-dichlorophenyl) -4-methylthiazole-5-carboxylate (S18): chemical combination The synthesized reference T7 of object S18.Yield 90%, white solid are unfolded system PE/EA 20:1 (Rf=0.5, PE/EA=10:1),¹H NMR(400MHz,CDCl₃) 7.82 (d, J=1.8Hz, 2H), 7.42 (t, J=1.8Hz, 1H), 4.38 (q, J=7.1Hz, 2H), 2.76 (s, 3H), 1.40 (t, J=7.1Hz, 3H)；¹³C NMR(150MHz,CDCl₃)δ166.3,161.8,161.1, 135.8,135.5,130.5,125.0,123.1,61.5,17.4,14.3；HR-ESI-MS(positive mode)m/z: 315.9970[M+H]⁺(Calcd for C₁₃H₁₂Cl₂NO₂S:315.9966),m/z:337.9777[M+Na]⁺(Calcd for C₁₃H₁₁Cl₂NO₂SNa:337.9785).

2-hydroxyethyl 2-(3,5-dichlorophenyl)-4-methylthiazole-5-carboxylate (S19): the synthesized reference T10 of compound S19.System PE/EA 1:1 (Rf=0.5, PE/ is unfolded in yield 88%, white solid ), EA=1:1¹H NMR(400MHz,CDCl₃) 7.85 (d, J=1.8Hz, 2H), 7.45 (t, J=1.8Hz, 1H), 4.46 (t, J =4.6Hz, 2H), 3.96 (t, J=4.7Hz, 2H), 2.79 (s, 3H), 1.81 (s, 1H)；¹³C NMR(150MHz,CDCl₃)δ 166.7,162.1,161.9,135.9,135.4,130.7,125.1,122.3,66.9,61.2,17.5；HR-ESI-MS (positive mode)m/z:331.9906[M+H]⁺(Calcd for C₁₃H₁₂Cl₂NO₃S:331.9915),m/z: 353.9720[M+Na]⁺(Calcd for C₁₃H₁₁Cl₂NO₃SNa:353.9734).

Ethyl 2- (4-ethoxyphenyl) -4-methylthiazole-5-carboxylate (S20): compound The synthesized reference T7 of S20.Yield 89%, white solid are unfolded system PE/EA 20:1 (Rf=0.5, PE/EA=10:1),¹H NMR(400MHz,CDCl₃) 7.90 (d, J=8.8Hz, 2H), 6.94 (d, J=8.8Hz, 2H), 4.37 (q, J=7.1Hz, 2H), 4.11 (q, J=7.1Hz, 2H), 2.76 (s, 3H), 1.46 (t, J=7.0Hz, 3H), 1.40 (t, J=7.0Hz, 3H)；¹³C NMR (150MHz,CDCl₃)δ169.9,162.4,161.4,160.9,128.4,125.7,120.8,114.9,63.7,61.1, 17.5,14.7,14.3；HR-ESI-MS(positive mode)m/z:292.1003[M+H]⁺(Calcd for C₁₅H₁₈NO₃S: 292.1007),m/z:314.0816[M+Na]⁺(Calcd for C₁₅H₁₇NO₃SNa:314.0827).

2-hydroxyethyl 2-(4-ethoxyphenyl)-4-methylthiazole-5-carboxylate (S21): the synthesized reference T10 of compound S21.System PE/EA 1:1 (Rf=0.5, PE/ is unfolded in yield 90%, white solid ), EA=1:1¹H NMR(400MHz,CDCl₃) 7.90 (d, J=8.8Hz, 2H), 6.95 (d, J=8.8Hz, 2H), 4.44 (t, J =4.6Hz, 2H), 4.12 (q, J=7.0Hz, 2H), 3.95 (t, J=4.7Hz, 2H), 2.76 (s, 3H), 2.10 (s, 1H), 1.46 (t, J=7.0Hz, 3H)；¹³C NMR(150MHz,CDCl₃)δ170.4,162.6,161.7,161.5,128.5,125.5, 120.0,114.9,66.7,63.7,61.3,17.6,14.7；HR-ESI-MS(positive mode)m/z:308.0955[M+ H]⁺(Calcd for C₁₅H₁₈NO₄S:308.0957),m/z:330.0770[M+Na]⁺(Calcd for C₁₅H₁₇NO₄SNa: 330.0776).

Ethyl 4-methyl-2- (m-tolyl) thiazole-5-carboxylate (S22): the synthesis of compound S22 With reference to T7.Yield 86%, white solid are unfolded system PE/EA 20:1 (Rf=0.5, PE/EA=10:1),¹H NMR (400MHz,CDCl₃) 7.76 (s, 1H), 7.71 (d, J=7.6Hz, 1H), 7.29 (t, J=7.6Hz, 1H), 7.23 (d, J= 7.6Hz, 1H), 4.34 (q, J=7.1Hz, 2H), 2.76 (s, 3H), 2.37 (s, 3H), 1.38 (t, J=7.1Hz, 3H)；¹³C NMR(100MHz,CDCl₃)δ170.0,162.2,160.9,138.8,132.8,131.7,128.8,127.2,124.0, 121.6,61.1,21.3,17.5,14.3；HR-ESI-MS(positive mode)m/z:262.0900[M+H]⁺(Calcd for C₁₄H₁₆NO₂S:262.0902),m/z:284.0718[M+Na]⁺(Calcd for C₁₄H₁₅NO₂SNa:284.0721).

2-hydroxyethyl 4-methyl-2- (m-tolyl) thiazole-5-carboxylate (S23): compound The synthesized reference T10 of S23.Yield 84%, white solid are unfolded system PE/EA 1:1 (Rf=0.5, PE/EA=1:1),¹H NMR(400MHz,CDCl₃) 7.75 (s, 1H), 7.71 (d, J=7.4Hz, 1H), 7.34-7.28 (m, 2H), 4.42 (t, J= 4.4Hz, 2H), 3.94 (t, J=4.7Hz, 2H), 3.01 (s, 1H), 2.76 (s, 3H), 2.40 (s, 3H)；¹³C NMR(100MHz, CDCl₃)δ170.7,162.4,161.5,138.9,132.6,129.0,127.3,124.1,121.0,66.8,61.0,21.3, 17.5；HR-ESI-MS(positive mode)m/z:278.0845[M+H]⁺(Calcd for C₁₄H₁₆NO₃S:278.0851), m/z:300.0661[M+Na]⁺(Calcd for C₁₄H₁₅NO₃SNa:300.0670).

Ethyl 2- (3,4-dimethylphenyl) -4-methylthiazole-5-carboxylate (S24): chemical combination The synthesized reference T7 of object S24.Yield 88%, white solid are unfolded system PE/EA 20:1 (Rf=0.5, PE/EA=10:1),¹H NMR(400MHz,CDCl₃) 7.73 (s, 1H), 7.66 (dd, J=1.6Hz, 1.6Hz, 1H), 7.17 (d, J=7.8Hz, 1H), 4.36 (q, J=7.1Hz, 2H), 2.76 (s, 3H), 2.29 (d, J=7.5Hz, 6H), 1.39 (t, J=6.4Hz, 3H)；¹³C NMR (100MHz,CDCl₃)δ170.3,162.4,160.9,140.2,137.4,130.6,130.3,127.7,124.4,121.2, 61.4,19.9,19.7,17.6,14.4；HR-ESI-MS(positive mode)m/z:276.1046[M+H]⁺(Calcd for C₁₅H₁₈NO₂S:276.1058),m/z:298.0863[M+Na]⁺(Calcd for C₁₅H₁₇NO₂SNa:298.0878).

2-hydroxyethyl 2-(3,4-dimethylphenyl)-4-methylthiazole-5-carboxylate (S25): the synthesized reference T10 of compound S25.System PE/EA 1:1 (Rf=0.5, PE/ is unfolded in yield 82%, white solid ), EA=1:1¹H NMR(400MHz,CDCl₃) 7.54 (s, 1H), 7.48 (d, J=7.6Hz, 1H), 7.04 (d, J=7.8Hz, 1H), 4.30 (t, J=5.9Hz, 2H), 3.92 (s, 1H), 3.84 (t, J=4.5Hz, 2H), 2.63 (s, 3H), 2.18 (d, J= 6.9Hz,6H)；¹³C NMR(100MHz,CDCl₃)δ170.8,162.3,161.2,140.4,137.4,130.2,127.7, 124.3,120.6,66.7,60.6,19.8,19.7,17.4；HR-ESI-MS(positive mode)m/z:292.1001[M+ H]⁺(Calcd for C₁₅H₁₈NO₃S:292.1007),m/z:314.0817[M+Na]⁺(Calcd for C₁₅H₁₇NO₃SNa: 314.0827).

Ethyl 2- (3,5-dimethoxyphenyl) -4-methylthiazole-5-carboxylate (S26): change Close the synthesized reference T7 of object S26.Yield 84%, white solid, expansion system PE/EA 20:1 (Rf=0.5, PE/EA=10: 1),¹H NMR(400MHz,CDCl₃) 7.11 (d, J=2.2Hz, 2H), 6.56 (t, J=2.2Hz, 2H), 4.37 (q, J= 7.1Hz, 2H), 3.85 (s, 6H), 2.77 (s, 3H), 1.40 (t, J=7.1Hz, 3H)；¹³C NMR(150MHz,CDCl₃)δ 169.7,162.2,161.2,160.8,134.7,121.9,104.7,103.5,61.2,55.6,17.5,14.3；HR-ESI-MS (positive mode)m/z:308.0951[M+H]⁺(Calcd for C₁₅H₁₈NO₄S:308.0957),m/z:330.0769[M +Na]⁺(Calcd for C₁₅H₁₇NO₄SNa:330.0776).

2-hydroxyethyl 2-(3,5-dimethoxyphenyl)-4-methylthiazole-5-carboxylate (S27): the synthesized reference T10 of compound S27.System PE/EA 1:1 (Rf=0.5, PE/ is unfolded in yield 78%, white solid ), EA=1:1¹H NMR(400MHz,CDCl₃) 7.08 (d, J=2.2Hz, 2H), 6.55 (t, J=2.2Hz, 1H), 4.42 (t, J =4.6Hz, 2H), 3.94 (t, J=4.6Hz, 2H), 3.84 (s, 6H), 2.76 (s, 3H), 2.36 (s, 1H)；¹³C NMR (150MHz,CDCl₃)δ170.2,162.4,161.5,161.2,134.4,121.2,104.7,103.7,66.8,61.1, 55.6,17.5；HR-ESI-MS(positive mode)m/z:324.0894[M+H]⁺(Calcd for C₁₅H₁₈NO₅S: 324.0906),m/z:346.0707[M+Na]⁺(Calcd for C₁₅H₁₇NO₅SNa:346.0725).

Ethyl 4-methyl-2-(3,4,5-trimethoxyphenyl)thiazole-5-carboxylate(S28): The synthesized reference T7 of compound S28.Yield 79%, white solid, expansion system PE/EA 20:1 (Rf=0.5, PE/EA=10: 1),¹H NMR(400MHz,CDCl₃) 7.17 (s, 2H), 4.35 (q, J=7.1Hz, 2H), 3.92 (s, 6H), 3.88 (s, 3H), 2.74 (s, 3H), 1.38 (t, J=7.1Hz, 3H)；¹³C NMR(150MHz,CDCl₃)δ169.6,162.2,160.9,153.6, 140.7,128.4,121.5,104.0,61.2,60.9,56.3,17.5,14.3；HR-ESI-MS(positive mode)m/z: 338.1048[M+H]⁺(Calcd for C₁₆H₂₀NO₅S:338.1062),m/z:360.0868[M+Na]⁺(Calcd for C₁₆H₁₉NO₅SNa:360.0882).

2-hydroxyethyl 4-methyl-2-(3,4,5-trimethoxyphenyl)thiazole-5- Carboxylate (S29): compound S29 synthesized reference T10.System PE/EA 1:1 is unfolded in yield 83%, white solid (Rf=0.5, PE/EA=1:1),¹H NMR(400MHz,CDCl₃) 7.18 (s, 2H), 4.44 (t, J=4.6Hz, 2H), 3.95- 3.93(m,8H),3.90(s,3H),2.77(s,3H),2.12(s,1H)；¹³C NMR(150MHz,CDCl₃)δ170.1,162.4, 161.6,153.6,140.9,128.2,120.8,104.1,66.8,61.2,61.0,56.3,17.6；HR-ESI-MS (positive mode)m/z:354.1003[M+H]⁺(Calcd for C₁₆H₂₀NO₆S:354.1011),m/z:376.0821[M +Na]⁺(Calcd for C₁₆H₁₉NO₆SNa:376.0831).

2-((5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl) Pentanoyl) oxy) ethyl2- (4-methoxyphenyl) -5-methyloxazole-4-carboxylate (T58): D-Biotin (D-Biotin) (244.3mg, 1mmol) is added in the round-bottomed flask of 25mL, compound 3b (261.3mg, 1mmol), dicyclohexylcarbodiimide (DCC) (412.6mg, 2mmol), 4-dimethylaminopyridine (DMAP) (24.4mg, It 0.2mmol) with DMF (20mL), stirs at normal temperature for 24 hours, removes solvent after completion of the reaction, through column chromatographic isolation and purification, obtain Compound T56.Yield 49%, white solid are unfolded system DCM/EtOH 30:1 (Rf=0.5, DCM/EtOH=20:1),¹H NMR(600MHz,DMSO-d₆) 7.90 (d, J=8.8Hz, 2H), 7.08 (d, J=8.8Hz, 2H), 5.55 (d, J=7.9Hz, 2H), 4.89 (t, J=5.6Hz, 1H), 4.26 (t, J=4.9Hz, 2H), 3.82 (s, 3H), 3.69 (q, J=4.7Hz, 2H), 2.64(s,3H),1.71-1.69(m,4H),1.61-1.58(m,4H),1.50-1.47(m,2H)；¹³C NMR(150MHz, DMSO-d₆)δ162.2,161.8,159.2,157.1,156.4,128.4,128.3,119.2,115.1,66.5,59.5, 55.9,48.0,33.8,25.8,24.9,12.4；HR-ESI-MS(positive mode)m/z:504.1804[M+H]⁺ (Calcd for C₂₄H₃₀N₃O₇S:504.1810),m/z:526.1622[M+Na]⁺(Calcd for C₂₄H₂₉N₃O₇SNa: 526.1624).

The synthesis of reference examples 1, control compound 3b

Ethyl 2- (4-methoxyphenyl) -5-methyloxazole-4-carboxylate (3b): 25mL's 4- methoxybenzylamine (274.4mg, 2mmol) and DMF (10mL) are added in round-bottomed flask, then sequentially adds I₂ (304.6mg, 1.2mmol), ethyl acetoacetate (130.1mg, 1mmol), TBHP (180.2mg, 2mmol), copper acetate (39.9mg,0.2mmol)；Reaction system stirs 4 hours at normal temperature.10mL water quenching reaction is added after stopping in reaction, is added Saturated sodium thiosulfate (20mL) removes unreacted iodine, is extracted with ethyl acetate, and organic phase is cleaned with saturated salt solution, merges Organic phase, anhydrous sodium sulfate are dried and concentrated, and crude product is again through silica gel column chromatography separating purification (petrol ether/ethyl acetate=10/1) Obtain target compound 3b.Yield 87%, white solid are unfolded system PE/EA 10:1 (Rf=0.6, PE/EA=3:1),¹H NMR(400MHz,CDCl₃) 8.00 (d, J=9.0Hz, 2H), 6.96 (d, J=9.0Hz, 2H), 4.43 (q, J=7.2Hz, 2H), 3.85 (s, 3H), 2.67 (s, 3H), 1.42 (t, J=7.1Hz, 2H)；¹³C NMR(100MHz,CDCl₃)δ162.6,161.6, 159.8,155.6,128.6,128.3,119.4,114.1,61.0,55.4,14.4,12.2；HR-ESI-MS(positive mode)m/z:262.1068[M+H]⁺(Calcd for C₁₄H₁₆NO₄:262.1079),m/z:284.0890[M+Na]⁺(Calcd for C₁₄H₁₅NO₄Na:284.0899).

The beneficial effect of compound prepared by the present invention is proved below by way of experimental example.

Experimental example 1, compound analyze the inhibitory activity of miRNA-21

1, experimental method:

(1) experimental material and instrument

A. main agents: DMEM/ high glucose medium, PBS buffer solution, EDTA-0.25% pancreatin, penicillin streptomycin are (double It is anti-), BIOMYC-3Antibiotic Solution etc. be purchased from Hyclone company；Fetal calf serum is purchased from Gibco company； Coenzyme A sodium salt hydrate, D-Luciferin sodium salt are purchased from Sigma company；DMSO,G418 It is purchased from Amresco company.

B. key instrument: CO₂Constant temperature cell incubator, superclean bench, the multi-functional readout instrument of full wavelength scanner formula, cell Station, optics inverted microscope, pressure steam sterilizer, micro centrifuge, electronic thermostatic water-bath, horizontal shaker etc..

(2) foundation of cell model

The cell screening model of miRNA-21 micromolecular inhibitor is constructed by luciferase reporter gene: compound passes through The expression for promoting (or inhibit) intracellular miRNA-21, make the miRNA-21 complementary series of transfection occur complementary pairing increase (or Reduce), to inhibit the expression of (or enhancing) luciferase, as shown in Figure 6.

(3) cell culture

Quick-thawing in water-bath is put into from the HeLa-luciferase-miRNA-21 cell frozen is taken out in liquid nitrogen container, Cell suspension sucking prepared culture medium (+1% penicillin of+10% fetal calf serum of DMEM high sugar cell culture medium has been shifted to an earlier date into Streptomysin+1%BIOMYC-3Antibiotic Solution+G418 (300 μ g/mL)) culture dish in, then place 5% CO₂37 DEG C of cell incubators in.

When cell density reaches 80-90%, cell culture medium is outwelled, is cleaned cell 2 times with PBS buffer solution, outwells PBS 0.25% pancreatin that 1mL contains EDTA is added afterwards, is put into 37 DEG C of digestion 3-5min, to be seen arrive in cell incubator and digests completely After Fresh cell culture medium is added, with piping and druming pipe softly blow and beat repeatedly, cell is blown and beaten into cell suspension from wall, is taken 100ul 1×10⁴A cell is uniformly inoculated into 96 orifice plates, is placed in 5%CO₂, overnight incubation in 37 DEG C of cell incubators.Wherein 96 orifice plate periphery holes are not used in screening compound, to exclude the influence of edge effect.

(4) detection of luciferase reporter gene

After carrying out corresponding agent-feeding treatment for 24 hours to 96 orifice plate cells, sucks supernatant culture medium and cleaned with PBS buffer solution, so After be added 50 μ L cell lysis buffer lysates and on shaking table concussion cracking 15-20 minutes after, take 50 μ L lysates (50 hole μ L/) is added in sample to be tested into 96 orifice plate of black to be measured, then by the luciferase substrate to thaw in advance, by more Function readout instrument detects the fluorescent value in every hole.

(5) screening active ingredients

Recovery and secondary culture are carried out to Hela-miRNA-21 stable transfection type cell strain；By logarithmic phase cell inoculation in 96 orifice plates (1 × 10⁴) in, it is adherent to cell, then (primary dcreening operation of all compounds is dense for addition reactive compound in superclean bench Degree is 10 μM).Three multiple holes of each compound, and 96 orifice plates are placed cell incubator 24 hours, it is thin then to measure each hole The fluorescence signal intensity of cellular lysate liquid: RFS (Relative Fluorescence Signal), relative intensity of fluorescence are greater than early period The miRNA-21 inhibitor 3b compound having verified that then carries out next step experiment.(RFS=RFS sample/RFS control)

(6) cytotoxicity assay

Compound is carried out to Hela-luciferase- using AM-Blue cell Proliferation and activity detection kit The influence of miRNA-21 cell line vigor.

The testing principle of the kit: the main component of SunBio Am-Blue is the indigo-blue blue indicator of oxidized form, The blue indicator can generate stable pink fluorescent material after being reduced.The excitation wavelength of the fluorescent material is in 530- Between 560nm, wavelength of transmitted light 590nM.Cell in proliferation its into the cell with extracellular compared in reducing condition, The water solubility of SunBio Am-Blue reagent very well, is easier to enter cell interior, powder is finally reduced in mitochondria Red fluorescent material is then released into extracellular and is dissolved in culture medium, make culture medium become to have from non-blooming indigo-blue indigo plant it is glimmering The pink of light.By with common spectrophotometer or fluophotometer carry out detection absorbance or fluorescence intensity variation to The case where analyzing cell Proliferation.It is thin that Hela-luciferase-miRNA-21 stable transfected cells are inoculated with 100 L1 × 104 μ Born of the same parents are in 96 orifice plates, and 8 concentration gradients are arranged: 0,0.01 μM, 0.1 μM, 1 μM, 5 μM, 10 μM, 25 μM, 50 μM, compound handles 24 After hour, its fluorescent value is measured, the IC of institute's screening compounds is then calculated by GraphPad Prism5 software₅₀Value.

2, experimental result:

Relative intensity of fluorescence of the compound prepared by the present invention in screening active ingredients experiment is as shown in Figure 1, 2, with compound 3b is control.

Find out from Fig. 1,2, compound T24, T30 prepared by the present invention, T32, T35, T40, T2, T10, S11, S17, S20, S21, S25 have apparent inhibitory activity to miRNA-21 biosynthesis, and inhibitory activity is higher than 3b.

It is living that we have further carried out Hela-luciferase-miRNA-21 cell toxicant to the compound of above-mentioned high activity Property detection test, found out according to Fig. 3, the EC of S21₅₀Having reached is 0.093 μM, and activity is compared to 3b (EC₅₀=1.38 μM) it mentions It is 15.3 times high, similarly, the EC of compound T24, T30, T32, T35, T40, T2, T10, S11, S17, S20, S25₅₀Also below Compound 3b has higher activity compared to compound 3b.

Affinity analysis 1 that experimental example 2, compound are docked with TRBP Binding proteins in miRNA biosynthetic process, Experimental method:

In research in our prior, we by 7 kinds it has been reported that miRNA biosynthetic process in TRBP correlation combine Albumen (deriving from PDB database, 4 kinds of sources are mammal and 2 kinds of sources are that drosophila gene and a kind derive from plant) point Do not carried out molecular docking with compound 3b, as a result, it has been found that only from mammal two kinds of albumen crystal models (4WYQ: Dicer-TRBP and 3LLH:TRBP (dsRBD2)) active pocket can be formed with 3b.

TRBP is as Dicer companion in zooblast, TRBP can by influence miRNA that Dicer and Ago2 is mediated at It is ripe, and then the pernicious transfer of cancer cell is influenced, TRBP is proved during RNA interference (RNAi) and can not Or lack；TRBP interacts with Dicer or Ago2 and dsRNA is combined to combine to form induction RISC complex (RNA Induced silencing complex) gene silencing function is played, processable Pre-miRNA forms mature miRNA.

In order to further study the compounds of this invention to the inhibiting effect of miRNA, we are by both albumin crystal models The high activity micromolecular inhibitor that obtains is screened with above by activity experiment respectively, carries out molecule affinity pair on computers It connects.

(1) target protein and device information

The receptor protein of reading: 4WYQ (interface Dicer-TRBP), 3LLH (the dsRBD2 structure of TRBP), 5N8M (with The composite structure of TRBP dsRBD 1 and 2 (complex A) that 19bp siRNA is combined), 5N8L is (in conjunction with 19bp siRNA The composite structure of TRBP dsRBD 1 and 2 (complex B)), 3ADL (TRBP2 structure (plant source)), 5NPA (drosophila dsRBD2 knot Structure), 5NPG (drosophila dsRBD1 structure)；

Receptor protein derives from PDB database:https://www.rcsb.org/。

Software platform: Discovery Studio 4.5.

(2) experimental implementation

1) protein receptor downloaded from PDB database is imported into molecule window, assigns CHARMM to protein receptor Position adds hydrogen, removes hydrone, corrects etc. unreasonable conformation.

2) small molecule is imported in another window, small molecule is named, sequence assigns ChARMM position and energy most Smallization operation.

3) possible active site is selected according to cavity in protein receptor, at 10 groups, taking first group of site is pair for symbiosis Connect active pocket.

4) it is carried out between receptor and small molecule with the CDOCKER module under receptor-ligand interaction module Docking semi-flexible, individual molecule conformation collection generate up to 10, molecular dynamics maximum step number be 1000.

5) after the completion of docking, score evaluation is carried out to all docking conformations, selecting highest scoring is Optimum configuration, is carried out into one The observation of step.

2, experimental result

The albumin crystal molecular docking result of reactive compound and Dicer-TRBP prepared by the present invention is as shown in Figure 4.This The albumin crystal molecular docking result for inventing the reactive compound and TRBP (dsRBD2) of preparation is as shown in Figure 5.

Compound prepared by the present invention and two kinds of albumen to dock energy balane structure as shown in table 1.From small molecule and egg The conjugation of amino acid residue on white, can be in conjunction with inhibitor on Dicer-TRBP albumen in conjunction with conformation from the point of view of energy Amino acid residue quantity is more, and the docking energy ratio TRBP (dsRBD2) of best conformation is almost low 1 times (table 1), therefore In Dicer-TRBP interface protein model, affinity is much higher than TRBP (dsRBD2).Pass through Computer-aided Molecular pair Energy balane is connect it is found that active higher compound, best conformation energy value is lower, illustrates the combination of compound and target protein It spends closer.

As seen from Table 1, the best conformation energy value that compound T10, S21, S25 is docked with 4WYQ albumen is lower than compound 3b and 4WYQ albumen；The best conformation energy value that compound T10, T32, S21, S25 are docked with 3LLH albumen is lower than compound 3b and 3LLH albumen.That is, above compound and the conjugation of corresponding target protein ratio 3b are closer.

The reactive compound prepared by the present invention of table 1 docks energy with albumen^a

^aBest conformation energy value；^bTotal energy value evaluation function is negative, and value is bigger, and energy is smaller.

Small molecule is able to suppress the biosynthesis of miRNA-21, and then reach by docking with TRBP Binding proteins Inhibit the generation of malignant tumour and blocks the effect of malignant tumour migration.And compound prepared by the present invention, compared to controlization Object 3b is closed, there is closer affine combination ability with TRBP Binding proteins, is had more to the biosynthesis of miRNA-21 There is strong inhibiting effect, therefore, the compound of the present invention has superior therapeutic effect to malignant tumour.

It, can be in conclusion the present invention provides a kind of I compound represented of formula or its pharmaceutically acceptable salt Binding proteins in miRNA biosynthetic process closely combine, and can effectively inhibit the synthesis of miRNA-21.This The reactive compound of invention preparation can be used as miRNA-21 inhibitor, be further used as the potential drug for the treatment of malignant tumour.

Claims

1. Formulas I compound represented or its conformer or its optical isomer or its pharmaceutically acceptable salt:

Wherein: X is selected from O, S；Y is selected from N；

R₁、R₂、R₃、R₄、R₅It is each independently selected from hydrogen, substituted or unsubstituted C1-2 alkyl, substituted or unsubstituted C1-2 alkane Oxygroup, halogen, cyano, nitro, substituted or unsubstituted phenyl ,-COOR₉；

Or, R₁、R₂、R₅It is each independently selected from hydrogen, substituted or unsubstituted C1-2 alkyl, substituted or unsubstituted C1-2 alcoxyl Base, halogen, cyano, nitro, substituted or unsubstituted phenyl ,-COOR₉, R₃、R₄Two carbon atoms connected to it are formed together Phenyl ring；Above-mentioned substituent group is selected from halogen, C1-4 alkyl, hydroxyl, carboxyl, nitro, amino, carboxyl, C₂-C₄Alkenyl, C₁-C₄Alkyl, C₂-C₄Alkynyl, carbocylic radical, heterocycle；

R₆、R₈Be each independently selected from hydrogen, hydroxyl, halogen, C1-4 alkyl, C3-6 cycloalkyl group, phenyl,- COOR₉、-CONHR₁₀, wherein R₉、R₁0 be each independently selected from C1-2 alkyl, Wherein L₀、L₁、L₂、L₃It is each independently selected from 1-4 methylene, R₁₁Selected from halogen or hydroxyl；

And when X is O, Y N, R₁、R₂、R₄、R₅For hydrogen, R₃For methoxyl group, R₆For methyl, R₈For-COOR₉When, R₉It is not ethyl.

2. compound according to claim 1 or its conformer or its optical isomer or its can pharmaceutically connect The salt received, it is characterised in that:

X is selected from O, S；Y is selected from N；

R₁、R₂、R₃、R₄、R₅It is each independently selected from hydrogen, substituted or unsubstituted C1-2 alkyl, substituted or unsubstituted C1-2 alkane Oxygroup, halogen, substituted or unsubstituted phenyl；

Or, R₁、R₂、R₅It is each independently selected from hydrogen, substituted or unsubstituted C1-2 alkyl, substituted or unsubstituted C1-2 alcoxyl Base, halogen, substituted or unsubstituted phenyl, R₃、R₄Two carbon atoms connected to it are formed together phenyl ring；Above-mentioned substituent group choosing From halogen；

R₆、R₈It is each independently selected from C1-2 alkyl ,-COOR₉, wherein R₉Selected from C1-2 alkyl,Wherein L₀Choosing From 1-4 methylene.

3. compound according to claim 2 or its conformer or its optical isomer or its can pharmaceutically connect The salt received, it is characterised in that:

X is selected from O；Y is selected from N；

R₁、R₂、R₃、R₄、R₅It is each independently selected from hydrogen, substituted or unsubstituted C1-2 alkyl, substituted or unsubstituted C1-2 alkane Oxygroup, halogen, phenyl substituted or unsubstituted；The substituent group is selected from halogen；

R₆、R₈It is each independently selected from C1-2 alkyl ,-COOR₉, wherein R₉Selected from C1-2 alkyl,Wherein L₀Choosing From 2 methylene.

4. compound according to claim 3 or its conformer or its optical isomer or its can pharmaceutically connect The salt received, it is characterised in that: the structure of the compound is as shown in Formulas I -1:

Wherein:

R₆Selected from C1-2 alkyl；

R₇Selected from C1-2 alkyl,Wherein L₀Selected from 2 methylene；

5. compound according to claim 2 or its conformer or its optical isomer or its can pharmaceutically connect The salt received, it is characterised in that:

X is selected from S；Y is selected from N；

R₁、R₂、R₃、R₄、R₅It is each independently selected from hydrogen, substituted or unsubstituted C1-2 alkyl, substituted or unsubstituted C1-2 alkane Oxygroup, halogen, substituted or unsubstituted phenyl；Or, R₁、R₂、R₅It is each independently selected from hydrogen, substituted or unsubstituted C1-2 alkane Base, substituted or unsubstituted C1-2 alkoxy, halogen, substituted or unsubstituted phenyl, R₃、R₄Two carbon atoms connected to it It is formed together phenyl ring；The substituent group is selected from halogen；

6. compound according to claim 5 or its conformer or its optical isomer or its can pharmaceutically connect The salt received, it is characterised in that:

X is selected from S；Y is selected from N；

R₁、R₂、R₄、R₅For hydrogen, R₃Selected from substituted or unsubstituted C1-2 alkyl, substituted or unsubstituted C1-2 alkoxy, halogen, Substituted or unsubstituted phenyl, the substituent group are selected from halogen；

Or, R₁、R₂、R₅For hydrogen, R₃、R₄Two carbon atoms connected to it are formed together phenyl ring；

R₆、R₈It is each independently selected from C1-2 alkyl ,-COOR₉, wherein R₉Selected from C1-2 alkyl,L₀For 2 Asias Methyl.

7. compound according to claim 6 or its conformer or its optical isomer or its can pharmaceutically connect The salt received, it is characterised in that: its structure such as Formulas I -2:

Wherein:

R₆、R₈In, one is C1-2 alkyl, another is-COOR₉, wherein R9 be selected from C1-2 alkyl,L₀It is 2 A methylene；

R₃Selected from halogenated or not halogenated C1-2 alkyl, C1-2 alkoxy, halogen, phenyl, preferably trifluoromethyl, methoxyl group, ethoxy Base, methyl；R₄Selected from hydrogen, methyl；

Or, R₃、R₄Two carbon atoms connected to it are formed together phenyl ring.

8.-the 7 any compound or its conformer or its optical isomer or its pharmacy according to claim 1 Upper acceptable salt, it is characterised in that: the compound are as follows:

9. claim the 1-8 any compound or its conformer or its optical isomer or its pharmaceutically may be used Purposes of the salt of receiving as miRNA-21 biosynthesis inhibitor.

10. purposes according to claim 9, it is characterised in that: the inhibitor is the drug for treating tumour, is preferably controlled Treat the drug of malignant tumour.