Compound, preparation method and application
阅读说明：本技术 一种化合物以及制备方法和应用 (Compound, preparation method and application ) 是由 王振亚 徐海伟 牟晓东 胡晓宁 李媛媛 刘宏民 于 2018-09-03 设计创作，主要内容包括：本申请公开了一种化合物,其结构式如下式所示：<Image he="359" wi="414" file="DDA0001787467790000011.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>其中,R<Sup>1</Sup>选自F、Cl、Br、C<Sub>1</Sub>～C<Sub>5</Sub>的烷基、具有式(1)所示结构式的基团中的任意一种；n＝0、1、2、3、4或5；R<Sup>2</Sup>选自具有式(1)所示结构式的基团、具有式(2)所示结构式的基团、具有式(3)所示结构式的基团中的任意一种。所述化合物制备方法简单,可作为神经氨酸酶抑制剂,具有良好的抗病毒活性。(The application discloses a compound, the structural formula of which is shown as the following formula: wherein R is 1 Selected from F, Cl, Br, C 1 ～C 5 Any one of an alkyl group of (1) and a group having a structural formula shown in formula (1); n is 0, 1,2.3, 4 or 5; r 2 Is selected from any one of a group having a structural formula shown in a formula (1), a group having a structural formula shown in a formula (2) and a group having a structural formula shown in a formula (3). The compound has simple preparation method, can be used as a neuraminidase inhibitor and has good antiviral activity.)
1. A compound having a structural formula according to formula I:
wherein R is1Selected from F, Cl, Br, C1～C5Any one of an alkyl group of (1) and a group having a structural formula shown in formula (1); n is 0, 1, 2, 3,4 or 5;
R2any one selected from the group having a structural formula shown in formula (1), the group having a structural formula shown in formula (2), and the group having a structural formula shown in formula (3);
*-O-M11formula (1)
Wherein M is11Is selected from H or C1～C5Alkyl groups of (a);
wherein M is21、M22、M23、M24、M25、M26、M27、M28Independently selected from H or C1～C5Alkyl groups of (a);
wherein M is31Selected from H, F, Cl, Br, C1～C5Alkyl group of (2), having a structural formula represented by the formula (1)Any one of the groups; m32、M33、M34Independently selected from H or C1～C5Alkyl group of (1).
2. The compound of claim 1, wherein in formula In＝0；
Preferably, R in formula I1One selected from F, Cl, Br, methyl, ethyl, methoxy and ethoxy; n is 1 or 2;
preferably, R in formula I1Selected from 4-F, 4-Cl, 4-Br, 4-CH3、4-OCH33-F, 2-F, 3, 4-2F;
preferably, M in formula (1)11Selected from methyl or ethyl;
preferably, M in formula (2)21、M22、M23、M24、M25、M26、M27、M28Independently selected from H, methyl or ethyl;
preferably, M in the formula (3)31Selected from H, F, Cl, Br, methyl, ethyl, methoxy or ethoxy;
M32、M33、M34independently selected from hydrogen, methyl or ethyl;
preferably, R in formula I2Is selected from: -OCH3、
3. The compound of claim 1, having a structural formula according to formula I-5:
preferably, R1Selected from 4-F, 4-Cl, 4-Br, 4-CH3、4-OCH33-F, 2-F or 3,4-2F, and n is 1; or n ═ 0.
4. The compound of claim 1, having a structural formula according to formula I-1:
preferably, said R is1Selected from F, Cl, — CH3、*-OCH3And n ═ 1; or n is 0;
preferably, said R is1Selected from 4-F, 4-Cl, 4-OCH3、4-CH3Or 3-F.
5. The compound of claim 1, having a structural formula according to formula I-2:
preferably, the structural formula is shown in formula I-3:
preferably, said M31Selected from H, 4-F, 4-Cl, 4-Br, 4-OCH3、4-CH33-F or 2-F;
preferably, the structural formula is shown in formula I-4:
preferably, said R is1Selected from Cl,. about. -OCH3At least one of;
preferably, said R is1Is selected from 4-Cl or 4-OCH3。
6. A process for the preparation of a compound according to any one of claims 1 to 5, comprising:
(I) carrying out esterification reaction on a mixture I containing substituted phenylacetic acid and halogenated ethyl acetate in the presence of a catalyst to obtain an intermediate I, wherein the structural formula of the intermediate I is shown as a formula II-1:
(II) carrying out Dieckmann reaction on a raw material I containing an intermediate I to obtain an intermediate II, wherein the structural formula of the intermediate II is shown as a formula II-2:
preferably, the mixture I in the step (I) also comprises a solvent I and an acid-binding agent;
the molar ratio of the substituted phenylacetic acid to the halogenated ethyl acetate to the acid-binding agent is 1-2: 1-2: 1-2;
the esterification reaction conditions are as follows: carrying out reflux reaction at the temperature of 60-80 ℃;
preferably, the solvent I comprises one of tetrahydrofuran, dichloromethane, DMF, ethyl acetate, acetone, methanol and ethanol;
the acid-binding agent comprises at least one of triethylamine, pyridine, anhydrous potassium carbonate, sodium acetate and anhydrous sodium carbonate;
the molar ratio of the substituted phenylacetic acid to the halogenated ethyl acetate to the acid-binding agent is 1:1.2: 1.2;
the esterification reaction conditions are as follows: heating and refluxing at 75 ℃ under the condition of stirring;
preferably, the raw material I in the step (II) also comprises a solvent II and a base I;
the molar ratio of the intermediate I to the base I is 1: 1.5 to 3;
the Dieckmann reaction conditions are as follows: stirring in an ice bath for 20-50 min, and then continuing to react at room temperature;
preferably, the solvent II comprises DMF, THF, t-BuOH, acetone, EtOAc, CH2Cl2At least one of DMSO and methanol;
the base I comprises potassium tert-butoxide and anhydrous Na2CO3、CH3ONa、C2H5ONa、NaOH、NaHCO3NaH, anhydrous K2CO3At least one of;
the molar ratio of the intermediate I to the base I is 1: 2;
the Dieckmann reaction conditions are as follows: stirring in ice bath for 30min, and then continuing to react for 4-6 hours at room temperature.
7. The method of claim 6, wherein the method of preparing the compound further comprises:
(III-1) carrying out methylation reaction on a mixture II containing an intermediate II and dimethyl sulfate to obtain an intermediate III-1, wherein the structural formula of the intermediate III-1 is shown as a formula II-3-1:
(IV-1) reacting a mixture III containing the intermediate III-1, sodium methoxide and paraformaldehyde to obtain a compound shown as a formula I-5;
preferably, the mixture II in the step (III-1) also comprises a solvent III and a base II;
the molar ratio of the intermediate II to the alkali II to the dimethyl sulfate is 1: 1-2: 1-2;
the methylation reaction conditions are as follows: reacting at room temperature under the condition of keeping out of the light;
preferably, the solvent III comprises acetone, methanol, CH2Cl2At least one of THF, DMF, EtOAc;
the base II comprises anhydrous K2CO3Anhydrous Na2CO3、CH3ONa、NaOH、NaHCO3At least one of NaH;
the molar ratio of the intermediate II to the alkali II to the dimethyl sulfate is 1:1.2: 1.2;
the methylation reaction conditions are as follows: reacting for 8-10 hours at room temperature under the condition of keeping out of the sun;
preferably, the mixture III in the step (IV-1) also comprises a solvent IV;
the molar ratio of the intermediate III-1 to the sodium methoxide to the paraformaldehyde is 1: 0.1-1: 0.1 to 0.5;
the reaction conditions are as follows: carrying out reflux reaction at 60-100 ℃;
preferably, the solvent IV comprises methanol, acetone, CH2Cl2At least one of THF, DMF, EtOAc;
the reaction conditions are as follows: feeding the intermediate III-1, sodium methoxide and paraformaldehyde according to the feeding molar ratio of 1:0.6:0.3, refluxing and stirring at 75 ℃, adding paraformaldehyde and sodium methoxide at intervals, and reacting for 70-75 hours;
preferably, during the reaction: 0.3 mol of paraformaldehyde and sodium methoxide are added at intervals of 8 hours, and the reaction is carried out for 72 hours.
8. The method of claim 6, wherein the method of preparing the compound further comprises:
(III-2) carrying out chloro nucleophilic reaction on the raw material II containing the intermediate II to obtain an intermediate III-2; the structural formula of the intermediate III-2 is shown as a formula II-3-2:
(IV-2) carrying out nucleophilic substitution on the mixture IV containing the intermediate III-2 and morpholine under alkaline conditions to obtain an intermediate IV-1; the structural formula of the intermediate IV-1 is shown as a formula II-4-1:
(V-2) reacting a mixture V containing the intermediate IV-1, sodium methoxide and paraformaldehyde to obtain a compound shown in the formula I-1;
preferably, the raw material II in the step (III-2) further comprises: solvent V and nucleophile I;
the molar ratio of the intermediate III-2 to the nucleophilic reagent I is 1: 1-4;
the conditions of the chloro nucleophilic reaction are as follows: stirring in an ice bath for 20-50 min, and then reacting at room temperature;
preferably, the solvent V is selected from the group consisting of solvents having a volume ratio of 1:1 of DMF and CH2Cl2At least one of THF, DMF, EtOAc, methanol and acetone;
the nucleophilic reagent I is selected from oxalyl chloride and POCl3At least one of thionyl chloride;
the molar ratio of the intermediate III-2 to the nucleophile I is 1: 2;
the conditions of the chloro nucleophilic reaction are as follows: stirring in ice bath for 30min, and then reacting at room temperature for 6 hours;
preferably, the mixture IV in the step (IV-2) also comprises a solvent VI and a base III;
the molar ratio of the intermediate III-2 to the base III to the morpholine is 1: 1-4: 1-4;
the reaction time of the nucleophilic substitution is 3-6 hours;
preferably, the solvent VI comprises THF, methanol, acetone, CH2Cl2At least one of DMF and EtOAc;
the base III comprises: anhydrous K2CO3Anhydrous Na2CO3、CH3ONa、NaOH、NaHCO3At least one of NaH;
the molar ratio of the intermediate III-2 to the base III to the morpholine is 1:2: 2;
the reaction time of the nucleophilic substitution is 4 h;
preferably, the mixture V in step (V-2) further comprises a solvent VII;
the molar ratio of the intermediate IV-1 to the sodium methoxide to the paraformaldehyde is 1: 0.1-1: 0.1 to 0.5;
the reaction conditions are as follows: carrying out reflux reaction at 60-100 ℃;
preferably, the solvent VIIComprises methanol, acetone and CH2Cl2At least one of THF, DMF, EtOAc;
the reaction conditions are as follows: feeding the intermediate IV-1, sodium methoxide and paraformaldehyde according to the feeding molar ratio of 1:0.6:0.3, refluxing and stirring at 75 ℃, adding paraformaldehyde and sodium methoxide at intervals, and reacting for 70-75 hours;
preferably, during the reaction: 0.3 mol of paraformaldehyde and sodium methoxide are added at intervals of 8 hours, and the reaction is carried out for 72 hours.
9. The method of claim 6, wherein the method of preparing the compound further comprises:
(III-2) carrying out chloro nucleophilic reaction on the raw material II containing the intermediate II to obtain an intermediate III-2; the structural formula of the intermediate III-2 is shown as a formula II-3-2:
(IV-3) carrying out nucleophilic substitution on the mixture VI containing the intermediate III-2 and benzylamine or substituted benzylamine under alkaline conditions to obtain an intermediate IV-2; the structural formula of the intermediate IV-2 is shown as a formula II-4-2:
(V-3) reacting a mixture VII containing the intermediate IV-2, sodium methoxide and paraformaldehyde to obtain a compound shown as a formula I-2;
preferably, the mixture VI in the step (IV-3) further comprises a solvent VIII and a base IV;
the molar ratio of the intermediate III-2 to the base IV to the benzylamine or the substituted benzylamine is 1: 1-4: 1-4;
the reaction time of the nucleophilic substitution is 3-6 hours;
preferably, the solvent VIII comprises THF, methanol, acetone, CH2Cl2At least one of DMF and EtOAc;
the above-mentionedThe base IV comprises: anhydrous K2CO3Anhydrous Na2CO3、CH3ONa、NaOH、NaHCO3At least one of NaH;
the molar ratio of the intermediate III-2 to the base IV to the benzylamine or the substituted benzylamine is 1:2: 2;
the reaction time of the nucleophilic substitution is 4 h;
preferably, the mixture VII in step (V-3) further comprises a solvent IX;
the molar ratio of the intermediate IV-2 to the sodium methoxide to the paraformaldehyde is 1: 0.1-1: 0.1 to 0.5;
the reaction conditions are as follows: carrying out reflux reaction at 60-100 ℃;
preferably, the solvent IX comprises methanol, acetone, CH2Cl2At least one of THF, DMF, EtOAc;
the reaction conditions are as follows: feeding the intermediate IV-2, sodium methoxide and paraformaldehyde according to the feeding molar ratio of 1:0.6:0.3, refluxing and stirring at 75 ℃, adding paraformaldehyde and sodium methoxide at intervals, and reacting for 70-75 hours;
preferably, the reaction conditions further comprise: 0.3 mol of paraformaldehyde and sodium methoxide are added at intervals of 8 hours, and the reaction is carried out for 72 hours.
10. A neuraminidase inhibitor comprising at least one compound according to any one of claims 1 to 6, prepared according to the method of any one of claims 7 to 9.
Technical Field
The application relates to a compound, a preparation method and application thereof, belonging to the field of chemistry.
Background
Influenza virus has become one of the most serious diseases threatening human life and health, causes hundreds of thousands of deaths each year, and causes enormous loss to social property each year. At present, although many drugs are on the market, the development of new anti-influenza virus drugs with high efficacy and low toxicity is urgent because influenza viruses are very likely to produce resistant virus strains.
Neuraminidase has attracted a great deal of medical attention as one of the targets for treating influenza virus. Neuraminidase is closely related to virus replication, and inhibition of neuraminidase can inhibit influenza virus replication to a certain extent. Therefore, designing and synthesizing a novel neuraminidase inhibitor is very important for preventing and treating influenza virus.
Disclosure of Invention
According to one aspect of the present application, there is provided a compound useful as a neuraminidase inhibitor.
The compound is characterized in that the structural formula is shown as a formula I:
wherein R is1Selected from F, Cl, Br, C1～C5Any one of an alkyl group of (1) and a group having a structural formula shown in formula (1); n is 0, 1, 2, 3,4 or 5;
R2any one selected from the group having a structural formula shown in formula (1), the group having a structural formula shown in formula (2), and the group having a structural formula shown in formula (3);
*-O-M11formula (1)
Wherein M is11Is selected from H or C1～C5Alkyl groups of (a);
wherein M is21、M22、M23、M24、M25、M26、M27、M28Independently selected from H or C1～C5Alkyl groups of (a);
wherein M is31Selected from H, F, Cl, Br, C1～C5Any one of an alkyl group of (1) and a group having a structural formula shown in formula (1); m32、M33、M34Independently selected from H or C1～C5Alkyl group of (1).
Optionally, wherein R1Any one selected from F, Cl, Br, methyl, ethyl and a group having a structural formula shown in formula (1); n is 0, 1, 2, 3,4 or 5.
Optionally, n ═ 0 in formula I.
Alternatively, R in formula I1One selected from F, Cl, Br, methyl, ethyl, methoxy and ethoxy; n is 1 or 2.
Alternatively, R in formula I1Selected from 4-F, 4-Cl, 4-Br, 4-CH3、4-OCH33-F, 2-F, 3, 4-2F.
Optionally, M in formula (1)11Is selected from C1～C4Alkyl group of (1).
Optionally, M in formula (1)11Selected from methyl or ethyl.
Optionally, M in formula (2)21、M22、M23、M24、M25、M26、M27、M28Are all hydrogen.
Optionally, M in formula (2)21、M22、M23、M24、M25、M26、M27、M28Independently selected from C1～C4Alkyl group of (1).
Optionally, M in formula (2)21、M22、M23、M24、M25、M26、M27、M28Independently selected from hydrogen, methyl or ethyl.
Optionally, M in formula (3)31Selected from H, F, Cl, Br, C1～C4Any one of the alkyl group of (1) and a group having a structural formula shown in formula (1).
Optionally, M in formula (3)31Selected from any one of H, F, Cl, Br, methyl, ethyl and a group with a structural formula shown in a formula (1).
Optionally, M in formula (3)31Selected from any one of H, F, Cl, Br, methyl, ethyl, methoxy and ethoxy.
Optionally, M in formula (3)32、M33、M34Independently selected from H, methyl or ethyl.
Alternatively, R1Selected from F, Cl, Br, — CH3、*-OCH3At least one of (1).
Alternatively, R1Selected from F, Cl, Br, — CH3、*-OCH3N is 2.
Alternatively, R1Selected from F, Cl, Br, — CH3、*-OCH3Wherein n is 1.
Alternatively, R in formula I2Is selected from: -OCH3、
Alternatively, R in formula I2is-OCH3。
As a specific embodiment, the structural formula of the compound is shown in formula I-5:
alternatively, R1Selected from 4-F, 4-Cl, 4-Br, 4-CH3、4-OCH33-F, 2-F or 3,4-2F, and n-1 or n-0.
Alternatively, the structural formula is shown in formula I-1:
alternatively, the R is1Selected from F, Cl, — CH3、*-OCH3And n ═ 1; or n ═ 0.
Alternatively, the R is1Selected from 4-F, 4-Cl, 4-OCH3、4-CH3Or 3-F.
Alternatively, the structural formula is shown in formula I-2:
alternatively, the structural formula is shown in formula I-3:
optionally, the M31Selected from H, 4-F, 4-Cl, 4-Br, 4-OCH3、4-CH33-F or 2-F.
Alternatively, the structural formula is shown in formula I-4:
alternatively, R in formula I-41Selected from Cl,. about. -OCH3At least one of (1).
Alternatively, R in formula I-41Is selected from 4-Cl or 4-OCH3。
In another aspect of the present application, there is provided a method for preparing a compound according to any one of the above, comprising:
(I) carrying out esterification reaction on a mixture I containing substituted phenylacetic acid and halogenated ethyl acetate in the presence of a catalyst to obtain an intermediate I, wherein the structural formula of the intermediate I is shown as a formula II-1:
(II) carrying out Dieckmann reaction on a raw material I containing an intermediate I to obtain an intermediate II, wherein the structural formula of the intermediate II is shown as a formula II-2:
optionally, the mixture I in the step (I) further comprises a solvent I and an acid-binding agent;
the molar ratio of the substituted phenylacetic acid to the halogenated ethyl acetate to the acid-binding agent is 1-2: 1-2: 1-2;
the esterification reaction conditions are as follows: and carrying out reflux reaction at the temperature of 60-80 ℃.
Optionally, the solvent I comprises at least one of tetrahydrofuran, dichloromethane, DMF, ethyl acetate, acetone, methanol, ethanol;
the acid-binding agent comprises at least one of triethylamine, pyridine, anhydrous potassium carbonate, sodium acetate and anhydrous sodium carbonate;
the molar ratio of the substituted phenylacetic acid to the halogenated ethyl acetate to the acid-binding agent is 1:1.2: 1.2;
the esterification reaction conditions are as follows: heated to reflux at 75 ℃ with stirring.
As a specific embodiment, the synthesis route of the intermediate I is as follows:
a：(Et)3N，THF，75℃，98％。
as a specific embodiment, the synthesis route of the intermediate I is as follows:
a：(Et)3N，THF，75℃，98％。
as a specific embodiment, the synthesis route of the intermediate I is as follows:
wherein, the reaction conditions and yield are as follows:
a：(Et)3N，THF，75℃，98％。
optionally, the synthesis method of the intermediate I comprises: heating and refluxing a mixture containing substituted phenylacetic acid, halogenated ethyl acetate, a solvent I and an acid-binding agent under the condition of stirring; TCL monitors the extent of reaction; and after the reaction is finished, cooling, removing triethylamine salt, extracting, washing, drying and filtering to obtain a target product.
As a specific implementation mode, commercially available substituted phenylacetic acid and ethyl chloroacetate are used as raw materials, tetrahydrofuran is used as a solvent, triethylamine is used as an acid-binding agent, and the raw materials are fed in a ratio of n (substituted phenylacetic acid): n (ethyl chloroacetate): n (triethylamine) ═ 1:1.2:1.2, stirring, and heating at 75 ℃ under reflux. The reaction was monitored by TLC as a single point quantitative reaction, which was complete in about 6 hours. After the reaction is finished, cooling to room temperature, and filtering to remove triethylamine salt generated in the reaction. Concentrated under reduced pressure, extracted with ethyl acetate, washed three times with saturated brine, and dried over anhydrous sodium sulfate overnight. And (4) carrying out suction filtration and reduced pressure concentration to obtain dark red liquid, which is directly used for the next reaction without purification.
Optionally, the raw material I in the step (II) further comprises a solvent II and a base I;
the molar ratio of the intermediate I to the base I is 1: 1.5 to 3;
the Dieckmann reaction conditions are as follows: stirring in an ice bath for 20-50 min, and then continuing to react at room temperature.
Alternatively, the solvent II comprises DMF, THF, t-BuOH, methanol, acetone, CH2Cl2At least one of DMSO, EtOAc;
the base I comprises potassium tert-butoxide and anhydrous K2CO3Anhydrous Na2CO3、CH3ONa、C2H5ONa、NaOH、NaHCO3At least one of NaH;
the molar ratio of the intermediate I to the base I is 1: 2;
the Dieckmann reaction conditions are as follows: stirring in ice bath for 30min, and then continuing to react for 4-6 hours at room temperature.
Alternatively, the intermediate I has an active methylene structure and is synthesized by a Dieckmann reaction.
As a specific embodiment, the synthetic route of the intermediate II is as follows:
b: t-BuOK, DMF, 0-room temperature, 80-90%.
As a specific embodiment, the synthetic route of the intermediate II is as follows:
wherein, the reaction conditions and the yield are as follows:
b: t-BuOK, DMF, 0-room temperature, 80-90%.
As a specific embodiment, the synthetic route of the intermediate II is as follows:
b: t-BuOK, DMF, 0-room temperature, 80-90%.
Optionally, the preparation method of the intermediate II comprises: adding a base I into a mixture containing the intermediate I and the base I under ice bath conditions, stirring, reacting at room temperature, and stopping the reaction after the reaction is completely detected by thin layer chromatography.
Mixing a mixture containing an intermediate I, a base I and a solvent II
As a specific embodiment, the synthesis method of the intermediate II comprises: DMF (or t-BuOH and THF) is taken as a solvent, and the feeding ratio n (an intermediate I): n (potassium tert-butoxide) ═ 1:2, under the ice-bath condition, the potassium tert-butoxide in the reaction system is added in 6 times and a half hours. And (3) stirring and reacting for 30min under an ice bath condition, and then placing the system at room temperature for further reaction for 4-6 hours. The reaction is a single-point quantitative reaction, and the reaction is stopped after the thin-layer chromatography detection reaction is completed.
Alternatively, in the post-treatment, 1-fold amount of water was added to the reaction system to dilute the reaction system. And (3) dropwise adding 5% dilute hydrochloric acid into the system under the ice-bath stirring condition to adjust the pH to about 2-3, continuously separating out solids in the pH adjusting process, and adding a large amount of water into the system until no solids are separated out when the pH is adjusted. And (5) placing the mixture in a refrigerator for cooling for about 2 hours, carrying out suction filtration, and recrystallizing dichloromethane to obtain an intermediate 3. However, this method of work-up is not suitable for the substrates used, and some products do not precipitate as solids during the pH adjustment. If no solid is formed, saturated NaCl extraction and anhydrous Na2SO4Drying overnight and purifying by column chromatography.
Optionally, the method of preparing the compound further comprises:
(III-1) carrying out methylation reaction on a mixture II containing an intermediate II and dimethyl sulfate to obtain an intermediate III-1, wherein the structural formula of the intermediate III-1 is shown as a formula II-3-1:
(IV-1) reacting a mixture III containing the intermediate III-1, sodium methoxide and paraformaldehyde to obtain a compound shown in the formula I; wherein R in the formula I2is-OCH3. Namely the compound shown as the formula I-5.
Optionally, the mixture II in the step (III-1) further comprises a solvent III and a base II;
the molar ratio of the intermediate II to the alkali II to the dimethyl sulfate is 1: 1-2: 1-2;
the methylation reaction conditions are as follows: the reaction was carried out at room temperature under protection from light.
Optionally, the solvent III comprises acetone, methanol, CH2Cl2At least one of THF, DMF, EtOAc;
the base II comprises anhydrous K2CO3Anhydrous Na2CO3、CH3ONa、NaOH、NaHCO3At least one of NaH;
the molar ratio of the intermediate II to the alkali II to the dimethyl sulfate is 1:1.2: 1.2;
the methylation reaction conditions are as follows: and reacting for 8-10 hours at room temperature under the condition of keeping out of the light.
As a specific embodiment, the synthetic route of the intermediate III-1 is as follows:
wherein, the reaction conditions and the yield are as follows:
c1：Me2SO4，K2CO3and 70-80% of acetone.
As a specific embodiment, the synthetic route of the intermediate III-1 is as follows:
c1：Me2SO4，K2CO3and 70-80% of acetone.
As a specific embodiment, the synthetic route of the intermediate III-1 is as follows:
c1：Me2SO4，K2CO3and 70-80% of acetone.
Alternatively, the preparation method of the intermediate III-1 comprises the following steps: under the condition of keeping out of the sun, dripping dimethyl sulfate into a mixed solution containing an intermediate II, alkali II and a solvent III, and reacting at room temperature; completion of the reaction was checked by thin layer chromatography. And concentrating, extracting, washing, drying, performing suction filtration and rotary evaporation, and recrystallizing the product obtained after the reaction to obtain an intermediate III-1.
As a specific embodiment, the preparation method of the intermediate III-1 comprises the following steps: using acetone as solvent, anhydrous K2CO3The reaction needs to slowly dropwise add an acetone solution of dimethyl sulfate by using a constant-pressure dropping funnel under the condition of keeping out of the sun because dimethyl sulfate is easy to decompose under the action of light, and the material ratio n (an intermediate II): n (anhydrous K)2CO3): n (dimethyl sulfate) ═ 1:1.2: 1.2. Reacting at room temperature in dark for about 9h, detecting by thin layer chromatography, concentrating under reduced pressure to remove acetone, extracting with ethyl acetate, washing with saturated NaCl, and adding anhydrous Na2SO4Dry overnight. And (3) carrying out suction filtration and rotary evaporation to obtain a solid, wherein the solid is ethyl acetate: recrystallizing with petroleum ether at a ratio of 1:1 to obtain the intermediate compound III-1.
Optionally, the mixture III in the step (IV-1) further comprises a solvent IV;
the molar ratio of the intermediate III-1 to the sodium methoxide to the paraformaldehyde is 1: 0.1-0.5;
the reaction conditions are as follows: reflux reaction at 60-100 ℃.
Optionally, the reaction conditions are 60-80 ℃.
Optionally, the solvent IV comprises methanol, acetone, CH2Cl2At least one of THF, DMF, EtOAc;
the reaction conditions are as follows: and (3) feeding the intermediate III-1, sodium methoxide and paraformaldehyde according to the molar ratio of 1:0.6:0.3, refluxing and stirring at 75 ℃, adding paraformaldehyde and sodium methoxide at intervals, and reacting for 70-75 hours.
Optionally, during the reaction: 0.3 mol of paraformaldehyde and sodium methoxide are added at intervals of 8 hours, and the reaction is carried out for 72 hours.
Alternatively, the synthetic route of step (IV-1) is as follows:
d：(CH2O)n，CH3ONa，75℃，45～50％。
alternatively, the synthetic route of step (IV-1) is as follows:
d：(CH2O)n，CH3ONa，75℃，45～50％。
alternatively, the synthetic route of step (IV-1) is as follows:
d：(CH2O)n，CH3ONa，75℃，45～50％。
optionally, the step (IV-1) comprises: stirring a mixture containing a solvent IV, sodium methoxide and an intermediate III-1 for reflux reaction, and adding paraformaldehyde and sodium methoxide at intervals; and (3) after TLC monitoring that the reactant completely disappears, removing methanol, extracting, washing, drying, carrying out suction filtration, concentrating and purifying to obtain the target product.
As a specific embodiment, the step (IV-1) includes: taking methanol as a solvent and sodium methoxide as a base, and feeding the mixture according to a feeding ratio n (an intermediate III-1): n (sodium methoxide): n (paraformaldehyde) ═ 1:0.6:0.3, and the mixture was stirred under reflux at 75 ℃. 0.3N paraformaldehyde and sodium methoxide are added every 8 hours for reaction for about 48 hours, after TLC monitors that the reactants completely disappear, the methanol is removed by decompression and concentration, ethyl acetate extraction is carried out, the mixture is washed by saturated saline solution, and dried by anhydrous sodium sulfate overnight. Filtering, concentrating, and performing column chromatography to obtain the target compound.
Optionally, the gamma position of butenolide in the intermediate III-1 can attack carbonyl positive ions in paraformaldehyde to perform a condensation reaction, so as to synthesize a target compound.
As a specific embodiment, the synthetic route of the compound represented by formula I-5 is as follows:
Alternatively, R in the synthetic route of the compound represented by the above formula I-51And H is substituted.
Optionally, the method of preparing the compound further comprises:
(III-2) carrying out chloro nucleophilic reaction on the raw material II containing the intermediate II to obtain an intermediate III-2; the structural formula of the intermediate III-2 is shown as a formula II-3-2:
(IV-2) carrying out nucleophilic substitution on the mixture IV containing the intermediate III-2 and morpholine under alkaline conditions to obtain an intermediate IV-1; the structural formula of the intermediate IV-1 is shown as a formula II-4-1:
(V-2) reacting the mixture V containing the intermediate IV-1, sodium methoxide and paraformaldehyde to obtain the compound shown in the formula I-1.
Optionally, the raw material II in step (III-2) further includes: solvent V and nucleophile I;
the molar ratio of the intermediate III-2 to the nucleophilic reagent I is 1: 1-4;
the conditions of the chloro nucleophilic reaction are as follows: stirring in ice bath for 20-50 min, and then reacting at room temperature.
Alternatively, the solvent V is selected from 1:1 of DMF and CH2Cl2At least one of methanol, acetone, THF, DMF, EtOAc;
the nucleophilic reagent I is selected from oxalyl chloride and POCl3At least one of thionyl chloride;
the molar ratio of the intermediate III-2 to the nucleophile I is 1: 2;
the conditions of the chloro nucleophilic reaction are as follows: the mixture was stirred in an ice bath for 30min, and then reacted at room temperature for 6 hours.
Alternatively, the chloro nucleophilic reaction may be obtained by other means, such as: in POCl3With NaOH and H under the conditions of2Taking O mixed solvent as reaction solvent to perform chlorination and Me3SiCl as chloro reagent, DMSO as catalyst, POCl3As chlorinating reagent in the reaction, POCl is used in the reaction3Also as a reaction solvent, diisopropylamine is used as a base, and acid is generated in the neutralization reaction.
Alternatively, the synthetic route of the intermediate III-2 is as follows:
c 2: oxalyl chloride, DMF CH2Cl2(volume ratio 1:1), 95%.
Alternatively, the synthetic route of the intermediate III-2 is as follows:
c 2: oxalyl chloride, DMF CH2Cl2(volume ratio 1:1), 95%.
Alternatively, the preparation method of the intermediate III-2 comprises the following steps: and under the ice bath condition, adding the nucleophilic reagent I into a mixture containing the intermediate II and the solvent V, stirring, reacting at room temperature, completely detecting by TLC, extracting, washing, drying, performing suction filtration, and purifying to obtain an intermediate III-2.
As a specific embodiment, DMF/CH is used2Cl2(1:1) is used as a solvent, oxalyl chloride is slowly dropped under the ice bath condition, and the feeding ratio n (intermediate II): n (oxalyl chloride) ═ 1: 2. After stirring in an ice bath for about half an hour, the reaction was continued for about 6h with the system brought to room temperature. After TLC detection of reaction completion, saturated NaHCO was used3Adjusting the pH of the system to be neutral, and then using CH2Cl2Extracting for three times, combining organic phases, washing with 15ml of saturated saline solution, drying over night through anhydrous sodium sulfate, filtering, and carrying out column chromatography to obtain a pure intermediate compound III-2.
Optionally, the mixture IV in the step (IV-2) further comprises a solvent VI and a base III;
the molar ratio of the intermediate III-2 to the base III to the morpholine is 1: 1-4: 1-4;
the reaction time of the nucleophilic substitution is 3-6 hours.
Alternatively, the solvent VI comprises THF, methanol, acetone, CH2Cl2At least one of DMF and EtOAc;
the base III comprises: anhydrous K2CO3Anhydrous Na2CO3、CH3ONa、NaOH、NaHCO3At least one of NaH;
the molar ratio of the intermediate III-2, the base III and the morpholine is 1:2: 2;
the reaction time for the nucleophilic substitution was 4 h.
Alternatively, the synthetic route of the intermediate IV-1 is as follows:
d 1: morpholine, K2CO3，THF。
Alternatively, the synthetic route of the intermediate IV-1 is as follows:
wherein, the reaction conditions are as follows:
d 1: morpholine, K2CO3，THF。
Alternatively, the preparation method of the intermediate IV-1 comprises the following steps: adding alkali III into a solution containing an intermediate III-2 and a solvent VI, then dropwise adding morpholine, stirring and reacting; and (3) after the TLC spot plate detection reaction is completed, carrying out suction filtration/concentration, extraction, washing, drying and purification to obtain the product.
As a specific embodiment, the preparation method of the intermediate IV-1 comprises the following steps: THF as solvent, anhydrous K2CO3Adding intermediate compound III-2 into 2ml THF solution as alkali to obtain a completely soluble solution, and adding anhydrous K into the system2CO3Slowly dropwise adding morpholine under the stirring condition, wherein the feeding ratio n (an intermediate III-2): n (anhydrous K)2CO3): (morpholine) ═ 1:2: 2. During the dropping process, the system gradually turns into yellow, and a yellow solid is continuously separated out during the stirring process, and the reaction lasts for about 4 hours. And after TLC plate counting detection reaction is completed, carrying out suction filtration to obtain an
Optionally, the mixture V in the step (V-2) further comprises a solvent VII;
the molar ratio of the intermediate IV-1 to the sodium methoxide to the paraformaldehyde is 1: 0.1-1: 0.1 to 0.5;
the reaction conditions are as follows: reflux reaction at 60-100 ℃.
Alternatively, the solvent VII comprises methanol, THF, acetone, CH2Cl2At least one of DMF and EtOAc;
the reaction conditions are as follows: feeding the intermediate IV-1, sodium methoxide and paraformaldehyde according to the molar ratio of 1:0.6:0.3, refluxing and stirring at 75 ℃, adding paraformaldehyde and sodium methoxide at intervals, and reacting for 70-75 hours.
Optionally, during the reaction: 0.3 mol of paraformaldehyde and sodium methoxide are added at intervals of 8 hours, and the reaction is carried out for 72 hours.
Alternatively, the synthetic route of the step (V-2) is as follows:
e1：(CH2O)n，CH3ONa，75℃，30～40％。
alternatively, the synthetic route of the step (V-2) is as follows:
e1：(CH2O)n，CH3ONa，75℃，30～40％。
alternatively, the preparation method of step (V-2) comprises: stirring a mixture containing a solvent VII, sodium methoxide and an intermediate IV-1 for reflux reaction, and adding paraformaldehyde and sodium methoxide at intervals; and (3) after TLC monitoring that the reactant completely disappears, removing methanol, extracting, washing, drying, carrying out suction filtration, concentrating and purifying to obtain the target product.
As a specific embodiment, the step (V-2) specifically includes: taking methanol as a solvent and sodium methoxide as a base, and feeding the mixture according to a feeding ratio n (an intermediate IV-1): n (sodium methoxide): n (paraformaldehyde) ═ 1:0.6:0.3, and the mixture was stirred under reflux at 75 ℃. 0.3N paraformaldehyde and sodium methoxide are added every 8 hours for reaction for about 72 hours, after the reaction is monitored by a thin layer, the mixture is concentrated under reduced pressure to remove methanol, extracted by ethyl acetate (3X 15ml), washed by saturated saline solution and dried over night by anhydrous sodium sulfate. Filtering, concentrating, and performing column chromatography to obtain the target compound.
As a specific embodiment, the synthetic route of the compound represented by formula I-1 is as follows:
a：(Et)3n, THF, 75 ℃, 98%; b: 80-90% of t-BuOK, DMF, 0-room temperature; c 2: oxalyl chloride, DMF CH2Cl2(volume ratio 1:1), 95%; d 1: morpholine, K2CO3，THF；e1：(CH2O)n，CH3ONa，75℃，30～40％。
Alternatively, R in the synthetic route of the compound represented by the above formula I-11And H is substituted.
Optionally, the method of preparing the compound further comprises:
(III-2) carrying out chloro nucleophilic reaction on the raw material II containing the intermediate II to obtain an intermediate III-2; the structural formula of the intermediate III-2 is shown as a formula II-3-2:
(IV-3) carrying out nucleophilic substitution on the mixture VI containing the intermediate III-2 and benzylamine or substituted benzylamine under alkaline conditions to obtain an intermediate IV-2; the structural formula of the intermediate IV-2 is shown as a formula II-4-2:
(V-3) reacting a mixture VII containing the intermediate IV-2, sodium methoxide and paraformaldehyde to obtain the compound shown in the formula I-2.
Optionally, the mixture VI in step (IV-3) further comprises a solvent VIII and a base IV;
the molar ratio of the intermediate III-2 to the base IV to the benzylamine or the substituted benzylamine is 1: 1-4: 1-4;
the reaction time of the nucleophilic substitution is 3-6 hours.
Alternatively, the solvent VIII comprises THF, methanol, acetone, CH2Cl2At least one of DMF and EtOAc;
the base IV comprises: anhydrous K2CO3Anhydrous Na2CO3、CH3ONa、NaOH、NaHCO3NaH, inAt least one of (a);
the molar ratio of the intermediate III-2 to the base IV to the benzylamine or the substituted benzylamine is 1:2: 2;
the reaction time for the nucleophilic substitution was 4 h.
Alternatively, the synthetic route of the intermediate IV-2 is as follows:
wherein, the reaction conditions are as follows:
d 3: benzylamine or substituted benzylamine (e.g. 4-methylbenzylamine or benzylamine), K2CO3，THF。
Alternatively, the preparation method of the intermediate IV-2 comprises the following steps: and (2) dropwise adding benzylamine or substituted benzylamine into a mixture containing the intermediate III-2, the solvent VIII and the alkali IV, stirring at room temperature for reaction, performing suction filtration and washing after TLC detection reaction is complete, thus obtaining the compound.
As a specific embodiment, the preparation method of the intermediate IV-2 comprises the following steps: THF is added into an intermediate III-2 container, and anhydrous K is added under stirring2CO3Slowly adding benzylamine or substituted benzylamine (such as 4-methylbenzylamine) dropwise, continuously generating yellow solid in the dropwise adding process, stirring at room temperature for 3h after dropwise adding is finished, performing suction filtration after detection reaction is completed, and washing with saturated salt solution to obtain the product.
Optionally, the mixture VII in step (V-3) further comprises a solvent IX;
the molar ratio of the intermediate IV-2 to the sodium methoxide to the paraformaldehyde is 1: 0.1-1: 0.1 to 0.5;
the reaction conditions are as follows: reflux reaction at 60-100 ℃.
Optionally, the solvent IX comprises methanol, acetone, CH2Cl2At least one of THF, DMF, EtOAc;
the reaction conditions are as follows: and (3) feeding the intermediate IV-2, sodium methoxide and paraformaldehyde according to the molar ratio of 1:0.6:0.3, refluxing and stirring at 75 ℃, adding paraformaldehyde and sodium methoxide at intervals, and reacting for 70-75 hours.
Optionally, the reaction conditions further comprise: 0.3 mol of paraformaldehyde and sodium methoxide are added at intervals of 8 hours, and the reaction is carried out for 72 hours.
Alternatively, the synthetic route of the step (V-3) is as follows:
e2：(CH2O)n，CH3ONa，75℃，30～40％。
alternatively, the preparation method of step (V-2) comprises: stirring a mixture containing a solvent IX, sodium methoxide and an intermediate IV-2 for reflux reaction, and adding paraformaldehyde and sodium methoxide at intervals; and (3) after TLC monitoring that the reactant completely disappears, removing methanol, extracting, washing, drying, carrying out suction filtration, concentrating and purifying to obtain the target product.
As a specific embodiment, the step (V-3) specifically includes: taking methanol as a solvent and sodium methoxide as a base, and feeding the mixture according to a feeding ratio n (an intermediate IV-2): n (sodium methoxide): n (paraformaldehyde) ═ 1:0.6:0.3, and the mixture was stirred under reflux at 75 ℃. 0.3N paraformaldehyde and sodium methoxide are added every 8 hours for reaction for about 72 hours, after the reaction is monitored by a thin layer, the mixture is concentrated under reduced pressure to remove methanol, extracted by ethyl acetate (3X 15ml), washed by saturated saline solution and dried over night by anhydrous sodium sulfate. Filtering, concentrating, and performing column chromatography to obtain the target compound.
Alternatively, the synthetic route for the compound of formula I-2 is as follows:
a：(Et)3n, THF, 75 ℃, 98%; b: 80-90% of t-BuOK, DMF, 0-room temperature; c 2: oxalyl chloride, DMF CH2Cl2(volume ratio is 1:1), 95%; d 2: benzylamine or substituted benzylamine, K2CO3，THF；e2：(CH2O)n，CH3ONa，75℃，30-40％。
Alternatively, as shown in the above formula I-2Synthetic route to the Compounds R1And H is substituted.
Alternatively, the synthetic route for the compound of formula I-2 is as follows:
wherein, the reaction conditions and yield are as follows:
d 2': benzylamine, K2CO3，THF；e：(CH2O)n，CH3ONa，75℃，30-40％。
d 2': 4-methylbenzylamine, K2CO3，THF；e：(CH2O)n，CH3ONa，75℃，42.6％。
d2' ″: 4-methylbenzylamine, K2CO3，THF；e：(CH2O)n，CH3ONa，75℃，44.3％。
In yet another aspect of the present application, there is provided a neuraminidase inhibitor characterized by comprising at least one compound of any one of the compounds described above, or a compound prepared according to any one of the methods described above.
Optionally, the neuraminidase inhibitor is at least one of the compound described in any of the above, or a compound prepared according to the method described in any of the above.
In-vivo animal experiments of the neuraminidase inhibitor also prove that the compound has a better treatment effect and has significance for further research.
In the present application, the alkyl group is a group formed by an alkane compound having any one hydrogen atom removed. The alkane compound comprises straight-chain alkane, branched-chain alkane and cycloalkane.
In this application, C1～C5Represents the number of carbon atoms comprising the group.
In the present application, "4-F", "3-F", etc. are substitution positions of the respective groups in the compounds described in the present application; wherein, the number of the substitution position is the relative position of the substituent on the benzene ring, and the specific structure is as follows:
In the present application, "CC50"refers to the ability of a drug to inhibit viral replication, and is the concentration of drug that inhibits 50% of viral replication.
In the present application, "EC50By "is meant the half-effect concentration of a dose of drug that causes 50% of the subjects to produce a particular effect.
The beneficial effects that this application can produce include:
1) the compound provided by the application takes butenolide as a mother nucleus and can be used as a neuraminidase inhibitor; the compound has no cytotoxicity, has a higher safety index, and has the same inhibition rate on neuraminidase as oseltamivir.
2) EC of Compounds provided herein50Can reach 6.68 +/-1.32 mu M and is superior to a positive medicament ribavirin (EC)50105 ± 51 μ M) and also has a higher safety index SI of 85.6.
3) The preparation method of the compound provided by the application is simple, and raw materials are easy to obtain.
Drawings
FIG. 1 is a drawing of compound Ia in one embodiment of the present application1HNMR spectrogram;
FIG. 2 is a drawing of compound Ia in one embodiment of the present application13CNMR spectrogram;
FIG. 3 shows the preparation of compound Ib according to an embodiment of the present application1H NMR spectrum;
FIG. 4 shows an embodiment of the present application of compound Ib13C NMR spectrum;
FIG. 5 shows an embodiment of the present applicationOf the Compound Ic1H NMR spectrum;
FIG. 6 shows the preparation of compound Ic in one embodiment of the present application13C NMR spectrum;
FIG. 7 shows one embodiment of the present application for compound Id1H NMR spectrum;
FIG. 8 shows one embodiment of the present application for compound Id13C NMR spectrum;
FIG. 9 is a drawing of compound Ie in one embodiment of the present application1H NMR spectrum;
FIG. 10 is a drawing of compound Ie in one embodiment of the present application13C NMR spectrum;
FIG. 11 is a 1H NMR spectrum of compound If according to one embodiment of the disclosure;
FIG. 12 is a drawing of compound If in one embodiment of the present application13C NMR spectrum;
FIG. 13 shows the preparation of compound Ig in one embodiment of the present application1H NMR spectrum;
FIG. 14 shows the preparation of compound Ig in one embodiment of the present application13C NMR spectrum;
FIG. 15 is a drawing of compound Ih in one embodiment of the present application1H NMR spectrum;
FIG. 16 is a drawing of compound Ih in one embodiment of the present application13C NMR spectrum;
FIG. 17 is a representation of compound Ii in one embodiment of the present application1H NMR spectrum;
FIG. 18 shows one embodiment of the present application of Compound Ii13C NMR spectrum;
FIG. 19 is a drawing of compound II-1 in one embodiment of the present application1H NMR spectrum;
FIG. 20 is a drawing of compound II-1 in one embodiment of the present application13C NMR spectrum;
FIG. 21 is a drawing of compound II-2 in one embodiment of the present application1H NMR spectrum;
FIG. 22 is a drawing of compound II-2 in one embodiment of the present application13C NMR spectrum;
FIG. 23 shows an embodiment of the present applicationOf Compound II-3 in the scheme1H NMR spectrum;
FIG. 24 is a drawing of compound II-3 in one embodiment of the present application13C NMR spectrum;
FIG. 25 is a drawing of compound II-4 in one embodiment of the present application1H NMR spectrum;
FIG. 26 is a drawing of compound II-4 in one embodiment of the present application13C NMR spectrum;
FIG. 27 is a drawing of compound II-5 in one embodiment of the present application1H NMR spectrum;
FIG. 28 is a drawing of compound II-5 in one embodiment of the present application13C NMR spectrum;
FIG. 29 is a drawing of compound II-6 in one embodiment of the present application1H NMR spectrum;
FIG. 30 is a drawing of compound II-7 in one embodiment of the present application1HNMR spectrogram;
FIG. 31 is a drawing of compound II-8 in one embodiment of the present application1H NMR spectrum;
FIG. 32 is a drawing of compound II-9 in one embodiment of the present application1H NMR spectrum;
FIG. 33 is a drawing of compound II-10 in one embodiment of the present application1H NMR spectrum;
FIG. 34 is a drawing of compound II-11 in one embodiment of the present application1H NMR spectrum;
FIG. 35 is a drawing of compound II-12 in one embodiment of the present application1H NMR spectrum;
FIG. 36 is a drawing of compound II-13 in one embodiment of the present application1H NMR spectrum;
FIG. 37 is a drawing of compound II-14 in one embodiment of the present application1H NMR spectrum;
FIG. 38 is a drawing of compound II-15 according to one embodiment of the present application1H NMR spectrum;
FIG. 39 is the EC for α -aryl- β -methoxy- γ -methoxy compounds (Ia, Ib, Id, If, Ig) in this application50；
FIG. 40: a is EC of compound II-1-450(ii) a B is a compound II-3 pairThe rate of viral inhibition varies with concentration dependence;
FIG. 41 is a graph showing the inhibition rate of neuraminidase by Compound II-3;
FIGS. 42 and 43 show the results of molecular docking of Compound II-3 (Gray: oseltamivir, Violet: II-3), in which FIG. 42 shows the results of molecular docking of Compound II-3 (Gray: oseltamivir, Violet: II-3) and FIG. 43 shows the results of molecular docking of Compound II-3 (Violet: II-3).
FIG. 44 shows the results of in vivo experiments with II-3; wherein, the body temperature of the mice of the drug group and the control group is changed, and the body weight of the mice of the drug group and the control group is changed.
Detailed Description
The present application will be described in detail with reference to examples, but the present application is not limited to these examples.
Unless otherwise specified, the raw materials and catalysts in the examples of the present application were all purchased commercially.
The analysis method in the examples of the present application is as follows:
melting point analysis was performed using an X5 model microscopic digital melting point apparatus (Beijing Cork instruments electro-optical apparatus).
NMR analysis was carried out using a model DPX-400 superconducting NMR spectrometer (Bruker, Sweden).