CN109867636B - Compound for resisting CVA16 type hand-foot-and-mouth disease and synthetic method thereof - Google Patents
Compound for resisting CVA16 type hand-foot-and-mouth disease and synthetic method thereof Download PDFInfo
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Abstract
The invention discloses a compound for resisting CVA16 type hand-foot-and-mouth disease and a synthesis method thereof, belonging to the technical field of drug synthesis, wherein the chemical formula of the compound is shown as I, and antiviral experiments show that the compound has good activity of resisting CVA16 type virus and can be used as a candidate drug for resisting the hand-foot-and-mouth disease virus.
Description
Technical Field
The invention relates to a medicine synthesis technology, in particular to a compound for resisting CVA16 type hand-foot-and-mouth disease and a synthesis method agent thereof.
Technical Field
Hand-foot-and-mouth disease (HFMD) is a global infectious disease, especially prevalent in asia-pacific areas. In recent years, the disease is the third infectious disease with the highest morbidity and mortality in China, and the patients with the disease mainly comprise children. The disease is discovered in Shanghai in 1981, later, more than ten provinces (cities) such as Beijing and Hebei are reported every year, and epidemic situation report data in recent years show that the hand-foot-and-mouth disease in China is in a spreading and rising trend. The prevention, control and treatment of the hand-foot-and-mouth disease are regarded as important public health problems and are paid much attention to by people.
The hand-foot-and-mouth disease is an infectious disease caused by enteroviruses, and the enteroviruses causing the hand-foot-and-mouth disease are more than 20 types, wherein the coxsackievirus A16 type (CVA 16) and the enterovirus 71 type (EV 71) are the most common. Clinical manifestations are fever, rash and herpes in the mouth and hands and feet, severe children can cause myocarditis, pulmonary edema, aseptic meningoencephalitis and other fatal complications, and individual severe children can rapidly develop the disease and die. At present, exact and effective antiviral drugs are lacked, and the clinical treatment mainly aims at symptomatic treatment, relieves the symptoms of patients and prevents the occurrence and development of serious complications.
At present, the first EV 71 vaccine reaches a third-stage clinical test in China, the protection rate of the vaccine on the hand-foot-and-mouth disease infected by the clinical EV 71 reaches 90%, but the vaccine has no obvious protection effect on other pathogens causing the hand-foot-and-mouth disease, such as CVA16 and the like. The dominant genotype of CVA16, the first pathogen found to be related to hand-foot-and-mouth disease, is prevalent at home and abroad for many years, but the social attention is far less than that of EV 71, and the research on the etiology is insufficient.
The capsid protein of the CVA16 virus is a symmetrical spherical dodecahedral solid structure with a diameter of about 30 nm. The nucleocapsid consists of single positive strand RNA and protein, with no processes and envelopes. The genome consists of approximately 7410 nucleotides. The viral genome consists of 5 'non-coding regions (UTRs), P1, P2 and P3 regions, and 3' non-coding regions (UTRs). The 5 'UTRs contain a virion protein (VPg1) involved in the synthesis and assembly of viral RNA, and the 3' UTRs have the ability to enhance viral infection. The viral genome has only one open reading frame of a polyprotein of 2193 amino acids, which can be cleaved into three precursor proteins, P1, P2 and P3, by hydrolysis with proteases. P1 can be degraded further, and finally four structural proteins VP1-VP4 are formed, three of which are located on the outer shell surface of the virus, i.e. VP1, VP2 and VP3, wherein most of the antigenic determinants are contained, and VP4 is embedded inside the virus and linked with the viral RNA. These surface epitopes play an important role in the recognition of invasion by the virus into the host. P2 and P3 encode 2A, 2B, 2C, 3A, 3B, 3C and 3D 7 nonstructural proteins, which are important for replication and post-translational modification of the virus, and also in the efficient infection of host cells by the virus.
With the development of new methods and new technologies for drug research, more and more protein three-dimensional structures are confirmed, more and more drug targets are generated, and a large number of target proteins have been selected as potential therapeutic targets for research. The molecular docking method is widely applied to drug research and design, and mainly applies computer technology to simulate the complex reaction between a small molecular ligand and a large molecular receptor so as to realize computer-aided drug virtual screening. The virtual screening method based on the molecular docking technology can greatly save the research cost of the drugs and effectively shorten the drug development period. The molecular docking technology is an important method for drug design according to receptor structures, and is an important means for innovating and developing drugs.
Disclosure of Invention
In order to solve the problems in the prior art, the invention aims to provide a novel efficient CVA16 type hand-foot-and-mouth disease resistant medicament with an accurate molecular target according to molecular docking operation between a small molecular compound and a drug target.
The technical scheme of the invention is as follows:
a compound of the formula I,
derivatives of the compounds of formula I above have the formula:
The application of the compound shown in the formula I or the derivative thereof in preparing the medicine is used for treating or preventing the hand-foot-and-mouth disease, and the medicine takes the compound shown in the formula I or the derivative thereof as a medicine active ingredient.
The application of the compound shown as the formula I or the derivative thereof in preparing the medicine for resisting CVA16 is characterized in that the compound shown as the formula I or the derivative thereof is used as an active ingredient for resisting CVA 16.
In the pharmaceutical application, the compound of the formula I is added with pharmaceutically acceptable auxiliary materials.
The synthesis method of the compound shown in the formula I is characterized in that the synthesis route is as follows:
preferably, the synthesis steps are as follows:
and (1).
Adding compound 1-o-hydroxybenzoic acid, succinic anhydride and pyridine into a round-bottom flask, heating to 85-95 ℃ with 4-dimethylaminopyridine as a catalyst, and reacting for 4-6 h; after the reaction is finished, removing the solvent by reduced pressure distillation, and performing column chromatography separation by using a mixed solution of dichloromethane and methanol as a mobile phase to obtain a compound 2;
and 2. step 2.
Adding the compound 2, diethylene glycol monomethyl ether, a catalyst dicyclohexylcarbodiimide and dichloromethane into a round-bottom flask, heating to reflux, and reacting for 10-14 h; after completion of the reaction, the solvent was distilled off under reduced pressure, and then column chromatography was performed using a mixed solution of dichloromethane and methanol as a mobile phase to obtain compound 4.
And 3. step 3.
Adding a compound 4 and DMF (dimethyl formamide) into a round-bottom flask, dropwise adding thionyl chloride under the protection of nitrogen, reacting for 2-4h at room temperature, adding 2-amino-5-nitrothiazole, continuing to react for 4-6h, carrying out reduced pressure distillation to remove a solvent, and carrying out column chromatography separation by using a mixed solution of dichloromethane and methanol as a mobile phase to obtain a light yellow solid compound I.
Preferably, the synthesis method comprises the following steps:
step 1.50ml of round-bottom flask is added with 1.38g of compound 1-o-hydroxybenzoic acid, 1.46g of succinic anhydride, 15ml of pyridine and 2.47g of catalyst 4-dimethylaminopyridine, and the mixture is heated to 90 ℃ to react for 5 hours; after the reaction was completed, the solvent was distilled off under reduced pressure, and then the reaction mixture was stirred at a volume ratio of 2: 1, taking dichloromethane-methanol mixed solution as mobile phase column chromatography to obtain a compound 2;
step 2, adding 2.38g of compound 2, 1.4g of diethylene glycol monomethyl ether, 2.47g of catalyst dicyclohexylcarbodiimide and 5ml of dichloromethane into a 50ml round-bottom flask, heating to reflux, and reacting for 12 hours; after the reaction was completed and the solvent was distilled off under reduced pressure, the reaction mixture was stirred at a volume ratio of 3: 1 dichloromethane: separating the methanol mixed solution as a mobile phase column chromatography to obtain a compound 4;
step 3, adding 3.40g of compound 4 and 15ml of DMF (dimethyl formamide) into a 50ml round-bottom flask, dropwise adding thionyl chloride under the protection of nitrogen, reacting for 3 hours at room temperature, adding 1.56g of 2-amino-5-nitrothiazole, continuing to react for 5 hours, distilling under reduced pressure to remove the solvent, and then using a volume ratio of 2: 1, and performing column chromatography on a mixed solution of dichloromethane and methanol as a mobile phase to obtain a light yellow solid compound I.
A process for the synthesis of derivatives of the compound of formula I, compound 1 position in step 1
Wherein R is methyl, ethyl, carboxymethyl, nitro, methoxy, formyl, acetyl, formamido, acetamido or phenyl.
The invention takes coxsackievirus A16 type (CVA 16) as a target, utilizes a molecular docking technology to design and screen a small molecular compound medicament with good inhibitory effect on coxsackievirus A16 type (CVA 16), and antiviral efficacy experiment results show that the small molecular compound has excellent antiviral activity on the CVA16 type. The invention further provides a synthetic method of the small molecule medicament.
The dosages of the starting materials indicated in the above methods are to be understood as ratios, and in practical examples, the synthetic processes are carried out with each starting material being added in equal multiples, according to the different scales of the preparation process.
Detailed Description
Preferred embodiments of the present invention will be described in detail with reference to the following examples. It is to be understood that the following examples are given for illustrative purposes only and are not intended to limit the scope of the present invention. Various modifications and alterations of this invention will become apparent to those skilled in the art without departing from the spirit and scope of this invention.
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Example 1 Synthesis routes and conditions for Compounds of formula (I) according to the invention
The synthetic route is as follows:
and (1).
A50 mL round-bottom flask was charged with Compound 1-hydroxybenzoic acid (1.38g, 10mmol), succinic anhydride (1.2g, 12mmol), pyridine (15mL), DMAP (4-dimethylaminopyridine, 1.46g,12mmol) as a catalyst, and heated to 90 deg.C for 5 h.
After the reaction was completed, the solvent was distilled off under reduced pressure and methylene chloride was used: methanol (volume ratio 2: 1) was used as mobile phase column chromatography to obtain compound 2.
And 2. step 2.
A50 ml round bottom flask was charged with Compound 2(2.38g, 10mmol), Compound 3(Cas #111-77-3, diethylene glycol monomethyl ether, 1.4g, 12mmol), catalyst DCC (dicyclohexylcarbodiimide, 2.47g, 12mmol) and dichloromethane (5 ml). Heated to reflux and reacted for 12 h. After the reaction was completed and the solvent was distilled off under reduced pressure, the reaction mixture was purified by using methylene chloride: methanol (volume ratio 3: 1) was used as mobile phase column chromatography to give compound 4.
And 3. step 3.
A50-mL round-bottom flask was charged with Compound 4(3.40g, 10mmol) and DMF (15mL), and thionyl chloride was added dropwise under nitrogen protection, and after reacting at room temperature for 3 hours, Compound 5(cas #121-66-4, 2-amino-5-nitrothiazole 1.56g, 12mmol) was added, and after further reacting for 5 hours, the solvent was distilled off under reduced pressure, and then the mixture was purified with dichloromethane: methanol (volume ratio 2: 1) was used as mobile phase column chromatography to give a pale yellow solid.
The product detection method comprises the following steps:
the instrument comprises the following steps: the name model of the instrument adopted in nuclear magnetic detection is Bruker 400Hz, and the name model of the instrument used in mass spectrometry is Agilent 6520;
detection result data:
MS：m/z＝468.01[M+H]+
1HNMR(CDCl3，δppm):8.69(1H,s),7.85(1H,dd),7.7(1H,td),7.47(1H,td),7.32(1H,dd),4.20(2H,m),3.82-3.74(4H,m),3.394(3H,s),3.85(2H,m),2.42(4H,m)
the detection result shows that the light yellow solid is represented by the chemical formula C19H21O9N3S is a compound represented by formula I.
Example 2 derivatives of the Compounds of formula (I) according to the invention
Derivatives of Compound I
The synthetic route is the same as example 1.
And (1).
A50 mL round-bottom flask was charged with Compound 1(1.38g, 10mmol), succinic anhydride (1.2g, 12mmol), pyridine (15mL), DMAP (4-dimethylaminopyridine, 1.46g,12mmol) as a catalyst, and heated to 90 deg.C for 5 h.
After the reaction was completed, the solvent was distilled off under reduced pressure and methylene chloride was used: methanol (volume ratio 2: 1) was used as mobile phase column chromatography to obtain compound 2.
Compound 1 is
Wherein R is methyl, ethyl, carboxymethyl, nitro, methoxy, formyl, acetyl, formamido, acetamido or phenyl.
And 2. step 2.
A50 ml round bottom flask was charged with Compound 2(2.38g, 10mmol), Compound 3(Cas #111-77-3, diethylene glycol monomethyl ether, 1.4g, 12mmol), catalyst DCC (dicyclohexylcarbodiimide, 2.47g, 12mmol) and dichloromethane (5 ml). Heated to reflux and reacted for 12 h. After the reaction was completed and the solvent was distilled off under reduced pressure, the reaction mixture was purified by using methylene chloride: methanol (volume ratio 3: 1) was used as mobile phase column chromatography to give compound 4.
And 3. step 3.
Adding compound 3(3.40g, 10mmol) and DMF (15ml) into a 50ml round bottom flask, dropwise adding thionyl chloride under the protection of nitrogen, reacting at room temperature for 3h, adding compound 5(1.56g, 12mmol), continuing to react for 5h, distilling under reduced pressure to remove the solvent, and adding dichloromethane: methanol (volume ratio 2: 1) was used as mobile phase column chromatography to give compound I as a pale yellow solid.
The detection method is the same as that of example 1, and the detection result shows that the light yellow solid is the compound I.
Experimental example 1 the compounds of formula (I) according to the invention were tested for their antiviral activity.
In order to detect the in vitro antiviral activity of the test compound, CVA16 was isolated from clinical specimens of the hands, feet and mouths and subjected to in vitro antiviral artificial re-certification by plaque assay.
Will be 1 × 106RD cells (human malignant embryonal rhabdomyoma cells, Cat No.: CL-0193, tissue source: muscle, rhabdomyosarcoma) of each well were inoculated in 6-well plates, after overnight culture, RD cells were infected by dilution of a virus solution to be tested at 10-fold ratio, 3 wells were repeated for each dilution, the supernatant was removed after infection at 37 ℃ for 1h, semisolid medium (DMEM high-sugar (Cat No.: PM150210) + 10% FBS (Cat No.: 164210-500) + 1% P/S (Cat No.: PB180120)) containing 1.2% methylcellulose and 2% serum was added, and the cells were cultured in an incubator for 5-6 days until viral plaques were formed.
And counting plaques after fixing by paraformaldehyde, staining by crystal violet, rinsing by clear water and airing, and calculating the virus titer.
RD cells inoculated in 6-well plates were infected with a test virus solution having an MOI of 0.001/0.01, and after infection, a semisolid medium containing 5 μmol/L of compound i was added, incubated at 37 ℃ in an incubator, and plaque count was performed. The effect of test compound i on viral titer was examined with the addition of semi-solid medium containing the solvent DMSO as a control.
The result of the plaque assay shows that the number of plaque formation is significantly reduced in the presence of test compound i compared to the DMSO solvent control, indicating that the test compound inhibits replication of CVA16 virus to some extent and has antiviral activity.
EXAMPLE 2 determination of the Virus-inhibiting Rate of the Compounds of formula (I) according to the invention.
The CVA16 suspension is diluted by a cell maintenance solution containing 2 percent serum (preparation method: adding the inactivated fetal bovine serum into a DMEM culture solution to a final concentration of 2 percent, respectively adding penicillin and streptomycin to a final concentration of 100 mu g/mL and glutamine to a final concentration of 2mmol/L) from 10 < -1 > to 10 < -10 > in different concentration gradients, respectively titrating the diluted solutions on RD cells attached to a wall and growing full of a 96-well plate monolayer, and continuously culturing in a 5 percent CO2 incubator at 37 ℃ by using a new cell maintenance solution.
Incubating virus-infected 96-well plate cells in an incubator for 48h, visually observing cytopathic effect (CPE) under a microscope, observing the virus titration concentration and the number of diseased wells with cytopathic effect, and calculating the cell culture half infectious dose of virus solution according to Reed-Muench formula (TCID50) The virus concentration adopted in the subsequent experiment is 100TCID50。
Cells in logarithmic growth phase were seeded in 96-well plates at 37 ℃ with 5% CO2After growth in an incubator to a monolayer, the culture medium was discarded, washed twice with PBS, and 100TCID mixed with the test compound was added50100. mu.l of virus suspension (5% CO at 37 ℃)2After incubation for 1.5h in the incubator, the infectious agent was discarded, washed three times with PBS, and cultured by adding DMEM cell maintenance medium containing 20. mu.g/mL and 10. mu.g/mL of test compound, respectively.
After 48h, when about 90% of the cytopathic effect of the virus control wells appeared, the pathological effect of the drug group cells was visually observed by a microscope.
The whole experimental process is set as a cell control group without any treatment, and a virus control group without test drugs.
Cell CPE determination criteria: no cytopathic effect is noted-less than 25% cytopathic effect is noted +, 25-50% cytopathic effect is noted + +, 50-75% cytopathic effect is noted + + +, and more than 75% cytopathic effect is noted + + +.
Removing the supernatant of the cell culture plate after the cytopathic effect is observed, washing the cell culture plate twice by PBS, and calculating the virus inhibition rate by using an MTT colorimetric method:
the TCID of the test strain CVA16 was determined50Is 10-5.9The virus concentration used in the test was 100TCID50. After the virus solution is diluted according to the dilution gradient, the inhibition effect of the drug treatment on the virus is visually observed by a microscope when the CPE of the cells of the virus control group is +++ after the cells are infected, and the virus inhibition rate is 58% by the MTT method.
Assays on derivatives of the compounds of formula (I) show that similar results are obtained.
EXAMPLE 3 CC50, EC50 and SI determination of the compounds of formula (I) according to the invention.
CC50, the median toxic concentration, indicates the concentration of drug that elicits 50% drug toxicity in cells. Testing the sampleThe compound was prepared as a stock solution at 10mg/mL in DMSO (dimethyl sulfoxide) and diluted to the corresponding concentration in the cell maintenance solution during the experiment. The RD cells grown in a monolayer in a 96-well plate were removed from the culture and replaced with cell maintenance solutions containing different concentrations of the diluted drug. Each concentration was 3 replicates, while normal cells were set as a control. 5% CO at 37 ℃2Culturing in an incubator for 48 h. And (4) observing the cytotoxicity condition of the drug treated at different concentrations under an inverted microscope, and determining the cell survival rate by using an MTT colorimetric method. Drug CC50 was calculated by Probit regression of SPSS software.
EC50, the median effective concentration, refers to the concentration of drug that is 50% effective in inhibiting the virus. In substantial agreement with the cytopathic index observation procedure of example 2, the drug concentration was set to a plurality of different concentration gradients. When the CPE disease of the virus control group is +++ and the cell disease condition of the drug treatment group with different concentrations is observed under a microscope, the MTT colorimetric method is used for determining the respective virus inhibition rate. The Probit regression method of the SPSS software calculated drug EC 50.
SI is the therapeutic index of the drug CC50/EC 50. The greater the SI value, the greater the antiviral potential.
The results are given in the following table:
the cytotoxic and anti-CVA 16 activity of formula (I) is shown
Assays on derivatives of the compounds of formula (I) show that similar results are obtained.
The application of the small molecule pharmaceutical compound of the present invention as an anti-CVA 16 virus agent has been described by way of specific examples, and it will be obvious to those skilled in the art that the content of the present invention can be used as a reference, and the raw materials, process conditions and the like can be appropriately changed to achieve other corresponding purposes, without departing from the content of the present invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (10)
1. A compound of the formula I,
2. a derivative of the compound of claim 1, of the formula:
3. Use of a compound represented by formula I or a derivative thereof for the preparation of a medicament for the treatment or prevention of hand-foot-and-mouth disease, said medicament comprising a compound represented by formula I as defined in claim 1 or a derivative as defined in claim 2 as a pharmaceutically active ingredient.
4. Use of a compound of formula I or a derivative thereof for the manufacture of a medicament against CVA16 type, comprising a compound of formula I as claimed in claim 1 or a derivative thereof as claimed in claim 2 as an active ingredient against CVA16 type virus.
5. The pharmaceutical use according to claim 3 or 4, further comprising a pharmaceutically acceptable excipient.
6. A process for the synthesis of compounds of formula i according to claim 1, characterized in that the following synthetic route is used:
7. the method of synthesis according to claim 6, comprising the steps of:
and (1).
Adding compound 1-o-hydroxybenzoic acid, succinic anhydride and pyridine into a round-bottom flask, heating to 85-95 ℃ with 4-dimethylaminopyridine as a catalyst, and reacting for 4-6 h; after the reaction is finished, removing the solvent by reduced pressure distillation, and performing column chromatography separation by using a mixed solution of dichloromethane and methanol as a mobile phase to obtain a compound 2;
and 2. step 2.
Adding the compound 2, diethylene glycol monomethyl ether, a catalyst dicyclohexylcarbodiimide and dichloromethane into a round-bottom flask, heating to reflux, and reacting for 10-14 h; after the reaction is finished, removing the solvent by reduced pressure distillation, and performing column chromatography separation by using a mixed solution of dichloromethane and methanol as a mobile phase to obtain a compound 4;
and 3. step 3.
Adding a compound 4 and DMF (dimethyl formamide) into a round-bottom flask, dropwise adding thionyl chloride under the protection of nitrogen, reacting for 2-4h at room temperature, adding 2-amino-5-nitrothiazole, continuing to react for 4-6h, carrying out reduced pressure distillation to remove a solvent, and carrying out column chromatography separation by using a mixed solution of dichloromethane and methanol as a mobile phase to obtain a light yellow solid compound I.
8. The method of synthesis according to claim 7, the steps of synthesis being as follows:
the step 1 is as follows: 1.38g of compound 1-o-hydroxybenzoic acid, 1.46g of succinic anhydride, 15ml of pyridine and 2.47g of catalyst 4-dimethylaminopyridine are added into a 50ml round-bottom flask, and the mixture is heated to 90 ℃ to react for 5 hours; after the reaction was completed, the solvent was distilled off under reduced pressure, and then the reaction mixture was stirred at a volume ratio of 2: 1, taking dichloromethane-methanol mixed solution as mobile phase column chromatography to obtain a compound 2;
the step 2 is as follows: adding 2.38g of compound 2, 1.4g of diethylene glycol monomethyl ether, 2.47g of catalyst dicyclohexylcarbodiimide and 5ml of dichloromethane into a 50ml round-bottom flask, heating to reflux, and reacting for 12 hours; after the reaction was completed and the solvent was distilled off under reduced pressure, the reaction mixture was stirred at a volume ratio of 3: 1 dichloromethane: separating the methanol mixed solution as a mobile phase column chromatography to obtain a compound 4;
the step 3 is as follows: adding 3.40g of compound 4 and 15ml of DMF into a 50ml round bottom flask, dropwise adding thionyl chloride under the protection of nitrogen, reacting at room temperature for 3 hours, adding 1.56g of 2-amino-5-nitrothiazole, continuing to react for 5 hours, removing the solvent by reduced pressure distillation, and then using a volume ratio of 2: 1, and performing column chromatography on a mixed solution of dichloromethane and methanol as a mobile phase to obtain a light yellow solid compound I.
9. A process for the synthesis of derivatives of the compounds of formula i as claimed in claim 2, comprising the synthetic steps of:
step 1, adding a compound 1, succinic anhydride and pyridine into a round-bottom flask, heating to 85-95 ℃ by taking 4-dimethylaminopyridine as a catalyst, and reacting for 4-6 hours; after the reaction is finished, removing the solvent by reduced pressure distillation, and performing column chromatography separation by using a mixed solution of dichloromethane and methanol as a mobile phase to obtain a compound 2;
step 2, adding the compound 2, diethylene glycol monomethyl ether, a catalyst dicyclohexylcarbodiimide and dichloromethane into a round-bottom flask, heating to reflux, and reacting for 10-14 hours; after the reaction is finished, removing the solvent by reduced pressure distillation, and performing column chromatography separation by using a mixed solution of dichloromethane and methanol as a mobile phase to obtain a compound 4;
step 3, adding a compound 4 and DMF (dimethyl formamide) into a round-bottom flask, dropwise adding thionyl chloride under the protection of nitrogen, reacting for 2-4h at room temperature, adding 2-amino-5-nitrothiazole, continuing to react for 4-6h, distilling under reduced pressure to remove a solvent, and performing column chromatography separation by using a mixed solution of dichloromethane and methanol as a mobile phase to obtain a light yellow solid derivative of the compound shown in the formula I;
the compound 1 is
Wherein R is methyl, ethyl, carboxymethyl, nitro, methoxy, formyl, acetyl, carboxamido, acetylamino or phenyl.
10. The method of synthesis according to claim 9, the steps of synthesis being as follows:
the step 1 is as follows: adding 10mmol of compound 1, 1.46g of succinic anhydride, 15ml of pyridine and 2.47g of catalyst 4-dimethylaminopyridine into a 50ml round-bottom flask, and heating to 90 ℃ for reaction for 5 hours; after the reaction was completed, the solvent was distilled off under reduced pressure, and then the reaction mixture was stirred at a volume ratio of 2: 1, taking dichloromethane-methanol mixed solution as mobile phase column chromatography to obtain a compound 2;
the step 2 is as follows: adding 2.38g of compound 2, 1.4g of diethylene glycol monomethyl ether, 2.47g of catalyst dicyclohexylcarbodiimide and 5ml of dichloromethane into a 50ml round-bottom flask, heating to reflux, and reacting for 12 hours; after the reaction was completed and the solvent was distilled off under reduced pressure, the reaction mixture was stirred at a volume ratio of 3: 1 dichloromethane: separating the methanol mixed solution as a mobile phase column chromatography to obtain a compound 4;
the step 3 is as follows: adding 3.40g of compound 4 and 15ml of DMF into a 50ml round bottom flask, dropwise adding thionyl chloride under the protection of nitrogen, reacting at room temperature for 3 hours, adding 1.56g of 2-amino-5-nitrothiazole, continuing to react for 5 hours, removing the solvent by reduced pressure distillation, and then using a volume ratio of 2: 1, dichloromethane and methanol mixed solution is used as a mobile phase column chromatography to obtain light yellow solid derivatives of the compound shown in the formula I.
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| CN107281148A (en) * | 2017-05-08 | 2017-10-24 | 天津国际生物医药联合研究院 | A kind of preparation method of solid dispersions and its solid pharmaceutical preparation |
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