CN112707874A - Antiviral compound and preparation method thereof - Google Patents
Antiviral compound and preparation method thereof Download PDFInfo
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- CN112707874A CN112707874A CN202011590753.4A CN202011590753A CN112707874A CN 112707874 A CN112707874 A CN 112707874A CN 202011590753 A CN202011590753 A CN 202011590753A CN 112707874 A CN112707874 A CN 112707874A
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Classifications
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- C07D277/02—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings
- C07D277/20—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
- C07D277/32—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
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- A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
- C07F9/02—Phosphorus compounds
- C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
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Abstract
The invention discloses an antiviral compound and a preparation method thereof. The structural formula of the compound is shown as formula I, wherein in the formula I, R1Selected from: mono-or poly-substituted H, F, methyl, trifluoromethyl; r2Selected from: H. straight or substituted alkanes (C1-C6); r3Selected from: mono-or polysubstituted H, Cl, Br, F; when X is NH, R4Selected from: H. acyl including sulfonyl, phosphoryl or alkanoyl; when X ═ O, R4Is composed of
Wherein n is an integer of 0 to 6, such as 2, 3; y is selected from O or N. Experiments prove that the compound has a good inhibitory effect on H1N1 influenza A virus and coronavirus, does not have toxicity to normal human cells, and can inhibit the virus and the inflammatory reaction.
Description
Technical Field
The invention belongs to the field of medicines, and particularly relates to an antiviral compound and a preparation method thereof.
Background
Influenza virus is called influenza virus for short. It is classified into types A (A), B (B) and C (C), and influenza viruses discovered in recent years are classified into types D (D). The influenza virus can cause infection and morbidity of various animals such as human, poultry, pigs, horses, bats and the like, and is a pathogen of epidemic diseases of human and animals such as human influenza, avian influenza, swine influenza, horse influenza and the like.
The typical clinical symptoms of these diseases are acute hyperpyrexia, general pain, marked debilitation and respiratory symptoms. Influenza viruses are transmitted primarily by airborne droplets, contact between a susceptible and infected person, or contact with contaminated items. The autumn and winter season is the high-incidence period. Human influenza is mainly caused by influenza a and influenza b viruses. Influenza a viruses often have antigenic variation and can be further divided into subtypes H1N1, H3N2, H5N1, H7N9 and the like (wherein H and N represent two surface glycoproteins of influenza viruses, respectively). Influenza viruses are not very resistant to the environment. Animal influenza viruses do not normally infect humans, human influenza viruses do not normally infect animals, but swine is the exception. Pigs can be infected with both human and avian influenza viruses, but they are also predominantly infected with swine influenza virus. A small number of animal influenza viruses, after adaptation to humans, can cause a human influenza pandemic.
Human coronaviruses can cause the common cold, Severe Acute Respiratory Syndrome (SARS) and Middle East Respiratory Syndrome (MERS) in humans, with some differences in epidemiological characteristics.
Globally, 10% to 30% of upper respiratory infections are caused by the four groups of coronaviruses HCoV-229E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1, which account for the second place in the etiology of the common cold, second only to rhinoviruses. Infections appear seasonal, with high disease rates in the spring and winter of each year. The incubation period is 2-5 days, and the population is generally susceptible. Mainly by human-to-human contact. SARS is caused by infection of human beings by SARS-CoV, and appears in part of China Guangdong province first, and then reaches 24 provinces, autonomous regions, direct prefecture cities and other 28 countries and regions in the world. In the first global SARS epidemic from 11 months to 7 months in 2002, 8096 clinical diagnosis cases, 774 deaths and 9.6 percent of fatality rate are reported globally. The latent period of SARS is usually limited to 2 weeks, usually about 2 to 10 days. The crowd is common and easy to feel. SARS is the most important source of infection, with the patient with obvious symptoms having strong infectivity, and the patient in latent period or cured period has no infectivity. MERS is a viral respiratory disease caused by MERS-CoV and was first identified in saudi arabia in 2012. Since 2012 MERS co-propagated in the middle east, asia, europe and 27 countries and regions worldwide, 80% of cases were from saudi arabia with approximately 35% mortality. The incubation period is 14 days at most, and the population is generally susceptible. Dromedary camels are a large host of MERS-CoV and are a major source of infection in interpersonal cases, with limited interpersonal transmission.
Influenza a H1N1 is a highly contagious acute respiratory disease in pigs caused by one or more swine influenza a viruses. The morbidity is often high, but the mortality is low (1-4%). The virus is spread in the herd by aerosols, direct and indirect contact and asymptomatic pigs carrying the virus. Herd epidemic may occur throughout the year. In the fall and winter in temperate zones, the incidence of disease rises. Human influenza A H1N1 usually comes from infected pigs, but some human cases do not have a history of exposure to pigs or the environment in which they are located. Interpersonal transmission occurs in some cases, but is limited to close contacts and people in an enclosed environment.
Coronavirus (HCoV-229E) is a species of coronavirus. Coronaviruses belong to the order of the nested viruses, the family of coronaviruses, the genus of coronaviruses, a family of large viruses, and are widely found in nature. Coronaviruses only infect vertebrates, are associated with a variety of diseases in humans and animals, and can cause diseases in the respiratory, digestive and nervous systems of humans and animals.
Therefore, the research of an effective antiviral drug has important practical significance.
Disclosure of Invention
The invention aims to provide a compound shown as a formula I and pharmaceutically acceptable salts, esters and solvates thereof.
The structural general formula of the compound provided by the invention is shown as formula I:
in the formula (I), the compound has the following structure,
R1selected from: mono-or poly-substituted H, F, methyl, trifluoromethyl, preferably H;
R2selected from: H. straight or substituted alkanes (C1-C6), preferably methyl, isopropyl;
R3selected from: mono-or polysubstituted H, Cl, Br, F, preferably Cl;
when X is NH, R4Selected from: H. acyl including sulfonyl (e.g. methanesulfonyl, ethanesulfonyl), phosphoryl (e.g. O, O-diisopropylphosphoryl) or alkanoyl (e.g. butyryl), preferably H or methanesulfonyl;
when X ═ O, R4Is composed of
Wherein n is an integer from 0 to 6, such as 2, 3 or 4;
y is selected from O or N;
R5、R6independently selected from any one of the following groups: H. (C)1-C6) Alkyl, (C)3-C8) Carbocyclylalkyl group, (C) containing substituent1-C18) Alkane of (C)2-C8) Alkenyl group, (C) having substituent2-C8) Alkenyl, (C)2-C8) Alkynyl group, substituent-containing (C)2-C8) Alkynyl, (C)6-C20) Aryl group, substituted group-containing (C)6-C20) Aryl group, (C)2-C20) Heterocyclic group, (C) containing substituent2-C20) A heterocyclic group; or R5、R6Form a ring with each other to form (C)3-C8) Heterocycloalkyl or substituted (C)3-C8) Heterocycloalkyl group, (C)6-C20) Heteroaryl or substituted (C)6-C20) A heteroaryl group.
The R is5、R6Wherein said substituent is selected from at least one of: h, methyl, ethyl, isopropyl, pyrrolyl, piperidinyl, morpholinyl and piperazinyl.
In some embodiments of the present invention, the compounds of formula I according to the present invention may be exemplified by, but are not limited to, the structures shown below:
the invention also provides a preparation method of the compound shown in the formula I.
According to Med. chem. Commun. 2016,7, 1441-1448, the reaction of alkylhydrazine of compound 1 (formula a) with sodium thiocyanate gives thiourea compound 2 (formula b), which is then condensed with R1-substituted o-benzoic acid benzaldehyde to give compound 3 (formula c), which is finally reacted with R1-substituted o-benzoic acid benzaldehyde3Substituted bromoacetophenone for ring closure, thiazole is easily obtainedCompound 4 (formula d); under the action of a condensing agent, the benzamide derivative shown in the formula I is obtained. The method comprises the following specific steps:
1) reacting the compound shown in the formula a with sodium thiocyanate to obtain a compound shown in a formula b;
wherein R in the formula a2Is as defined in formula I; in the formula b, R2Is as defined in formula a;
2) reacting a compound represented by the formula b with R represented by the formula e1Carrying out condensation reaction on substituted o-benzoic acid benzaldehyde to obtain a compound shown as a formula c;
wherein R in the formula e1Is as defined in formula I; in the formula c, R1Is defined by the formula e, R2Is as defined in formula b;
3) reacting a compound represented by the formula c with R represented by the formula f3Substituting bromoacetophenone for carrying out a ring closure reaction to obtain a compound shown as a formula d;
wherein R in the formula f3Is as defined in formula I, formula d wherein R1、R2Is defined by the formula c, R3Is as defined in formula f;
4) carrying out condensation reaction on the compound shown in the formula d and the compound shown in the formula g under the action of a condensing agent to obtain a compound shown in the formula I;
H-X-R4(formula g)
Wherein, X, R in the formula g4Is as defined in formula I.
In the step 1), the reaction conditions of the reaction are as follows: the reaction temperature is 50-100 ℃, and the reaction time is 24-72 hours; the reaction is carried out in a solvent which may be methanol, ethanol, tetrahydrofuran, acetonitrile, etc., preferably ethanol.
In the step 2), the reaction conditions of the condensation reaction are as follows: the reaction temperature is 50-100 ℃, and the reaction time is 1-3 hours; the reaction is carried out in a solvent which may be methanol, ethanol, tetrahydrofuran, acetonitrile, etc., preferably ethanol.
In the step 3), the reaction conditions of the ring closure reaction are as follows: the reaction temperature is 50-100 ℃, and the reaction time is 3-6 hours; the reaction is carried out in a solvent which may be methanol, ethanol, tetrahydrofuran, acetonitrile, etc., preferably ethanol.
In the step 4), the reaction conditions of the condensation reaction are as follows: the reaction temperature is 0-25 ℃, and the reaction time is 2-8 hours; the reaction is carried out in a solvent which can be dichloromethane, tetrahydrofuran, acetonitrile and the like, and dichloromethane is preferred; .
Other compounds claimed in the claims of the present invention can be obtained by reference to the preparation process of the examples of the present invention.
Another object of the present invention is to provide the use of the compounds of formula I as described above.
The application provided by the invention is that the compound shown in the formula I or the pharmaceutically acceptable salt, ester and solvate thereof is applied to the following (a) and/or (b) and/or (c):
(a) the application of the compound shown in the formula I or the pharmaceutically acceptable salt, ester and solvate thereof in preparing products for treating diseases caused by viruses or viral infection;
(b) the application of the compound shown in the formula I or the pharmaceutically acceptable salt, ester and solvate thereof in preparing products for preventing diseases caused by viruses or viral infection;
(c) the application of the compound shown in the formula I or pharmaceutically acceptable salts, esters and solvates thereof in preparing a virus inhibitor.
The product may be a medicament or a pharmaceutical formulation.
The viral inhibitor is capable of inhibiting viral replication.
The virus includes influenza virus and coronavirus.
The influenza virus may be specifically influenza a virus (H1N 1);
the coronavirus may be an alpha coronavirus and/or a beta coronavirus, and is specifically selected from HCoV-229E.
In the present invention, the disease caused by the virus may be a respiratory infectious disease.
The respiratory system infection is respiratory tract infection and/or lung infection; the respiratory tract infection can be nasopharyngitis, rhinitis, pharyngolaryngitis, tracheitis and/or bronchitis; the pulmonary infection may be pneumonia.
In the present invention, the diseases caused by influenza virus generally include acute respiratory infectious diseases caused by influenza virus and the like.
In the present invention, the diseases caused by coronavirus generally include viral pneumonia, severe acute respiratory syndrome, and the like.
In the present invention, the coronavirus infection usually causes viral pneumonia, severe acute respiratory syndrome and the like.
The compound of the invention has the inhibition effect on coronavirus and H1N1 influenza A virus, has no toxicity on normal cells of human, can inhibit the generation degree of inflammatory reaction, reduces the damage of pneumonia to organisms and promotes the recovery of the organisms.
The antiviral drug prepared by using the compound shown in the formula I as an active ingredient also belongs to the protection scope of the invention.
The antiviral drug can be introduced into body such as muscle, intradermal, subcutaneous, intravenous, mucosal tissue by injection, spray, nasal drop, eye drop, penetration, absorption, physical or chemical mediated method; or mixed or coated with other materials and introduced into body.
If necessary, one or more pharmaceutically acceptable carriers can be added into the medicine. The carrier includes diluent, excipient, filler, binder, wetting agent, disintegrating agent, absorption enhancer, surfactant, adsorption carrier, lubricant, etc. which are conventional in the pharmaceutical field.
The above medicine can be made into tablet, powder, granule, capsule, oral liquid, unguent, cream, injection, etc.; the medicaments in various dosage forms can be prepared according to the conventional method in the pharmaceutical field.
The invention also provides a medicine or a medicine composition, and the active ingredient of the medicine or the medicine composition is a compound shown in the formula I or pharmaceutically acceptable salt, ester and solvate thereof.
The medicament or the medicament composition has at least one of the following effects:
1) treating a disease or viral infection caused by a virus;
2) preventing a disease or viral infection caused by a virus;
3) inhibiting viruses.
The above-mentioned drugs or pharmaceutical compositions can be prepared into dosage forms such as solutions, tablets, capsules or injections according to conventional methods known to those skilled in the art.
When the compound shown in the formula I or the pharmaceutically acceptable salt thereof provided by the invention is used for preventing and/or treating infection caused by virus, an effective amount of the compound shown in the formula I or the pharmaceutically acceptable salt thereof is administered to a subject organism.
Experiments prove that the compound has a good inhibitory effect on H1N1 influenza A virus and coronavirus, does not have toxicity to normal human cells, and can inhibit the virus and the inflammatory reaction.
Drawings
FIG. 1 is a scheme showing the synthesis of the compound of formula I according to the present invention.
Detailed Description
The present invention will be further illustrated with reference to the following specific examples, but the present invention is not limited to the following examples. The method is a conventional method unless otherwise specified. The starting materials are commercially available from the open literature unless otherwise specified.
Examples 1 to 13
1. Synthesis of (E) -2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) benzamide (ZONK2003-1)
1) 2-Methylaminothiourea (1-2)
Methylhydrazine (23.0g, 0.5mol), ammonium thiocyanate (38.0g, 0.5mol) and ethanol (200mL) were added to a single-neck round-bottom flask, and the mixture was heated under reflux for 72 hours. The reaction solution was cooled, concentrated and purified by column chromatography to give 2-methylaminothiourea (44.1g, 84.0%) as an off-white solid.1H NMR(DMSO-d6 400MHz)δ7.24(s,2H),6.85(s,2H),3.14(s,3H).ESI-MS m/z:106.1[M+H]+.
2) (E) -2- ((2-aminomethylthiono-2-methylhydrazono) methyl) benzoic acid (1-3)
2-Methylaminothiourea (40.0g, 0.38mol), 2-carbobenzoic acid (57.0g, 0.38mol) and ethanol (300mL) were added to a single-neck round-bottom flask, and the mixture was refluxed for 2 hours. The reaction solution was cooled, concentrated and purified by column chromatography to give (E) -2- ((2-aminomethylthiaacyl-2-methylhydrazono) methyl) benzoic acid (85.6g, 95.0%) as a pale yellow solid.1H NMR(DMSO-d6 400MHz)δ13.0(s,1H),8.12-7.23(m,7H),2.47(s,3H).ESI-MS m/z:238.1[M+H]+.
3) (E) -2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) benzoic acid (1-4)
(E) -2- ((2-aminomethylthiaacyl-2-methylhydrazono) methyl) benzoic acid (80.0g, 0.34mol), 2-bromo-1- (2-chlorophenyl) ethanone (79.2g, 0.34mol) and ethanol (400mL) were added to a single-neck round-bottom flask, and the mixture was refluxed for 3 hours. The reaction solution was cooled, concentrated and purified by column chromatography to give (E) -2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) benzoic acid (124g, 98.5%) as a pale yellow solid.1H NMR(DMSO-d6 400MHz)δ13.23(s,1H),8.60(s,1H),8.01-7.34(m,9H),3.66(s,3H).ESI-MS m/z:372.1[M+H]+.
4) (E) -2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) benzamide
(E) -2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) benzoic acid (3.71g, 10mmol) was dissolved in dichloromethane (30mL), N-hydroxysuccinimide (1.15g, 10mmol), dicyclohexylcarbodiimide (2.27g, 11mmol) were added, respectively, and the mixture was stirred at room temperature for 3 h. After TLC detection of disappearance of the starting material, concentrated ammonia (3mL) was added and the reaction was continued at room temperature for 5 h. The reaction was concentrated and purified by column chromatography to give (E) -2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) benzamide (3.16g, 85.5%) as a pale yellow solid.1H NMR(DMSO-d6 400MHz)δ8.24(s,1H),8.00-7.34(m,11H),3.64(s,3H).ESI-MS m/z:371.1[M+H]+.
2. Synthesis of (E) -2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) -N- (methylsulfonyl) benzamide (ZONK2003-2)
(E) -2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) benzoic acid (3.71g, 10mmol), 4-dimethylaminopyridine (1.22g, 10mmol), aminomethanesulfonic acid (1.11g, 10mmol) were dissolved in dichloromethane (30mL), followed by addition of dicyclohexylcarbodiimide (2.27g, 11mmol) and stirring at room temperature for 5 h. The reaction was concentrated and purified by column chromatography to give (E) -2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) -N- (methylsulfonyl) benzamide (3.41g, 76.2%) as a pale yellow solid.1H NMR(DMSO-d6 400MHz)δ8.10(s,1H),8.00-7.43(m,9H),3.67(s,3H),3.42(s,3H).ESI-MS m/z:449.1[M+H]+.
3. Synthesis of (E) -diisopropyl (2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) benzoyl) phosphoramidate (ZONK2003-3)
(E) -2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) benzamide (3.70g, 10mmol), triethylamine (2.0g, 20mmol) were dissolved in dichloromethane (20mL), respectively, and a solution of diisopropyl chlorophosphate (2.20g, 11mmol) in dichloromethane (10mL) was added dropwise and stirred at room temperature for 3 h. The reaction was concentrated and purified by column chromatography to give (E) -diisopropyl (2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) benzoyl) phosphoramidate (3.41g, 76.2%) as a pale yellow solid.1H NMR(DMSO-d6400MHz)δ8.15(s,1H),8.00-7.43(m,9H),4.11(m,2H),3.67(s,3H),1.28(s,12H).ESI-MS m/z:535.1[M+H]+.
4. Synthesis of (E) -2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) -N- (ethylsulfonyl) benzylamine (ZONK2003-4)
By combining the procedures of examples 1 to 3, (E) -2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) -N- (ethylsulfonyl) benzylamine was synthesized.1H NMR(DMSO-d6 400MHz)δ8.10(s,1H),8.00-7.33(m,9H),3.45(m,2H),3.27(s,3H),1.23(m,3H).ESI-MS m/z:463.1[M+H]+.
5. Synthesis of (E) -N-butyryl-2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) benzamide (ZONK2003-5)
By combining the procedures of examples 1 to 3, (E) -N-butyryl-2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) benzamide was synthesized.1H NMR(DMSO-d6 400MHz)δ8.10(s,1H),8.00-7.33(m,9H),3.27(s,3H),2.25(m,2H),1.23(m,2H),0.90(m,3H).ESI-MS m/z:441.1[M+H]+.
6. Synthesis of (E) -2-hydroxyethyl 2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) benzoate (ZONK2003-6)
By combining the preparation methods of examples 1 to 3, (E) -2-hydroxyethyl 2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) benzoate was synthesized.1H NMR(DMSO-d6 400MHz)δ8.20(s,1H),8.01-7.34(m,9H),5.54(s,1H),4.39-3.87(m,4H),3.54(s,3H).ESI-MS m/z:416.1[M+H]+.
7. Synthesis of (E) -2- (methylamino) ethyl 2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) hydrazono) methyl) -5-fluorobenzoate (ZONK2003-7)
By combining the procedures of examples 1 to 3, (E) -2- (methylamino) ethyl 2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) hydrazono) methyl) -5-fluorobenzoic acid ester was synthesized.1H NMR(DMSO-d6 400MHz)δ8.20(s,1H),8.01-7.34(m,8H),4.54(m,2H),3.56(m,2H),3.24(s,3H).ESI-MS m/z:433.1[M+H]+.
8. Synthesis of (E) -3- (dimethylamino) propyl 2- ((2- (4- (4-chlorophenyl) thiazol-2-yl) hydrazono) methyl) -5-fluorobenzoate (ZONK2003-8)
By combining the preparation methods of examples 1 to 3, (E) -3- (dimethylamino) propyl 2- ((2- (4- (4-chlorophenyl) thiazol-2-yl) hydrazono) methyl) -5-fluorobenzoic acid ester was synthesized.1H NMR(DMSO-d6 400MHz)δ8.20(s,1H),8.01-7.34(m,8H),4.55(m,2H),3.52(m,2H),3.24(s,6H),1.86(m,2H).ESI-MS m/z:461.1[M+H]+.
9. Synthesis of (E) -2- (diethylamino) ethyl 2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) benzoate (ZONK2003-9)
By combining the preparation methods of examples 1 to 3, (E) -2- (diethylamino) ethyl 2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) benzoate was synthesized.1H NMR(DMSO-d6 400MHz)δ8.20(s,1H),8.01-7.34(m,9H),4.55(m,2H),3.52(s,3H),3.24-3.01(m,6H),1.12(m,6H).ESI-MS m/z:471.1[M+H]+.
10. Synthesis of (E) -2- (pyrrolidin-1-yl) ethyl 2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) benzoate (ZONK2003-10)
By combining the procedures of examples 1 to 3, (E) -2- (pyrrolidin-1-yl) ethyl 2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) -2-methylhydrazono) methyl) benzoate was synthesized.1H NMR(DMSO-d6 400MHz)δ8.20(s,1H),8.01-7.34(m,9H),4.55(m,2H),3.52(s,3H),3.24-3.01(m,6H),1.68(m,4H).ESI-MS m/z:469.1[M+H]+.
11. Synthesis of (E) -2- (piperidin-1-yl) ethyl 2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) hydrazono) methyl) benzoate (ZONK2003-11)
By combining the procedures of examples 1 to 3, (E) -2- (piperidin-1-yl) ethyl 2- ((2- (4- (2-chlorophenyl) thiazol-2-yl) hydrazono) methyl) benzoate was synthesized.1H NMR(DMSO-d6 400MHz)δ8.20(s,1H),8.01-7.34(m,9H),4.55-3.54(m,8H),1.86-1.55(m,6H).ESI-MS m/z:469.1[M+H]+.
12. Synthesis of (E) -2-morpholinoethyl 2- ((2- (4- (2-fluorophenyl) thiazol-2-yl) hydrazono) methyl) benzoate (ZONK2003-12)
By combining the preparation methods of examples 1 to 3, (E) -2-morpholinoethyl 2- ((2- (4- (2-fluorophenyl) thiazol-2-yl) hydrazono) methyl) benzoate was synthesized.1H NMR(DMSO-d6 400MHz)δ8.20(s,1H),8.01-7.34(m,9H),4.55-3.54(m,12H).ESI-MS m/z:455.1[M+H]+.
13. Synthesis of (E) -2- (piperazin-1-yl) ethyl 2- ((2-methyl-2- (4-phenylthiazol-2-yl) hydrazono) methyl) benzoate (ZONK2003-13)
By combining the preparation methods of examples 1 to 3, (E) -2- (piperazin-1-yl) ethyl 2- ((2-methyl-2- (4-phenylthiazol-2-yl) hydrazono) methyl) benzoate was synthesized.1H NMR(DMSO-d6 400MHz)δ8.20(s,1H),8.01-7.11(m,10H),4.55-3.54(m,12H),3.31(s,3H).ESI-MS m/z:450.1[M+H]+.
EXAMPLE 14 series of Compounds in vitro antiviral assay
The toxicity of ZONK2003 series compounds (compounds shown in formula I) on virus-cultured MRC-5 and MDCK cells is determined by MTT method, and antiviral test is carried out by designing proper concentration. In vitro antiviral pharmacodynamic tests were performed by cytopathic effect (CPE) method to evaluate the effect of ZONK2003 series compounds on viruses causing respiratory diseases.
And (3) test results: the ZONK2003 series compounds have no obvious toxicity to MRC-5 and MDCK cells; has different degrees of antiviral effects on two viruses, and has half effective rate (EC)50) The antiviral effect was evaluated. EC for coronavirus (HCoV-229E)5012.83 mu mol/L, 13.70 mu mol/L and 15.00 mu mol/L; EC for influenza A virus (H1N1)5011.75. mu. mol/L, 10.00. mu. mol/L, 18.19. mu. mol/L.
And (4) test conclusion: within the concentration range of 10-20 mu mol/L, the ZONK2003 series compounds have different degrees of inhibition effects on two strains of viruses, and can promote the recovery of cells after the cells are infected by the viruses.
1. Test materials
1.1 test substances
The test substance: ZONK2003-4, batch number: 20200521, purity 98.70%, molecular weight 371.8; ZONK2003-1, batch number: 20200525, purity 99.61%, molecular weight 448.9, ZONK2003-2, batch number: 20200513, purity 99.01%, molecular weight 370.8, supplied by Guangdong Zhongke pharmaceutical research Co., Ltd. Prepared into 50.0, 25.0, 12.5, 6.25, 3.13 and 1.56 mu mol/L by DMEM medium, and stored for later use at the temperature of 2-8 ℃.
1.2 Positive control
Oseltamivir phosphate granules with the specification of 15mg multiplied by 10 are bagged, and the product of Yangtong sunshine Yangtze river pharmaceutical industry Limited company with the batch number: 0371912115, expiration date to 2021.12.11, for a positive control against influenza a virus; recombinant human interferon alpha 2b gel with specification of 1.0 × 105IU/g, 10 g/count, Megaku pharmaceuticals (fertilizer combination) Co., Ltd, for use as a positive control against coronavirus.
1.3 Virus strains
Coronavirus (HCoV-229E), accession number: ATCC-VR-740, available from ATCC in the United states; influenza A virus (H1N1), accession number ATCC-VR-95, was purchased from ATCC in the United states. All cultured and stored in the company II level biological safety laboratory.
1.4 cell lines
Human embryonic lung cells (MRC-5), canine kidney cells (MDCK), were obtained from Wuhan Protech Life technologies, Inc.
1.5 Primary reagents
1.6 Main instruments
BSCIIB2-1101 type biological safety cabinet, manufactured by Shanghai Ruiyang purification equipment Limited, center number: 019, YXQ-50A model vertical pressure steam sterilizer, manufactured by Shanghai Bochwisdom medical Bio-instruments GmbH, center number: 584;3111 type CO2Incubator, manufactured by thermo fisher corporation, usa, center number: 147.
1.7 test facility
Changsha pathogenic microorganism laboratory, laboratory grade: biosafety class II (BSL-II), docket number: changwaijie Ji Zi (2019) No. B001. The test was conducted in the central biosafety class II laboratory (BSL-II-1), performed strictly in accordance with regulations and the central SOPs.
2. Test method
2.1 cell culture
Human embryonic lung cells (MRC-5) are anchorage-dependent growing cells. The culture medium is DMEM containing 10% FBS, and when the growth state is good, passage can be performed every 2-3 d. Removing the culture medium in a purification workbench, washing with 1 × PBS for 2-3 times, then adding an appropriate amount of 0.25% Trypsin-EDTA for digestion, after about 1-3 min, adding an appropriate amount of DMEM culture medium containing 10% FBS to stop the digestion of pancreatin, blowing and beating into a single cell suspension, transferring into an EP tube, and centrifuging at 1000rpm for 5 min. Discarding the culture medium, adding fresh culture medium for re-suspension, and proportionally (the cell density is about 10)5/mL) was inoculated into a new flask and placed at 37 ℃ with 5% CO2Culturing in an incubator.
Canine kidney cells (MDCK) are anchorage-dependent growing cells. The culture medium is DMEM containing 10% FBS, and when the growth state is good, passage can be performed every 2-3 d. Removing the culture medium in a purification workbench, washing with 1 × PBS for 2-3 times, then adding an appropriate amount of 0.25% Trypsin-EDTA for digestion, after about 2-5 min, adding an appropriate amount of DMEM culture medium containing 10% FBS to stop the digestion of pancreatin, blowing and beating into a single cell suspension, transferring into an EP tube, and centrifuging at 1000rpm for 5 min. Discarding the culture medium, adding fresh culture medium for re-suspension, and proportionally (the cell density is about 10)5/mL) was inoculated into a new flask and placed at 37 ℃ with 5% CO2Culturing in an incubator.
2.2 amplification of viruses
2.2.1 coronavirus (HCoV-229E): MRC-5 cell expansion
MRC-5 cells were seeded at 75cm2CulturingIn a bottle, when the cell density reaches 80-90%, removing part of the culture medium, covering the rest cells, adding appropriate amount of HCoV-229E virus, after the virus is adsorbed on the cell surface (about 3h, gently shaking the culture plate every 30min to make the virus adsorbed uniformly), replacing the fresh culture medium without FBS, placing at 35 ℃ and 5% CO2Culturing in a humidified constant-temperature incubator. And (3) observing that the cells start to generate lesions until the cells do not generate lesions (generally 5-7 days), performing repeated freeze thawing method, separating at 3000rpm for 10min to remove cell residues, collecting supernate, subpackaging in a freezing storage tube, labeling, and then performing medium-short term storage at-80 ℃ or medium-long term storage in liquid nitrogen for later use. 2.2.2 influenza A Virus (H1N 1): MDCK cell expansion
MDCK cells were seeded at 75cm2In a culture bottle, when the cell density reaches 70-80%, removing part of the culture medium, covering the rest cells, adding a proper amount of H1N1 virus, after the virus is adsorbed on the cell surface (about 3H, gently shaking the culture plate every 30min to make the virus adsorbed uniformly), replacing the fresh culture medium without FBS, placing at 33 ℃ and 5% CO2Culturing in a humidified constant-temperature incubator. Observing that the cells start to generate lesions until the cells do not generate lesions (generally 2-3 days), adopting a repeated freeze thawing method, separating at 3000rpm for 10min to remove cell residues, collecting supernate, subpackaging in a freezing storage tube, labeling, and then storing at-80 ℃ for a medium-short period or in liquid nitrogen for a long period for later use.
2.3 Virus half Tissue culture infection concentration (TCID)50) Measurement of (2)
TCID was performed on the virus solutions collected at 2.2.1 and 2.2.250The determination of (1): inoculating 100 μ L of corresponding cell suspension with appropriate density into 96-well cell culture plate, culturing for 24 hr, sucking out culture solution, adding 100 μ L of virus solution diluted with cell maintenance solution (virus is carried out 10 times)-1、10-2、10-3、10-4、10-5、10-6Gradient dilution) were performed in duplicate, 10 wells for each dilution, 33 ℃, 5% CO2Adsorbing and culturing for 3h in an incubator, adsorbing unadsorbed virus liquid, supplementing 100 mu L of cell vitamin growth liquid to each hole, and continuing culturing. On a daily basisObserving cytopathic effect (CPE, the characteristic of CPE caused by virus in cultured cells, namely cell rounding, stronger refractivity, fused cell protrusion, partial wall separation, filamentous protrusion or pseudo-podiform cytoplasm, irregular map of the whole shape, and formation of large and round fused multinuclear giant cells) under an inverted microscope. And the number of wells with CPE was recorded, ending with the highest dilution no longer presenting with lesions, and the extent of cytopathic effect is indicated by "- + + + + + + +": no cytopathy "-", < 25% cytopathy "+", 25% -50% cytopathy "+", 50% -75% cytopathy "+ + +",>75% cytopathic "+++". Calculating TCID according to Reed-Muench formula50：
TCID50Log (dilution of virus with CPE less than 50% + distance ratio x dilution spacing
Wherein the distance ratio is (percentage higher than 50% -50) ÷ (percentage higher than 50% -percentage lower than 50%)
2.4 concentration profiling of test substances
2.4.1 cytotoxicity assays
ZONK2003 series compound (concentration gradient of 50.0, 25.0, 12.5, 6.25, 3.13, 1.56. mu. mol/L) and oseltamivir phosphate (concentration gradient of 20.0, 10.0, 5.0, 2.5, 1.25, 0.625. mu.g/mL) and interferon alpha 2b (concentration gradient of 1 × 10) were prepared in DMEM medium in series3、5×102、2.5×102、1.25×102、6.25×101、3.1×101IU/mL), added to the cultured MRC-5 and MDCK cells, respectively, at 37 deg.C and 5% CO2Culturing in humidified incubator for 72h, adding MTT, culturing for 4h, measuring OD value of each well at 492nm wavelength, and calculating EC of ZONK2003 series compounds on each cell50。
2.4.2 when the compound is cytotoxic 1/2IC is used50For the highest concentration, 3 test concentrations were set diluted down at 2-fold intervals; when the compounds are not toxic to cells, the highest concentration of 20 μmol/L is used, and 3 concentrations tested are set by dilution down at 2-fold intervals.
2.5 detection of antiviral Effect
2.5.1 cell seeding: 24h before virus infection, corresponding cell strains which grow well are used for controlling the appropriate density (about 10)5One/well) were inoculated in a 96-well plate at 100. mu.L/well in 5% CO at 37 ℃2Culturing in an incubator;
2.5.2 after the cells reach about 70% -80%, sucking out part of the culture medium, covering the cells (making the viruses and the cells better adsorbed), and inoculating 100TCID50After culturing the virus solution (50. mu.L/well) in an incubator for 3 hours, the medium in a 96-well plate was aspirated, and a sample solution (set according to cytotoxicity) of 200. mu.L/well prepared with a serum-free cell maintenance solution and having different concentrations was added thereto at 33 ℃ and 5% CO2Culturing in an incubator.
2.5.3 grouping
Normal control group: group not infected with virus;
model control group: a virus-infected group;
positive control group: a commercially available control drug;
test group: infection group + different concentrations of test substance.
2.5.4 evaluation of antiviral Activity (CPE method)
Cytopathic effect was observed day by day, and the number of cytopathic and non-cytopathic wells was recorded for each concentration, with continuous observation that the cytopathic effect did not increase.
2.6 evaluation of results
The rate of cell lesion inhibition (%) was 100% based on the normal control cell lesion-free rate of 100% (1-each group of cells lesion-free wells/8) × 100%.
The test data significant figure rounding was done by rounding and the software used was counted as SPSS 16.0. The measured data is averaged + -SD
Showing that the method of Leven's test is used for checking the normality and the homogeneity of variance. If it has no statistical significance (P)>0.05), statistical analysis was performed using one-way analysis of variance (ANOVA). If ANOVA has statistical significance (P is less than or equal to 0.05), the LSD method is used for comparative analysis. If the variance is not uniform (P is less than 0.05), the test is performed by Kruskal-WallisAnd (6) testing. If the Kruskal-Wallis Test is statistically significant (P.ltoreq.0.05), a comparative analysis is performed using Dunnett's Test (nonparametric method). The statistical results are tested with α ≦ 0.05, where P ≦ 0.05 indicates statistical significance and P ≦ 0.01 indicates that the difference tested was of significant significance.
3. Test results
3.1 Effect of series of Compounds on proliferation of individual cells
As shown in tables 1-1 and 1-2, the three compounds have no obvious cytotoxicity to MRC-5 and MDCK within the tested concentration range (1.56-50.0 mu mol/L), so the highest concentration is set to be 20.0 mu mol/L in antiviral research. As shown in Table 2, the positive drug had interferon alpha 2b (6.25X 10) in the range of concentration tested1～1×103IU/mL) has no obvious cytotoxicity to MRC-5 cells and oseltamivir phosphate (0.625-20.0 mu mol/L) to MDCK cells, so the highest concentration is respectively set to be 1 multiplied by 10 in antiviral research3IU/mL、20.0μmol/L。
TABLE 1-1 Effect of three Compounds on MRC-5 cell proliferation
Tables 1-2 Effect of three Compounds on MDCK cell proliferation
TABLE 2 Effect of Positive drugs on proliferation of individual cells
3.2 Virus titer results detection
As shown in tables 3 and 4, the crownHalf of the viral infectivity TCID of the Toxovirus (HCoV-229E) and influenza A virus (H1N1)50Are respectively 10-3.5/0.1mL、10-3.850.1mL, 3.16X 10 virus doses respectively4Multiple, 7.08 × 104When diluted twice, 0.1mL of inoculated cells can cause lesion of 50% of cells. Get 100 TCIDs50Viral load, i.e., 316-fold and 708-fold dilutions, respectively, were tested for in vitro antiviral assays.
TABLE 3 amount of HCoV-229E infected with half of the viruses of MRC-5 cells
TABLE 4 amount of infection of MDCK cells with half of the virus by H1N1
3.3 Effect of ZONK2003 series Compounds on cell viability following Virus infection
EC of compounds ZONK2003-4, ZONK2003-1, ZONK2003-2 against coronavirus (HCoV-229E)50Respectively 12.83 mu mol/L, 13.70 mu mol/L and 15.00 mu mol/L; anti-influenza A virus (H1N1) EC for compounds ZONK2003-4, ZONK2003-1, ZONK2003-25011.75. mu. mol/L, 10.00. mu. mol/L and 18.19. mu. mol/L, respectively.
EC of interferon alpha 2b against coronavirus (HCoV-229E)50Is 2.42 multiplied by 102IU/mL; EC of Oseltamivir phosphate against influenza A virus (H1N1)50It was 5.06. mu. mol/L.
TABLE 5-1 Effect of test/control on HCoV-229E infection of MRC-5 cells
TABLE 6-1 Effect of three Compounds on H1N1 infection of MDCK cells
4. Conclusion and evaluation
Under the present test conditions:
the three compounds ZONK2003-4, ZONK2003-1 and ZONK2003-2 have no obvious cytotoxicity to MRC-5 and MDCK cells within the tested concentration range (1.56-50.0 mu mol/L). The positive control drug has no obvious toxicity to MRC-5 and MDCK cells within the test concentration range (0.625-20 mmol/L).
The three compounds ZONK2003-4, ZONK2003-1 and ZONK2003-2 have different degrees of inhibitory effects on two strains of viruses in the test, and have EC (inhibitory activity) on coronavirus (HCoV-229E)50Respectively 12.83 mu mol/L, 13.70 mu mol/L and 15.00 mu mol/L; EC for influenza A virus (H1N1)5011.75. mu. mol/L, 10.00. mu. mol/L and 18.19. mu. mol/L, respectively.
The positive control drug has different degrees of inhibition on two strains of virus in the experiment, and has EC on coronavirus (HCoV-229E)50Is 2.42 multiplied by 102IU/mL; EC of Oseltamivir phosphate against influenza A virus (H1N1)50It was 5.06. mu. mol/L.
EXAMPLE 15 protective Effect of Compounds on influenza A Virus A/FM/1/47(H1N1) infected mice
The test substance: ZONK2003-1 to 10 (which in turn correspond to the compounds of examples 1-10); supplied by Guangdong Chinese medicine research Co.
Oseltamivir phosphate granules with the specification of 15mg multiplied by 10 are bagged, and the product of Yangtong sunshine Yangtze river pharmaceutical industry Limited company with the batch number: 0371912115, expiration date to 2021.12.11, for a positive control against influenza a virus;
experimental Material: influenza A virus mouse lung adapted strain A/FM/1/47(H1N1), inoculated with chick embryo, collected allantoic fluid and stored. ICR mice, weight 18 ~ 22 g. During the administration period, food and water are taken freely, the light is 12 hours a day, the dark is 12 hours a day, the temperature is 22 +/-2 ℃, and the humidity is 55-70%. The experimental method comprises the following steps: after 3 days of acclimatization, the experiment was started. Except for uninfected control group, mice in each group were lightly anesthetized with ether, and inoculated intranasally with a solution of 8 × LD diluted with physiological saline5050 μ L/mouse of allantoic fluid of chick embryo of influenza A/FM/1/47(H1N1), mice of the positive control oseltamivir group and the example compound group were first gavaged 2H after infection, and each compound was orally administered at a dose of 10 μmol/kg, 20 μmol/kg, and 30 μmol/kg twice daily for 5 days. The survival of the mice was observed within 14 days,
and the mortality protection rate of the drug for mice was calculated (mortality protection rate ═ model group mortality rate — experimental group mortality rate).
Influenza A virus (H1N1 influenza A virus)
Example 16: relieving effect of drug on mouse lung inflammation caused by influenza virus H1N1 infection
The experimental method comprises the following steps: after 3 days of acclimatization, the experiment was started. Except for uninfected control group, mice in each group were lightly anesthetized with ether, and inoculated intranasally with a solution of 8 × LD diluted with physiological saline5050 mu L/mouse of allantoic fluid of chick embryo of influenza A virus/FM/1/47 (H1N1), mice of positive control oseltamivir group and test administration group were first gavaged with 80mg/kg 24H after virus infection, and then 1 time a day, and the virus control group and uninfected control group were orally administered with physiological saline by the same method 1 time a day in an administration volume of 0.1mL/10g body weight. For a total of 5 days. Weighing 3 mice in each group on day 6, removing eyeballAnd (4) killing blood, taking out the whole lung, weighing, and calculating the lung index and the inhibition rate of the lung index.
Percent weight loss-weight before dosing-weight after dosing/weight before dosing x 100%
Lung index ═ lung weight of mouse/body weight of mouse × 100
Pulmonary index inhibition (%) is mean pulmonary index of virus control group-mean pulmonary index of administration group/mean pulmonary index of virus control group × 100%
| Group of | Percentage of weight loss% | Index of lung | Lung index inhibition% |
| Normal control group | 0.57 | ||
| Model control group | 2.09 | ||
| Oseltamivir group | 15.12 | 1.66 | 20.57 |
| ZONK2003-1 | 10.23 | 1.16 | 45.14 |
| ZONK2003-2 | 11.45 | 1.23 | 41.14 |
| ZONK2003-3 | 10.87 | 1.05 | 49.76 |
| ZONK2003-4 | 9.56 | 1.18 | 43.54 |
| ZONK2003-5 | 9.89 | 1.20 | 42.58 |
| ZONK2003-6 | 11.58 | 1.16 | 44.49 |
| ZONK2003-7 | 12.03 | 1.01 | 51.67 |
| ZONK2003-8 | 9.78 | 1.25 | 40.19 |
| ZONK2003-9 | 12.45 | 1.19 | 43.06 |
| ZONK2003-10 | 11.02 | 1.08 | 48.32 |
The experimental results are as follows: therefore, the compound of the embodiment has obvious protection and inhibition effects on lung inflammation caused by influenza virus, and the effect is better than that of an oseltamivir control group.
Claims (10)
1. A compound with a structural general formula shown in formula I or pharmaceutically acceptable salts, esters and solvates thereof:
in the formula (I), the compound has the following structure,
R1selected from: mono-or poly-substituted H, F, methyl, trifluoromethyl;
R2selected from: H. linear or substituted C1-C6 alkanes;
R3selected from: mono-or polysubstituted H, Cl, Br, F;
when X is NH, R4Selected from: H. an acyl group; the acyl group comprises a sulfonyl group, a phosphoryl group or an alkanoyl group;
when X ═ O, R4Is composed of
Wherein n is an integer of 0 to 6,
y is selected from O or N;
R5、R6independently selected from any one of the following groups: H. (C)1-C6) Alkyl, (C)3-C8) Carbocyclylalkyl group, (C) containing substituent1-C18) Alkane of (C)2-C8) Alkenyl group, (C) having substituent2-C8) Alkenyl, (C)2-C8) Alkynyl group, substituent-containing (C)2-C8) Alkynyl, (C)6-C20) Aryl group, substituted group-containing (C)6-C20) Aryl group, (C)2-C20) Heterocyclic group, (C) containing substituent2-C20) A heterocyclic group; or R5、R6Form a ring with each other to form (C)3-C8) Heterocycloalkyl or substituted (C)3-C8) Heterocycloalkyl group, (C)6-C20) Heteroaryl or substituted (C)6-C20) A heteroaryl group.
The R is5、R6Wherein said substituent is selected from at least one of: h, methyl, ethyl, isopropyl, pyrrolyl, piperidinyl, morpholinyl, piperazinyl.
2. The compound according to claim 1, or a pharmaceutically acceptable salt, ester, solvate thereof, wherein:
the R is1Is selected from H;
or, said R2Selected from methyl or isopropyl;
or, said R3Selected from Cl;
or, when said X ═ NH, R4Selected from H or methylsulfonyl.
3. A process for the preparation of a compound of formula I as claimed in claim 1 or 2, comprising the steps of:
1) reacting the compound shown in the formula a with sodium thiocyanate to obtain a compound shown in a formula b;
wherein R in the formula a2Is as defined in formula I; in the formula b, R2Is as defined in formula a;
2) reacting a compound represented by the formula b with R represented by the formula e1Carrying out condensation reaction on substituted o-benzoic acid benzaldehyde to obtain a compound shown as a formula c;
wherein R in the formula e1Is as defined in formula I; in the formula c, R1Is defined by the formula e, R2Is as defined in formula b;
3) reacting a compound represented by the formula c with R represented by the formula f3Substituting bromoacetophenone for carrying out a ring closure reaction to obtain a compound shown as a formula d;
wherein R in the formula f3Is as defined in formula I, formula d wherein R1、R2Is defined by the formula c, R3Is as defined in formula f;
4) carrying out condensation reaction on the compound shown in the formula d and the compound shown in the formula g under the action of a condensing agent to obtain a compound shown in the formula I;
H-X-R4(formula g)
Wherein, X, R in the formula g4Is as defined in formula I.
4. The method of claim 3, wherein:
in the step 1), the reaction conditions of the reaction are as follows: the reaction temperature is 50-100 ℃, and the reaction time is 24-72 hours; the reaction is carried out in a solvent selected from any one of the following: methanol, ethanol, tetrahydrofuran, acetonitrile, etc., preferably ethanol;
in the step 2), the reaction conditions of the condensation reaction are as follows: the reaction temperature is 50-100 ℃, and the reaction time is 1-3 hours; the reaction is carried out in a solvent selected from any one of the following: methanol, ethanol, tetrahydrofuran, acetonitrile, preferably ethanol;
in the step 3), the reaction conditions of the ring closing reaction are as follows: the reaction temperature is 50-100 ℃, and the reaction time is 3-6 hours; the reaction is carried out in a solvent selected from any one of the following: methanol, ethanol, tetrahydrofuran, acetonitrile, preferably ethanol;
in the step 4), the reaction conditions of the condensation reaction are as follows: the reaction temperature is 0-25 ℃, and the reaction time is 2-8 hours; the reaction is carried out in a solvent selected from any one of the following: dichloromethane, tetrahydrofuran, acetonitrile, preferably dichloromethane.
5. The compound of formula I according to claim 1, or a pharmaceutically acceptable salt, ester or solvate thereof, wherein the compound is used as (a) and/or (b) and/or (c):
(a) the application of the compound shown in the formula I or the pharmaceutically acceptable salt, ester and solvate thereof in preparing products for treating diseases caused by viruses or viral infection;
(b) the application of the compound shown in the formula I or the pharmaceutically acceptable salt, ester and solvate thereof in preparing products for preventing diseases caused by viruses or viral infection;
(c) the application of the compound shown in the formula I or pharmaceutically acceptable salts, esters and solvates thereof in preparing a virus inhibitor.
6. Use according to claim 5, characterized in that:
the product is a medicament or pharmaceutical formulation; the viral inhibitor is capable of inhibiting viral replication;
the virus includes influenza virus and coronavirus.
7. Use according to claim 6, characterized in that: the influenza virus is influenza A virus; the coronavirus is HCoV-229E.
8. A medicine or pharmaceutical composition comprises compound shown in formula I or its pharmaceutically acceptable salt, ester, and solvate as active ingredient;
the medicament or the medicament composition has at least one of the following effects:
1) treating a disease or viral infection caused by a virus;
2) preventing a disease or viral infection caused by a virus;
3) inhibiting viruses.
9. The medicament or pharmaceutical composition of claim 8, wherein:
the product is a medicament or pharmaceutical formulation; the viral inhibitor is capable of inhibiting viral replication;
the virus includes influenza virus and coronavirus.
10. The medicament or pharmaceutical composition of claim 9, wherein: the influenza virus is influenza A virus; the coronavirus is HCoV-229E.
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|---|---|---|---|---|
| CN113773275A (en) * | 2020-12-29 | 2021-12-10 | 广东中科药物研究有限公司 | Antiviral compound and preparation method thereof |
| WO2022142727A1 (en) * | 2020-12-29 | 2022-07-07 | 广东中科药物研究有限公司 | Antiviral compound and method for preparation thereof |
| WO2023011312A1 (en) * | 2021-08-02 | 2023-02-09 | 上海汇伦医药股份有限公司 | Multi-substituted thiazole derivative and application thereof in disease treatment |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113773275A (en) | 2021-12-10 |
| WO2022142731A1 (en) | 2022-07-07 |
| CN113416172B (en) | 2022-06-21 |
| CN113773275B (en) | 2023-04-25 |
| WO2022142727A1 (en) | 2022-07-07 |
| CN113416172A (en) | 2021-09-21 |
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