WO2022142731A1 - Antiviral compound and preparation method therefor - Google Patents

Abstract

Disclosed are an antiviral compound and a preparation method therefor. The structural formula of the compound is as represented by formula I. In formula (I), R1 is selected from: monosubstituted or polysubstituted H, F, methyl, and trifluoromethyl; R2 is selected from: H and straight-chain or substituted alkanes (C1-C6); R3 is selected from: monosubstituted or polysubstituted H, Cl, Br, and F. Experiments prove that the compound of the present invention not only has a good inhibition effect on H1N1 influenza A virus, but also has a good inhibition effect on coronavirus. No toxicity to human normal cells is observed, and the degree of an inflammatory response can be inhibited while resisting viruses.

Description

A kind of antiviral compound and preparation method thereof technical field

The invention belongs to the field of medicine, and in particular relates to an antiviral compound and a preparation method thereof.

Background technique

Influenza virus is referred to as influenza virus. It is divided into three types: A (A), B (B), and C (C). Influenza viruses discovered in recent years will be classified as type D (D). Influenza virus can cause infection and disease in humans, birds, pigs, horses, bats and other animals, and is the pathogen of human and animal diseases such as human influenza, avian influenza, swine influenza, and equine influenza.

The typical clinical symptoms of these diseases are acute high fever, generalized pain, marked fatigue, and respiratory symptoms. Influenza viruses are mainly spread through airborne droplets, contact between susceptible and infected persons, or contact with contaminated items. Generally, autumn and winter are the high incidence period. Human influenza is mainly caused by influenza A and B viruses. Influenza A virus frequently undergoes antigenic variation and can be further divided into subtypes such as H1N1, H3N2, H5N1, and H7N9 (where H and N represent the two surface glycoproteins of influenza virus, respectively). Influenza viruses are not very resistant to the outside world. Animal influenza viruses do not usually infect humans, and human influenza viruses do not usually infect animals, with the exception of pigs. Pigs can be infected with both human and avian influenza viruses, but they are mainly infected with swine influenza viruses. A small number of animal influenza viruses can cause a human influenza pandemic after being adapted to humans.

Human coronaviruses can cause common cold, severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS) in humans, and there are certain differences in epidemiological characteristics.

Globally, 10% to 30% of upper respiratory tract infections are caused by four types of coronaviruses, HCoV-229E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1, which occupy the second place in the cause of common cold, second only to nasal Virus. Infections are seasonal, with peak disease in spring and winter each year. The incubation period is 2-5 days, and the population is generally susceptible. It is mainly spread through person-to-person contact.

Influenza A (H1N1) is a highly contagious acute respiratory disease of swine caused by one or more swine influenza A viruses. Morbidity tends to be high, but mortality is low (1 to 4%). The virus spreads in herds through aerosols, direct and indirect contact, and asymptomatic pigs that carry the virus. Swine outbreaks can occur throughout the year. Incidence increases during autumn and winter in temperate zones. Influenza A (H1N1) in humans usually comes from infected pigs, but some human cases have no history of contact with pigs or their environment. Human-to-human transmission has occurred in some cases, but limited to close contacts and people in closed settings.

Coronavirus (HCoV-229E) is a type of coronavirus. Coronaviruses belong to the order Nesteviridae, the family Coronaviridae, and the genus Coronaviridae. They are a large virus family that exists widely in nature. Coronaviruses only infect vertebrates and are associated with a variety of diseases in humans and animals, and can cause respiratory, gastrointestinal and nervous system diseases in humans and animals.

Therefore, it is of great practical significance to study an effective antiviral drug.

SUMMARY OF THE INVENTION

The object of the present invention is to provide a compound represented by formula I and pharmaceutically acceptable salts and solvates thereof.

In the formula (I), n=an integer of 0-5, such as 1, 2, 3;

R ¹ is selected from: mono- or poly-substituted H, F, methyl, trifluoromethyl, preferably H;

R ² is selected from: H, straight-chain or substituted alkane (C1-C6), preferably methyl, isopropyl;

R ³ is selected from: mono- or poly-substituted H, Cl, Br, F, preferably Cl;

R ⁴ and R ⁵ are independently selected from any one of the following groups: (C ₁ -C ₆ ) alkyl, (C ₃ -C ₈ ) carbocyclyl alkyl, substituted (C ₁ -C 8 ) C ₁₈ ) alkane, (C ₂ -C ₈ ) alkenyl, substituted (C ₂ -C ₈ ) alkenyl, (C ₂ -C ₈ ) alkynyl, substituted (C ₂ -C ₈ ) ) alkynyl, (C ₆ -C ₂₀ ) aryl, substituted (C ₆ -C ₂₀ ) aryl, (C ₂ -C ₂₀ ) heterocyclic, substituted (C ₂ -C ₂₀ ) Heterocyclyl; or R ⁴ and R ⁵ form a ring with each other to form (C ₃ -C ₈ )heterocycloalkyl or substituted (C ₃ -C ₈ )heterocycloalkyl, (C ₆ -C ₂₀ )heteroaryl or Substituted (C ₆ -C ₂₀ )heteroaryl.

The substituents described in the R ⁴ and R ⁵ are selected from at least one of the following: methyl, ethyl, isopropyl, pyrrolyl, piperidinyl, morpholinyl and piperazinyl.

The (C ₃ -C ₈ )heterocycloalkyl may in particular be pyrrolyl, piperidinyl, morpholinyl or piperazinyl.

In some embodiments of the present invention, the compound shown in formula I of the present invention can be enumerated as the structure shown below, but is not limited to the following structure:

Pharmacodynamic tests show that when R ⁴ and R ⁵ are not H, the compound of general formula (I) can obtain a higher blood concentration of the metabolite ZONK2003-0, with excellent bioavailability and a much higher AUC. When the plasma concentration of ZONK2003-0 was equimolar by gavage, and when the R ⁴ substituent was H, such as ZONK2003-22, a very low plasma concentration of ZONK2003-0 was obtained after gavage, suggesting that it has a very low plasma concentration. Bioavailability or conversion to other substances.

The present invention also provides a preparation method of the compound represented by the above formula I.

According to the literature Med.Chem.Commun., 2016,7,1441–1448, compound 1 (formula a) alkylhydrazine reacts with sodium thiocyanate to obtain thiourea compound 2 (formula b), which is then reacted with R ¹ substituted o-formic acid Benzaldehyde is condensed to obtain compound 3 (formula c), which is finally closed with ^R substituted bromoacetophenone to obtain thiazole compound 4 (formula d); under the action of a condensing agent, the benzamide shown in formula I is obtained derivative. Specific steps are as follows:

1) react the compound shown in formula a with sodium thiocyanate to obtain the compound shown in formula b;

Wherein, the definition of R ² in formula a is the same as formula I; the definition of R ² in formula b is the same as formula a;

² ) the o-formic acid benzaldehyde that the compound shown in formula b is substituted with R shown in formula e carries out condensation reaction to obtain compound shown in formula c;

Wherein, the definition of R ¹ in formula e is the same as formula I; the definition of R ¹ in formula c is the same as formula e, and the definition of R ² is the same as formula b;

³ ) compound shown in formula c and R shown in formula f substituted bromoacetophenone carry out ring closure reaction, obtain compound shown in formula d;

Wherein, the definition of R ³ in formula f is the same as formula I, the definitions of R ¹ and R ² in formula d are the same as formula c, and the definition of R ³ is the same as formula f;

4) under the action of the condensing agent, make the compound shown in formula d and the compound shown in formula g to carry out condensation reaction, obtain the compound shown in formula I;

Wherein, the definitions of n, R ⁴ and R ⁵ in formula g are the same as those of formula I.

In the above step 1), the reaction conditions of the reaction are: the reaction temperature is 50-100 ° C, and the reaction time is 24-72 hours; the reaction is carried out in a solvent, and the solvent can be methanol, ethanol, tetrahydrofuran, acetonitrile, etc., preferably Ethanol.

In the above step 2), the reaction conditions of the condensation reaction are: the reaction temperature is 50-100 ° C, the reaction time is 1-3 hours; the reaction is carried out in a solvent, and the solvent can be methanol, ethanol, tetrahydrofuran, acetonitrile, etc., Ethanol is preferred.

In the above step 3), the reaction conditions of the ring-closing reaction are: the reaction temperature is 50-100 ° C, and the reaction time is 3-6 hours; the reaction is carried out in a solvent, and the solvent can be methanol, ethanol, tetrahydrofuran, acetonitrile, etc. , preferably ethanol.

In the above step 4), the reaction conditions of the condensation reaction are: the reaction temperature is 0-25°C, and the reaction time is 2-8 hours; the reaction is carried out in a solvent, and the solvent can be dichloromethane, tetrahydrofuran, acetonitrile, etc., Dichloromethane is preferred;

Other compounds protected in the claims of the present invention can be obtained by referring to the preparation methods of the examples of the present invention.

Another object of the present invention is to provide the application of the compound represented by the above formula I.

The application provided by the present invention is that the application of the compound represented by formula I or its pharmaceutically acceptable salt, ester and solvate is the following (a) and/or (b) and/or (c):

(a) the application of the compound shown in formula I or its pharmaceutically acceptable salt, ester, solvate in the preparation of the product for the treatment of virus-induced disease or virus infection;

(b) the application of the compound shown in formula I or its pharmaceutically acceptable salt, ester, solvate in the preparation of the product for preventing virus-induced disease or virus infection;

(c) Use of the compound represented by formula I or a pharmaceutically acceptable salt, ester or solvate thereof in the preparation of a virus inhibitor.

The product may be a drug or a pharmaceutical formulation.

The viral inhibitor is capable of inhibiting viral replication.

The virus includes influenza virus, coronavirus.

The influenza virus can specifically be influenza A virus (H1N1);

The coronavirus can be alpha coronavirus and/or beta coronavirus, specifically selected from HCoV-229E.

In the present invention, the disease caused by the virus may be an infectious disease of the respiratory system.

The respiratory infection is a respiratory infection and/or a pulmonary infection; the respiratory infection can be nasopharyngitis, rhinitis, pharyngitis, tracheitis and/or bronchitis; the pulmonary infection can 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 the coronavirus usually include viral pneumonia, severe acute respiratory syndrome and the like.

In the present invention, the coronavirus infection usually causes diseases such as viral pneumonia and severe acute respiratory syndrome.

The compound of the present invention has inhibitory effect on coronavirus and H1N1 influenza A virus at the same time, and has no toxicity to normal human cells, can inhibit the degree of inflammatory reaction, reduce the damage of pneumonia to the body, and promote the recovery of the body.

The antiviral drug prepared with the compound represented by formula I as the active ingredient also belongs to the protection scope of the present invention.

The antiviral drug can be introduced into the body such as muscle, intradermal, subcutaneous, intravenous, mucosal tissue by injection, spray, nasal instillation, eye instillation, penetration, absorption, physical or chemical mediation; or mixed with other substances or Introduce into the body after wrapping.

When necessary, one or more pharmaceutically acceptable carriers can also be added to the above-mentioned drugs. The carrier includes conventional diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorption carriers, lubricants and the like conventional in the pharmaceutical field.

The above-mentioned medicines can be made into various forms such as tablets, powders, granules, capsules, oral liquids, ointments, creams, injections, etc. The medicines of the above-mentioned various dosage forms can be prepared according to conventional methods in the pharmaceutical field.

The present invention also provides a medicine or a pharmaceutical composition, the active ingredient of which is the compound represented by formula I or a pharmaceutically acceptable salt, ester or solvate thereof.

The medicament or pharmaceutical composition has at least one of the following effects:

1) Treatment of viral diseases or viral infections;

2) Prevention of viral diseases or viral infections;

3) Inhibit virus.

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 using the compound of formula I or a pharmaceutically acceptable salt thereof provided by the present invention to prevent and/or treat infection caused by a virus, an effective amount of the compound of formula I or a pharmaceutically acceptable salt thereof is administered to a subject organism.

The compounds described in the present invention have been confirmed by experiments that they not only have a good inhibitory effect on the H1N1 influenza A virus, but also have a good inhibitory effect on the coronavirus. The extent to which the virus simultaneously suppresses the inflammatory response.

Description of drawings

Fig. 1 is the synthetic route diagram of the compound represented by formula I of the present invention.

Detailed ways

The present invention will be further described below in conjunction with specific embodiments, but the present invention is not limited to the following embodiments. The methods are conventional methods unless otherwise specified. The raw materials can be obtained from open commercial sources unless otherwise specified.

Examples 1-10

1. (E)-2-(methylamino)ethyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2-methylhydrazono)methyl)benzyl Synthesis of acid esters (ZONK2003-22)

1) 2-Methylthiosemicarbazide (1-2)

Methyl hydrazine (23.0 g, 0.5 mol), ammonium thiocyanate (38.0 g, 0.5 mol), and ethanol (200 mL) were added to a single-neck round-bottomed flask respectively, and the reaction was heated under reflux for 72 h. The reaction solution was cooled and concentrated, and purified by column chromatography to obtain 2-methylthiosemicarbazide (44.1 g, 84.0%) as an off-white solid. ¹ H NMR(DMSO-d ₆ 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-carbamoyl-2-methylhydrazono)methyl)benzoic acid (1-3)

2-Methylthiosemicarbazide (40.0 g, 0.38 mol), 2-carbonylbenzoic acid (57.0 g, 0.38 mol), and ethanol (300 mL) were added to a single-neck round-bottomed flask respectively, and the reaction was heated under reflux for 2 h. The reaction solution was cooled and concentrated, and purified by column chromatography to obtain (E)-2-((2-carbamoyl-2-methylhydrazono)methyl)benzoic acid (85.6 g, 95.0%) as a pale yellow solid. ¹ H NMR(DMSO-d ₆ 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) (ZONK2003- 0)

Combine (E)-2-((2-carbamoyl-2-methylhydrazono)methyl)benzoic acid (80.0 g, 0.34 mol), 2-bromo-1-(2-chlorophenyl) ) ethyl ketone (79.2 g, 0.34 mol) and ethanol (400 mL) were respectively added to the single-neck round-bottomed flask, and the reaction was heated under reflux for 3 h. The reaction solution was cooled and concentrated, and purified by column chromatography to obtain a pale yellow solid (E)-2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2-methylhydrazono)methyl ) benzoic acid (124 g, 98.5%). ¹ H NMR(DMSO-d ₆ 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-((tert-butoxycarbonyl)(methyl)amino)ethyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2-methyl hydrazono)methyl)benzoate (1-5)

(E)-2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2-methylhydrazono)methyl)benzoic acid (3.71 g, 10 mmol) was dissolved in In methyl chloride (30 mL), N-hydroxysuccinimide (1.15 g, 10 mmol) and dicyclohexylcarbodiimide (2.27 g, 11 mmol) were added respectively, and the mixture was stirred at room temperature for 3 h. After TLC detected the disappearance of the raw materials, 2-(N-Boc-N-methylamino)ethanol (1.75 g, 10 mmol) was added, and the reaction was continued at room temperature for 5 h. The reaction solution was concentrated and purified by column chromatography to obtain off-white solid (E)-2-((tert-butoxycarbonyl)(methyl)amino)ethyl 2-((2-(4-(2-chlorophenyl)thiazole) -2-yl)-2-methylhydrazono)methyl)benzoate (4.52 g, 85.5%). ¹ H NMR(DMSO-d ₆ 400MHz)δ8.24(s,1H),8.00-7.34(m,9H),4.68(m,2H),3.64(s,3H),3.42(m,2H),3.26 (s,3H),1.36(s,9H).ESI-MS m/z:529.5[M+H] ⁺ .

5) (E)-2-(methylamino)ethyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2-methylhydrazono)methyl)benzyl Synthesis of acid esters

(E)-2-((tert-butoxycarbonyl)(methyl)amino)ethyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2-methylidene Hydrazino)methyl)benzoate (4.52 g, 8.54 mmol) was dissolved in dichloromethane (20 mL), concentrated hydrochloric acid (10 mL) was added, and the reaction was refluxed for 3 h. After TLC detected the disappearance of the raw materials, the reaction solution was concentrated and purified by column chromatography to obtain (E)-2-(methylamino)ethyl 2-((2-(4-(2-chlorophenyl)thiazole-2-) as an off-white solid (3.22 g, 88.1%). ¹ H NMR(DMSO-d ₆ 400MHz)δ8.24(s,1H),8.00-7.34(m,9H),4.68(m,2H),3.64(s,3H),3.42(m,2H),3.26 (s,3H).ESI-MS m/z: 429.5[M+H] ⁺ .

2. (E)-2-(dimethylamino)ethyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2-methylhydrazono)methyl) Synthesis of Benzoate (ZONK2003-23)

(E)-2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2-methylhydrazono)methyl)benzoic acid (3.71 g, 10 mmol) was dissolved in In methyl chloride (30 mL), N-hydroxysuccinimide (1.15 g, 10 mmol) and 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (2.11 g) were added respectively. , 11 mmol), and stirred at room temperature for 3 h. After TLC detected the disappearance of the raw materials, 2-(dimethylamino)ethanol (0.89 g, 10 mmol) was added, and the reaction was continued at room temperature for 5 h. The reaction solution was concentrated and purified by column chromatography to obtain off-white solid (E)-2-(dimethylamino)ethyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2 - Methylhydrazono)methyl)benzoate (3.68 g, 83.3%). ¹ H NMR(DMSO-d ₆ 400MHz)δ8.20(s,1H),8.01-7.34(m,9H),4.55(m,2H),4.07(m,2H),3.52(s,3H),2.68 (s,6H).ESI-MS m/z: 443.5[M+H] ⁺ .

3. (E)-2-(diethylamino)ethyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2-methylhydrazono)methyl)benzene Synthesis of Formate (ZONK2003-9)

(E)-2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2-methylhydrazono)methyl)benzoic acid (3.71 g, 10 mmol) was dissolved in In methyl chloride (30 mL), N-hydroxysuccinimide (1.15 g, 10 mmol) and dicyclohexylcarbodiimide (2.27 g, 11 mmol) were added respectively, and the mixture was stirred at room temperature for 3 h. After TLC detected the disappearance of the raw materials, 2-(diethylamino)ethanol (1.17 g, 10 mmol) was added, and the reaction was continued at room temperature for 5 h. The reaction solution was concentrated and purified by column chromatography to obtain off-white solid (E)-2-(diethylamino)ethyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2- Methylhydrazono)methyl)benzoate (3.85 g, 81.8%). ¹ H NMR(DMSO-d ₆ 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] ⁺ .

Example 4, (E)-2-(pyrrolidin-1-yl)ethyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2-methylhydrazono ) methyl) benzoate synthesis (ZONK2003-10)

(E)-2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2-methylhydrazono)methyl)benzoic acid (3.71 g, 10 mmol) was dissolved in In methyl chloride (30 mL), N-hydroxysuccinimide (1.15 g, 10 mmol) and 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (2.11 g) were added respectively. , 11 mmol), and stirred at room temperature for 3 h. After TLC detected the disappearance of the raw materials, 2-(pyrrolidin-1-yl)ethanol hydrochloride (1.52 g, 10 mmol) was added, and the reaction was continued at room temperature for 5 h. The reaction solution was concentrated and purified by column chromatography to obtain off-white solid (E)-2-(pyrrolidin-1-yl)ethyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl) -2-Methylhydrazono)methyl)benzoate (3.64 g, 77.6%). ¹ H NMR(DMSO-d ₆ 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:469.5[M+H] ⁺ .

Example 5, (E)-2-(piperidin-1-yl)ethyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl)methylhydrazono)methyl ) Synthesis of benzoate (ZONK2003-24)

(E)-2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2-methylhydrazono)methyl)benzoic acid (3.71 g, 10 mmol) was dissolved in In methyl chloride (30 mL), N-hydroxysuccinimide (1.15 g, 10 mmol) and dicyclohexylcarbodiimide (2.27 g, 11 mmol) were respectively added, and the mixture was stirred at room temperature for 3 h. After TLC detected the disappearance of the raw materials, N-hydroxyethyl piperidine (1.29 g, 10 mmol) was added, and the reaction was continued at room temperature for 5 h. The reaction solution was concentrated and purified by column chromatography to obtain off-white solid (E)-2-(piperidin-1-yl)ethyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl) Methylhydrazono)methyl)benzoate (3.86 g, 80.1%). ¹ H NMR(DMSO-d ₆ 400MHz)δ8.20(s,1H),8.01-7.34(m,9H),4.55-3.54(m,11H),1.86-1.55(m,6H).ESI-MS m /z:483.5[M+H]+.

6. Synthesis of (E)-2-morpholinoethyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl)methylhydrazono)methyl)benzoate ( ZONK2003-25)

(E)-2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2-methylhydrazono)methyl)benzoic acid (3.71 g, 10 mmol) was dissolved in In methyl chloride (30 mL), N-hydroxysuccinimide (1.15 g, 10 mmol) and 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (2.11 g) were added respectively. , 11 mmol), and stirred at room temperature for 3 h. After TLC detected the disappearance of the raw materials, 2-morpholinoethanol (1.31 g, 10 mmol) was added, and the reaction was continued at room temperature for 5 h. The reaction solution was concentrated and purified by column chromatography to obtain off-white solid (E)-2-morpholinoethyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl)methylhydrazono) Methyl)benzoate (3.70 g, 76.5%). ¹ H NMR(DMSO-d ₆ 400MHz)δ8.20(s,1H),8.01-7.34(m,9H),4.55-3.54(m,15H).ESI-MS m/z:485.5[M+H] ⁺ .

Example 7, (E)-2-(piperazin-1-yl)ethyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2-methylhydrazono ) methyl) benzoate synthesis (ZONK2003-26)

(E)-2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2-methylhydrazono)methyl)benzoic acid (3.71 g, 10 mmol) was dissolved in In methyl chloride (30 mL), N-hydroxysuccinimide (1.15 g, 10 mmol) and dicyclohexylcarbodiimide (2.27 g, 11 mmol) were added respectively, and the mixture was stirred at room temperature for 3 h. After TLC detected the disappearance of the raw materials, 1-piperazine ethanol (1.30 g, 10 mmol) was added, and the reaction was continued at room temperature for 5 h. The reaction solution was concentrated and purified by column chromatography to obtain off-white solid (E)-2-(piperazin-1-yl)ethyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl) -2-Methylhydrazono)methyl)benzoate (3.58 g, 74.1%). ¹ H NMR(DMSO-d ₆ 400MHz)δ8.20(s,1H),8.01-7.11(m,9H),4.55-3.54(m,12H),3.31(s,3H).ESI-MS m/z :484.5[M+H] ⁺ .

Example 8, (E)-2-(piperazin-1-yl)ethyl 2-((2-(4-(2,4-dichlorophenyl)thiazol-2-yl)-2-methyl Synthesis of hydrazono)methyl)-4-fluorobenzoate (ZONK2003-27)

By combining the preparation method of Example 7, (E)-2-(piperazin-1-yl)ethyl 2-((2-(4-(2,4-dichlorophenyl)thiazol-2-yl) was synthesized )-2-methylhydrazono)methyl)-4-fluorobenzoate. ¹ H NMR(DMSO-d ₆ 400MHz)δ8.20(s,1H),8.01-7.11(m,7H),4.55-3.54(m,12H),3.31(s,3H).ESI-MS m/z :536.5[M+H] ⁺ .

Example 9, (E)-2-morpholinoethyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl)hydrazono)methyl)-4-methylbenzyl Synthesis of acid esters (ZONK2003-28)

Combining the preparation method of Example 6, (E)-2-morpholinoethyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl)hydrazono)methyl)- 4-methylbenzoate. ¹ H NMR(DMSO-d ₆ 400MHz)δ8.20(s,1H),8.01-7.34(m,8H),4.55-3.54(m,12H),2.68(s,3H).ESI-MS m/z :485.5[M+H]+.

Example 10, (E)-3-(diethylamino)propyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2-methylhydrazono)methane Synthesis of base) benzoate (ZONK2003-29)

Synthesizing the preparation method of Example 3, (E)-3-(diethylamino)propyl 2-((2-(4-(2-chlorophenyl)thiazol-2-yl)-2-methyl) was synthesized hydrazono)methyl)benzoate ¹ H 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, 2H), 1.12(m, 6H). ESI-MS m/z: 485.5[M+H] ⁺ .

Example 11, (E)-3-(dimethylamino)propyl 2-((2-(4-(4-chlorophenyl)thiazol-2-yl)hydrazono)methyl)-5-fluoro Synthesis of Benzoate (ZONK2003-8)

Synthesizing the preparation method of Example 3, (E)-3-(dimethylamino)propyl 2-((2-(4-(4-chlorophenyl)thiazol-2-yl)hydrazono)methan was synthesized base)-5-fluorobenzoate. ¹ H 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]+.

Example 12, (E)-2-(piperazin-1-yl)ethyl 2-((2-methyl-2-(4-phenylthiazol-2-yl)hydrazono)methyl)benzene Synthesis of Formate (ZONK2003-13)

Synthesizing the preparation method of Example 3, (E)-2-(piperazin-1-yl)ethyl 2-((2-methyl-2-(4-phenylthiazol-2-yl)hydrazide) was synthesized group) methyl) benzoate. ¹ H 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 13. Protective effect of a series of compounds on mice infected with influenza A virus A/FM/1/47 (H1N1)

Subject:

ZONK2003-0; ZONK2003-22; ZONK2003-9; ZONK2003-25; Provided by Research Ltd.

Oseltamivir Phosphate Granules, 15mg×10 bags, product of Yichang Dongguang Changjiang Pharmaceutical Co., Ltd., batch number: 0371912115, valid until 2021.12.11, positive control drug for anti-influenza A virus;

Experimental materials: Influenza A virus mouse lung-adapted strain A/FM/1/47 (H1N1), inoculated with chicken embryos, and collected allantoic fluid for preservation. ICR mice, weighing 18-22 g. During the administration period, the patients were free to eat and drink, with 12 hours of light and 12 hours of darkness every day, the temperature was 22±2°C, and the humidity was 55-70%. Experimental method: After 3 days of adaptive feeding, the experiment was started. Except for the uninfected control group, mice in other groups were lightly anesthetized with ether, and intranasally inoculated with chick embryo allantoies of influenza virus A/FM/1/47 (H1N1) equivalent to 8×LD ₅₀ diluted with normal saline. The mice in the positive control oseltamivir group and the example compound group were administered intragastrically for the first time 2 h after infection, and each compound was administered by intragastric administration at the doses of 10 μmol/kg, 20 μmol/kg and 30 μmol/kg. , administered twice a day for 5 consecutive days. Observe the survival of the mice within 14 days,

And calculate the death protection rate of the drug for the mice (death protection rate=mortality rate of model group-mortality rate of experimental group).

Table 1. Protective effects of compounds on mice infected with influenza A virus (H1N1 influenza A virus)

Example 14: Alleviating effect of drugs on pulmonary inflammation in mice caused by influenza virus H1N1 infection

Experimental method: After 3 days of adaptive feeding, the experiment was started. Except for the uninfected control group, mice in other groups were lightly anesthetized with ether, and intranasally inoculated with chick embryo allantoies of influenza virus A/FM/1/47 (H1N1) equivalent to 8×LD ₅₀ diluted with normal saline. The mice in the positive control oseltamivir group and the test administration group were intragastrically administered with 80 mg/kg for the first time 24 hours after virus infection, and then once a day, the same as the virus control group and the uninfected control group. Oral saline, once a day, the administration volume is 0.1mL/10g body weight. 5 days in total. On the 6th day, 3 mice in each group were weighed, the eyeballs were enucleated and exsanguinated, and the whole lung was taken out, weighed, and the lung index and lung index inhibition rate were calculated.

Percentage of weight loss = pre-dose body weight - post-dose body weight / pre-dose body weight × 100%

Lung index = mouse lung weight / mouse body weight × 100

Lung index inhibition rate (%) = mean lung index in virus control group - mean lung index in drug administration group/mean lung index in virus control group × 100%

group	Weight loss %	Lung index	Lung Index Inhibition %
normal control group		0.57
model control group		2.09
Oseltamivir group	15.12	1.66	20.57
ZONK2003-8	9.78	1.25	40.19
ZONK2003-9	12.45	1.19	43.06
ZONK2003-10	11.02	1.08	48.32

Experimental results: Therefore, the compounds of the examples have obvious protective and inhibitory effects on pulmonary inflammation caused by influenza virus, and the effect is better than that of the oseltamivir control group.

Example 15: Pharmacokinetic experiment of ZONK2003 in rats by gavage

There were 3 male rats in each group, ZONK2003-0 group, ZONK2003-22 group, ZONK2003-9 group, and ZONK2003-25 group, respectively. Each rat was given 10 discrete time points.

An LC-MS/MS method was established to determine the concentrations of ZONK2003-0, ZONK2003-22, ZONK2003-9 and ZONK2003-25 in whole blood of ICR mice. The obtained plasma drug concentration data were also used to calculate the relevant pharmacokinetic parameters using the pharmacokinetic processing software Pharsight Phoenix WinNonlin 8.0 non-compartmental model.

Rats were given ZONK2003-0 by gavage at a dose of 2.51 mg/kg;

Rats were given an equimolar dose of ZONK2003-22 at a dose of 3.14 mg/kg;

Rats were given equimolar ZONK2003-9 at a dose of 3.27 mg/kg;

Rats were given an equimolar dose of ZONK2003-25 at a dose of 3.42 mg/kg;

Solvent: 20% Solutol HS-15/saline;

The plasma concentration of ZONK2003-0 was detected, and the pharmacokinetic parameters were calculated.

The detailed results are shown in the table below.

Table 2 After intragastric administration of 2.51 mg/kg of ZONK2003-0 to rats, the blood concentration of the prototype drug in rats (ng/mL)

Table 3 After intragastric administration of 2.51 mg/kg ZONK2003-0 to rats, the main pharmacokinetic parameters of the prototype drug

Table 4 After intragastric administration of 3.14 mg/kg of ZONK2003-22 to rats, the detection results of the plasma concentration of the metabolite ZONK2003-0 (ng/mL)

ND: Not detected, that is, the measured value is lower than the lower limit of quantification after the blood concentration reaches the peak, —: The value cannot be calculated

Table 5 After intragastric administration of 3.14 mg/kg of ZONK2003-22 to rats, the main pharmacokinetic parameters of the metabolite ZONK2003-0

Table 6 After intragastric administration of 3.27 mg/kg of ZONK2003-9 to rats, the detection results of the plasma concentration of the metabolite ZONK2003-0 (ng/mL)

Table 7 After intragastric administration of 3.27 mg/kg of ZONK2003-9 to rats, the main pharmacokinetic parameters of the metabolite ZONK2003-0

Table 8 After intragastric administration of 3.42 mg/kg of ZONK2003-25 to rats, the detection results of the plasma concentration of the metabolite ZONK2003-0 (ng/mL)

Table 9 After intragastric administration of 3.42 mg/kg of ZONK2003-25 to rats, the main pharmacokinetic parameters of the metabolite ZONK2003-0

After intragastric administration of equimolar ZONK2003-9 and ZONK2003-25, the plasma concentration of the metabolite ZONK2003-0 was detected, and the AUC was 2-3 times that of equimolar ZONK2003-0, suggesting that ZONK2003-9 and ZONK2003-25 The bioavailability of ZONK2003-0 is 2-3 times higher than that of ZONK2003-0, ZONK2003-9 and ZONK2003-25 are excellent prodrugs of ZONK2003-0, and the metabolite ZONK2003-0 is detected after gavage of equimolar ZONK2003-22. The AUC of ZONK2003-9 and ZONK2003-5 groups was much lower than that of ZONK2003-9 and ZONK2003-5 groups, and also lower than that of ZONK2003-0 gavage group, suggesting that ZONK2003-22 had low bioavailability or was converted into other substances after gavage, and failed to obtain Higher plasma concentrations of the metabolite ZONK2003-0.

Claims (10)

1. The compound whose general structural formula is shown in formula I or a pharmaceutically acceptable salt or solvate thereof:

   

   In the formula (I), n is an integer of 0-5;

   R ¹ is selected from: mono- or poly-substituted H, F, methyl, trifluoromethyl;

   R ² is selected from: H, straight chain or substituted alkane (C1-C6);

   R ³ is selected from: mono- or poly-substituted H, Cl, Br, F;

   R ⁴ and R ⁵ are independently selected from any one of the following groups: (C ₁ -C ₆ ) alkyl, (C ₃ -C ₈ ) carbocyclyl alkyl, substituted (C ₁ -C 8 ) C ₁₈ ) alkane, (C ₂ -C ₈ ) alkenyl, substituted (C ₂ -C ₈ ) alkenyl, (C ₂ -C ₈ ) alkynyl, substituted (C ₂ -C ₈ ) ) alkynyl, (C ₆ -C ₂₀ ) aryl, substituted (C ₆ -C ₂₀ ) aryl, (C ₂ -C ₂₀ ) heterocyclic, substituted (C ₂ -C ₂₀ ) Heterocyclyl; or R ⁴ and R ⁵ form a ring with each other to form (C ₃ -C ₈ )heterocycloalkyl or substituted (C ₃ -C ₈ )heterocycloalkyl, (C ₆ -C ₂₀ )heteroaryl or Substituted (C ₆ -C ₂₀ )heteroaryl;

   The substituents described in the R ⁴ and R ⁵ are selected from at least one of the following: methyl, ethyl, isopropyl, pyrrolyl, piperidinyl, morpholinyl and piperazinyl.
2. The compound according to claim 1 or a pharmaceutically acceptable salt, ester or solvate thereof, characterized in that: the n is 1, 2 or 3;

   Or, the R ¹ is selected from H;

   Or, described R ² is selected from methyl or isopropyl;

   Alternatively, the R ³ is selected from Cl.
3. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt, ester or solvate thereof, is characterized in that: the compound represented by the formula I is selected from any of the following compounds:

   
4. The preparation method of the compound shown in the formula I described in any one of claim 1-3, comprises the steps:

   1) react the compound shown in formula a with sodium thiocyanate to obtain the compound shown in formula b;

   

   Wherein, the definition of R ² in formula a is the same as formula I; the definition of R ² in formula b is the same as formula a;

   ² ) the o-formic acid benzaldehyde that the compound shown in formula b is substituted with R shown in formula e carries out condensation reaction to obtain compound shown in formula c;

   

   Wherein, the definition of R ¹ in formula e is the same as formula I; the definition of R ¹ in formula c is the same as formula e, and the definition of R ² is the same as formula b;

   ³ ) compound shown in formula c and R shown in formula f substituted bromoacetophenone carry out ring closure reaction, obtain compound shown in formula d;

   

   Wherein, the definition of R ³ in formula f is the same as formula I, the definitions of R ¹ and R ² in formula d are the same as formula c, and the definition of R ³ is the same as formula f;

   4) under the action of the condensing agent, make the compound shown in formula d and the compound shown in formula g to carry out condensation reaction, obtain the compound shown in formula I;

   

   Wherein, the definitions of n, R ⁴ and R ⁵ in formula g are the same as those of formula I.
5. The method according to claim 4, wherein:

   In the step 1), the reaction conditions of the reaction are: the reaction temperature is 50-100 ° C, and the reaction time is 24-72 hours; the reaction is carried out in a solvent, and the solvent is 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: the reaction temperature is 50-100° C., and the reaction time is 1-3 hours; the reaction is carried out in a solvent, and the solvent is 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: the reaction temperature is 50-100 ° C, and the reaction time is 3-6 hours; the reaction is carried out in a solvent, and the solvent is 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: the reaction temperature is 0-25°C, and the reaction time is 2-8 hours; the reaction is carried out in a solvent, and the solvent is selected from any one of the following: dichloride Methane, tetrahydrofuran, acetonitrile, preferably dichloromethane.
6. The application of the compound shown in the formula I of any one of claims 1-3 or its pharmaceutically acceptable salt, solvate is the following (a) and/or (b) and/or (c):

   (a) the application of the compound shown in formula I or its pharmaceutically acceptable salt, ester, solvate in the preparation of the product for the treatment of virus-induced disease or virus infection;

   (b) the application of the compound shown in formula I or its pharmaceutically acceptable salt, ester, solvate in the preparation of the product for preventing virus-induced disease or virus infection;

   (c) Use of the compound represented by formula I or a pharmaceutically acceptable salt, ester or solvate thereof in the preparation of a virus inhibitor.
7. The application according to claim 6, wherein:

   The product is a medicine or a pharmaceutical preparation; the virus inhibitor can inhibit the replication of the virus;

   The virus includes influenza virus, coronavirus.
8. The application according to claim 7, wherein the influenza virus is influenza A virus; and the coronavirus is HCoV-229E.
9. A kind of medicine or pharmaceutical composition, its active component comprises the compound shown in formula I described in any one of claim 1-3 or its pharmaceutically acceptable salt, ester, solvate;

   The medicament or pharmaceutical composition has at least one of the following effects:

   1) Treatment of viral diseases or viral infections;

   2) Prevention of viral diseases or viral infections;

   3) Inhibit virus.
10. medicine or pharmaceutical composition according to claim 9, is characterized in that:

    The product is a medicine or a pharmaceutical preparation; the virus inhibitor can inhibit the replication of the virus;

    The virus includes influenza virus, coronavirus;

    The influenza virus is influenza A virus; the coronavirus is HCoV-229E.

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