CN100584828C

CN100584828C - New 5-hydroxy indole-3- carboxylic esters derivative

Info

Publication number: CN100584828C
Application number: CN200510005454A
Authority: CN
Inventors: 宫平; 吴春福; 裴钢; 左建平; 郭海燕; 王钝; 赵燕芳
Original assignee: Shenyang Pharmaceutical University
Current assignee: Shenyang Pharmaceutical University
Priority date: 2004-01-07
Filing date: 2005-01-01
Publication date: 2010-01-27
Anticipated expiration: 2025-01-01
Also published as: CN1660807A

Abstract

The present invention is a new 5-hydroxy indole-3-carboxylic esters ramification which includes an acceptable salt pharmaceutically and a drug combination which takes this compounds as its activity components,also includes the application of preparing the coronavirus infection drug used to treat and/or prevent SARS,among which,R[1],R[2],R[3],R[4] and X is multi substituent.This ramification can be made by the common method of pharmacy field and the combination of this invention can be made into various formulation including the common excipient of pharmacy field.

Description

5-hydroxyindole-3-carboxylic ester derivatives

The technical field is as follows:

the invention relates to the technical field of medicines, in particular to a novel 5-hydroxyindole-3-carboxylic ester derivative, which comprises a racemate or an optical isomer or a pharmaceutically acceptable salt and/or hydrate thereof, and a pharmaceutical composition taking the compound as an active ingredient, and application of the derivative in preparing medicines for treating and/or preventing SARS coronavirus infection.

Background art:

severe Acute Respiratory Syndrome (SARS), also known as "infectious atypical pneumonia", first appeared in local areas of guangdong province in our country since 2002 in 11 th of the month, and then rapidly spread to 33 countries and regions in five continents including hong kong, taiwan, vietnam, singapore, and germany, local spread occurred in at least 6 countries, and accumulated over 8200 patients and over 700 patients died of illness. The disease has high propagation speed and high death rate, thereby causing great attention to the international society. Based on the global influenza surveillance network of WHO, SARS working groups of 13 laboratories in 10 countries are participated in a multi-center research plan, and a pathogenic virus of SARS, namely a new coronavirus, is found. The WHO formally announced that the pathogen of SARS is this new Coronavirus at 16.4.2003 and was named SARS Coronavirus (SARS Coronavir, SARS-Cov).

Coronavirus (Coronavir) belongs to the order Nidovirales, family Coronaviridae, genus Coronaviridae. Coronavirus can infect birds and mammals and cause respiratory, intestinal, cardiovascular and nervous system diseases. The SARS coronavirus causing SARS epidemic is different from coronavirus found before, and is a new coronavirus, which is probably originated from an ancient ancestor of coronavirus and is formed after repeated passage of animals. SARS is characterized by acute onset, fast course, strong infectivity and high fatality rate, and is a serious infectious disease seriously harming human life and health. Clinically, there is a lack of antiviral drugs against SARS-Cov. Research and development of antiviral medicine with SARS-Cov inhibiting effect and capable of preventing and treating SARS is significant.

The research of 5-hydroxyindole-3-carboxylic ester derivatives as novel anti-influenza virus medicaments starts in the early eighties of the twentieth century. Grinev a.h., et al, khim-Farm Zh, 1987, 21(1), 52; parshevae.k.et al. khim Farm Zh, 1988, 22(5), 565; mezentseva m.v.et al.khim FarmZh, 1990, 24(10), 52; otova s.a., et al, khim Farm Zh, 1992, 26(1), 52; the synthesis and pharmacological activity research of some 5-hydroxyindole-3-carboxylate compounds are reported in ZotovoAs.A.et al, Khim Farm Zh, 1995, 29(1), 51 and other documents, and experiments show that some of the compounds have the activity of resisting influenza viruses, and have the pharmacological effects of inducing the production of interferon and enhancing the immunity of human bodies.

In 1993, the structural derivative, 1-methyl-4- (dimethylamino) methyl-2-phenylthiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride (Arbidol, pct appl.wo 9008135(RUSS), 1990-6-26), developed by the former soviet company VNIKhFI, was marketed in russia for the treatment and prevention of influenza a, b and acute viral respiratory infections, and it also has interferon-inducing and immunomodulating effects.

The inventor conducts extensive research on 5-hydroxyindole-3-carboxylic ester compounds, modifies and reforms a plurality of structural sites to synthesize a series of new derivatives, and in vitro antiviral activity tests show that the compounds have good antiviral effect. The inventor applies Chinese patent application No. 03110803.2 to the compound.

The inventor adopts the established model of the pathological effect of African green monkey kidney cells (VERO-E6) caused by SARS-Cov to carry out in vitro screening on the synthesized 5-oxindole-3-carboxylic ester derivatives, so as to obtain the compound with strong inhibition effect on SARS-Cov from the derivatives, and develop the compound into the clinical medicine for preventing and treating SARS.

The invention content is as follows:

the invention aims to provide a new application of 5-hydroxy indole-3-carboxylic ester derivatives including racemates or optical isomers or pharmaceutically acceptable salts and/or hydrates thereof in preparing medicaments for treating and/or preventing SARS coronavirus infection.

Wherein,

R₁is C₁～C₈Alkyl radical, C₃-C₈A cycloalkyl group;

R₂is C₁～C₈Alkyl radical, C₃-C₈A cycloalkyl group;

x represents H, halogen;

z is CH₂SR₅，CH₂NR₆R₇；

R₃、R₄Independently selected from H, amino, C₁-C₁₀Alkyl radical, C₃～C₈Cycloalkyl radical, C₂-C₁₀Alkenyl radical, C₂-C₁₀Alkynyl, which may be substituted by 1 to 3R₈Optionally substituted;

or R₃、R₄And N together form a guanidino group, or a 4-to 10-membered monocyclic or polycyclic saturated ring or a 5-to 10-membered heteroaromatic ring, wherein said saturated ring and heteroaromatic ring optionally include 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur, except R₃And R₄The saturated ring optionally includes 1 or 2 carbon-carbon double or triple bonds in addition to the nitrogen atom to which it is attached, and the saturated ring and the heteroaromatic ring may be substituted with 1 to 3R₈Optionally substituted;

R₅is unsubstituted or substituted by 1 to 3R₈Optionally substituted phenyl;

or R₅Is a 4-to 10-membered monocyclic or polycyclic saturated ring or a 5-to 10-membered heteroaromatic ring having 1 to 4 heteroatoms selected from nitrogen, oxygen and sulfur, wherein the saturated ring and the heteroaromatic ring may be substituted with 1 to 3R₈Optionally substituted;

R₆、R₇independently selected from H, amino, C₁-C₁₀Alkyl radical, C₃～C₈Cycloalkyl radical, C₂-C₁₀Alkenyl radical, C₂-C₁₀Alkynyl, which may be substituted by 1 to 3R₈Optionally substituted

Or R₆、R₇And N together form a 4-to 10-membered monocyclic or polycyclic saturated ring or a 5-to 10-membered heteroaromatic ring, wherein the saturated ring and heteroaromatic ring optionally include 1-4 heteroatoms selected from nitrogen, oxygen, and sulfur, except R₃And R₄The saturated ring optionally includes 1 or 2 carbon-carbon double or triple bonds in addition to the nitrogen atom to which it is attached, and the saturated ring and the heteroaromatic ring may be substituted with 1 to 3R₈Optionally substituted;

R₈is represented by C₁-C₆Alkyl radical, C₃～C₈Cycloalkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, hydroxy, C₁-C₆Alkoxy radical, C₂-C₆Alkenyloxy radical, C₂-C₆Alkynyloxy, mercapto, C₁-C₆Alkylthio radical, C₂-C₆Alkenylthio, phenyl, benzyl, free, salified, esterified and amidated carboxyl, halogen, trifluoromethyl, C₁-C₆Alkanoyl, nitro, cyano, amino, C₁-C₆Alkylamido, or mono-or di (C)₁-C₆Alkyl) substituted amino;

according to the invention, in the definition of the substituents:

preferred R₁Is methyl, ethyl, propyl, isopropyl, cyclopropyl;

preferred Z is CH₂SR₅；

Preferred R₅Is unsubstituted or substituted by 1 to 3 groups selected from C₁-C₄Straight or branched alkyl, C₁-C₄Alkoxy and halogenPhenyl substituted with an aryl group.

Preferred R₆、R₇And N together is a 5-or 6-membered monocyclic or polycyclic saturated ring, wherein said saturated ring optionally comprises 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur.

Preferred R₃And R₄Independently selected from C₁～C₈Alkyl radical, C₃～C₈A cycloalkyl group.

Preferred R₃、R₄And N together are 1-pyrrolidinyl, 1-piperidinyl, 4-carboxylic acid-1-piperidinyl, 1-piperazinyl, 4-methyl-1-piperazinyl, 4-morpholinyl.

Preferred X is H, a bromine atom.

Particularly preferred 5-hydroxyindole-3-carboxylic acid ester derivatives of formula I above according to the present invention include:

1-methyl-4- (1-piperidinyl) methyl-2-phenylthiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

1-methyl-4- (N-methyl-N-butyl-amino) methyl-2- (3-methylphenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

1-methyl-4- (N-methyl-N-octylamino) methyl-2- (3-methylphenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

1-methyl-4- (N-methyl-N-cyclopropylamino) methyl-2- (4-methoxyphenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

4- (4-methyl-1-piperazinyl) methyl-1-cyclopropyl-2- (3, 4-difluorophenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester dihydrochloride;

4- (4-methyl-1-piperazinyl) methyl-1-cyclopropyl-2- (3-methoxyphenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester dihydrochloride;

4- (4-methyl-1-piperazinyl) methyl-1-cyclopropyl-2- (2-fluorophenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester dihydrochloride;

4- (1-pyrrolidinyl) methyl-1-cyclopropyl-2-phenylthiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

1-ethyl-4- (1-pyrrolidinyl) methyl-2- (4-methylphenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

1-ethyl-4- (1-pyrrolidinyl) methyl-2- (3-methylphenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

1-ethyl-4- (1-pyrrolidinyl) methyl-2- (2-fluorophenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

1-ethyl-4- (1-pyrrolidinyl) methyl-2-phenylthiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

4- (1-pyrrolidinyl) methyl-1-cyclopropyl-2- (4-methylphenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

4- (4-methyl-1-piperazinyl) methyl-1-cyclopropyl-2- (4-fluorophenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester dihydrochloride;

1-ethyl-4- (diethylamino) methyl-2- (4-fluorophenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

1-ethyl-4- (4-morpholinyl) methyl-2- (4-fluorophenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

1-methyl-4- (diethylamino) methyl-2- (4-methylphenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

1-ethyl-4- (1-pyrrolidinyl) methyl-2- (2, 6-dichlorophenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

1-ethyl-4- (dimethylamino) methyl-2- (2, 6-dichlorophenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

4- (4-morpholinyl) methyl-1-isopropyl-2-phenylthiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

4- (dimethylamino) methyl-1-cyclopropyl-2- (4-fluorophenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

4- (dimethylamino) methyl-2- (1-piperidinyl) methyl-1-cyclopropyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

4- (dimethylamino) methyl-2- (4-morpholinyl) methyl-1-cyclopropyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

1-methyl-4- (dimethylamino) methyl-2-phenylthiomethyl-5-hydroxy-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

1-methyl-4- (1-pyrrolidinyl) methyl-2-phenylthiomethyl-5-hydroxy-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

4- (N-methyl-N-cyclopropylamino) methyl-1-cyclopropyl-2-phenylthiomethyl-5-hydroxy-1H-indole-3-carboxylic acid ethyl ester oxalate;

4- (N-methyl-N-isobutylamino) methyl-1-cyclopropyl-2-phenylthiomethyl-5-hydroxy-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

1-methyl-4- (4-carboxylic acid-1-piperidinyl) methyl-2-phenylthiomethyl-5-hydroxy-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

1-methyl-4- (2-methyl-1H-1-imidazolyl) methyl-2-phenylthiomethyl-5-hydroxy-1H-indole-3-carboxylic acid ethyl ester;

furthermore, the 5-hydroxy-3-carboxylate indole derivatives of formula I of the present invention may be formed into pharmaceutically acceptable salts with acids according to conventional methods in the art. The acid may include inorganic or organic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, formic acid, acetic acid, propionic acid, trifluoroacetic acid, maleic acid, tartaric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid and the like.

The term "alkyl" as used above refers to a straight or branched chain alkyl group, "cycloalkyl" refers to a substituted or unsubstituted cycloalkyl group, "alkenyl" refers to a straight or branched chain alkenyl group, and "alkynyl" refers to a substituted or unsubstituted alkynyl group.

The 5-hydroxy indole-3-carboxylic ester compound of the above formula I according to the present invention has the activity of inhibiting SARS-Cov and protecting infected cells, so that it can be clinically used for preparing anti-SARS-Cov virus agent and medicine for treating and/or preventing SARS-Cov infection.

The compounds according to the invention can be used as active ingredients for the preparation of medicaments for the treatment or prophylaxis of SARS-Cov infection, and the invention also provides methods for the treatment or prophylaxis of the abovementioned diseases, which comprise administering a therapeutically effective amount of a compound according to the invention to a patient suffering from or susceptible to such a disease.

The invention comprises a pharmaceutical composition, which contains the 5-oxindole-3-carboxylic ester compound shown in the formula I or pharmaceutically acceptable salt thereof as an active ingredient, and pharmaceutically acceptable excipients. The pharmaceutically acceptable excipient refers to any diluent, adjuvant and/or carrier that can be used in the pharmaceutical field. The compounds of the present invention may be used in combination with other active ingredients as long as they do not produce other adverse effects, such as allergic reactions.

The pharmaceutical composition of the present invention can be formulated into several dosage forms containing some excipients commonly used in the pharmaceutical field; for example, oral formulations (e.g., tablets, capsules, lozenges, solutions or suspensions); injectable formulations (e.g., injectable solutions or suspensions, or injectable dry powders, which are immediately ready for use by addition of water for injection prior to injection); topical formulations (e.g. ointments or solutions).

Carriers for the pharmaceutical compositions of the present invention are of the usual type available in the pharmaceutical art and include: binders, lubricants, disintegrants, solubilizing agents, diluents, stabilizers, suspending agents, non-coloring agents, flavoring agents, etc. for oral preparations; preservatives, solubilizers, stabilizers and the like for injectable preparations; bases for topical formulations, diluents, lubricants, preservatives, and the like. Pharmaceutical formulations may be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or topically), and if certain drugs are unstable under gastric conditions, they may be formulated as enteric coated tablets.

Furthermore, the clinical dosage of the 5-hydroxyindole-3-carboxylate compounds of formula I above for a patient may be based on: the therapeutic efficacy and bioavailability of the active ingredients in vivo, their metabolic and excretory rates and the age, sex, disease stage of the patient are suitably adjusted, although the daily dose for an adult should generally be 10 to 500mg, preferably 50 to 300 mg. Therefore, when the pharmaceutical composition of the present invention is formulated into a unit dosage form, each unit dosage form should contain 10 to 500mg of the 5-hydroxy-3-carboxylate indole compound of the above formula I, preferably 50 to 300mg, in view of the above effective dose. These formulations may be administered in several doses (preferably one to six times) at regular intervals, according to the guidance of a doctor or pharmacist.

The following synthetic schemes A-C describe the preparation of the compounds of formula I of the present invention, all starting materials prepared by the methods described in these schemes, by methods well known to those of ordinary skill in the art of organic chemistry or commercially available. All of the final compounds of the present invention are prepared by the methods described in these schemes or by methods analogous thereto, which are well known to those of ordinary skill in the art of organic chemistry. All variable factors applied in these diagrams are as defined below or in the claims.

Compounds of formula I according to the invention, in scheme A, Z is CH₂SR₅Or CH₂NR₆R₇Substituent R₁、R₂、R₅、R₆And R₇As defined above, wherein R₃、R₄Independently selected from H, amino, C₁～C₁₀Alkyl radical, C₃～C₈Cycloalkyl radical, C₂～C₁₀Alkenyl radical, C₂～C₁₀Alkynyl, which may be substituted by 1 to 3R₈Optionally substituted; or R₃、R₄And N together form a 4-to 10-membered monocyclic or polycyclic saturated ring, wherein the saturated ring optionally includes 1-4 heteroatoms selected from nitrogen, oxygen, and sulfur, except that R₃And R₄The saturated ring optionally includes 1 or 2 carbon-carbon double or triple bonds other than the nitrogen atom to which it is attached, and may be substituted with 1 to 3R₈Optionally substituted;

route A

R is to be₁Adding substituted amine into acetoacetic ester, heating to react to generate a compound 3-substituted amino-2-butenoic ester (A-1), refluxing the compound A-1 and p-benzoquinone in 1, 2-dichloroethane to obtain a compound 1-alkyl substituted-2-methyl-5-hydroxyindole-3-carboxylate (A-2), acylating with acetic anhydride or acetyl chloride to obtain 1-alkyl substituted-2-methyl-5-acetoxy indole-3-carboxylate (A-3), heating and refluxing the A-3 and liquid bromine in carbon tetrachloride under the catalysis of benzoyl peroxide to obtain 1-substituted-5-acetoxy-6-bromo-2-bromomethyl-1H-indole-3-carboxylate (A-4), under alkaline conditions, with suitable HSR₅Or HNR₆R₇Substitution reaction is carried out to obtain a compound A-5, and then the compound A-5, formaldehyde and R are added₃R₄The NH undergoes a Mannich reaction to produce compound A-6 of formula I.

Compounds of formula I according to the invention have in scheme B the substituents X as hydrogen, fluorine or chlorine atoms, the other substituents being as defined for the substituents of the compounds of scheme A.

Route B

Chloroacetoacetates are commercially available as a commercial product, which is reacted with HSR₅Or HNR₆R₇Reacting (substituted thiophenol or amine) at room temperature under an alkaline condition to obtain a compound B-1, then performing reflux reaction on the compound B-1 and substituted amine in 1, hot 2-dichloroethane for 12-24 h to obtain a compound B-2, performing reflux reaction on the compound B-2 and p-benzoquinone in 1, 2-dichloroethane for 6-12 h, cooling to separate out a compound B-3, and performing Mannich reaction to obtain a compound B-4 in a formula I.

Route C

Compounds of formula I according to the invention, in scheme C, Z is as defined for compounds of formula I, R₃、R₄And N together is a guanidino group or a 5-to 10-membered heteroaromatic ring, wherein the heteroaromatic ring optionally includes 1 to 4 heteroatoms selected from nitrogen, oxygen, and sulfur, and may be substituted with 1 to 3R₈Optionally substituted; other substituents are as defined for compounds of formula I.

Preparation of Compound C-1 As described in scheme A or B above, Compound C-1 is reacted with a compound of formula HNR₃R₄Of compounds of (2) wherein NR₃R₄As defined above, the organic solvent used in the reaction is C₁～C₄And (3) heating and refluxing the linear chain or branched chain alcohol and acetonitrile and other polar organic solvents at the reaction temperature for 4-6 h to obtain the compound C-2 shown in the formula I.

The invention has the advantages that: provides a new 5-hydroxy indole-3-carboxylic ester derivative which can be prepared into a medicine for treating or preventing SARS virus, and provides a new medicine for treating infectious atypical pneumonia patients.

The specific implementation mode is as follows:

the compounds of the present invention and their preparation will be better understood in connection with the following examples, which are intended to illustrate, but not to limit, the scope of the invention.

Example 1:

1-methyl-4- (1-piperidinyl) methyl-2-phenylthiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride

Step A: preparation of 3-methylamino-3-butenoic acid ethyl ester (1)

Under stirring and slight heating, 274mL (1.4mol) of methylamine solution is dropped into 300mL of 50% sodium hydroxide solution, generated methylamine gas is introduced into 169mL (1.3mol) of ethyl acetoacetate, the reaction is released heat, stirring is carried out, the reaction solution is cooled in a water bath, the reaction temperature is maintained at 35-40 ℃ until the reaction solution is turbid, light green turbid solution is obtained, the ventilation is stopped, and the reaction solution is stirred at room temperature overnight. The next day, stirring and pumping for 2h, adding 300mL of ether for layering, washing the organic layer with water until the pH value is 8, drying the organic layer with anhydrous sodium sulfate, and concentrating the organic layer under reduced pressure to obtain 180.6g of transparent liquid, wherein the yield is as follows: 97.1%, the concentrate obtained was used directly in the next reaction.

And B: preparation of 1, 2-dimethyl-5-hydroxy-1H-indole-3-carboxylic acid ethyl ester (2)

Dissolving 117.5g (1.05mol) of p-benzoquinone in 1200mL of 1, 2-dichloroethane, dropwise adding the compound (1) under stirring, controlling the reaction temperature to be 40-45 ℃, reacting for 1h, naturally cooling to room temperature, standing overnight, separating out a solid, performing suction filtration, washing with cold acetone, drying, and recrystallizing with acetone to obtain 165.0g of the compound (2), wherein the yield is as follows: 57.8 percent and mp 207-208 ℃.

And C: preparation of 1, 2-dimethyl-5-acetoxy-1H-indole-3-carboxylic acid ethyl ester (3)

Adding 82.7g (0.355mol) of the compound (2) into 500mL of acetone, adding 98.6mL (0.71mol) of triethylamine, stirring until the mixture is completely dissolved, dropwise adding 52mL (0.71mol) of acetyl chloride, controlling the reaction temperature to be 20-25 ℃, stirring at room temperature for reacting for 4 hours, pouring the reaction liquid into ice water while stirring, separating out a solid, filtering, washing with water, drying, and recrystallizing 1, 4-dioxane to obtain 87.4g of a white solid compound (3), wherein the yield is 89.6%.

Step D: preparation of 1-methyl-2- (bromomethyl) -5-acetoxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester (4)

82.5g (0.30mol) of the compound (3) is added into 400mL of carbon tetrachloride, 3 pieces of benzoyl peroxide are added, 43mL (0.84mol) of bromine is added dropwise, the mixture is refluxed for 4 hours after the dropwise addition, and after the reaction, the mixture is kept stand, cooled, filtered, and recrystallized by ethyl acetate to obtain 109g of the compound (4), wherein the yield is 84%.

Step E: preparation of 1-methyl-2-phenylthiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester (5)

Adding 11.5g (0.29mol) of sodium hydroxide and 30.4g (0.24mol) of thiophenol into 400mL of methanol, stirring for 2h at room temperature, adding 94.0g (0.24mol) of the compound (4) in portions, stirring for 6h at room temperature, neutralizing the reaction solution with dilute acetic acid to be neutral, precipitating a large amount of solid, carrying out suction filtration, washing with water, drying, and recrystallizing with ethyl acetate to obtain 92.7g of the compound (5), wherein the yield is as follows: 92.0 percent.

Step F: preparation of 1-methyl-4- (1-piperidinyl) methyl-2-phenylthiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride

Sequentially adding 8.3mL (0.029mol) of piperidine, 3.8mL (0.024mol) of 37% formaldehyde solution and 10g (0.024mol) of compound (5) into 40mL of ethanol, adding a proper amount of glacial acetic acid to adjust the pH value to 4-5, reacting at 40-45 ℃ for 6h, evaporating part of ethanol under reduced pressure, pouring the concentrated solution into 20mL of water, adding 50% sodium hydroxide solution to adjust the pH value to 10-12, extracting diethyl ether, drying an organic layer with anhydrous magnesium sulfate, evaporating to dryness, dissolving the obtained oily substance into 30mL of acetone, dropwise adding hydrochloric ether solution under reflux until the pH value of the reaction solution is 2-3, continuously refluxing for 0.5h, naturally cooling, standing for crystallization, and recrystallizing acetone-ethanol to obtain 9.8g of a solid with the yield of 74%.

¹H-NMR(CDCl₃)：δppm：1.35(t，3H)，1.38(m，1H)，1.75(m，2H)，1.87(m，1H)，2.27(m，2H)，2.91(m，2H)，3.32(m，2H)，3.58(s，3H)，4.21(q，2H)，4.54(s，2H)，5.20(s，2H)，7.22～7.32(m，5H)，7.63(s，1H)，10.25(br s，2H)

The preparation of the compounds of examples 2 to 24 is the same as in example 1.

Example 2

1-methyl-4- (N-methyl-N-butylamino) methyl-2- (3-methylphenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

¹H-NMR(CDCl₃)：δppm：0.98(t，3H)，1.35(t，3H)，1.41(m，2H)，1.96(m，2H)，2.27(s，3H)，2.64(d，2H)，3.14(m，2H)，3.59(s，3H)，4.20(q，2H)，4.48(d，1H)，4.60(d，2H)，5.01(m，1H)，5.48(m，1H)，7.05～7.16(m，4H)，7.63(s，1H)，10.48(s，1H)

example 3

¹H-NMR(CDCl₃)：δppm：0.88(t，3H)，1.27(m，13H)，1.96(m，2H)，2.26(s，3H)，2.63(d，3H)，3.13(m，2H)，3.58(s，3H)，4.19(q，2H)，4.47(d，1H)，4.59(d，1H)，4.98(m，1H)，5.47(m，1H)，7.05～7.16(m，4H)，8.29(s，1H)，10.50(br s，1H)

example 4

1-methyl-4- (N-methyl-N-octylamino) methyl-2- (4-methoxyphenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

¹H-NMR(CDCl₃)：δppm：0.88(t，3H)，1.27～1.33(m，10H)，1.38(t，3H)，1.96(m，2H)，2.63(d，3H)，3.16(m，2H)，3.50(s，3H)，3.7(s，3H)，4.20(q，2H)，4.17(d，2H)，4.22(d，2H)，4.54(m，1H)，5.47(m，1H)，6.75(d，2H)，7.16(d，2H)，7.60(s，1H)，8.26(brs，1H)，10.29(br s，1H)

example 5

¹H-NMR(CDCl₃)：δppm：0.81(t，3H)，3.19(s，1H)，3.21(m，1H)，3.53(d，3H)，3.78(s，3H)，3.89(m，2H)，4.22(q，2H)，4.29(m，2H)，4.45(s，2H)，5.28(d，2H)，6.74(d，2H)，7.17(d，2H)，7.63(s，1H)，11.18(br s，1H)

example 6

4- (4-methyl-1-piperazinyl) methyl-1-cyclopropyl-2- (3, 4-difluorophenyl) thiomethyl-6-bromo-5-hydroxy-1H-indole-3-carboxylic acid ethyl ester dihydrochloride;

¹H-NMR(DMSO)：δppm：1.06(m，2H)，1.26～1.28(m，5H)，2.79(d，3H)，3.17(m，1H)，3.57(m，4H)，3.93(m，4H)，4.23(q，2H)，4.79(s，2H)，5.0(br s，2H)，7.22(m，1H)，7.13(q，1H)，7.53(m，1H)，7.95(s，1H)，9.88(br s，2H)，11.95(br s，1H)

example 7

¹H-NMR(CDCl₃)：δppm：1.11(m，2H)，1.22(m，2H)，1.35(t，3H)，2.87(s，3H)，2.95(m，1H)，3.46(m，2H)，3.64(m，2H)，3.70(s，3H)，4.16(m，2H)，4.32(m，5H)，4.78(s，2H)，5.387(s，2H)，6.82(m，2H)，6.91(d，1H)，7.19(m，1H)，7.90(s，1H)，11.66(br s，1H)，13.61(br s，1H)

example 8

¹H-NMR(DMSO)：δppm：1.07(m，2H)，1.26(m，5H)，2.77(s，3H)，3.14(m，3H)，3.56(m，2H)，3.74(m，4H)，4.21(q，2H)，4.76(s，2H)，5.00(br s，2H)，7.20(m，1H)，7.27(m，1H)，7.39～7.46(m，2H)，7.93(s，1H)，9.95(br s，2H)，12.00(br s，1H)

example 9

¹H-NMR(CDCl₃)：δppm：1.08(m，2H)，1.24(m，2H)，1.35(t，3H)，2.07(m，2H)，2.15(m，2H)，2.91(m，1H)，3.13(m，2H)，3.53(m，2H)，4.21(q，2H)，4.73(s，2H)，5.21(d，2H)，7.21～7.26(m，5H)，7.89(s，1H)，10.78(br s，1H)

example 10

¹H-NMR(CDCl₃)：δppm：1.34(t，3H)，1.38(t，3H)，2.08(m，2H)，2.15(m，2H)，2.32(s，3H)，3.17(m，2H)，3.54(m，2H)，4.09(q，2H)，4.18(，2H)，5.24(d，2H)，5.26(s，2H)，7.05(d，2H)，7.17(d，2H)，8.10(s，1H)，8.10(br s，1H)，10.76(br s，1H)

example 11

H-NMR(CDCl₃)：δppm：1.36(t，3H)，2.09(m，2H)，2.16(m，2H)，2.28(s，3H)，3.15(m，2H)，3.52(m，2H)，4.11(q，2H)，4.21(q，2H)，4.53(s，2H)，5.26(d，2H)，7.05～7.17(m，4H)，7.63(s，1H)，10.77(br s，1H)

example 12

1-ethyl-4- (1-pyrrolidinyl) methyl-2- (2-fluorophenyl) thiomethyl-6-bromo-5-hydroxy-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

¹H-NMR(CDCl₃)：δppm：1.37(t，3H)，1.45(t，3H)，2.07(m，2H)，2.18(m，2H)，3.13(m，2H)，3.52(m，2H)，4.17～4.26(m，4H)，4.56(s，2H)，5.22(d，2H)，6.98～7.11(m，2H)，7.22～7.33(m，2H)，7.64(s，1H)，10.74(s，1H)

example 13

¹H-NMR(CDCl₃)：δppm：1.33(t，3H)，1.40(t，3H)，2.07(m，2H)，2.16(m，2H)，3.14(m，2H)，3.53(m，2H)，4.10(q，2H)，4.18(q，2H)，4.54(s，2H)，5.25(d，2H)，7.22～7.26(m，5H)，7.63(s，1H)，10.79(br s，1H)

example 14

¹H-NMR(CDCl₃)：δppm：1.07(m，2H)，1.21(m，2H)，1.35(t，3H)，2.08(m，2H)，2.16(m，2H)，2.32(s，3H)，2.87(m，1H)，3.14(m，2H)，3.53(m，2H)，4.21(q，2H)，4.68(s，2H)，5.22(d，2H)，7.04(d，2H)，7.16(d，2H)，8.00(s，1H)，8.01(br s，1H)，10.80(br s，1H)

example 15

1-ethyl-4- (4-methyl-1-piperazinyl) methyl-2- (4-fluorophenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester dihydrochloride;

¹H-NMR(DMSO)：δppm：1.22(t，3H)，1.25(t，3H)，2.77(d，3H)，3.30～3.48(m，8H)，4.18(q，2H)，4.24(q，2H)，4.65(s，2H)，5.02(br s，2H)，7.18(m，2H)，7.39(m，2H)8.00(s，1H)，9.69(br s，1H)，11.70(br s，1H)

example 16

1-ethyl-4- (diethylamino) methyl-bromo-2- (4-fluorophenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylic acid ethyl ester hydrochloride;

¹H-NMR(CDCl₃)：δppm：1.33(t，3H)，1.41(t，6H)，1.44(t，3H)，3.10～3.15(m，4H)，4.10(q，2H)，4.17(q，2H)，4.48(s，2H)，5.23(d，2H)，6.94(m，2H)，7.23(m，2H)，7.66(s，1H)，10.04(br s，1H)

example 17

¹H-NMR(CDCl₃)：δppm：1.36(t，3H)，1.41(t，3H)，3.18～3.21(m，4H)，3.87～3.91(m，2H)，4.11(q，2H)，4.21(q，2H)，4.27～4.36(m，2H)，4.50(s，2H)，5.27(d，2H)，6.94(m，2H)，7.23(m，2H)，7.67(s，1H)，9.46(br s，1H)，11.20(br s，1H)

example 18

¹H-NMR(CDCl₃)：δppm：1.35(t，3H)，1.43(t，6H)，2.33(s，3H)，3.14(m，4H)，3.54(s，3H)，4.19(q，2H)，4.49(s，2H)，5.26(d，2H)，7.06(d，2H)，7.17(d，2H)，7.61(s1H)，9.98(br s，1H)

example 19

¹H-NMR(CDCl₃)：δppm：1.35(t，3H)，1.46(t，3H)，2.05(m，2H)，2.17(m，2H)，3.12(m，2H)，3.49(m，2H)，4.15(q，2H)，4.24(q，2H)，4.54(s，2H)，5.14(d，2H)，7.18～7.33(m，3H)，7.63(s，1H)，10.80(br s，1H)

example 20

¹H-NMR(CDCl₃)：δppm：1.36(t，3H)，1.46(t，3H)，2.80(d，6H)，4.16(q，2H)，4.24(q，2H)，4.55(s，2H)，5.11(d，2H)，7.20～7.33(m，3H)，7.65(s，1H)，10.71(br s，1H)

example 21

¹H-NMR(CDCl₃)：δppm：1.31(t，3H)，1.67(d，2H)，3.18(br s，4h)，3.87(m，2H)，4.19(q，2H)，432(m，2H)，4.59(s，2H)，4.81(m，1H)，5.19(d，2H)，7.26(m，5H)，7.91(s，1H)，11.18(br s，1H)

example 22

¹H-NMR(CDCl₃)：δppm：1.07(m，2H)，1.24(m，2H)，1.33(t，3H)，2.79(d，6H)，2.91(m，1H)，4.20(q，2H)，4.67(s，2H)，5.16(d，2H)，6.92(t，2H)，7.21(t，2H)，7.89(s，1H)，10.81(s，1H)

example 23

¹H-NMR(DMSO)：δppm：1.05(m，2H)，1.41(m，5H)，1.65～1.78(m，3H)，1.91～2.01(m，3H)，2.72(s，6H)，3.20(m，2H)，3.46(m，2H)，3.73(s，1H)，4.41(q，2H)，4.87(s，2H)，4.99(s，2H)，8.07(s，1H)，9.21(br s，1H)，9.61(s，1H)，9.93(br s，1H)

example 24

¹H-NMR(DMSO)：δppm：1.05(m，2H)，1.41(m，5H)，2.75(s，6H)，3.77(s，1H)，4.42(q，2H)，4.90(br s，2H)，4.98(s，2H)，8.08(s，1H)，9.16(br s，1H)，9.62(b rs，1H)，10.85(br s，1H)

example 25:

1-methyl-4- (dimethylamino) methyl-2-phenylthiomethyl-5-hydroxy-1H-indole-3-carboxylic acid ethyl ester hydrochloride

Step A: preparation of ethyl phenylthioacetacetate:

dissolving 4.7g (0.12mol) of sodium hydroxide in 80mL of absolute ethanol, dropwise adding 12g (0.11mol) of thiophenol, stirring at room temperature for 1h, dropwise adding 16.4g (0.1mol) of ethyl chloroacetoacetate, reacting at room temperature for 6h, concentrating the reaction solution after the reaction is finished, extracting with diethyl ether, washing with 10% sodium carbonate solution, drying, and evaporating to dryness to obtain 20g of ethyl phenylthioacetoacetate, wherein the yield is 67%.

And B: preparation of 3-methylamino-4-thiophenyl-2-butenoic acid ethyl ester

Introducing methylamine gas (about 0.14mol) into 150mL of 1, 2-dichloroethane solution containing 16.7g (0.07mol) of ethyl phenylthioacetoacetate, reacting at 50-60 ℃ for 18h, washing with water, drying, and evaporating to dryness to obtain 15g of ethyl 4-phenylthio-3-methylamino-2-butenoate with the yield of 86%.

And C: preparation of 1-methyl-2-phenylthiomethyl-5-hydroxy-1H-indole-3-carboxylic acid ethyl ester

Dripping 100mL of 1, 2-dichloroethane solution containing 25g (0.1mol) of 4-thiophenyl-3-methylamino-2-butenoic acid ethyl ester into 60mL of 1, 2-dichloroethane solution containing 11.9g (0.11mol) of p-benzoquinone, keeping the solution slightly boiling in the dripping process, carrying out reflux reaction for 8H after finishing the dripping, cooling for crystallization, carrying out suction filtration and drying after the reaction is finished, thus obtaining 20.1g of 5-hydroxy-1-methyl-2- (phenylthio) methyl-1H-indole-3-carboxylic acid ethyl ester, wherein the yield is 59%.

Step D: preparation of 1-methyl-4- (dimethylamino) methyl-2-phenylthiomethyl-5-hydroxy-1H-indole-3-carboxylic acid ethyl ester hydrochloride

According to the procedure of step E of example 1, the compound ethyl 1-methyl-2-phenylthiomethyl-5-hydroxy-1H-indole-3-carboxylate was reacted with dimethylamine and formaldehyde to give ethyl 1-methyl-4- (dimethylamino) methyl-2-phenylthiomethyl-5-hydroxy-1H-indole-3-carboxylate, which was then reacted with ethyl hydrochloride to give hydrochloride.

¹H-NMR(DMSO)：1.25(t，3H)，2.76(d，6H)，3.69(s，3H)，4.20(m，2H)，4.73(s，2H)，4.75(s，2H)，7.04(d，1H)，7.29～7.41(m，5H)，7.56(d，1H)，8.69(s，1H)

The preparation of the compounds of examples 26 to 29 is carried out in the same manner as in example 24.

Example 26

¹H-NMR(DMSO)：δppm：0.72(m，2H)，0.85(m，2H)，1.09(m，2H)，1.23(m，2H)，1.30(t，2H)，2.61(s，3H)，2.74(m，1H)，3.12(m，1H)，4.24(q，2H)，4.77(s，2H)，4.84(d，2H)，7.00(d，2H)，7.32～7.43(m，5H)，7.65(d，1H)，8.50(br s，2H).

example 27

¹H-NMR(DMSO)：δppm：0.97(s，3H)，0.99(s，3H)，1.04(m，2H)，1.22～1.27(m，5H)，2.13(m，1H)，2.63(s，3H)，3.10(m，3H)，4.20(m，2H)，4.61(m，2H)，4.81(s，2H)，7.02(d，1H)，7.31～7.41(m，5H)，7.67(d，1H)，10.08(s，1H)

example 28

¹H-NMR(DMSO)：δppm：1.24(t，3H)，1.77(m，2H)，2.06(m，2H)，3.15(m，3H)，3.70(s，3H)，4.19(q，2H)，4.77(m，4H)，7.02(d，1H)，7.30～7.42(m，4H)，7.56(d，1H)，8.46(br s，1H)，8.65(br s，1H)，10.02(d，1H)，12.46(br s，1H)

example 29

1-methyl-4- (1-pyrrolidinyl) methyl-2-phenylthiomethyl-5-hydroxy-1H-indole-3-carboxylic acid ethyl ester hydrochloride

¹H-NMR(DMSO)：1.24(t，3H)，1.89(m，2H)，2.03(s，2H)，3.25(m，2H)，3.39(m，4H)，3.69(s，3H)，4.20(m，2H)，4.76(s，2H)，4.83(d，2H)7.04(d，1H)，7.30～7.42(m，5H)，7.54(d，1H)，9.07(s，1H)

Example 30

1-methyl-6-bromo-4- (1H-1-imidazolyl) methyl-2- (4-chlorophenyl) thiomethyl-5-hydroxy-1H-indole-3-carboxylic acid ethyl ester hydrochloride

Ethyl 1-methyl-4- (dimethylamino) methyl-2- (4-chlorophenyl) thiomethyl-5-hydroxy-6-bromo-1H-indole-3-carboxylate prepared according to the procedure of example 1, 10.2g (0.02mol) of this compound was added to 80mL of ethanol, stirred to dissolve it, 3.4g (0.05mol) of imidazole was added thereto, and the mixture was refluxed for 4 hours, concentrated under reduced pressure to give an oily substance, reacted with ethyl ether hydrochloride to give a solid, and recrystallized to give 9.7g of the objective compound, with a yield: 91 percent.

¹H-NMR(DMSO)：1.14(t，3H)，3.72(s，3H)，4.08(q，2)，4.65(s，2H)，5.73(s，2H)，7.28～7.39(m，5H)，7.89(s，1H)，9.05(s，1H)

Example 31

1-methyl-4- (2-methyl-1H-1-imidazolyl) methyl-2-phenylthiomethyl-5-hydroxy-1H-indole-3-carboxylic acid ethyl ester

Prepared according to the method of example 30.

¹H-NMR(DMSO)：1.07(t，3H)，2.33(s，3H)，3.71(s，3H)，3.99(q，2H)，4.65(s，2H)，5.52(s，2H)，6.27(s，1H)，6.52(s，1H)，7.29(m，5H)，7.88(s，1H)，8.95(br s，1H)

Pharmacological study of the products of the invention

Next, the anti-SARS-Cov activity of the 5-hydroxyindole-3-carboxylic acid ester derivative of the above formula I according to the present invention was evaluated in vitro. The Vero-E6 is used as virus host cell, the test sample resists the effect of virus infection cell, the test index is cell denaturation reaction (CPE) and the protection rate of infected cell, the virus strain adopts BJ-01.

The experimental method comprises the following steps:

the samples were dissolved in culture medium or DMSO to make up the appropriate initial concentration, 5-fold dilution, 3 dilutions. Vero-E6 cells are inoculated on a 96-well culture plate, cultured at 37 ℃ under 5% CO, and after SARS virus infects cells, samples with different dilution concentrations are respectively added, and virus control, cell control and sample control are set. The results were observed daily under the mirror, CPE recorded and OD values determined by neutral red staining, and the effect of the samples against SARS virus was calculated and evaluated with reference to the control.

The results of the assay of the partial compounds of the invention are as follows:

protection rate of infected cells: calculating the protective activity of the sample on virus-infected cells by comparing the OD values of the virus control, the cell control and the sample control, wherein the protective rate is more than EC₂₀It was preliminarily thought that the sample had some protective effect on cells infected with the virus and had antiviral activity.

^*Sample cytotoxicity: if the sample has a cytotoxicity to cell control ratio of greater than 50 (> CC)₅₀) CPE evaluation and protection ratio calculation were not given.

Claims

1. The application of 5-hydroxy indole-3-carboxylic ester derivative or racemate or optical isomer or pharmaceutically acceptable salt or hydrate thereof in the preparation of drugs for treating or preventing SARS coronavirus infection,

wherein,

R₁is C₁-C₃Alkyl, cyclopropyl；

R₂Is ethyl;

x represents H, halogen;

z is CH₂SR₅，CH₂NR₆R₇；

R₃、R₄And N together form a guanidino group, or form a 5-or 6-membered saturated or heteroaromatic ring, wherein said saturated or heteroaromatic ring is other than R₃And R₄Optionally 1 to 4 heteroatoms selected from nitrogen, oxygen and sulfur, in addition to the nitrogen atom to which it is attached, said saturated and heteroaromatic rings being substituted with 1 to 3R₈Substitution;

R₅is unsubstituted or substituted by 1 to 3R₈Optionally substituted phenyl;

or R₅Is a 4-10 membered monocyclic or polycyclic saturated ring or a 5-10 membered heteroaromatic ring having 1-4 heteroatoms selected from nitrogen, oxygen, sulfur, wherein said saturated ring and heteroaromatic ring may be substituted with 1-3R₈Optionally substituted;

R₆、R₇and N together is a 5-or 6-membered monocyclic or polycyclic saturated ring, wherein said saturated ring optionally contains 1 to 3 heteroatoms selected from nitrogen, oxygen and sulfur;

R₈is represented by C₁-C₆Alkyl radical, C₃-C₈Cycloalkyl radical, C₂-C₆Alkenyl radical, C₂-C₆Alkynyl, hydroxy, C₁-C₆Alkoxy radical, C₂-C₆Alkenyloxy radical, C₂-C₆Alkynyloxy, mercapto, C₁-C₆Alkylthio radical, C₂-C₆Alkenylthio, phenyl, benzyl, free, salified, esterified or amidated carboxyl, halogen, trifluoromethyl, C₁-C₆Alkanoyl, nitro, cyano, amino, C₁-C₆Alkylamido, or mono-or di (C)₁-C₆Alkyl) substituted amino.

2. The use as claimed in claim 1, wherein Z is CH₂SR₅。

3. The use of claim 2, wherein R₅Is unsubstituted or substituted by 1 to 3 substituents selected from C₁-C₄Alkyl radical, C₁-C₄Alkoxy and halo groups substituted phenyl.

4. The use of claim 1, wherein R₃、R₄And N together are 1-pyrrolidinyl, 1-piperidinyl, 4-carboxylic acid-1-piperidinyl, 1-piperazinyl, 4-methyl-1-piperazinyl, 4-morpholinyl.

5. The use according to claim 1, wherein X is H, a bromine atom.

6. The use according to claim 1, wherein the derivative of formula I is:

1-methyl-4- (2-methyl-1H-1-imidazolyl) methyl-2-phenylthiomethyl-5-hydroxy-1H-indole-3-carboxylic acid ethyl ester.

7. Use of a pharmaceutical composition comprising a derivative of formula I according to claim 1 and pharmaceutically acceptable salts thereof as active ingredient in combination with pharmaceutically acceptable excipients for the manufacture of a medicament for the treatment or prevention of SARS coronavirus infection.