CN115518058A - Application of N-cycloalkyl substituted aromatic methylamine compound in preparation of antiviral drug, structure and preparation method - Google Patents

Abstract

The invention relates to application of N-cycloalkyl substituted aromatic methylamine compounds in preparation of antiviral drugs, and experiments show that the compounds have inhibitory effect on virus infection.

Description

Application of N-cycloalkyl substituted aromatic methylamine compound in preparation of antiviral drug, structure and preparation method

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to application of an N-cycloalkyl substituted aromatic methylamine compound in preparation of an antiviral medicine, a novel compound structure and a preparation method of the compound.

Background

Viral infections have been a significant threat to human health, and humans have historically developed multiple serious viral infection events. With the advance of medicine and the health of public health systems, many viral infectious diseases have been effectively prevented and treated, and vaccination and treatment with antiviral drugs are common means for human beings against viral infectious diseases. Coronavirus is highly variable and vaccines may fail as the virus varies. With the global prevalence of new coronavirus pneumonia (COVID-19), the diagnostic and therapeutic protocol of COVID-19 is continuously updated and perfected, and clinical studies on SARS-CoV-2 show that: antiviral drugs such as the drugs of the rituxivir, the lopinavir/ritonavir, the hydroxychloroquine, the Favipiravir, the Abiradol, the Danorvir and the like do not show clinical treatment advantages to the COVID-19 in clinical tests, but in the aspect of safety, the lopinavir/ritonavir can cause serious gastrointestinal adverse reactions, and the hydroxychloroquine can cause the abnormality of liver and kidney functions and cardiac functions; the Reinecke is suitable for early virus infection, and serious adverse reaction possibly occurs when the Reinecke is used in the middle and later stages, so that the illness condition is further aggravated; the clinical curative effects of Favipiravir, abidol and Danory are not clear; traditional Chinese medicine plays an important role in the treatment process of the novel coronavirus infection, but most of the treatment processes are only suitable for light and common cases; at present, the clinical treatment still takes supportive therapy and improvement of the immunity of patients as main means, and no specific SARS-CoV-2 medicine is available. Aiming at the infection epidemic situation of the novel coronavirus, a medicine with definite treatment effect is searched, the death rate is reduced to the maximum extent, the cure rate is improved, and the effect is not slow at all.

The latest authoritative research in foreign countries confirms that the key action target of the novel coronavirus for infecting the human body is angiotensin converting enzyme ACE2, the novel coronavirus can infect epithelial cells through ACE2 in host cells, and the screening of the novel coronavirus related prevention and treatment medicines is also concentrated on ACE2 combined medicines. A newly published research result of a German research team in 31 days in 1 month and 1 month in 2020 clarifies a new coronavirus SARS-CoV-2 infection pathway, namely a possible virus infection pathway of SARS-CoV-2-ACE2-TMPRSS2, transmembrane serine protease TMPRSS2 starts SARS-CoV-2 spike protein (the spike protein of coronavirus is helpful for the virus to enter target cells), and then the spike protein is combined with ACE2 to enter host cells, and simultaneously, the possibility of clinical treatment of novel coronavirus infection by a TMPRSS2 inhibitor is presumed. The development of targeted TMPRSS2 inhibitors may be an effective means of treating SARS-CoV-2 and may also be a "gold key" to solve the puzzle of all coronavirus infections. In addition, large amounts of mucus were found in anatomical lung specimens from deaths due to SARS-CoV-2 infection, which mucus blocks breathing in critically ill patients, and hypoxic asphyxia may be a significant cause of death in critically ill patients. Based on the above, the compound of the invention can realize the double-overlapping effect of antivirus function and phlegm-reducing and toxin-expelling function, and is undoubtedly more suitable for the treatment of SARS-CoV-2 infection.

Disclosure of Invention

One of the objects of the present invention is to provide the use of a series of N-cycloalkyl substituted aromatic methylamine type compounds, their pharmaceutically acceptable salts or esters, solvates, isomers, polymorphs, isotopically labelled compounds, metabolites or prodrugs for the preparation of antiviral medicaments.

It is still another object of the present invention to provide the N-cycloalkyl substituted aromatic methylamine compound, a pharmaceutically acceptable salt or ester thereof, a solvate, an isomer, a polymorph, an isotopically labeled compound, a metabolite or a prodrug thereof.

The invention further aims to provide a synthesis method of the N-cycloalkyl substituted aromatic methylamine compound.

Another object of the present invention is to provide a pharmaceutical composition or a pharmaceutical preparation of the compound, its pharmaceutically acceptable salts or esters, solvate compounds, isomers, polymorphs, isotopically labeled metabolites or prodrugs as an active ingredient, and the use of the pharmaceutical composition in the preparation of an antiviral medicament.

Definition of terms

The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Laboratory procedures in organic chemistry, pharmaceutical chemistry, biology, the nomenclature used herein and the descriptions herein are those well known and commonly employed in the art. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

As used in the description of the embodiments of the present invention and the appended claims, the singular forms "a," "an," "the," and "the" are intended to refer to both the singular and the plural of the article, unless the context clearly dictates otherwise. For example, a compound includes one or more than one compound.

As used herein, "and/or" refers to and includes any and all possible combinations of one or more of the associated listed items.

As used herein, the term "disease" or "disorder" refers to any change in the state of the body or some organ, interrupting or interfering with its functioning and/or causing symptoms.

As used herein, the term "treatment" is intended to reduce or eliminate the disease state or condition for which it is directed. A subject is successfully "treated" if the subject, following the methods described herein, receives a therapeutic amount of a compound, or a pharmaceutically acceptable salt, isomer, polymorph, solvate, isotopically labeled compound, metabolite or prodrug thereof, or a pharmaceutical composition thereof, that exhibits an observable and/or detectable reduction or improvement in one or more of the subject's indications and symptoms. It is also understood that treatment of the disease state or condition described includes not only complete treatment, but also less than complete treatment, but achieves some biologically or medically relevant result.

As used herein, the term "subject" may refer to a patient or other animal receiving a composition of the invention for the treatment, prevention, alleviation and/or alleviation of a disease or disorder described herein, particularly in humans and mammals.

Technical subject 1

The invention provides an application of a compound with a structure shown as a formula I, a medicinal salt or ester thereof, a solvent compound, an isomer, a polymorphic substance, an isotope labeled compound, a metabolite or a prodrug in preparing an anti-tumor medicament, wherein the formula I comprises the following components:

wherein the content of the first and second substances,

ring A represents a 5-6 membered aromatic or heteroaromatic ring;

n represents 1 to 4.

R ₁ Represents one or more independent substitutions of the following groups: halogen, cyano, nitro, hydroxyl, C1-C5 alkyl, halogen substituted C1-C5 alkyl, amino, C1-C5 alkyl mono-substituted or di-substituted amino, substituted pyranyl amino, C1-C5 oxyl, halogen substituted C1-C5 oxyl and amino substituted C1-C5 acyl.

R ₂ Represents one or more independent substitutions of the following groups: hydrogen, chlorine, cyano-group, nitro-group, hydroxyl, C1-C5 alkyl, amino, C1-C5 alkyl mono-substituted or di-substituted amino;

R ₃ represents H, C1-C5 hydrocarbyl, C1-C5 acyl, or: r ₁ And R ₃ The A ring and the N ring which are connected and respectively connected with the A ring and the N ring form a double-ring structure together;

in some embodiments of the invention, ring a represents a benzene ring, a pyridine ring, a furan ring, a thiophene ring, a pyrrole ring, or a pyrimidine ring.

In some embodiments of the invention, R ₁ Represents one or more independent substitutions of the following groups: halogen, cyano, nitro, hydroxyl, C1-C5 alkyl, amino, C1-C5 alkyl mono-substituted or di-substituted amino, and amino-substituted C1-C5 acyl.

In some embodiments of the invention, R ₁ Represents one or more independent substitutions of the following groups: halogen, cyano, nitro, C1-C5 alkyl, amino, carbamoyl.

In some embodiments of the invention, n represents 1 or 2.

In some embodiments of the invention, the following compounds are included:

n- (2-amino-3, 5-dibromobenzyl) -N-methylcyclohexylamine hydrochloride (bromhexine hydrochloride);

N-methyl-N-cyclohexyl-2-aminobenzamide;

N-methyl-N-cyclohexyl-2-amino-5-bromobenzenemethamine;

n-cyclohexyl-2-amino-3, 5-dibromobenzylamine;

N-methyl-N-cyclohexyl-2-amino-3-bromobenzenemethamine;

N-methyl-N-cyclohexyl-2-amino-3-chloro-5-bromobenzenemethamine;

2-bromo-4-chloro-6- ((cyclohexyl (methyl) amino) methyl) aniline hydrochloride;

(2R, 3S,4S, 5R) -2- (hydroxymethyl) -6- (((2, 4-dibromo-6- ((cyclohexyl (methyl) amino) methyl) phenyl) amino) tetrahydro-2H-pyran-3, 4, 5-triol

2-cyano-5-bromo-N-cyclohexyl-N-methylbenzylamine hydrochloride;

2-cyano-5-fluoro-N-cyclohexyl-N-methylbenzylamine hydrochloride;

2-cyano-5-chloro-N-cyclohexyl-N-methylbenzylamine hydrochloride;

2-amino-3, 5-difluoro-N-cyclohexyl-N-methylbenzylamine;

2-amino-3-bromo-5-nitro-N-cyclohexyl-N-methylbenzylamine hydrochloride;

n- (((3, 5-dibromothien-2-yl) methyl) -N-methylcyclohexylamine hydrochloride;

2-amino-5-nitro-N-cyclohexyl-N-methylbenzylamine hydrochloride;

4- ((cyclohexyl (methyl) amino) methyl) benzamidine hydrochloride;

2-amino-3, 5-dichloro-N-cyclohexyl-N-methylbenzylamine hydrochloride.

As used herein, "hydrocarbyl" may be, but is not limited to, straight, cyclic, or branched-chain hydrocarbyl, and may be saturated or unsaturated, e.g., methyl, ethyl, isopropyl, n-propyl, n-butoxy, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, n-hexyl, isohexyl, cyclopropylmethyl, allyl, and the like.

As used herein, "hydrocarbyloxy" can be, but is not limited to, straight, cyclic, or branched hydrocarbyloxy groups, which can be saturated or unsaturated, such as, for example, methoxy, ethoxy, isopropoxy, n-propoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, n-pentoxy, isopentoxy, cyclopropylmethoxy, allyloxy, and the like.

As used herein, "halo" or "halogen" may be fluoro, chloro, bromo, or iodo.

As used herein, "pharmaceutically acceptable salt" refers to a salt that retains the desired biological activity of the subject compound and exhibits minimal undesirable toxicological effects. When compounds of the invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of the compounds of the invention with a sufficient amount of a base in neat solution or in a suitable inert solvent. The pharmaceutically acceptable base comprises salts prepared from inorganic bases and organic bases, and the salts of the inorganic bases comprise aluminum salts, ammonium salts, calcium salts, copper salts, ferric salts, ferrous salts, lithium salts, magnesium salts, manganese salts, manganous salts, potassium salts, sodium salts, zinc salts and the like. The salts of the organic nontoxic base comprise salts of primary amine, secondary amine and tertiary amine, and comprise substituted amine and cyclic amine. For example: n, N' -dibenzylethylenediamine, diethylamine, 2-dimethylaminoethanol, aminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, histidine, hydroxycobalamin, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine-piperidine solution, polyamine resin, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine and the like. When compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include salts with inorganic acids, including, for example, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, nitric acid, or the like; organic acids such as formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, caproic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2- (4-hydroxybenzoyl) -benzoic acid, camphoric acid, cinnamic acid, cyclopentanepropionic acid, diglucosic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, pamoic acid, pectinic acid, 3-phenylpropionic acid, picric acid, pivalic acid, 2-hydroxyethanesulfonic acid, itaconic acid, sulfamic acid, trifluoromethanesulfonic acid, dodecylsulfuric acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalenedisulfonic acid, camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid, ascorbic acid, glucoheptonic acid, glycerophosphoric acid, aspartic acid, sulfosalicylic acid, and the like include salts of sodium, potassium, magnesium, lithium, aluminum, calcium, zinc, N' -dibenzylethylenediamine, choline, ethylenediamine, N-methyl chloride, and dimethylene.

As used herein, "pharmaceutically acceptable ester" refers to an ester of-OH present in a compound provided herein with an appropriate acid (e.g., a carboxylic acid or an oxygen-containing inorganic acid). Suitable ester groups include, but are not limited to, formate, acetate, propionate, butyrate, acrylate, ethylsuccinate, stearate or palmitate.

As used herein, "isomer" means that where a compound of formula I contains one or more asymmetric centers and/or double bonds, the compounds of the present invention can exist as racemates, racemic mixtures, single enantiomers, diastereomeric mixtures, single diastereomers, geometric isomers, and the like. These compounds may be represented by the symbols "R" or "S", depending on the configuration of the substituents around the stereogenic carbon atom, and possibly by the symbols "Z" or "E", depending on the arrangement of the substituents around the carbon-carbon double bond, or the substituents around the carbon-carbon double bond may be referred to as "cis" or "trans". The compounds disclosed herein may exist as tautomers and both tautomeric forms are intended to be included within the scope of the invention even though only one tautomeric structure is depicted, e.g., keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, imine-enamine tautomers, and the like.

As used herein, "polymorph" means that the compound of formula I may also exist in various crystal forms, by recrystallizing the compound or a pharmaceutically acceptable salt thereof in a solvent to obtain different single crystal forms as well as mixtures of polymorphs thereof.

As used herein, "solvate" means that a compound of formula I may exist as a solvate (e.g., hydrate), wherein the compounds of the invention comprise a polar solvent as a structural element of the crystal lattice of the compound, particularly, for example, water, methanol, or ethanol. The amount of polar solvent, particularly water, may be present in stoichiometric or non-stoichiometric proportions.

As used herein, "isotopic" means that the compound of formula I is also intended to include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the structure of the present invention, but replacing hydrogen with deuterium (2H) or tritium (3H), or replacing carbon with a 13C-or 14C-carbon atom, are within the scope of the present invention. Such compounds are useful, for example, as analytical tools, probes in bioassays, or as therapeutics.

As used herein, "prodrug" means that the compound of formula I may also be in the form of a prodrug or a form that releases the active ingredient upon metabolic change in vivo. The selection and preparation of suitable prodrug derivatives is well known to those skilled in the art.

Subject matter two

A compound having the structure of formula I, a pharmaceutically acceptable salt or ester thereof, a solvate, an isomer, a polymorph, an isotopically labeled compound, a metabolite, or a prodrug thereof:

wherein, the first and the second end of the pipe are connected with each other,

ring A represents a 5-6 membered aromatic or heteroaromatic ring;

n represents 1 to 4.

R ₁ Represents one or more independent substitutions of the following groups: halogen, cyano, nitro, hydroxyl, C1-C5 alkyl, halogen substituted C1-C5 alkyl, amino, C1-C5 alkyl mono-substituted or di-substituted amino, substituted pyranyl amino, C1-C5 oxyl, halogen substituted C1-C5 oxyl and amino substituted C1-C5 acyl.

R ₂ Represents one or more independent substitutions of the following groups: hydrogen, chlorine, cyano-group, nitro-group, hydroxyl, C1-C5 alkyl, amino, C1-C5 alkyl mono-substituted or di-substituted amino;

R ₃ represents H, C1-C5 hydrocarbyl, C1-C5 acyl, or: r ₁ And R ₃ The A ring and the N ring which are connected with each other and are respectively connected with each other form a double-ring structure together;

in some embodiments of the invention, ring a represents a benzene, pyridine, furan, thiophene, pyrrole, or pyrimidine ring.

In some embodiments of the invention, R ₁ Represents one or more independent substitutions of the following groups: halogen, cyano, nitro, hydroxyl, C1-C5 alkyl, amino, C1-C5 alkyl mono-substituted or di-substituted amino, substituted pyranyl amino and amino-substituted C1-C5 acyl.

In some embodiments of the invention, R ₁ Represents one or more independent substitutions of the following groups: halogen, cyano, nitro, C1-C5 alkyl, amino, carbamoyl.

In some embodiments of the invention, n represents 1 or 2.

In some embodiments of the invention, the following compounds are included:

N-methyl-N-cyclohexyl-2-amino-3-chloro-5-bromobenzenemethamine;

2-bromo-4-chloro-6- ((cyclohexyl (methyl) amino) methyl) aniline hydrochloride;

(2R, 3S,4S, 5R) -2- (hydroxymethyl) -6- (((2, 4-dibromo-6- ((cyclohexylamino) methyl) phenyl) amino) tetrahydro-2H-pyran-3, 4, 5-triol

N-cyclohexyl-N-methyl-2-cyano-5-bromobenzenemethamine hydrochloride;

N-cyclohexyl-N-methyl-2-cyano-5-fluorobenzylamine hydrochloride;

N-cyclohexyl-N-methyl-2-cyano-5-chlorobenzylamine hydrochloride;

N-cyclohexyl-N-methyl-2-amino-3, 5-difluorobenzylamine;

N-cyclohexyl-N-methyl-2-amino-3-bromo-5-nitrobenzamide hydrochloride;

n- (((3, 5-dibromothien-2-yl) methyl) -N-methylcyclohexylamine hydrochloride;

N-cyclohexyl-N-methyl-2-amino-5-nitrobenzamide hydrochloride;

4- ((cyclohexyl (methyl) amino) methyl) benzamide hydrochloride.

Subject three

The invention also provides a synthetic method of the compound shown in the formula I.

Route 1:

when R1 is mono-or polysubstituted and A is an aromatic ring, and the other groups are as defined in claim 1, the process shown in scheme 1 is employed:

reducing the compound 1 in an organic solvent by borane to obtain a compound 2, obtaining a compound 3 by the compound 2 in the presence of thionyl chloride, and reacting the compound 3 with cyclohexylamine or nitrogen methyl cyclohexylamine to generate a compound 4;

route 2:

when a = phenyl ring or thiophene, R1 is a disubstituted compound, 2-CN, 5-halogen, and the other groups are as defined in claim 1, the method shown in scheme 2 is employed:

reacting the compound 5 with N-bromosuccinimide in an organic solvent to obtain a compound 6, reacting the compound 6 with N-methyl cyclohexylamine in the presence of the organic solvent to generate a compound 7, and salifying the compound 7 with hydrochloric acid under the action of the organic solvent to obtain a compound 8;

route 3:

when a = phenyl ring, R1 is a disubstituted compound, the 5-position is a formamidinyl group, and the other groups are as defined in claim 1, characterized by the method shown in scheme 2: (1-28 preparation method)

Reacting the compound 9 with NBS in the presence of an organic solvent to obtain a compound 10, reacting the compound 10 with N-methyl-cyclohexylamine or cyclohexylamine to obtain a compound 11, reacting the compound 11 with bis (trimethylsilyl) amido lithium, and salifying the reaction product with hydrochloric acid to obtain a compound 12.

Subject four

The invention provides a pharmaceutical composition, which comprises an N-alkyl substituted aromatic methylamine compound shown as a formula I, a medicinal salt or ester, a solvate, an isomer, a polymorphic substance, an isotope labeled compound, a metabolite or a prodrug thereof, and a pharmaceutically acceptable carrier or excipient.

As used herein, a "pharmaceutical composition" contains a therapeutically effective amount of a compound of formula I, a pharmaceutically acceptable salt or ester, solvate, isomer, polymorph, isotopically labeled compound, metabolite or prodrug thereof, and one or more pharmaceutically acceptable carriers, formulated in the form of tablets, capsules, granules, powders, suspensions, emulsions, powders, solutions, gels, syrups, pills, tinctures, vinum, electuaries, lozenges, mixtures, suppositories, injections, inhalants or sprays. The pharmaceutical composition preferably contains 0.1-99.5% by weight of a compound of formula (la) or a pharmaceutically acceptable salt thereof of the present invention as an active ingredient, more preferably 0.5-99.5% by weight of the active ingredient.

As used herein, "pharmaceutically acceptable carrier or excipient" includes: diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifiers, sweeteners, flavoring agents, taste masking agents, colorants, anti-caking agents, humectants, chelating agents, plasticizers, tackifiers, antioxidants, preservatives, stabilizers, surfactants, and buffers, it will be understood by those skilled in the art that certain pharmaceutically acceptable excipients may be used in more than one function and in alternative functions depending on how much of the excipient is present in the formulation and what other ingredients are present in the formulation. For example: when used orally, it can be formulated into oral preparations such as tablets (including plain tablets, enteric tablets, troches, dispersible tablets, chewable tablets, effervescent tablets, orally disintegrating tablets), capsules (including hard capsules, soft capsules, enteric capsules), granules, pills and the like, containing fillers (e.g., saccharide derivatives such as lactose, sucrose, glucose, mannitol and sorbitol, starch derivatives such as corn starch, potato starch, dextrin and carboxymethyl starch, cellulose derivatives such as crystalline cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, calcium carboxymethyl cellulose, sodium carboxymethyl cellulose, acacia, dextran, silicate derivatives such as magnesium aluminum metasilicate, phosphate derivatives such as calcium phosphate, carbonate derivatives such as calcium carbonate, sulfate derivatives such as calcium sulfate and the like, binders (e.g., gelatin, polyvinylpyrrolidone and polyethylene glycol), disintegrants (e.g., cellulose derivatives such as sodium carboxymethyl cellulose, polyvinylpyrrolidone), lubricants (e.g., talc, calcium stearate, magnesium stearate, sodium cetyl formate, boric acid, benzene, leucine), stabilizers (e.g., methyl paraben, propyl paraben and the like), flavoring agents (e.g., sweeteners, sour agents, and flavors which are commonly used, and the like). When used parenterally, it may be prepared into injection, including sterile powder for injection and solvent for injection, with carrier or excipient including sterile water, ringer's solution and isotonic sodium chloride solution, and with proper additives such as antioxidant, buffering agent, bacteriostat, solubilizer, cosolvent, pH regulator and osmotic pressure regulator. The solubilizer or cosolvent can be poloxamer, lecithin, hydroxypropyl-beta-cyclodextrin, etc.; the pH regulator can be phosphate, acetate, hydrochloric acid, sodium hydroxide, etc.; the osmotic pressure regulator can be sodium chloride, mannitol, glucose, phosphate, acetate, etc. For example, in the preparation of lyophilized powder for injection, mannitol and glucose can also be added as proppant. When used for rectal administration, the medicament may be formulated as a suppository or the like. For pulmonary administration, the medicament may be formulated as an inhalant or a spray. There are many sources available to those skilled in the art which describe pharmaceutically acceptable excipients and which can be used to select suitable pharmaceutically acceptable excipients, for example, books of the pharmaceutical universe of ramiden, the annual book of pharmacy of china, pharmacy, etc.

The compound and the pharmaceutically acceptable salt thereof can be prepared into common preparations, sustained release preparations, controlled release preparations, targeting preparations and various particle delivery systems.

The present invention may be administered by any suitable method known in the art, for example, oral, intravenous, intraperitoneal, intramuscular, topical, transdermal, ocular, nasal, inhalation, subcutaneous, intramuscular, buccal, sublingual, rectal administration and the like, suitable dosages per day ranging from 0.001 to 150mg/Kg body weight, preferably from 0.1 to 100mg/Kg body weight, more preferably from 1 to 60mg/Kg body weight, most preferably from 2 to 30mg/Kg body weight. The above-described dosage may be administered in one dosage unit or divided into several dosage units, depending on the clinical experience of the physician and the dosage regimen including the use of other therapeutic means.

The compounds of the present invention may be used alone, or in combination with one or more other active ingredients, for the treatment, prevention, inhibition, or amelioration of a disease or condition, where the combination of drugs is safer or more effective than either drug alone. Such other agents may be administered, by a route and in an amount commonly used therefor, either simultaneously or sequentially with a compound of the present invention. When a compound of the invention is used contemporaneously with one or more other drugs, a pharmaceutical composition containing the other drug and the compound of the invention in unit dosage form is preferred, particularly in combination with a pharmaceutically acceptable carrier. However, combination therapy may also include therapy in which a compound of the invention and one or more other drugs are administered on different overlapping schedules. It is also contemplated that when used in combination with one or more other active ingredients, the compounds of the present invention and the other active ingredients may be used in lower doses than when each is used alone. Thus, in addition to the compounds of the present invention, the pharmaceutical compositions of the present invention also include those compositions that contain one or more other active ingredients.

The invention has the following beneficial effects:

through a large amount of researches, the inventor discovers that a compound has an inhibiting effect on virus infection, and the presumed mechanism is probably related to TMPRSS2 inhibition, and meanwhile, some compounds in the compound can act on bronchial glands and release lysosomes of mucus secretory cells, so that mucopolysaccharides in mucus are depolymerized, and the viscosity of the mucus is reduced; it can also cause respiratory tract to secrete low-viscosity small molecule mucin, so that sputum becomes thin and is easy to expectorate, and has positive adjuvant therapy effect on asphyxia. The compound provided by the invention can realize double-superposition effect of antivirus action and phlegm-reducing and toxin-expelling action, and provides a new alternative for treating SARS-CoV-2 infection.

Drawings

FIG. 1 is a graph showing the inhibition of SARS-CoV-2 pseudovirus infection by a portion of the compounds of the present invention.

Detailed Description

The present invention is described below in conjunction with specific examples, which are not intended to limit the scope of the present invention, but rather to provide guidance to those skilled in the art in making and using the compounds, compositions, and methods of the present invention. The chemical names of the compounds described in this application are generally from ChemDraw Ultra (chambridge soft) and generated/or generally follow the principles of the IUPAC nomenclature.

The compounds of this example section were prepared by the following route:

example 1N-methyl-N-cyclohexyl-2-aminobenzamide

Adding o-aminobenzaldehyde (5g, 0.041mol) and anhydrous ethanol (25 mL) into a 50mL three-neck flask, cooling to 0-10 ℃, adding sodium borohydride (2g, 0.053 mol) in batches, heating to 25 ℃, stirring for 2h, adding purified water (25 mL) for dilution, stirring for 30min, adjusting the pH value to 6 by using 10% hydrochloric acid, and stirring for 30min. Filtering, washing with purified water, drying the filter cake at 60 ℃ for 5h to obtain the anthranilic alcohol, wherein the reaction yield range is about 90-95%.

Adding o-aminobenzol (4 g, 0.032mol) and dichloromethane (40 ml) into a 150ml three-neck flask, cooling to 0-10 ℃, dropwise adding thionyl chloride (11.42g, 0.096 mol), heating to 30 ℃, stirring for 3h, evaporating residual thionyl chloride at 35 ℃ under reduced pressure, adding petroleum ether (10 ml) into the residue, pulping at room temperature, and performing suction filtration to obtain 2- (chloromethyl) aniline, wherein the yield is 92-96%.

Adding N-methylcyclohexylamine (4.47g, 0.040mol) and ethyl acetate 5mL into a 100mL three-necked flask, cooling the mixed solution to 10-40 ℃, adding 2- (chloromethyl) aniline (4 g, 0.064mol) in batches, stirring for 2h at 25 ℃, adding anhydrous ethanol 10mL, heating to about 65 ℃, adding activated carbon 0.45g, refluxing and decoloring for 30min, performing heat filtration, drying the solvent by decompression of the filtrate, adding ethyl acetate 10mL, stirring for 10min, adjusting the pH value to 5-6 by using 6% HCl/ethanol solution, stirring and crystallizing for 3h at 25 ℃, filtering, adding 20mL purified water into the filter cake, pulping for 30min, filtering, washing the filter cake by using 20mL ethanol, and drying to obtain the compound 01-4, wherein the yield range is 60-65%.

¹ H NMR(400MHz,DMSO):δ7.628-7.609(d,J＝7.6Hz，1H),7.463-7.426(t, J＝14.8Hz，1H),7.324-7.306(d,J＝7.2Hz，1H),7.231-7.215(d,J＝6.4Hz， 1H),

4.678-4.597(t,J＝32.4Hz，1H),4.296(s,1H),3.336-3.277(t,J＝23.6Hz， 1H),2.586-2.504(t,J＝32.8Hz，3H),2.242-2.417(t,J＝10Hz，2H), 1.866-1.836(d,J＝12Hz，2H),1.648-1.508(m,3H),1.327-1.231(m,2H), 1.192-1.037(m,1H).

Example 2N-methyl-N-cyclohexyl-2-amino-5-bromobenzenemethanamine

LiAlH4 (2.2g, 0.058 mol) was added to a 100ml three-necked flask in an ice-water bath, 40ml of anhydrous tetrahydrofuran was added thereto, and a THF solution of methyl 2-amino-5-bromobenzoate (4 g in 10ml of THF, 0.017mol) was added dropwise to the reaction mixture under nitrogen protection, and after the addition, the mixture was warmed to room temperature and stirred for 1 hour. After the reaction is finished, dropwise adding water into the reaction liquid to quench the reaction, filtering, extracting the filtrate with ethyl acetate, combining organic phases, drying with anhydrous sodium sulfate, adding 0.2g of activated carbon for decolorization, filtering, and spin-drying the filtrate to obtain the 2-amino-5-bromo-benzyl alcohol with the yield range of 70-75%.

Adding 6ml of thionyl chloride into a 50ml reaction bottle, cooling to 10 ℃, adding 2-amino-5-bromo-benzyl alcohol (2g, 0.0099mol) in batches, controlling the feeding speed to ensure that the temperature of the reaction solution is 10-30 ℃, after the feeding is finished, stirring and reacting for 1.5 hours at 25 ℃. Removing thionyl chloride by evaporation, adding 5ml of petroleum ether, stirring for 10min, filtering, and drying a filter cake to obtain 2- (chloromethyl) 4-bromoaniline with the yield range of 90-95%.

Adding 2ml of ethyl acetate and N-methylcyclohexylamine (2.3g, 0.020mol) into a 50ml reaction flask, cooling to 15 ℃, adding 2-chloromethyl-4-bromoaniline (2g, 0.0091mol) in batches, controlling the temperature of feed liquid to 15-40 ℃, after the feeding is finished, stirring and reacting for 2 hours at 25 ℃. Adding 4ml of absolute ethyl alcohol, heating to 65 ℃, adding 0.1g of activated carbon, continuously heating and refluxing for 0.5h, filtering while hot, removing the solvent from the filtrate, adding 2ml of ethyl acetate, stirring for 10min, filtering, adjusting the pH to 5-6 by using a prepared 6% ethanol hydrochloride solution, stirring for 0.5h, crystallizing for 3h, filtering and drying to obtain the compound 01-5, wherein the yield is 85-90%.

¹ H NMR(400MHz,MeOD):δ7.691-7.686(d,J＝2.0Hz，1H),δ7.579-7.552(q, J＝10.8Hz，1H),δ7.109-7.088(d,J＝8.4Hz，1H),δ4.566-4.532(d,J＝13.6Hz， 1H),δ4.279-4.245(d,J＝13.6Hz，1H),δ3.454-3.393(m,1H),2.742(s,3H), 2.223-2.191(d,J＝12.8Hz，2H),2.031-1.997(d,J＝13.6Hz，2H), 1.779-1.628(m,3H),1.494-1.396(m,2H),1.330-1.240(m,1H).

Example 3N-cyclohexyl-2-amino-3, 5-dibromobenzylamine

3, 5-dibromo-o-aminobenzaldehyde (5g, 0.018mol), cyclohexylamine (1.95g, 0.020mol) and 50ml of absolute ethanol were charged into a 100ml three-necked flask, and heated under reflux for 5 hours under nitrogen protection. The reaction solution is naturally cooled to room temperature, sodium borohydride (1.36g, 0.038mol) is added into the reaction solution, the mixture is stirred for 18 hours at room temperature, 50ml of purified water is added, dichloromethane is used for extraction, an organic phase is collected, the organic phase is washed by saturated common salt, dried and evaporated to dryness to obtain a crude product. Adding 30ml of dichloromethane, dropwise adding 2N ethanol hydrochloride solution, separating out white solid, filtering, and drying to obtain a compound 01-7 with a yield range of 40-50%.

¹ H NMR(400MHz,MeOD):δ7.658-7.653(d,J＝2.0Hz，1H),7.454-7.448(d, J＝2.4Hz，1H),4.233(s,2H),3.225-3.170(m,1H),1.484-1.349(m,4H), 1.307-1.235(m,1H).

EXAMPLE 4N-methyl-N-cyclohexyl-2-amino-3-bromobenzylamine

Adding 2-amino-4-bromobenzoic acid (10g, 0.043mol) and tetrahydrofuran (150 ml) into a 250ml three-necked flask, cooling to below 0 ℃, adding triethylamine (9.35g, 0.092mol), slowly dropwise adding ethyl chloroformate (5 g, 0.046 mol), stirring for 1h at room temperature after adding is finished, filtering, cooling the filtrate to below 0 ℃, adding 1.5g of lithium aluminum hydride, stirring overnight at room temperature, dropwise adding 3.5ml of water and 15ml of THF solution at 0-10 ℃, then adding 10mL of 30-NaOH solution, stirring for 30min at room temperature, filtering, drying the filtrate, concentrating, and passing through a silica gel column to obtain 2-amino-3-bromobenzoic acid, wherein the yield ranges from 40 to 50%.

To a 100ml three-necked flask was added 2-amino 3-bromobenzyl alcohol (3.75g, 0.019mol). Reacting 50ml of thionyl chloride at room temperature for 2 hours, concentrating, adding dichloromethane into the residue to carry the residual thionyl chloride to obtain 2-bromo-6- (chloromethyl) aniline, wherein the yield range is 90-95%.

Adding 50ml of ethyl acetate and 10ml of N-methyl cyclohexylamine into the concentrate, reacting for 2h at room temperature, removing the solvent and the excessive N-methyl cyclohexylamine, and purifying by column chromatography to obtain the 2-bromo-6- ((cyclohexyl (methyl) amino) methyl) aniline, wherein the yield ranges from 30 to 35 percent.

2-bromo-6- ((cyclohexyl (methyl) amino) methyl) aniline (1.5 g, 0.005 mol) was added to a 100ml single-neck flask, 25ml ethyl acetate was added, 3ml of 2N ethyl acetate solution was added with stirring, stirred at room temperature for 1h, and filtered to give compounds 01-8 with yields ranging from 90-95%.

¹ H NMR(400MHz,DMSO)：δ7.506-7.7.483(q,J＝9.2Hz，1H), 7.397-7.7.378(d,J＝7.6Hz，1H),6.618-6.579(t,J＝15.6Hz，1H), 4.415-4.381(d,J＝11.2Hz，1H),4.239-4.205(d,J＝13.6Hz，1H), 3.296-3.236(t,J＝24.0Hz，1H),2.545-2.504(t,J＝16.4Hz，3H),2.150(s,2H), 1.833(s,2H),1.634-1.468(m,3H),1.292-1.108(m,3H).

Example 5N-methyl-N-cyclohexyl-2-amino-3-chloro-5-bromobenzylamine

Adding 2-amino-3-chlorobenzoic acid (10g, 0.058mol) into a 100ml three-necked bottle, dissolving in 50ml tetrahydrofuran, controlling the temperature to be 0-40 ℃, dropwise adding 5g of bromine, stirring at room temperature after dropwise adding for 24 hours, diluting with water after the reaction is finished, adjusting the pH value to 10 by using sodium carbonate, adding dichloromethane for extraction, washing by using purified water and a saturated sodium chloride solution in sequence, drying by using anhydrous sodium sulfate, filtering and evaporating filtrate to dryness to obtain the 2-amino-3-chloro-5-bromobenzoic acid, wherein the yield is 45-55%.

Adding 2-amino-3-chloro-5-bromobenzoic acid (7 g, 0.028mol) and 75mL of THF into a 150mL three-necked flask, cooling to below 0 ℃, adding triethylamine (5.5 g,0.054 mol), slowly and dropwise adding a solution of ethyl chloroformate (4 g, 0.037mol) and THF (5 mL), stirring at room temperature for 1h after adding, filtering, cooling the filtrate to 0 ℃, adding lithium aluminum hydride (2 g,0.053 mol), stirring at room temperature overnight, adding 2.5mL of water at 0-10 ℃, stirring at room temperature for 30min, filtering, drying the filtrate with anhydrous sodium sulfate, concentrating, and passing through a silica gel column to obtain 2-amino-3-chloro-5-bromobenzyl alcohol with the yield ranging from 55 to 65%.

Adding 2-amino-3-chloro-5-bromobenzyl alcohol (3.75g, 0.016 mol) and 30mL thionyl chloride into a 100mL three-neck flask, reacting for 2 hours at room temperature, concentrating, adding 15mL dichloromethane to the residue to drag residual thionyl chloride to obtain 2-chloro-4-bromo-6- (chloromethyl) aniline, wherein the yield ranges from 90% to 95%.

To the above concentrated residue were added 50mL of ethyl acetate and 10mL of N-methylcyclohexylamine, and the reaction was stirred at room temperature for 2 hours. The solvent and excess N-methylcyclohexylamine are removed and purified by column chromatography to obtain 2-chloro-4-bromo-6- ((cyclohexyl (methyl) amino) methyl) aniline with a yield in the range of 60-70%.

2-chloro-4-bromo-6- ((cyclohexyl (methyl) amino) methyl) aniline (3.5 g, 0.011 mol) was added to a 100ml single-neck flask, 25ml of ethyl acetate was added, 6ml of 2N ethyl hydrochloride solution was added with stirring, and the mixture was stirred at room temperature for 1 hour and filtered to obtain compounds 01 to 9 with a yield ranging from 90 to 95%.

¹ H NMR(400MHz,DMSO)：δ7.628-7.632(d,J＝2.0Hz，1H),7.549-7.544(d, J＝2.0Hz，1H),6.195-5.944(m,1H),4.386-4.352(d,J＝13.6Hz，1H), 4.269(s,1H),3.314-3.255(t,J＝23.6Hz，1H),2.537-2.505(d,J＝12.8Hz， 1H),2.163(s,2H),1.836-1.811(d,J＝10.0Hz，2H),1.635-1.473(m,3H), 1.265-1.107(m,3H).

Example 6-bromo-4-chloro-6- ((cyclohexylmethyl (amino) methyl) aniline hydrochloride

25ml of 2-amino-3-bromo-5-chlorobenzyl alcohol (5 g, 0.021mol) and 30ml of thionyl chloride are added into a 50ml three-necked bottle, the mixture reacts for 2 hours at room temperature, and the thionyl chloride is evaporated under reduced pressure to obtain 2- (chloromethyl) -4-chloro-6-bromoaniline, wherein the yield is 90-95%.

A50 ml three-necked flask was charged with 2- (chloromethyl) -4-chloro-6-bromoaniline (4.5g, 0.018mol), ethyl acetate 45ml, N-methylcyclohexylamine (2.03g, 0.018mol), stirred at room temperature for 2h, and the solvent and excess N-methylcyclohexylamine were removed to give the compound 2-bromo-4-chloro-6- ((cyclohexyl (methyl) amino) methyl) aniline in a yield ranging from 65 to 75%.

The compound 2-bromo-4-chloro-6- ((cyclohexyl (methyl) amino) methyl) aniline (4 g, 0.012mol), ethyl acetate 20ml were added to a 100ml single-neck flask, and 2N ethanol hydrochloride solution 6ml was added dropwise with stirring, stirred at room temperature for 1h, and filtered to obtain the compound 01-12 with a yield ranging from 90-95%.

¹ H NMR(400MHz,DMSO)：δ7.599-7.593(d,J＝2.4Hz，1H),7.563-7.557(d, J＝2.4Hz，1H),4.402-4.368(d,J＝13.6Hz，1H),4.277-4.242(d,J＝14.0Hz， 1H),3.321-3.261(t,J＝24.0Hz，1H),2.539-2.499(m,J＝16.0Hz，3H), 2.158-2.131(d,J＝10.8Hz，2H),1.862-1.812(t,J＝20.0Hz，2H), 1.636-1.554(q,J＝32.8Hz，1H),1.532-1.445(m,2H), 1.300-1.193(m,2H),1.175-1.140(m,1H),

Example 7 (2R, 3S,4R,5S, 6R) -2- ((2, 4-dibromo-6- ((cyclohexyl (methyl) amino) methyl) phenyl) amino) -6- (hydroxymethyl) tetrahydro-2H-pyran-3, 4, 5-triol

To a 500ml single-neck flask were added bromhexine hydrochloride (20g, 0.048 mol) and anhydrous D-glucose (17.57 g,0.033 mol), and methanol (200 ml) was added and stirred, followed by heating for 17 hours. Cooling to-10 deg.C, vacuum spin-drying the reaction solution, purifying the residue by column chromatography to obtain white solid, dissolving with 50ml of methanol, adding 5g of sodium carbonate solid, stirring for 60min, vacuum filtering, vacuum spin-drying the filtrate, purifying by column chromatography once, and vacuum spin-drying to obtain compound 01-13 with yield of 15-20%.

¹ H NMR(400MHz,MDOH)：δ7.615(s,1H),7.269(s,1H),4.718-4.697(d, J＝8.4Hz，1H),4.175-4.142(d,J＝13.2Hz，1H),3.811-3.777(d,J＝13.6Hz， 1H),3.655-3.612(q,J＝17.2Hz，1H),3.469-3.425(t,J＝17.6Hz，2H), 3.331-3.241(m,2H),3.220-3.183(m,2H),2.553-2.500(t,J＝21.2Hz，1H), 2.148(s,3H),1.962-1.934(d,J＝11.2Hz，1H), 1.890-1.834(t,J＝22.4Hz，3H),1.681-1.651(d,J＝12.0Hz，1H), 1.449-1.376(q,J＝29.2Hz，2H),1.343(s,3H),1.304-1.212(q,J＝36.8Hz， 1H).

Example 8-cyano-5-bromo-N-cyclohexyl-N-methylbenzylamine hydrochloride

Adding 4-bromo-2-methylbenzonitrile (5 g, 0.026mol), dichloromethane 50ml, N-bromosuccinimide (5.52g, 0.031mol), N-bromosuccinimide initiator benzoyl peroxide (0.25g, 5%) into a 100ml three-necked flask, stirring, heating to reflux, reacting for 4h while maintaining the temperature, washing the reaction solution with purified water for 3 times, drying the organic phase with anhydrous sodium sulfate, and spin-drying under reduced pressure to obtain the compound 4-bromo-2-methylbenzyl bromide (5.72g, 0.021mol), wherein the yield is 75-80%

4-bromo-2-methylbenzyl bromide (5.72g, 0.021mol), ethyl acetate 50ml, N-methylcyclohexylamine (2.38g, 0.021mol) were added to a 50ml three-neck flask, stirred at room temperature for 2 hours, and the solvent and excess N-methylcyclohexylamine were removed to give the compound 4-bromo-2- ((cyclohexyl (methyl) amino) methyl) benzonitrile in a yield range of 65-75%.

The compound 4-bromo-2- ((cyclohexyl (methyl) amino) methyl) benzonitrile (4.0 g, 0.013mol) and 40ml of ethyl acetate were added to a 100ml single-neck flask, and 7ml of a 2N ethanol hydrochloride solution was added dropwise with stirring, stirred at room temperature for 1 hour, and filtered to obtain compounds 01-14 with a yield ranging from 90 to 95%.

¹ H NMR(400MHz,DMSO)：δ11.009(s,1H),8.424(s,1H),7.924(s,2H), 4.637-4.594(q,J＝17.2Hz，1H),4.294-4.240(q,J＝21.6Hz，1H),3.258(s, 1H),2.645-2.633(d,J＝4.8Hz，3H),2.164(s,2H),1.871-1.817(t,J＝21.6Hz， 2H),1.636-1.518(m,3H),1.307-1.243(t,J＝25.6Hz，2H),1.193(s,1H)

Example 9-cyano-5-fluoro-N-cyclohexyl-N-methylbenzylamine hydrochloride

Adding 4-fluoro-2-methylbenzonitrile (5g, 0.037mol), dichloromethane 50ml, N-bromosuccinimide (7.30g, 0.041mol) and benzoyl peroxide (0.25g, 5%) as N-bromosuccinimide initiator into a 100ml three-necked bottle, stirring and heating to reflux, keeping the temperature for reaction for 4h, washing the reaction solution with purified water for 3 times, drying the organic phase with anhydrous sodium sulfate, and performing rotary drying under reduced pressure to obtain the compound 4-fluoro-2-methylbenzyl bromide (6.33g, 0.030mol), wherein the yield ranges from 75% to 80%

4-fluoro-2-methylbenzyl bromide (6.33g, 0.030mol), ethyl acetate 50ml, N-methylcyclohexylamine (3.40g, 0.030mol) were charged into a 50ml three-necked flask, stirred at room temperature for 2 hours, and the solvent and excess N-methylcyclohexylamine were removed to obtain the compound 4-fluoro-2- ((cyclohexyl (methyl) amino) methyl) benzonitrile in a yield range of 65-75%.

The compound 4-fluoro-2- ((cyclohexyl (methyl) amino) methyl) benzonitrile (5.17g, 0.021mol) and 40ml of ethyl acetate were added to a 100ml single-neck flask, and 12ml of a 2N ethanol hydrochloride solution was added dropwise with stirring, stirred at room temperature for 1 hour, and filtered to obtain the compound 01-15 with a yield ranging from 90 to 95%.

¹ H NMR(400MHz,DMSO)：δ11.252(s,1H),8.185-8.155(q,J＝12.0Hz， 1H),8.110-8.074(q,J＝14.4Hz，1H),7.609-7.560(m,J＝19.6Hz，1H), 4.650-4.605(q,J＝18.0Hz，1H),4.341-4.288(q,J＝21.2Hz，1H), 3.263-3.209(q,J＝21.6Hz，1H),2.654-2.642(d,J＝4.8Hz，3H),2.218-2.148(t, J＝28.0Hz，2H),1.865-1.809(t,J＝22.4Hz，2H)，1.630-1.495(m,3H), 1.322-1.290(d,J＝12.8Hz，2H),1.279-1.268(d,J＝4.4Hz，1H).

Example 10 2-cyano-5-chloro-N-cyclohexyl-N-methylbenzylamine hydrochloride

Adding 4-chloro-2-methylbenzonitrile (5g, 0.033mol), dichloromethane 50ml, N-bromosuccinimide (6.46g, 0.036 mol) and benzoyl peroxide (0.25g, 5%) as N-bromosuccinimide initiator into a 100ml three-necked flask, stirring and heating to reflux, keeping the temperature for reaction for 4h, washing the reaction solution with purified water for 3 times, drying the organic phase with anhydrous sodium sulfate, and performing rotary drying under reduced pressure to obtain the compound 4-chloro-2-methylbenzyl bromide (6.08g, 0.026mol), wherein the yield is 75-80%

4-chloro-2-methylbenzyl bromide (6.08g, 0.026mol), ethyl acetate 50ml, N-methylcyclohexylamine (2.99g, 0.026mol) were added to a 50ml three-necked flask, stirred at room temperature for 2 hours, and the solvent and excess N-methylcyclohexylamine were removed to give the compound 4-chloro-2- ((cyclohexyl (methyl) amino) methyl) benzonitrile in a yield ranging from 65 to 75%.

The compound 4-fluoro-2- ((cyclohexyl (methyl) amino) methyl) benzonitrile (4.78g, 0.018mol) and ethyl acetate (50 ml) were charged into a 100ml single-neck flask, and 2N ethanol hydrochloride solution (10 ml) was added dropwise with stirring, stirred at room temperature for 1 hour, and filtered to give the compound 01-16 in a yield range of 90-95%.

¹ H NMR(400MHz,DMSO)：δ11.3302(s,1H),8.365-8.360(d,J＝2.0Hz， 1H),8.021-8.000(d,J＝8.4Hz，1H),7.791-7.765(q,J＝10.4Hz，1H), 4.642-4.598(q,J＝17.6Hz，1H),4.315-4.262(q,J＝21.2Hz，1H), 3.271-3.217(t,J＝21.6Hz，1H),2.640-2.628(d,J＝4.8Hz，3H),2.222-2.163(t, J＝23.69Hz，2H),1.864-1.808(t,J＝22.4Hz，2H)，1.630-1.528(q,3H), 1.330-1.265(q,J＝26.0Hz，2H),1.239(s,1H).

Example 11 2-amino-3, 5-difluoro-N-cyclohexyl-N-methylbenzylamine

Adding 2-amino-3, 5 difluorobenzoic acid (5 g, 0.029mol) and tetrahydrofuran (20 ml) into a 100ml three-neck flask, stirring, cooling to 0-10 ℃, dropwise adding 1mol/L borane tetrahydrofuran solution (44 ml), reacting at room temperature overnight after dropwise adding, adding methanol to quench borane, and performing reduced pressure spin drying to obtain the compound 2-amino-3, 5 difluorobenzyl alcohol, wherein the yield range is 75-85%.

Adding 2-amino-3, 5-difluorobenzyl alcohol (3.7 g, 0.023mol), 20ml of dichloromethane and thionyl chloride (8.2g, 0.069mol) into a 100ml three-neck flask, reacting for 5h at 40 ℃, spin-drying under reduced pressure, evaporating off the solvent and the thionyl chloride, dragging the residue with dichloromethane, adding 15ml of ethyl acetate into the residue, cooling to 0-10 ℃, adding N-methylcyclohexylamine (2.6 g, 0.023mol) with stirring, reacting for 2h at room temperature, removing the solvent and the excess N-methylcyclohexylamine to obtain the compound 01-17 with the yield ranging from 65-75%.

¹ H NMR(400MHz,DMSO)：δ6.992-6.935(m,1H),6.786-6.763(d,J＝9.2Hz， 1H),5.098(s,2H),3.544(s,2H),2.405-2.350(m,1H),2.078(s,3H), 1.762-1.743(d,J＝7.6Hz，4H),1.589-1.559(d,J＝10.2Hz，1H), 1.321-1.050(m,5H)

Example 12-amino-3-bromo-5-nitro-N-cyclohexyl-N-methylbenzylamine hydrochloride

Adding 2-amino-3 bromo-5-nitrobenzoic acid (5 g, 0.019mol) and tetrahydrofuran (20 ml) into a 100ml three-necked bottle, stirring, cooling to 0-10 ℃, dropwise adding 1mol/L borane tetrahydrofuran solution (29 ml), reacting at room temperature overnight after dropwise adding, adding methanol to quench borane, and performing reduced pressure spin drying to obtain the compound 2-amino-3 bromo-5-nitrobenzyl alcohol, wherein the yield range is 75-85%.

Adding 2-amino-3 bromo-5-nitrobenzol (4.7 g, 0.019mol), dichloromethane (20 ml), thionyl chloride (6.78g, 0.057mol) into a 100ml three-necked flask, reacting for 4h at 40 ℃, spin-drying under reduced pressure, distilling off the solvent and thionyl chloride, carrying the residue with dichloromethane, adding ethyl acetate (20 ml) into the residue, cooling to 0-10 ℃, adding N-methylcyclohexylamine (2.2g, 0.019mol) with stirring, reacting for 2h at room temperature, removing the solvent and excess N-methylcyclohexylamine to obtain the compound 2-bromo-6- ((cyclohexyl (methyl) amino) methyl) -4-nitroaniline with the yield ranging from 65-75%.

The compound 2-bromo-6- ((cyclohexyl (methyl) amino) methyl) -4-nitroaniline (4.55g, 0.013mol) and 40ml of ethyl acetate were added to a 100ml single-neck flask, and 7ml of a 2N ethanol hydrochloride solution was added dropwise with stirring, stirred at room temperature for 1 hour, and filtered to obtain the compound 01-18 with a yield ranging from 90 to 95%.

¹ H NMR(400MHz,DMSO+D ₂ O)：δ8.352-8.345(d,J＝2.8Hz，1H), 8.321-8.315(d,J＝2.4Hz，1H),4.471-4.436(q,J＝14.0Hz，2H),3.420-3.358(t, J＝24.8Hz，1H),2.552-2.505(q,J＝18.8Hz，3H),2.142-2.114(d,J＝11.2Hz， 1H),2.045-2.019(d,J＝10.4Hz，1H),1.876-1.814(t,J＝24.8Hz， 2H),1.636-1.605(d,J＝12.4Hz，1H),1.498-1.437(t,J＝24.4Hz， 2H),1.324-1.228(q,J＝38.4Hz，2H),1.178-1.113(q,J＝26.0Hz，1H).

EXAMPLE 13N- (((3, 5-dibromothien-2-yl) methyl) -N-methylcyclohexylamine hydrochloride

Adding 3, 5-dibromothiophene-2-carboxylic acid (5g, 0.017mol) and tetrahydrofuran (20 ml) into a 100ml three-necked flask, stirring and cooling to 0-10 ℃, dropwise adding 26ml of borane tetrahydrofuran solution with the concentration of 1mol/L, reacting overnight at room temperature after dropwise adding, adding methanol to quench borane, and performing rotary drying under reduced pressure to obtain the compound (3, 5-dibromothiophene-2-yl) methanol with the yield range of 75-85%.

2-amino-3, 5-difluorobenzyl alcohol (3.7g, 0.014mol), dichloromethane (20 ml), thionyl chloride (1.67g, 0.042mol) were added to a 100ml three-necked flask, reacted at 40 ℃ for 3 hours, dried under reduced pressure, the solvent and thionyl chloride were evaporated off, the residue was taken up with dichloromethane, ethyl acetate (15 ml) was added to the residue, cooled to 0-10 ℃, N-methylcyclohexylamine (1.58g, 0.014mol) was added with stirring, reacted at room temperature for 2 hours, the solvent and excess N-methylcyclohexylamine were removed to give the compound N- ((3, 5-dibromothiophen-2-yl) methyl) -N-methylcyclohexylamine with a yield ranging from 65 to 75%.

Adding the compound N- ((3, 5-dibromothiophen-2-yl) methyl) -N-methylcyclohexylamine (3.59g, 0.0098mol) and 30ml of ethyl acetate into a 100ml single-neck bottle, dropwise adding 8ml of 2N hydrochloric acid ethanol solution while stirring, stirring for 1h at room temperature, and filtering to obtain the compound 01-24 with the yield range of 90-95%.

¹ H NMR(400MHz,DMSO)：δ11.161(s,1H),7.449(s,1H),4.551-4.503(q, J＝19.20Hz，1H),4.389-4.337(q,J＝20.8Hz，1H),3.170-3.117(q,J＝21.2Hz， 1H),2.639-2.627(d,J＝4.8Hz，3H),2.143-2.115(d,J＝11.2Hz，2H), 1.848-1.788(t,J＝24.0Hz，2H),1.619-1.441(m,3H),1.400-1.180(m,3H).

Example 14-amino-5-nitro-N-cyclohexyl-N-methylbenzylamine hydrochloride

Adding 2-amino-5-nitrobenzoic acid (5 g, 0.027mol) and tetrahydrofuran (20 ml) into a 100ml three-necked flask, stirring and cooling to 0-10 ℃, dropwise adding 1mol/L borane tetrahydrofuran solution (27 ml), reacting at room temperature overnight, adding methanol to quench borane, and performing reduced pressure spin drying to obtain the compound 2-amino-5-nitrobenzol, wherein the yield range is 75-85%.

Adding 2-amino-5-nitrobenzol (3.69g, 0.022mol), dichloromethane (20 ml) and thionyl chloride (7.84g, 0.066 mol) into a 100ml three-neck flask, reacting at 40 ℃ for 5h, performing rotary drying under reduced pressure, evaporating to remove the solvent and the thionyl chloride, dragging the residue with dichloromethane, adding ethyl acetate (15 ml) into the residue, cooling to 0-10 ℃, adding N-methylcyclohexylamine (2.5g, 0.022mol) with stirring, reacting at room temperature for 2h, removing the solvent and excessive N-methylcyclohexylamine to obtain the compound 2- ((cyclohexyl (methyl) amino) methyl) -4-nitroaniline, wherein the yield is 65-75%.

The compound 2- ((cyclohexyl (methyl) amino) methyl) -4-nitroaniline (4.06g, 0.015 mol) and 30ml of ethyl acetate are added into a 100ml single-neck bottle, 8ml of 2N hydrochloric acid ethanol solution is dropwise added under stirring, the mixture is stirred for 1h at room temperature and filtered to obtain the compound 01-26, and the yield ranges from 90 to 95%.

¹ H NMR(400MHz,CDCl ₃ )：δ11.670(s,1H),8.143-8.126(d,J＝6.8Hz，1H), 8.033(s,1H),6.811-6.792(d,J＝7.6Hz，1H),4.313(s,1H),4.113(s, 1H),3.194(s,1H), 2.672(s,3H),2.465(s,1H),2.273(s,1H),2.032-1.961(t,J＝28.4Hz，2H),1.791-1.760(d,J＝12.4Hz，1H),1.633-1.554(d,J＝31.6Hz，2H), 1.340-1.215(m,3H).

Example 15- ((cyclohexyl (methyl) amino) methyl) benzamidine hydrochloride

Adding 4-methylbenzonitrile (5 g, 0.026mol), dichloromethane (50ml), N-bromosuccinimide (5.16g, 0.029mol) and benzoyl peroxide (0.26 g, 5%) as N-bromosuccinimide initiator into a 100ml three-neck flask, stirring, heating to reflux, keeping the temperature for reaction for 4h, washing the reaction solution with purified water for 3 times, drying the organic phase with anhydrous sodium sulfate, and performing rotary drying under reduced pressure to obtain the compound 4- (bromomethyl) benzonitrile, wherein the yield ranges from 75 to 80 percent

4- (bromomethyl) benzonitrile (5.72g, 0.021mol), ethyl acetate 50ml, N-methylcyclohexylamine (2.38g, 0.0.021mol) were added to a 50ml three-neck flask, stirred at room temperature for 2h, and the solvent and excess N-methylcyclohexylamine were removed to give the compound 4- ((cyclohexyl (methyl) amino) methyl) benzonitrile in a yield range of 65-75%.

Adding compound 4- ((cyclohexyl (methyl) amino) methyl) benzonitrile (4.52 g,0.015 mol), 50ml of tetrahydrofuran and 15ml of lithium bis (trimethylsilyl) amide at 1mol/L into a 100ml three-necked flask, reacting for 8h in a water bath at 40 ℃, adjusting the pH to 1-3 by using concentrated hydrochloric acid, taking the lower layer liquid, performing reduced pressure spin drying, and performing column purification to obtain compound 1-28, wherein the yield is within 35-40%.

¹ H NMR(400MHz,MeOD):δ7.948-7.927(d,J＝8.4Hz，2H),7.878-7.857(d, J＝8.4Hz，2H),4.487(s,2H),3.365-3.289(m,J＝30.4Hz，1H),2.751(s,3H), 2.224-2.197(d,J＝10.8Hz，2H),2.002-1.969(d,J＝13.2Hz，2H), 1.762-1.603(m,3H),1.453-1.356(m,J＝38.8Hz，2H),1.330-1.283(m,J ＝18.8Hz，1H).

Example 16 2-amino-3, 5-dichloro-N-cyclohexyl-N-methylbenzylamine hydrochloride

Adding 2-amino 3, 5-dichlorobenzoic acid (5 g, 0.024mol) and tetrahydrofuran (20 ml) into a 100ml three-neck flask, stirring and cooling to 0-10 ℃, dropwise adding 1mol/L borane tetrahydrofuran solution (24 ml), reacting at room temperature overnight, adding methanol to quench borane, and performing reduced pressure rotary drying to obtain the compound 2-amino 3, 5-dichlorobenzoic acid, wherein the yield range is 75-85%.

2-amino-5-nitrobenzol (3.7g, 0.019mol), dichloromethane (20 ml), thionyl chloride (6.93g, 0.058mol) are added into a 100ml three-necked flask, reaction is carried out for 5h at 40 ℃, decompression and spin drying are carried out, the solvent and the thionyl chloride are evaporated, the residue is dragged by the dichloromethane, ethyl acetate (15 ml) is added into the residue, the temperature is reduced to 0-10 ℃, N-methylcyclohexylamine (2.15g, 0.013mol) is added with stirring and reaction is carried out for 2h at room temperature, the solvent and the excessive N-methylcyclohexylamine are removed, and the compound 2, 4-dichloro-6- ((cyclohexyl (methyl) amino) methyl) aniline is obtained, wherein the yield ranges from 65 to 75%.

The compound 2, 4-dichloro-6- ((cyclohexyl (methyl) amino) methyl) aniline (2.15g, 0.013mol) and 30ml of ethyl acetate were added to a 100ml single-neck flask, 8ml of a 2N ethanol hydrochloride solution was dropwise added with stirring, stirred at room temperature for 1 hour, and filtered to obtain the compound 01-26 with a yield range of 90-95%.

¹ H NMR(400MHz,DMSO+D ₂ O)：δ7.463(s,1H),308(s,1H),4.320(s, 1H),4.170(s,1H),3.256(s,1H),2.539-2.511(t,J＝11.2Hz，3H),2.016 (s,2H),1.819(s,1H),1.607-1.581(d,J＝10.4Hz，1H),1.446(s， 2H),1.259-1.233(d,J＝10.4Hz，2H),1.116(s,1H).

Test example 1 evaluation of inhibitory Effect against SARS-CoV-2 pseudovirus infection

SARS-CoV-2 is a highly pathogenic pathogen, and live virus operation needs to be carried out in a tertiary biosafety laboratory, which greatly limits the development of antiviral drugs. The pseudovirus technology can well solve the problem. Pseudoviruses are also known as chimeric virus particles, i.e., recombinant glycoproteins of a virus of interest are expressed on the surface of a replication-defective viral vector, thereby producing individual chimeric virus particles. The pseudovirus can reduce the biological safety level to the second level, has good consistency with the neutralization experiment of the true virus, has become a powerful tool for researching the pathogenesis and the prevention and treatment measures of the highly pathogenic virus, and can be used for screening the virus entry inhibitor.

The invention uses replication defective slow virus vector as frame, surface mosaic SARS-CoV-2 spike protein S, to construct coronavirus pseudo virus. The pseudovirus is provided with a luciferase reporter gene, and the fluorescence emitted under the action of a specific substrate can be quickly and quantitatively detected and analyzed by a specific detection instrument, so that an active drug with a potential antiviral effect can be quickly and efficiently screened in vitro. The Vero cells over expressing ACE2 are inoculated with SARS-CoV-2 pseudovirus at different time after adding medicine, and virus entrance inhibition curve is drawn according to luciferase detection result. Analyzing the action time of the antiviral drug, and clarifying the antiviral action mechanism of the high-activity compound. The inhibition curve of the compound for inhibiting SARS-CoV-2 pseudovirus infection in vitro is as follows:

the inhibition curve of some compounds on SARS-CoV-2 pseudovirus infection is shown in Table 1 by using bromhexine hydrochloride (1-01) as a control, the in vitro inhibition activity is shown in Table 1, and the inhibition rate of different concentrations of compounds on SARS-CoV-2 pseudovirus is shown in Table 2.

Table 1: compound in vitro inhibition efficiency of SARS-COV-2 pseudovirus

As can be seen from FIG. 1 and Table 1, the inhibition rates and curve fitting of the compounds 1-13 are better, the inhibition effects of 1-8 are the best, and the inhibition effects of 1-4 and 1-5 are the second best; 1-7, 1-9 dose-dependences were strong, and 1-12 dose-dependences were less.

TABLE 2 inhibition of SARS-CoV-2 pseudovirus by different concentrations of the compound

Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.

Claims (9)

1. Use of a compound having a structure as shown in formula I, a pharmaceutically acceptable salt or ester thereof, a solvate, an isomer, a polymorph, an isotopically labeled compound, a metabolite or a prodrug thereof in the preparation of an anti-tumor medicament:

wherein, the first and the second end of the pipe are connected with each other,

ring A represents a 5-6 membered aromatic or heteroaromatic ring;

n represents 1 to 4.

R ₁ Represents one or more independent substitutions of the following groups: halogen, cyano, nitro, hydroxyl, C1-C5 alkyl, halogen substituted C1-C5 alkyl, amino, C1-C5 alkyl mono-substituted or di-substituted amino, substituted pyranyl amino, C1-C5 oxyl, halogen substituted C1-C5 oxyl and amino substituted C1-C5 acyl.

R ₂ Represents one or more independent substitutions of the following groups: hydrogen, chlorine, cyano, nitro, hydroxyl, C1-C5 alkyl, amino, C1-C5 alkyl mono-substituted or di-substituted amino;

R ₃ represents H, C1-C5 hydrocarbyl, C1-C5 acyl, or: r ₁ And R ₃ The A rings and N, which are linked and linked to each other, together form a bicyclic structure.

2. The compound of formula I, a pharmaceutically acceptable salt or ester thereof, a solvate, an isomer, a polymorph, an isotopically labeled compound, a metabolite or a prodrug thereof according to claim 1, wherein ring a represents a benzene ring, a pyridine ring, a furan ring, a thiophene ring, a pyrrole ring or a pyrimidine ring.

3. A compound of formula I, pharmaceutically acceptable salts or esters thereof, solvates, isomers, polymorphs, isotopically labeled compounds, metabolites or prodrugs according to claim 1, wherein R is ₁ Represents one or more independent substitutions of the following groups: halogen, cyano, nitro, hydroxyl, C1-C5 alkyl, amino, C1-C5 alkyl mono-substituted or di-substituted amino, and amino-substituted C1-C5 acyl.

4. A compound of formula I, pharmaceutically acceptable salts or esters thereof, solvate, isomers, polymorphs, isotopically labeled compounds, metabolites or prodrugs according to claim 1 wherein R ₁ Represents one or more independent substitutions of the following groups:halogen, cyano, nitro, C1-C5 alkyl, amino, carbamoyl.

5. A compound of formula I, pharmaceutically acceptable salts or esters thereof, solvate compounds, isomers, polymorphs, isotopically labeled compounds, metabolites or pro-drugs according to claim 1 wherein n represents 1 or 2.

6. A compound of formula I, a pharmaceutically acceptable salt or ester thereof, a solvate, an isomer, a polymorph, an isotopically labeled compound, a metabolite or a prodrug thereof according to claim 1, comprising the following compounds:

n- (2-amino-3, 5-dibromobenzyl) -N-methylcyclohexylamine hydrochloride (bromhexine hydrochloride);

N-methyl-N-cyclohexyl-2-aminobenzamide;

N-methyl-N-cyclohexyl-2-amino-5-bromobenzenemethamine;

n-cyclohexyl-2-amino-3, 5-dibromobenzylamine;

N-methyl-N-cyclohexyl-2-amino-3-bromobenzenemethamine;

N-methyl-N-cyclohexyl-2-amino-3-chloro-5-bromobenzenemethamine;

2-bromo-4-chloro-6- ((cyclohexyl (methyl) amino) methyl) aniline hydrochloride;

(2R, 3S,4S, 5R) -2- (hydroxymethyl) -6- (((2, 4-dibromo-6- ((cyclohexyl (methyl) amino) methyl) phenyl) amino) tetrahydro-2H-pyran-3, 4, 5-triol

2-cyano-5-bromo-N-cyclohexyl-N-methylbenzylamine hydrochloride;

2-cyano-5-fluoro-N-cyclohexyl-N-methylbenzylamine hydrochloride;

2-cyano-5-chloro-N-cyclohexyl-N-methylbenzylamine hydrochloride;

2-amino-3, 5-difluoro-N-cyclohexyl-N-methylbenzylamine;

2-amino-3-bromo-5-nitro-N-cyclohexyl-N-methylbenzylamine hydrochloride;

n- (((3, 5-dibromothien-2-yl) methyl) -N-methylcyclohexylamine hydrochloride;

2-amino-5-nitro-N-cyclohexyl-N-methylbenzylamine hydrochloride;

4- ((cyclohexyl (methyl) amino) methyl) benzamidine hydrochloride;

2-amino-3, 5-dichloro-N-cyclohexyl-N-methylbenzylamine hydrochloride.

7. A compound having the structure shown in formula I, a pharmaceutically acceptable salt or ester thereof, a solvate compound, an isomer, a polymorph, an isotopically labeled compound, a metabolite or a prodrug thereof:

wherein the content of the first and second substances,

ring A represents a 5-6 membered aromatic or heteroaromatic ring;

n represents 1 to 4.

R ₁ Represents one or more independent substitutions of the following groups: halogen, cyano, nitro, hydroxyl, C1-C5 alkyl, halogen substituted C1-C5 alkyl, amino, C1-C5 alkyl mono-substituted or di-substituted amino, substituted pyranyl amino, C1-C5 oxyl, halogen substituted C1-C5 oxyl and amino substituted C1-C5 acyl.

R ₂ Represents one or more independent substitutions of the following groups: hydrogen, chlorine, cyano, nitro, hydroxyl, C1-C5 alkyl, amino, C1-C5 alkyl mono-substituted or di-substituted amino;

R ₃ represents H, C1-C5 hydrocarbyl, C1-C5 acyl, or: r is ₁ And R ₃ The A ring and N ring which are linked and linked to each other form a bicyclic structure.

8. A process for the synthesis of compounds of formula I according to claim 7, selected from the following routes:

route 1:

when R1 is mono-or polysubstituted and A is an aromatic ring, and the other groups are as defined in claim 1, the process shown in scheme 1 is employed:

reducing the compound 1 in an organic solvent by borane to obtain a compound 2, obtaining a compound 3 from the compound 2 in the presence of thionyl chloride, and reacting the compound 3 with cyclohexylamine or N-methyl cyclohexylamine to generate a compound 4;

route 2:

when a = phenyl ring or thiophene, R1 is a disubstituted compound, the substituents being-CN, halogen, respectively, and the other groups being as defined in claim 1, the method shown in scheme 2 is employed:

reacting the compound 5 with N-bromosuccinimide in an organic solvent to obtain a compound 6, reacting the compound 6 with N-methyl cyclohexylamine in the presence of the organic solvent to generate a compound 7, and salifying the compound 7 with hydrochloric acid under the action of the organic solvent to obtain a compound 8;

route 3:

when a = phenyl ring, R1 is a disubstituted compound, the 5-position is a formamidinyl group, and the other groups are as defined in claim 1, characterized by the method shown in scheme 2: (1-28 preparation method)

Reacting the compound 9 with NBS in the presence of an organic solvent to obtain a compound 10, reacting the compound 10 with N-methyl-cyclohexylamine or cyclohexylamine to obtain a compound 11, reacting the compound 11 with bis (trimethylsilyl) amido lithium, and salifying the reaction product with hydrochloric acid to obtain a compound 12.

9. A pharmaceutical composition comprising an N-alkyl substituted aromatic methylamine compound of formula I, pharmaceutically acceptable salt or ester thereof, solvate, isomer, polymorph, isotopically labeled compound, metabolite or prodrug as claimed in claim 7, and a pharmaceutically acceptable carrier or excipient.

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