Preparation of a nucleoside derivative and its use in the field of antiviral drugs

Abstract

This invention relates to a compound of formula I, its preparation method, and its uses. The compound of this invention, by virtue of its excellent antiviral activity, is suitable for development into novel antiviral drugs.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology. Specifically, the compounds of this invention enhance the antiviral activity of drug molecules by introducing carbon-sulfur double bond groups, and have good prospects for drug development. Background Technology

[0002] Orthomyxoviruses are viruses that have an affinity for the mucins on the surface of human or certain animal red blood cells. They possess a single-stranded RNA genome, and influenza viruses belong to this category. Orthomyxoviruses mainly include three types: influenza A, influenza B, and influenza C. Among them, influenza A is the most potent, infecting the largest number of people during flu season and causing severe respiratory infections, resulting in more than 300,000 deaths worldwide each year. In China, tens of millions of people are infected with influenza viruses annually, especially infants and the elderly, where morbidity and mortality rates are relatively high, and it can cause diseases such as pneumonia. Therefore, developing effective antiviral drugs to treat influenza A is particularly important. Currently, the main antiviral drugs on the market include amantadine, neuraminidase inhibitors oseltamivir and zanamivir. However, these compounds also exhibit poor physicochemical properties, such as low solubility and low bioavailability, making them unsuitable as ideal influenza treatments. Summary of the Invention

[0003] The first aspect of the present invention is to provide a method for preparing a nucleoside derivative with a novel structure and its use in the field of antiviral drugs.

[0004] A second aspect of the invention is that the compounds of the invention exhibit high antiviral activity in vitro and in vivo.

[0005] A third aspect of the invention is that the provided compounds can treat individuals infected with orthomyxoviridae viruses, paramyxoviridae viruses, coronaviruses, or filoviridae viruses.

[0006] A fourth aspect of the invention is that the provided compound exerts a strong antiviral effect in cells and in individuals infected with the aforementioned virus, achieving unexpected results.

[0007] Unless otherwise specified herein, the technical terms used in this invention have basic meanings generally understood by those skilled in the art.

[0008] This invention provides a compound, which is a compound of Formula I or a stereoisomer, tautomer, oxide, solvate, metabolite or pharmaceutically acceptable salt of a compound of Formula I:

[0009]

[0010] In structural formula I:

[0011] M represents 1, 2, 3, or 4;

[0012] R a It can be halogen or hydrogen;

[0013] R a When it is halogenated, -OR3 does not exist;

[0014] R b It can be halogen, methyl, or hydrogen;

[0015] R c It can be azide, cyano, or hydrogen;

[0016] R1 represents hydrogen, halogen, any alkyl group with 1-8 carbon atoms, any alkoxy group with 1-8 carbon atoms, any alkenyl group with 2-8 carbon atoms, any alkynyl group with 2-8 carbon atoms, any carbocyclic group with 3-8 carbon atoms, any heterocyclic group with 2-8 carbon atoms, any aryl group with 6-12 carbon atoms, and any heteroaryl group with 3-12 carbon atoms.

[0017] R2 represents hydrogen, deuterium, any alkyl group with 1-8 carbon atoms, any alkoxy group with 1-8 carbon atoms, -(CO)R6, -(CO)-OR6, -(CO)-NHR6;

[0018] R3 and R4 can independently represent hydrogen, deuterium, -COR6, and -(CO)-OR6, or R3 and R4 can be linked together to form a ring.

[0019] R5 represents , , ;

[0020] W represents O or S;

[0021] Q represents O, or -NH-;

[0022] R x R y Each of the following independently represents hydrogen, any alkyl group with 1-8 carbon atoms, any alkoxy group with 1-8 carbon atoms, or R. x R y Connected to form a ring;

[0023] R6 represents any alkyl group with 1-8 carbon atoms, any alkoxy group with 1-8 carbon atoms, any carbocyclic group with 3-8 carbon atoms, any heterocyclic group with 2-8 carbon atoms, any aryl group with 6-12 carbon atoms, or any heteroaryl group with 3-12 carbon atoms, which may optionally be substituted by one or more of the following groups: hydrogen, deuterium, halogen, hydroxyl, mercapto, cyano, methylthio, ethylthio, amino, trifluoromethyl, acetyl, carboxyl, alkyl group with 1-8 carbon atoms, alkoxy group with 1-8 carbon atoms, carbocyclic group with 3-8 carbon atoms, heterocyclic group with 2-8 carbon atoms, aryl group with 6-12 carbon atoms, or heteroaryl group with 3-12 carbon atoms;

[0024] R7 represents any alkyl group with 1-8 carbon atoms, any alkoxy group with 1-8 carbon atoms, any alkylamino group with 1-8 carbon atoms, any carbocyclic group with 3-8 carbon atoms, any heterocyclic group with 2-8 carbon atoms, any aryl group with 6-12 carbon atoms, or any heteroaryl group with 3-12 carbon atoms, optionally surrounded by one or more R7 groups. 12 replace;

[0025] R8 represents hydrogen, deuterium, any alkyl group with 1-8 carbon atoms, any alkoxy group with 1-8 carbon atoms, any alkylamino group with 1-8 carbon atoms, any carbocyclic group with 3-8 carbon atoms, any heterocyclic group with 2-8 carbon atoms, any aryl group with 6-12 carbon atoms, and any heteroaryl group with 3-12 carbon atoms, which may optionally be substituted by one or more of the following groups: hydrogen, deuterium, halogen, hydroxyl, mercapto, cyano, amino, trifluoromethyl, acetyl, carboxyl, alkyl group with 1-8 carbon atoms, alkoxy group with 1-8 carbon atoms, carbocyclic group with 3-8 carbon atoms, heterocyclic group with 2-8 carbon atoms, aryl group with 6-12 carbon atoms, and heteroaryl group with 3-12 carbon atoms;

[0026] R9 represents hydrogen, any alkyl group with 1-8 carbon atoms, any alkoxy group with 1-8 carbon atoms, any carbocyclic group with 3-8 carbon atoms, any heterocyclic group with 2-8 carbon atoms, any aryl group with 6-12 carbon atoms, and any heteroaryl group with 3-12 carbon atoms, which may optionally be substituted by one or more of the following groups: hydrogen, deuterium, halogen, hydroxyl, mercapto, cyano, amino, trifluoromethyl, acetyl, carboxyl, alkyl group with 1-8 carbon atoms, alkoxy group with 1-8 carbon atoms, carbocyclic group with 3-8 carbon atoms, heterocyclic group with 2-8 carbon atoms, aryl group with 6-12 carbon atoms, and heteroaryl group with 3-12 carbon atoms;

[0027] R 10 R 11Each of the following groups independently represents hydrogen, an alkyl group with 1-8 carbon atoms, an alkoxy group with 1-8 carbon atoms, an alkylamino group with 1-8 carbon atoms, a carbocyclic group with 3-8 carbon atoms, a heterocyclic group with 2-8 carbon atoms, an aryl group with 6-12 carbon atoms, or a heteroaryl group with 3-12 carbon atoms, and may optionally be represented by one or more R groups. 12 Replacement or R 10 R 11 Connected to form a ring;

[0028] R 12 Representing hydrogen, deuterium, halogen, hydroxyl, mercapto, cyano, methylthio, ethylthio, amino, trifluoromethyl, acetyl, propionyl, carboxyl, -COR6, alkyl with 1-8 carbon atoms, alkoxy with 1-8 carbon atoms, alkylamino with 1-8 carbon atoms, carbocyclic with 3-8 carbon atoms, heterocyclic with 2-8 carbon atoms, aryl with 6-12 carbon atoms, and heteroaryl with 3-12 carbon atoms, optionally represented by one or more R 13 replace;

[0029] R 13 Represents hydrogen, deuterium, halogen, hydroxyl, mercapto, cyano, methylthio, ethylthio, amino, trifluoromethyl, acetyl, propionyl, carboxyl, -COR6, alkyl with 1-8 carbon atoms, alkoxy with 1-8 carbon atoms, alkylamino with 1-8 carbon atoms, carbocyclic with 3-8 carbon atoms, heterocyclic with 2-8 carbon atoms, aryl with 6-12 carbon atoms, and heteroaryl with 3-12 carbon atoms;

[0030] The compound described has the following structure: Formula II

[0031]

[0032] The substituents in Formula II are defined as defined in Formula I as claimed in claim 1.

[0033] The compound described has the following structure, Formula III:

[0034]

[0035] The substituents in Formula III are defined as defined in Formula I as claimed in claim 1.

[0036] The compound described has the following structure:

[0037]

[0038] The substituents in Formula IV are defined as defined in Formula I as claimed in claim 1.

[0039] The compound has the following structure:

[0040]

[0041]

[0042]

[0043]

[0044] Or a stereoisomer, tautomer, oxide, solvate, metabolite, or pharmaceutically acceptable salt of the compound shown in Formula I.

[0045] The compounds described herein can be used to treat or prevent the activity of viruses in humans or other animals that are resistant to orthomyxoviridae, paramyxoviridae, coronaviruses, or filoviridae viruses.

[0046] The introduction of carbon-sulfur double bond groups into the compound significantly enhances its antiviral activity.

[0047] An antiviral pharmaceutical composition comprising the aforementioned compound.

[0048] Treatment of individuals infected with Orthomyxoviridae, Paramyxoviridae, Coronavirus, or Filoviridae viruses comprises administering an effective amount of the aforementioned compound or combinations thereof to the infected individual, and also includes pharmaceutically acceptable excipients.

[0049] The application described herein refers to the use of dosage forms for treating the viral infection, including injectable dosage forms, respiratory dosage forms, skin dosage forms, mucosal dosage forms, cavity dosage forms, and oral dosage forms.

[0050] The pharmaceutical composition comprises the compound in an amount ranging from 0.01% to 80% (w / w%).

[0051] Unless otherwise defined in the context, the technical terms used herein are intended to have the same meaning as commonly understood by those skilled in the art.

[0052] As used herein, “stereoisomer” refers to an enantiomer and diastereomer of a compound having one or more asymmetric centers. This invention also includes the use of optical isomers and stereoisomers of the compounds provided by this invention, or mixtures thereof in any proportion, as well as methods and uses of the compositions prepared therefrom.

[0053] In this article, "tautomer" refers to functional group isomers that arise from the rapid movement of an atom between two positions in a compound's structure. A pair of tautomers can interconvert, but the more stable isomer is usually the dominant form. Examples include enol-keto tautomers, amide-imino acid tautomers, lactam-lactamimide tautomers, amide-imino acid tautomers in heterocycles, and enamine-imine enamine tautomers.

[0054] In this article, "oxide" refers to a complex formed by the combination of oxygen and easily oxidized atoms in a compound. Easily oxidized atoms in the molecule include nitrogen (N) or sulfur (S), and the complexes formed include nitrogen oxides, sulfones, and sulfoxides.

[0055] In this article, "solvent" refers to a complex formed by a compound and a solvent molecule, such as water, ethanol, or acetic acid, which has low toxicity, through hydrogen bonding and has a specific crystal form.

[0056] In this document, "metabolite" refers to the intermediate or final metabolite produced by the degradation of a compound in vivo through oxidation, reduction, hydrolysis, esterification, defatting, enzymatic hydrolysis, or other methods. These intermediate or final metabolites may or may not possess physiological activity. The metabolites of the compounds provided in this invention are also included within the scope of protection of this invention.

[0057] The scope of this invention also includes metabolites of the compounds of this invention, i.e., substances formed in vivo by reactions such as oxidation, reduction, hydrolysis, amidation, deamidation, esterification, defatting, and enzymatic hydrolysis of the compounds of this invention administered in vivo. This invention also includes compounds obtained by exposing the compounds of this invention to mammals for a sufficient period of time to produce their metabolites.

[0058] In this document, "pharmaceutically acceptable salt" refers to acid salts formed by the combination of the compounds provided in this invention with pharmaceutically acceptable acids, such as hydrochlorides, hydrobroms, sulfates, nitrates, phosphates, citrates, tartrates, maleates, tartrates, succinates, acetates, methanesulfonates, ethanesulfonates, p-toluenesulfonates, etc.; it also refers to base salts formed by the combination of the compounds provided in this invention with pharmaceutically acceptable bases (including but not limited to pharmaceutically acceptable cations), such as potassium salts, sodium salts, calcium salts, ammonium salts, or addition salts with amines, such as ethylenediamine salts, triethylamine salts, diethylamine salts, histidine salts, arginine salts, lysine salts, N-methylglucosamine salts, etc.

[0059] In this article, "substitution" refers to a compound in which one or more atoms in a molecule are replaced by one or more other atoms or groups of atoms, but the substitution shall not exceed the valence of the atoms at the substituted positions.

[0060] In this article, "optionally replaced by one or more ----" means that the atoms or groups at the substituted atomic positions can be independently replaced by other atoms or groups, and the substituents can be connected at any reasonable position.

[0061] In this article, "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), or iodine (I).

[0062] In this article, "heteroatoms" refers to nitrogen (N), oxygen (O), phosphorus (P), or sulfur (S).

[0063] In this article, "any alkyl group with 1-8 carbon atoms" refers to a monovalent aliphatic hydrocarbon group having a straight chain or branched chain or combination thereof with 1-8 carbon atoms, such as methyl, ethyl, isopropyl, butyl, isobutyl, tert-butyl, isoheptyl, n-octyl, etc.

[0064] In this article, "any alkoxy group with 1-8 carbon atoms" refers to a substituent with one or more -O- or -OH groups inserted at any reasonable position in an alkyl group with 1-8 carbon atoms, such as methoxy, ethoxy, isopropoxy, tert-butoxy, pentoxy, 2-ethylhexoxy, etc.

[0065] In this article, "any alkylamino group with 1-8 carbon atoms" refers to a substituent with one or more -N-, -NH-, or -NH2 groups inserted at any reasonable position in an alkyl group with 1-8 carbon atoms, such as methylamino, ethylamino, propylamino, isopropylamino, ethylenediamino, di-n-propylamino, diisopropylamino, etc.

[0066] In this article, "any alkenyl group with 2-8 carbon atoms" refers to an aliphatic hydrocarbon group in a molecule composed of 2-8 carbon atoms that contains at least one carbon-carbon double bond, including straight-chain, branched, or cyclic alkenes, such as vinyl, propenyl, allyl, butenyl, 2-methyl-2-pentenyl, cyclohexenyl, 1-methyl-1-cyclohexenyl, 2,5-dimethyl-2,4-hexadienyl, etc.

[0067] In this article, "any 2-8 carbon atom alkynyl group" refers to an aliphatic hydrocarbon group in a molecule composed of 2-8 carbon atoms that contains at least one carbon-carbon triple bond, including straight-chain, branched-chain or cyclic alkynes, such as ethynyl, 2-butynyl, 1-butyn-4-yl, 5-methyl-1-hexynyl, 3-cyclopentyl-1-propynyl, etc.

[0068] In this article, "any carbon cyclic group with 3-8 carbon atoms" refers to a saturated or unsaturated cyclic aliphatic hydrocarbon group composed of 3-8 carbon atoms. It can be a monocyclic group or multiple carbon rings connected by spirocyclic or bridged carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, methylcyclopentyl, methylcyclohexyl, ethylcyclohexyl, (cis)1,2-dimethylcyclohexyl, cyclopentenyl, etc.

[0069] In this article, "any heterocyclic group with 2-8 carbon atoms" refers to a saturated or unsaturated cyclic group consisting of 2-8 carbon atoms containing at least one heteroatom (selected from N, O or S), such as ethylene oxide, propylene oxide, acridine, piperidinyl, homopiperazinyl, pyrrolidinyl, pyrazolyl, morpholinyl, 1,2-dihydropyridine-2-amino, etc.

[0070] In this article, "any aryl group with 6-12 carbon atoms" refers to one or more groups with an aromatic ring system consisting of 6-12 carbon atoms. Such groups can be fused or unfused, and the fused ring can be a saturated or unsaturated hydrocarbon group, such as phenyl, naphthyl, indanyl, biphenyl, tetrahydronaphthyl, etc.

[0071] In this document, "any heteroaryl group with 3-12 carbon atoms" refers to a monocyclic or polycyclic aromatic ring system composed of 3-12 carbon atoms and having at least one heteroatom (i.e., O, N, or S). This type of group can be fused or unfused, and the fused ring can be a saturated or unsaturated hydrocarbon group. Examples include pyridyl, pyrazinyl, pyridazinyl, imidazolyl, furanyl, thiophene, thiazolyl, pyrazolyl, triazinyl, purinyl, benzoxazolyl, benzofuranyl, benzothiazolyl, indole, and isoindoleyl. Furthermore, the heteroaryl groups also include nitrogen oxides and sulfur oxides containing nitrogen-containing heterocycles.

[0072] In this article, "orthomyxoviridae viruses" refers to influenza A, B, and C viruses that infect humans or other animals.

[0073] In this article, "paramyxoviridae viruses" refers to measles virus, mumps virus, respiratory syncytial virus, parainfluenza virus, etc., that infect humans or other animals.

[0074] In this article, "coronavirus" refers to enveloped, single-stranded, positive-sense RNA viruses that primarily infect vertebrates, such as human coronavirus (COV) NL63, long-winged bat coronavirus HKU1, long-winged bat coronavirus HKU8, porcine epidemic diarrhea virus (PEDV), porcine transmissible gastroenteritis virus (TGEV), feline coronavirus (FCOV), bovine coronavirus (BCOV), chicken infectious bronchitis virus (IBV), and thrush coronavirus.

[0075] In this article, "filoviridae viruses" refers to Marburg virus (MBV), Ebola virus (EBV), etc.

[0076] In this article, "individual" refers to a living organism that is sick or has obvious symptoms, including but not limited to humans and other mammals such as pigs, cattle, sheep, rats, dogs, and monkeys.

[0077] In this article, "pharmaceuticalally acceptable excipients" refers to diluents, excipients, and other similar substances that are administered together with therapeutic preparations and do not cause significant toxicity, irritation, or allergic reactions in human or other animal tissues. Examples include water, soybean oil, mineral oil, glucose, sucrose, maltose, silica gel, sodium stearate, glyceryl monostearate, sodium chloride, glycerin, propylene glycol, ethanol, pH buffers, colorants, starch, sodium saccharin, and cellulose.

[0078] In this document, "effective amount" refers to the mass content of the compound of the present invention or its pharmaceutically usable salt in the range of 0.01% to 50%.

[0079] The compounds of the present invention can be systematically administered via various routes, such as intramuscular injection, intravenous injection, subcutaneous injection, intraperitoneal injection, transdermal administration, oral administration, nasal administration, transmucosal administration, and inhalation administration.

[0080] The compounds of this invention can be prepared into various formulations with pharmaceutically acceptable excipients, such as tablets, capsules, powders, sprays, creams, gels, lotions, ointments, suspensions, tinctures, syrups, etc.

[0081] The compounds of this invention, when formulated with pharmaceutically acceptable excipients, can be administered in various ways to achieve optimal therapeutic outcomes, such as single or bolus administration, or dosage adjustments. The dosage varies depending on the severity of the individual's disease.

[0082] The present invention will be further described below with reference to embodiments, but this does not limit the invention in any way. The structures of all compounds have been analyzed by MS or... 1 H NMR confirmed. Detailed Implementation

[0083] The specific implementation method is as follows:

[0084] Example 1: N-2

[0085]

[0086] N-1:

[0087] The starting material Sm1 (26.5 g, 0.10 mol) was dissolved in dioxane (300 mL), and the mixture was cooled to 0 °C under nitrogen protection. HOBt (16.2 g, 0.12 mol), EDCI (22.9 g, 0.12 mol), and triethylamine (30.3 g, 0.3 mol) were slowly added. After the addition was complete, the mixture was stirred at room temperature for 2 hours. Sm2 (8.8 g, 0.10 mol) was then added, and the mixture was reacted at 50 °C for 6 hours. The mixture was then cooled, concentrated, and the residue was slurried with ice water (200 mL). The resulting solid was recrystallized from ethanol / water to give N-1 (17.8 g). MS (ESI, m / z): 337.2 [M+H] + .

[0088] N-2:

[0089] N-1 (16.8 g, 0.05 mol) was dissolved in dioxane (300 mL), and alumina-supported P2S5 (80.0 g) was added. The mixture was reacted at 60 °C for 16 hours, cooled to room temperature, filtered, concentrated, and separated by column chromatography to obtain N-2 (4.0 g). MS (ESI, m / z): 353.1 [M+H] + .

[0090] Example 2: PE-0

[0091]

[0092] N-3:

[0093] The starting material Sm3 (24.4 g, 0.10 mol) was dissolved in acetone (300 mL), cooled to 0 °C under nitrogen protection, and concentrated sulfuric acid (10.0 mL) was slowly added. After the addition was complete, the mixture was stirred at room temperature for 5 hours, cooled to 0 °C, and neutralized to a pH of 7–8 with potassium carbonate solution. The mixture was concentrated under reduced pressure, and the resulting solid was extracted with water (100 mL), ethyl acetate (100 mL × 2), washed with saturated brine (100 mL), concentrated, and separated by column chromatography to obtain N-3 (15.1 g). MS (ESI, m / z): 245.0 [M+H] + .

[0094] N-4:

[0095] N-2 (17.6 g, 0.05 mol) was dissolved in dioxane (300 mL), and the mixture was cooled to 0 °C under nitrogen protection. HOBt (8.1 g, 0.06 mol), EDCI (11.5 g, 0.06 mol), and triethylamine (15.2 g, 0.15 mol) were slowly added. After the addition was complete, the mixture was stirred at room temperature for 2 hours. N-3 (14.2 g, 0.05 mol) was then added, and the mixture was reacted at 50 °C for 6 hours. The mixture was cooled, concentrated, and the residue was slurried with ice water (200 mL). The resulting solid was recrystallized from ethanol / water to give N-4 (17.0 g). MS (ESI, m / z): 619.2 [M+H] + .

[0096] N-5:

[0097] 1,2,4-Triazole (0.7 g, 0.01 mol) was dissolved in anhydrous acetonitrile (15 mL). POCl3 (1.5 g, 0.01 mol) was slowly added under nitrogen protection, followed by the dropwise addition of triethylamine (3.0 g, 0.03 mol). After stirring at room temperature for 1 hour, N-4 (3.1 g, 5.0 mmol) in acetonitrile (25 mL) solution was slowly added dropwise. After the addition was complete, the mixture was reacted at room temperature for 16 hours, then concentrated. 50 mL of sodium bicarbonate solution (0.01 M) was added, resulting in the precipitation of a solid. The solid was filtered and recrystallized from ethanol / water to give N-5 (0.5 g). MS (ESI, m / z): 670.1 [M+H] + .

[0098] PE-01:

[0099] N-5 (0.5 g, 0.7 mmol) was dissolved in anhydrous acetonitrile (15 mL). Under nitrogen protection, 1.0 mL of 50% hydroxylamine aqueous solution was slowly added. The mixture was stirred at room temperature for 2 hours, concentrated, and then ice water (5 mL) was added. The mixture was filtered, and the resulting solid was added to acetonitrile (5 mL) and hydrochloric acid solution (5 mL, 0.01 M). The mixture was stirred at room temperature for 1 hour, and then sodium bicarbonate solution (10 mL, 0.01 M) was added. The mixture was concentrated, and the residue was separated by column chromatography to obtain PE-01 (0.04 g). MS (ESI, m / z): 494.2 [M+H] + . 1H-NMR (400 MHz, CDCl3) δ 9.00(1H, d, J 6.4), 7.22-7.20 (1H, m), 7.17-7.14 (4H, m), 5.90-5.88 (1H, m), 5.57(1H, d, J 6.4), 5.05-5.03 (1H, m), 4.60-4.58 (1H, m), 4.49-4.47 (1H, m),4.35-4.33 (1H, m), 4.11-4.09 (1H, m), 3.54-3.52 (1H, m), 3.41-3.39 (1H, m),3.19-3.17 (1H, m), 1.46 (3H, d, J 12.2).

[0100] Example 3: PE-02

[0101]

[0102] PE-02:

[0103] PE-01 (0.5 g, 1.0 mmol) was dissolved in anhydrous toluene (15 mL), and Lawson's reagent (0.4 g, 1.0 mmol) was slowly added under nitrogen protection. The mixture was heated to 90 °C and reacted for 3 hours. After cooling, the mixture was filtered, concentrated, and separated by column chromatography to obtain PE-02 (0.06 g). MS (ESI, m / z): 510.2 [M+H] + . 1 H-NMR (400 MHz, CDCl3) δ 9.04 (1H, d, J 6.6), 7.24-7.22 (1H, m), 7.19-7.15 (4H, m), 5.94-5.92 (1H, m), 5.59 (1H, d, J 6.6),5.08-5.05 (1H, m), 4.63-4.61 (1H, m), 4.50-4.47 (1H, m), 4.36-4.34 (1H, m), 4.13-4.11 (1H, m), 3.55-3.53 (1H, m), 3.47-3.45 (1H, m), 3.21-3.19 (1H, m),1.47 (3H, d, J 12.0).

[0104] Example 4: PE-05

[0105]

[0106] N-6:

[0107] N-5 (0.7 g, 1.0 mmol) was dissolved in anhydrous acetonitrile (15 mL). Under nitrogen protection, 1.0 mL of a 50% hydroxylamine aqueous solution was slowly added. The mixture was stirred at room temperature for 2 hours, concentrated, and then slurried with ice water (4 mL). The mixture was filtered, dried, and dissolved in anhydrous acetonitrile (10 mL). Under nitrogen protection, triethylamine (0.3 g, 3.0 mmol) was added. At 0 °C, a solution of isopropyl chloroformate (0.2 g, 1.5 mmol) in anhydrous acetonitrile (5 mL) was slowly added dropwise. The mixture was reacted at room temperature for 5 hours, concentrated, and then slurried with ice water (2 mL). The mixture was filtered, and the resulting solid was separated by column chromatography to obtain N-6 (0.3 g). MS (ESI, m / z): 720.4 [M+H] + .

[0108] PE-05:

[0109] N-6 (0.3 g, 0.4 mmol) was added to acetonitrile (5 mL) and hydrochloric acid solution (5 mL, 0.01 M), stirred at room temperature for 1 hour, then sodium bicarbonate solution (10 mL, 0.01 M) was added, and the mixture was concentrated. The residue was separated by column chromatography to obtain PE-05 (0.02 g), MS (ESI, m / z): 580.1 [M+H]. + . 1 H-NMR (400 MHz, CDCl3) δ 9.01 (1H, d, J 6.3), 7.20-7.18 (1H,m), 7.14-7.11 (4H, m), 5.87-5.85 (1H, m), 5.52 (1H, d, J 6.3), 5.06-5.04 (1H,m), 5.02-5.00 (1H, m), 4.58-4.56 (1H, m), 4.45-4.43 (1H, m), 4.32-4.30 (1H,m), 4.10-4.09 (1H, m), 3.51-3.49 (1H, m), 3.39-3.37 (1H,m), 3.14-3.12 (1H,m), 1.41 (3H, d, J 12.1), 1.23 (6H, d, J 11.2).

[0110] Example 5: N-8

[0111]

[0112] N-7:

[0113] Sm3 (24.4 g, 0.1 mol) was dissolved in anhydrous acetone (350 mL), and pyridine (25 mL) was added. Phosgene was slowly bubbled through the solution at room temperature until the starting material disappeared. The system was concentrated, and 40 mL of ice water was added. A solid precipitated out. The solid was filtered, washed with water, and recrystallized from acetonitrile / water to give N-7 (19.4 g). MS (ESI, m / z): 271.3 [M+H] + .

[0114] N-8:

[0115] Phosphorus oxychloride (15.3 g, 0.1 mol) was dissolved in anhydrous acetonitrile (350 mL), and pyridine (50 mL) was added. The mixture was cooled to 0 °C under nitrogen protection, and N-7 (13.5 g, 0.05 mol) in acetonitrile (200 mL) was slowly added dropwise. After the addition was complete, the mixture was reacted at room temperature for 12 hours, cooled to 0 °C, and then slowly stirred with sodium bicarbonate solution (50 mL, 0.01 M) for 1 hour. The mixture was concentrated under reduced pressure, and the residue was slurried with ice water (30 mL). The residue was recrystallized from the residue with water / acetone to give N-8 (1.6 g). MS (ESI, m / z): 351.0 [M+H] + .

[0116] Example 6: PE-31

[0117] N-9:

[0118] Sm1 (26.5 g, 0.10 mol) was dissolved in dioxane (500 mL), and the mixture was cooled to 0 °C under nitrogen protection. HOBt (16.2 g, 0.12 mol), EDCI (22.9 g, 0.12 mol), and triethylamine (30.4 g, 0.30 mol) were slowly added. After the addition was complete, the mixture was stirred at room temperature for 2 hours. Isopropanol (1.2 g, 0.20 mol) was then added, and the mixture was reacted at 50 °C for 6 hours. The mixture was then cooled, concentrated, and the residue was slurried with ice water (100 mL). The resulting solid was recrystallized from ethanol / water to give N-9 (16.9 g). MS (ESI, m / z): 308.1 [M+H] + .

[0119] N-10:

[0120] N-9 (15.3 g, 0.05 mol) was dissolved in dioxane (300 mL), and alumina-supported P2S5 (80.0 g) was added. The mixture was reacted at 60 °C for 16 hours, cooled to room temperature, filtered, concentrated, and separated by column chromatography to obtain N-10 (3.5 g). MS (ESI, m / z): 324.1 [M+H] + .

[0121] N-11:

[0122] Add N-10 (3.3 g, 0.01 mol) to hydrochloric acid (10 mL, 1.0 mM) solution, add methanol (10 mL), stir at room temperature for 2 hours, and then adjust the pH value to 7-8 with triethylamine for later use.

[0123] N-8 (3.5 g, 0.01 mol) was dissolved in anhydrous acetonitrile (80 mL), and the mixture was cooled to 0 °C under nitrogen protection. Thionyl chloride (2.4 g, 0.02 mol) was slowly added, and the mixture was heated to 50 °C and reacted for 2 hours. The mixture was then cooled to 0 °C, and the above-mentioned N-10 solution and triethylamine (3.0 g, 0.03 mol) were slowly added. After the addition was complete, the mixture was reacted at 50 °C for another 2 hours. The mixture was then cooled to room temperature and concentrated. The resulting residue was slurried with ice water (20 mL), filtered, and the resulting solid was separated by column chromatography to obtain N-11 (2.8 g). MS (ESI, m / z): 556.1 [M+H] + .

[0124] N-12:

[0125] 1,2,4-Triazole (0.7 g, 0.01 mol) was dissolved in anhydrous acetonitrile (15 mL). POCl3 (1.5 g, 0.01 mol) was slowly added under nitrogen protection, followed by the dropwise addition of triethylamine (3.0 g, 0.03 mol). After stirring at room temperature for 1 hour, a solution of N-11 (2.7 g, 5.0 mmol) in acetonitrile (25 mL) was slowly added dropwise. After the addition was complete, the mixture was reacted at room temperature for 16 hours. The solution was then concentrated, and 30 mL of sodium bicarbonate solution (0.01 M) was added. A solid precipitated, which was filtered and recrystallized from ethanol / water to give N-12 (0.6 g). MS (ESI, m / z): 607.2 [M+H] + .

[0126] PE-31:

[0127] N-12 (0.6 g, 1.0 mmol) was dissolved in anhydrous acetonitrile (15 mL). Under nitrogen protection, 1.0 mL of 50% hydroxylamine aqueous solution was slowly added. The mixture was stirred at room temperature for 2 hours, concentrated, and then filtration was performed. The resulting solid was dried and added to methanol (10 mL), followed by water (2 mL) and barium hydroxide (0.2 g). The mixture was heated to 60 °C and reacted for 6 hours. After cooling, the mixture was filtered, and the filtrate was concentrated. PE-31 (0.03 g) was obtained by column chromatography. MS (ESI, m / z): 545.3 [M+H] + . 1H-NMR (400 MHz, CDCl3) δ8.98 (1H, d, J 6.4), 7.16-7.14 (1H, m), 7.12-7.10 (4H, m), 5.83-5.81 (1H, m),5.50 (1H, d, J 6.4), 4.53-4.51 (2H, m), 4.44-4.42 (1H, m), 4.29-4.27 (1H, m), 4.00-3.98 (2H, m), 3.16-3.12 (2H, m), 2.95-2.93 (1H, m), 1.21 (6H, d, J11.4).

[0128] Example 7: PE-37

[0129]

[0130] N-13:

[0131] N-1 (5.6 g, 0.01 mol) was dissolved in anhydrous acetonitrile (80 mL), and the mixture was cooled to 0 °C under nitrogen protection. Thionyl chloride (2.4 g, 0.02 mol) was slowly added, and the mixture was heated to 50 °C and reacted for 2 hours. The mixture was then cooled to 0 °C, and phenol (1.9 g, 0.02 mol) and triethylamine (3.0 g, 0.03 mol) were slowly added. After the addition was complete, the mixture was reacted at 50 °C for another 2 hours. The mixture was then cooled to room temperature, concentrated, and the resulting residue was slurried with ice water (20 mL). The residue was filtered, and the resulting solid was separated by column chromatography to obtain N-13 (5.0 g). MS (ESI, m / z): 632.1 [M+H] + .

[0132] N-14:

[0133] 1,2,4-Triazole (0.7 g, 0.01 mol) was dissolved in anhydrous acetonitrile (15 mL). POCl3 (1.5 g, 0.01 mol) was slowly added under nitrogen protection, followed by the dropwise addition of triethylamine (4.0 g, 0.04 mol). After stirring at room temperature for 1 hour, a solution of N-13 (3.2 g, 5.0 mmol) in acetonitrile (25 mL) was slowly added dropwise. After the addition was complete, the mixture was reacted at room temperature for 16 hours. The solution was then concentrated, and 10 mL of sodium bicarbonate solution (0.01 M) was added. A solid precipitated, which was filtered and separated by column chromatography to obtain N-14 (0.6 g). MS (ESI, m / z): 683.3 [M+H] + .

[0134] PE-37:

[0135] N-14 (0.7 g, 1.0 mmol) was dissolved in anhydrous acetonitrile (15 mL). Under nitrogen protection, 1.0 mL of 50% hydroxylamine aqueous solution was slowly added. The mixture was stirred at room temperature for 2 hours, concentrated, and then filtration was performed. The resulting solid was dried and added to methanol (10 mL), followed by water (2 mL) and barium hydroxide (0.2 g). The mixture was heated to 60 °C and reacted for 4 hours. After cooling, the mixture was filtered, and the filtrate was concentrated. PE-37 (0.02 g) was obtained by column chromatography. MS (ESI, m / z): 621.1 [M+H] + . 1 H-NMR (400 MHz, CDCl3) δ8.94 (1H, d, J 6.6), 7.39-7.37 (2H, m), 7.17-7.14 (4H, m), 7.10-7.07 (4H, m),5.82-5.80 (1H, m), 5.49 (1H, d, J 6.6), 4.51-4.49 (1H, m), 4.43-4.41 (1H, m),4.39-4.37 (1H, m), 4.27-4.25 (1H, m), 4.01-3.99 (1H, m), 3.15-3.12 (2H, m),2.92-2.90 (1H, m), 1.22 (6H, d, J 10.4).

[0136] Example 8: PE-43

[0137]

[0138] N-15:

[0139] N-1 (5.6 g, 0.01 mol) was dissolved in anhydrous acetonitrile (80 mL), and the mixture was cooled to 0 °C under nitrogen protection. Thionyl chloride (2.4 g, 0.02 mol) was slowly added, and the mixture was heated to 50 °C and reacted for 2 hours. The mixture was then cooled to 0 °C, and aniline (1.9 g, 0.02 mol) and triethylamine (4.0 g, 0.04 mol) were slowly added. After the addition was complete, the mixture was reacted at 50 °C for another 2 hours. The mixture was then cooled to room temperature, concentrated, and the resulting residue was slurried with ice water (20 mL). The residue was filtered, and the resulting solid was separated by column chromatography to obtain N-15 (5.2 g). MS (ESI, m / z): 631.2 [M+H] + .

[0140] N-16:

[0141] 1,2,4-Triazole (0.7 g, 0.01 mol) was dissolved in anhydrous acetonitrile (15 mL). Under nitrogen protection, POCl3 (1.5 g, 0.01 mol) was slowly added, followed by the dropwise addition of triethylamine (4.0 g, 0.04 mol). After stirring at room temperature for 1 hour, N-15 (3.2 g, 5.0 mmol) in acetonitrile (25 mL) solution was slowly added dropwise. After the addition was complete, the mixture was reacted at room temperature for 16 hours, then concentrated. 10 mL of sodium bicarbonate solution (0.01 M) was added, resulting in the precipitation of a solid. The solid was filtered and separated by column chromatography to obtain N-16 (0.4 g). MS (ESI, m / z): 682.2 [M+H] + .

[0142] PE-43:

[0143] N-16 (0.7 g, 1.0 mmol) was dissolved in anhydrous acetonitrile (15 mL). Under nitrogen protection, 1.0 mL of 50% hydroxylamine aqueous solution was slowly added. The mixture was stirred at room temperature for 2 hours, concentrated, and then filtration was performed. The resulting solid was dried and added to methanol (10 mL), followed by water (2 mL) and barium hydroxide (0.2 g). The mixture was heated to 60 °C and reacted for 3 hours. After cooling, the mixture was filtered, and the filtrate was concentrated. PE-37 (0.02 g) was obtained by column chromatography. MS (ESI, m / z): 620.2 [M+H] + . 1 H-NMR (400 MHz, CDCl3) δ8.92 (1H, d, J 6.5), 7.36-7.34(2H, m), 7.15-7.12 (4H, m), 7.08-7.05 (4H, m),5.80-5.78 (1H, m), 5.44 (1H, d, J 6.4), 4.50-4.48 (1H, m), 4.40-4.39 (1H, m),4.35-4.33 (1H, m), 4.23-4.21 (1H, m), 4.00-3.98 (1H, m), 3.11-3.09 (2H, m),2.90-2.88 (1H, m), 1.20 (6H, d, J 11.2).

[0144] Following the synthesis methods provided in the above embodiments, and using similar means, the following compounds were synthesized using commercially available compounds or compounds that underwent reasonable protecting and deprotecting processes:

[0145]

[0146]

[0147]

[0148]

[0149]

[0150] Example 9: Effects of the compound on MDCK cell activity

[0151] 1×10 5 A homogeneous suspension of MDCK cells per mL was added to each well of a 96-well plate. 50 μL of cell culture medium (DMEM containing 10% fetal bovine serum) was added to each well. After 24 hours of incubation, the culture medium was discarded. 5.0 μM of the compound was added to each well. No drug was added to the blank control wells, only the corresponding volume of culture medium. Three replicates were set up for each compound. After adding the culture medium, the plates were incubated at 37°C with 5% CO2 for 48 hours. The culture medium was discarded, and 10.0 μL of 10.0 mg / mL MTT solution was added to each well. The plates were incubated for another 3 hours. The culture medium was discarded, and 100 μL of DMSO was added to each well. The plates were shaken for 15 minutes, and the absorbance (OD value) at 570 nm was measured using a microplate reader. The average absorbance value of the three replicates was calculated. Cell viability (%) was calculated as (OD value of experimental wells / OD value of blank control wells) × 100%. The results are shown in Table 1.

[0152]

[0153] Data show that the compounds of the present invention all maintained a cell survival rate of over 75% at a concentration of 5.0 μM, and the cell survival rates of compounds PE-01, PE-08, PE-28, PE-35, and PE-42 were all over 85%. Based on this, it can be inferred that the compounds of the present invention have low toxicity to MDCK cells, and compounds PE-01, PE-08, PE-28, PE-35, and PE-42 have even lower toxicity to MDCK cells.

[0154] Example 10: In vitro antiviral activity of the compound against H1N1 and H3N2 viruses

[0155] Experimental method: 1×10 5MDCK cells were homogeneously suspended at 1 / mL and added to 96-well plates. 50 μL of cell culture medium (DMEM containing 10% fetal bovine serum) was added to each well. After 24 hours of culture, the cells adhered, and the culture medium was discarded. The cells were then inoculated with virus culture medium containing H1N1 or H3N2 virus with a multiplicity of infection (MOI) of 0.1. Blank control wells were incubated with the appropriate volume of culture medium, without adding virus solution. After 2 hours of incubation, the culture medium was replaced with fresh medium, and compounds PE-01, PE-08, PE-28, PE-35, PE-42, and ribavirin (positive control) diluted in the culture medium were added. The final concentrations of each compound were set as follows: 0.01 μM, 0.05 μM, 0.2 μM, 1.0 μM, and 5.0 μM. Virus control wells were incubated with the appropriate volume of culture medium, without adding compound solution. Three wells were used for each concentration of each compound. The plates were then incubated for another 48 hours. After fixation and staining, the cells were measured at a wavelength of 570 nm. The absorbance value (OD value) at the treatment site was used to calculate the cytopathic effect inhibition rate (%). The inhibition rate was calculated as follows: [(OD value of drug-treated wells - OD value of virus control wells)] / [OD value of blank control wells - OD value of virus control wells] × 100%. Then, the EC50 of the compound against H1N1 or H3N2 viruses was calculated using GraphPad Prism 5.0 software. 50 The values ​​and calculation results are shown in Table 2:

[0156]

[0157] The results above show that the tested compounds PE-01, PE-08, PE-28, PE-35, and PE-42 all exhibit superior anti-H1N1 and H3N2 virus activity, with similar inhibitory activities, and are significantly higher than the inhibitory activity of ribavirin against H1N1 and H3N2 viruses. This suggests that the clinical administration of these compounds can achieve therapeutic effects at relatively low doses.

[0158] Example 11: In vivo anti-H1N1 virus activity of the compound

[0159] Eighty Kunming mice weighing 18.0-20.0g (half male and half female) were randomly divided into eight groups of ten mice each: blank control group, model control group, PE-01 group, PE-08 group, PE-28 group, PE-35 group, PE-42 group, and ribavirin group. Each group of mice was infected nasally with 50 μL of H1N1 virus solution. The blank control group was not infected nasally with H1N1 virus solution. All mice were allowed free access to food. Twenty-four hours after H1N1 virus infection, the compound was dissolved in 0.5% CMC-Na solution, uniformly suspended, and administered by gavage at a dose of 30 mg / kg once daily. The blank control group and model control group received no drug, only the corresponding volume of 0.5% CMC-Na solution. Administration continued for 10 days, once daily. The general physiological condition of each mouse was observed during this period, and mortality was recorded. On the last day after drug administration, mice were fasted for 12 hours. Surviving mice were weighed, dissected, and lung tissue was removed. The tissue was blotted dry with absorbent paper, weighed, and the lung index was calculated using Equation 1.

[0160] Formula 1: Lung Index = [Mouse lung tissue mass (g) / Mouse body weight (g)] × 100%

[0161] Simultaneously, Equation 2 was used to calculate the lung index inhibition rate:

[0162] Formula 2: Lung index inhibition rate = [(mean lung index of model control group - mean lung index of treatment group) / mean lung index of model control group] × 100%

[0163] The experimental results are shown in Table 3:

[0164]

[0165] The significantly increased lung index in the virus control mice indicates successful model establishment. Calculated data show that compounds PE-01, PE-08, PE-28, PE-35, and PE-42 all significantly reduced the lung index in mice in vivo, significantly exceeding the in vivo effect of ribavirin. Furthermore, regarding the lung index inhibition rate, ribavirin only achieved 28.2%, while compounds PE-01, PE-08, PE-28, PE-35, and PE-42 all exhibited inhibition rates exceeding 46% in vivo, demonstrating significantly superior inhibitory effects compared to ribavirin. This indicates that compounds PE-01, PE-08, PE-28, PE-35, and PE-42 possess significant anti-H1N1 virus activity in vivo.

[0166] The mortality rate of mice was observed and recorded for 10 consecutive days. The number of dead mice was recorded, and the mortality rate was calculated using Equation 3:

[0167] Formula 3: Mortality rate (%) = (Total number of mice that died in the group / Total number of mice in the group) × 100%

[0168] The calculation results are shown in Table 4:

[0169]

[0170] Data showed that after 10 days of continuous administration, the mortality rate in the ribavirin group was 50%, while the mortality rates in the PE-01, PE-08, PE-28, PE-35, and PE-42 groups were between 10% and 20%, significantly reducing the mortality rate and demonstrating better therapeutic efficacy than ribavirin in vivo.

[0171] During the experiment, mice in the model control group gradually developed loss of appetite, lethargy, loss of luster, and emaciation, with an increasing number of deaths. In contrast, mice in the PE-01, PE-08, PE-28, PE-35, and PE-42 groups showed good mental condition and no significant weight loss, indicating that compounds PE-01, PE-08, PE-28, PE-35, and PE-42 did not exhibit significant toxicity in vivo and were expected to have minimal toxic side effects.

[0172] This application describes several embodiments, but these descriptions are exemplary and not restrictive. There are many more embodiments and implementations that can be found within the scope of the embodiments described in this application.

Claims

1. A compound, which is a compound of Formula I or a stereoisomer of a compound of Formula I, or a pharmaceutically acceptable salt thereof: In structural formula I: m represents 1; R a It can be halogen or hydrogen; R a When it is halogenated, -OR3 does not exist; R b It is hydrogen; R c It is hydrogen; R1 represents hydrogen; R2 represents hydrogen, -(CO)R6, -(CO)-OR6, -(CO)-NHR6; R3 and R4 independently represent hydrogen, -COR6, and -(CO)-OR6, respectively; R5 represents , , ; W represents O or S; Q represents O, or -NH-; R x R y Each of the following independently represents hydrogen, any alkyl group with 1-8 carbon atoms, any alkoxy group with 1-8 carbon atoms, or R. x R y Connected to form a ring; R6 represents any alkyl group with 1-8 carbon atoms, any carbocyclic group with 3-8 carbon atoms, any heterocyclic group with 2-8 carbon atoms, any aryl group with 6-12 carbon atoms, or any heteroaryl group with 3-12 carbon atoms, which may optionally be substituted by one or more of the following groups: hydrogen, amino. R7 represents any alkyl group with 1-8 carbon atoms, any alkoxy group with 1-8 carbon atoms, any alkylamino group with 1-8 carbon atoms, any carbocyclic group with 3-8 carbon atoms, any heterocyclic group with 2-8 carbon atoms, any aryl group with 6-12 carbon atoms, or any heteroaryl group with 3-12 carbon atoms, optionally surrounded by one or more R7 groups. 12 replace; R8 represents hydrogen, deuterium, any alkyl group with 1 to 8 carbon atoms, or any aryl group with 6 to 12 carbon atoms; R9 represents hydrogen, any alkyl group with 1-8 carbon atoms, any alkoxy group with 1-8 carbon atoms, any carbocyclic group with 3-8 carbon atoms, any heterocyclic group with 2-8 carbon atoms, any aryl group with 6-12 carbon atoms, and any heteroaryl group with 3-12 carbon atoms, which may optionally be substituted by one or more of the following groups: hydrogen, deuterium, halogen, hydroxyl, mercapto, cyano, amino, trifluoromethyl, acetyl, carboxyl, alkyl group with 1-8 carbon atoms, alkoxy group with 1-8 carbon atoms, carbocyclic group with 3-8 carbon atoms, heterocyclic group with 2-8 carbon atoms, aryl group with 6-12 carbon atoms, and heteroaryl group with 3-12 carbon atoms; R 10 R 11 Each of the following can independently represent hydrogen, any alkyl group with 1-8 carbon atoms, or any aryl group with 6-12 carbon atoms.

2. The compound of claim 1, having the structure of formula II: The substituents in Formula II are defined as defined in Formula I as claimed in claim 1.

3. The compound of claim 1, having the structure of formula III: The substituents in Formula III are defined as defined in Formula I as claimed in claim 1.

4. The compound of claim 1, having the structure of formula IV: The substituents in Formula IV are defined as defined in Formula I as claimed in claim 1.

5. The compound of claim 1, having the following structure: 。 6. Use of the compound according to any one of claims 1 to 5 in the preparation of a drug for the treatment or prevention of orthomyxoviridae viruses, paramyxoviridae viruses, coronaviruses, or filoviridae viruses in humans or other animals.

7. An antiviral pharmaceutical composition comprising any one of the compounds of claims 1 to 5 and pharmaceutically acceptable excipients.

8. The antiviral pharmaceutical composition according to claim 7, wherein the dosage form is selected from injectable dosage forms, respiratory dosage forms, skin dosage forms, mucosal dosage forms, cavity dosage forms, and oral dosage forms.

9. The pharmaceutical composition according to claim 7, wherein the content of any one of the compounds according to claims 1 to 5 ranges from 0.01% to 80% (w / w%).

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