Antiviral and antibacterial N4-hydroxy cytidine derivative as well as application and preparation method thereof

Abstract

The invention discloses a compound derivative represented by a formula I or an isotope labeled compound thereof, or an optical isomer, a geometric isomer, a tautomer or an isomer mixture thereof, or a pharmaceutically acceptable salt thereof. According to the application of a double protection strategy, diversified carrier fragments are introduced to 5-hydroxyl and hydroxylamine in N4-hydroxycytidine molecules, the N4-hydroxycytidine prodrug is invented, the affinity of lung cell membranes (phospholipid membranes) is improved, and the physicochemical properties of the drug are made to meet the requirements of the physiological environment of lung tissue. Therefore, the compound disclosed by the invention has a wide application prospect in the aspect of broad-spectrum respiratory virus resistance. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of pharmacy, and specifically relates to nucleoside N4-hydroxycytidine derivatives or isotope-labeled compounds thereof, or optical isomers, geometric isomers, tautomers or isomer mixtures thereof, or pharmaceutically acceptable salts thereof, and uses and preparation methods thereof. Background Art

[0002] Respiratory virus infections pose a serious threat to human health. Looking back at major epidemics in human history, influenza A, avian influenza, SARS-COV-1, MERs-COV, etc. have caused huge social and economic losses. There is still a very real need to study broad-spectrum antiviral drugs.

[0003] N4-hydroxycytidine (NHC) is a broad-spectrum antiviral compound discovered in the middle of the last century. After entering the host cell, it is converted into a nucleotide analog in the form of triphosphate, acting as a substrate for cytidine triphosphate and uridine triphosphate in viral RNA, and inserting as an erroneous base during viral RNA replication. As mutations accumulate, viral protein translation is caused, and viral replication is eventually blocked, thereby exerting antiviral activity. However, N4-hydroxycytidine isobutyrate has a large polarity and poor membrane permeability. Therefore, in order to improve the lipophilic and hydrophilic balance of the compound and enhance oral absorption and utilization by increasing lipid solubility, NHC's isobutyric acid prodrug mopilavir (Molnupiravir, EIDD-2801) was launched in 2019. Its chemical name is ((2S,3R,4S,5S)-3,4-dihydroxy-5-(4-(hydroxyamino)-2-oxopyrimidine-1(2H)-yl)tetrahydrofuran-2-yl)methyl isobutyric acid. It has been shown to effectively inhibit the replication of MERS-CoV, SARS-CoV and SARS-CoV-2 (including remdesivir-resistant strains) in human cell culture and animal models, and has broad-spectrum antiviral activity against influenza viruses and multiple coronaviruses.

[0004]

[0005] In previous studies (CN 114805458 B), the inventors found that lauroyl prodrugs (compound 19), myristoyl prodrugs (compound 20) and palmitoyl prodrugs (compound 21) significantly improved their biomembrane permeability by introducing long-chain alkyl groups on the hydroxylamine in the N4-hydroxycytidine isobutyrate molecule of EIDD-2801. However, different carbon chains have different cytotoxicity, and the safety of carriers and prodrugs has not yet been clarified. Summary of the invention

[0006] Acute respiratory tract infections caused by coronavirus and influenza virus mainly occur in the respiratory tract. Oral administration has the disadvantages of slow onset, low bioavailability, and many adverse reactions. It is urgent to develop new dosage forms to solve this problem. Inhalation administration is an effective way to increase the drug concentration in lung tissue. However, lung tissue has a dual blood circulation system, rich blood flow, fast blood flow rate, and good lung cell permeability, so that after inhalation administration, the drug is easily diffused to the whole body, affecting the drug concentration in the target organ of respiratory cytovirus, thereby reducing the efficacy. Therefore, in order to achieve long-term drug retention in lung tissue, the present invention aims to design N4-hydroxycytidine isobutyrate into a prodrug, introduce a variety of carrier fragments on the hydroxylamine in the molecule, improve the affinity of the lung cell membrane (phospholipid membrane), and make the physicochemical properties of the drug suitable for the requirements of the physiological environment of lung tissue. Therefore, the compounds of the present invention have broad application prospects in broad-spectrum antiviral. It was also unexpectedly found that the prodrug compound provided by the present invention has unexpected antibacterial activity, and the difference in antibacterial activity is related to the introduction of a fat-soluble carrier of the prodrug.

[0007] In addition to the fact that the carrier of the N4-hydroxycytidine prodrug affects the lipid-water partition coefficient of the compound, thereby affecting the permeability and bioavailability of the drug, the present invention further found that different prodrug compounds have unexpected differences in antiviral activity, antibacterial activity and prodrug toxicity. Therefore, according to one aspect of the present invention, an object of the present invention is to provide a compound derivative or an isotope-labeled compound thereof, or an optical isomer, geometric isomer, tautomer or isomer mixture thereof, or a pharmaceutically acceptable salt thereof as shown in Formula I:

[0008]

[0009] Wherein R1 is substituted or unsubstituted C2-C 10 Alkyl, substituted or unsubstituted C 12 Alkyl, substituted or unsubstituted C 14 Alkyl, substituted or unsubstituted C 16 -C 27 Alkyl, substituted or unsubstituted C2-C 27 Alkenyl, substituted or unsubstituted C2-C 27 Alkynyl, substituted or unsubstituted C3-C6 cycloalkyl, wherein the "substituted" means that the group contains 1 to 3 R a The substituent, R a Selected from deuterium, hydroxyl, amino, cyano, nitro, halogen, C1-C5 alkoxy, C1-C5 alkoxycarbonyl, C3-C6 cycloalkyl, -NHCOOR a1 、C6-C 10Aryl, saturated or unsaturated three- to six-membered heterocyclic group containing 1-3 heteroatoms selected from N, O, and S, three- to six-membered heteroaryl group containing 1-3 heteroatoms selected from N, O, and S, unsubstituted or substituted with 1 to 2 R a2 Substituted steroid structure, the R a1 is selected from C1-C6 alkyl, said R a2 Selected from hydroxyl group, C2-C6 ester group;

[0010] R2 is a substituted or unsubstituted C1-C6 alkyl group, wherein the "substituted" means that the group contains 1 to 3 R b replaced by the R b is selected from deuterium, hydroxy, amino, cyano, nitro, halogen, C1-C5 alkoxy, C1-C5 alkyl substituted by halogen, C3-C6 cycloalkyl;

[0011] Preferably, R1 is a substituted or unsubstituted C4-C 10 Alkyl, substituted or unsubstituted C 12 Alkyl, substituted or unsubstituted C 14 Alkyl, substituted or unsubstituted C 16 -C 20 Alkyl, substituted or unsubstituted C3-C 17 Alkenyl, substituted or unsubstituted C3-C 17 Alkynyl, substituted or unsubstituted C5-C6 cycloalkyl, wherein the "substituted" means that the group contains 1 or 2 R a The substituent, R a Selected from deuterium, hydroxyl, amino, cyano, nitro, halogen, C1-C3 alkoxy, C1-C3 alkoxycarbonyl, C5-C6 cycloalkyl, -NHCOOR a1 、C6-C 10 Aryl, saturated or unsaturated five- to six-membered heterocyclic group containing 1 or 2 heteroatoms selected from N, O, and S, five- to six-membered heteroaryl group containing 1 or 2 heteroatoms selected from N, O, and S, unsubstituted or with 1 to 2 R a2 Substituted steroid structure, the R a1 is selected from C1-C4 alkyl, said R a2 Selected from hydroxyl group, C2-C6 ester group;

[0012] More preferably, R1 is a substituted or unsubstituted C7-C 10 Alkyl, substituted or unsubstituted C 12 Alkyl, substituted or unsubstituted C 14 Alkyl, substituted or unsubstituted C 16 -C 17 Alkyl, substituted or unsubstituted C8-C 17 Alkenyl, substituted or unsubstituted C8-C 17Alkynyl, substituted or unsubstituted C3-C6 cycloalkyl, wherein the "substituted" means that the group contains 1 or 2 R a The substituent, R a Selected from deuterium, hydroxyl, amino, cyano, nitro, halogen, methoxy, ethoxy, propoxy, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, cyclopentyl, cyclohexyl, -NHCOOR a1 , phenyl, naphthyl, a saturated or unsaturated five- to six-membered heterocyclic group containing 1 or 2 heteroatoms selected from N, O, and S, a five- to six-membered heteroaryl group containing 1 or 2 heteroatoms selected from N, O, and S, unsubstituted or replaced by 1 or 2 R a2 Substituted steroid structure, the R a1 is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, wherein R a2 Selected from hydroxyl, CH3COO-, CH3CH2COO-, CH3CH2CH2COO-;

[0013] More preferably, the substituent R a wherein the residue is unsubstituted or is substituted with 1 or 2 R a2 The substituted steroid is selected from the following structures:

[0014]

[0015]

[0016] Preferably, R2 is a substituted or unsubstituted C1-C4 alkyl group, wherein the "substituted" means that the group contains 1 or 2 R b replaced by the R b is selected from deuterium, hydroxy, amino, cyano, nitro, halogen, C1-C3 alkoxy, C1-C3 alkyl substituted by halogen, C3-C6 cycloalkyl;

[0017] More preferably, R2 is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, n-hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.

[0018] More preferably, R2 is isopropyl.

[0019] The compound represented by formula I according to the present invention or its isotope-labeled compound, or its optical isomer, geometric isomer, tautomer or isomer mixture, or its pharmaceutically acceptable salt is selected from the following structures:

[0020]

[0021]

[0022]

[0023] Another aspect of the present invention provides a method for preparing a compound of Formula I or an isotope-labeled compound thereof, or an optical isomer, geometric isomer, tautomer or isomer mixture thereof, or a pharmaceutically acceptable salt thereof. The preparation method can be carried out according to the contents disclosed in the prior art in combination with conventional techniques in the art. As an example, the method shown in Reaction Scheme 1 or Reaction Scheme 2 can be used:

[0024]

[0025] The method of reaction scheme 1 comprises the following steps:

[0026] i) uridine is protected, substituted, and deprotected to generate compound 5;

[0027] ii) Compound 5 is selectively esterified to generate compound 6;

[0028] iii) Compound 6 is hydroxylated to generate compound 9;

[0029] iv) Oxyacylation of the hydroxylamine group of compound 9 to generate a compound of formula I.

[0030] In a specific embodiment, in step i), trimethylsilyl chloride is used as a protecting group, chlorinated with phosphorus oxychloride in an ice bath and then substituted with triazole, and deprotected with glacial acetic acid, and the reaction temperature is 0-50°C.

[0031] In a specific embodiment, in step ii), the acyl chloride compound R2(=O)Cl is used as a base for selective esterification, and the reaction temperature is 0-50°C.

[0032] The definition of the substituent R2 in the acyl chloride compound R2(=O)Cl is the same as that in the above formula I.

[0033] In a specific embodiment, in step iii), hydroxylation is carried out using an aqueous solution prepared with hydroxylamine hydrochloride and sodium bicarbonate, and the reaction temperature is 0-50°C.

[0034] In a specific embodiment, in step iv), EDCI is 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, DMAP is p-dimethylaminopyridine, DCM is methyl chloride, EDCI is a condensing agent, and can also be replaced by DCC dicyclohexylcarbodiimide, DIC diisopropylcarbodiimide, or HATU, HBTU; the reaction temperature is 0-50°C.

[0035]

[0036] As shown in the above reaction flow diagram 2, the method comprises the following steps:

[0037] i) uridine is protected, substituted, and deprotected to generate compound 5;

[0038] ii) Compound 5 is subjected to acetone protection and esterification reaction to generate compound 7;

[0039] iii) Compound 7 is hydroxylated to generate compound 8;

[0040] iv) removing the protecting group of compound 8 to generate compound 9;

[0041] v) Oxyacylation of the hydroxylamine group of compound 9 to produce a compound of formula I.

[0042] In a specific embodiment, in step i), trimethylsilyl chloride is used as a protecting group, chlorinated with phosphorus oxychloride in an ice bath and then substituted with triazole, and the protecting group is deprotected with glacial acetic acid, and the reaction is carried out at room temperature.

[0043] In a specific embodiment, in step ii), acetone protection is carried out using 2,2-dimethoxypropane in the presence of sulfuric acid as a catalyst, and then esterification is carried out using an anhydride compound (R2(C=O))2O, and the reaction is carried out at room temperature.

[0044] The definition of the substituent R2 in the acid anhydride compound (R2(C=O))2O is the same as that in the above formula I.

[0045] In a specific embodiment, in step iii), hydroxylation is carried out using an aqueous solution prepared with hydroxylamine hydrochloride and sodium bicarbonate, and the reaction is carried out at room temperature.

[0046] In a specific embodiment, in step iv), aqueous sulfuric acid in methanol is used for deprotection.

[0047] In a specific embodiment, in step v), the condensing agent EDCI is 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, DMAP is p-dimethylaminopyridine, DCM is methyl chloride, and EDCI can also be replaced by DCC dicyclohexylcarbodiimide, DIC diisopropylcarbodiimide, or HATU, HBTU; the reaction temperature is 0-50°C.

[0048] In the reaction scheme, the substituent R1 in the carboxylic acid compound R1(C=O)OH has the same definition as that in the above formula I.

[0049] For example, the carboxylic acid compound Selected from octanoic acid, DL-lipoic acid, capric acid, octadecenoic acid, linoleic acid, 4-methyloctanoic acid, undecanoic acid, undecylenic acid, palmitoleic acid, monomethyl azelaic acid, cyclopentanepropionic acid, 6-phenylhexanoic acid, 10-undecynoic acid, 2-methylheptanoic acid, 4-ethyloctanoic acid, trans-4-butylcyclohexanecarboxylic acid, cyclohexylbutyric acid, BOC-11-aminoundecanoic acid prodrug, BOC-8-aminooctanoic acid, R-(+)-lipoic acid, chenodeoxycholic acid, ursodeoxycholic acid, lithocholic acid, deoxycholic acid, etc.

[0050] The chenodeoxycholic acid, ursodeoxycholic acid, lithocholic acid, deoxycholic acid, etc. are used to prepare prodrugs containing N4-hydroxycytidine isobutyrate with a steroid structure, such as compound 0939, compound 0941, compound 0943, and compound 0945.

[0051] The above preparation method may involve acetylation protection of the steroid 3-hydroxyl group, and the protection method is as follows:

[0052]

[0053] The raw material acid chenodeoxycholic acid, ursodeoxycholic acid, lithocholic acid or deoxycholic acid is dissolved in a solvent, 4-dimethylaminopyridine is added, acetic anhydride is added under stirring, and the mixture is stirred at room temperature for 3 hours. The reaction solution is washed with 1mol / L HCl (3×30ml), saturated sodium bicarbonate solution, and saturated sodium chloride solution, respectively, the organic layer is concentrated, and the obtained residue is purified by column chromatography to obtain a white solid. The white solid is directly condensed with N4-hydroxycytidine isobutyrate, and the target product is obtained by column chromatography.

[0054] According to one aspect of the present invention, an object of the present invention is to provide a compound represented by formula I or an isotope-labeled compound thereof, or an optical isomer, geometric isomer, tautomer or isomer mixture thereof, or a pharmaceutically acceptable salt thereof as a prodrug of cytidine isobutyrate N4-hydroxycytidine isobutyrate N4-hydroxycytidine isobutyrate.

[0055] Another aspect of the present invention is to provide a pharmaceutical composition, which comprises a therapeutically effective amount of the compound represented by Formula I or an isotope-labeled compound thereof, or an optical isomer, geometric isomer, tautomer or isomer mixture thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0056] Another aspect of the present invention is to provide the use of the compound represented by formula I or its isotope-labeled compound, or its optical isomers, geometric isomers, tautomers or isomer mixtures, or its pharmaceutically acceptable salts in the preparation of broad-spectrum antiviral or antibacterial drugs.

[0057] Preferably, the broad-spectrum antiviral or antibacterial drug is used to treat or prevent Gram-positive bacterial infection, and the Gram-positive bacteria include but are not limited to Staphylococcus, Streptococcus, Pneumococcus, Bacillus anthracis, Corynebacterium diphtheriae, and Corynebacterium tetanus.

[0058] Preferably, the broad-spectrum antiviral or antibacterial drug is prepared into an inhaler, a patch, a transdermal agent, an oral immediate-release preparation, a sustained-release or controlled-release preparation, a pill, a capsule, a granule, and the like.

[0059] Preferably, the inhalant includes a dry powder inhaler, a solution inhaler, a mixed inhaler, etc.

[0060] Another aspect of the present invention is to provide a method for treating or preventing diseases caused by viral or bacterial infection, the method comprising administering to a subject a therapeutically effective amount of the compound of formula I or its isotope-labeled compound, or its optical isomers, geometric isomers, tautomers or isomer mixtures, or its pharmaceutically acceptable salts or the pharmaceutical composition according to the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0061] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0062] Figure 1 The results of Beas-2B cell toxicity experiments of N4-hydroxycytidine isobutyrate and its prodrug. DETAILED DESCRIPTION

[0063] Hereinafter, the present invention will be described in detail. Before describing, it should be understood that the terms used in this specification and the appended claims should not be interpreted as being limited to the general meaning and dictionary meaning, but should be interpreted according to the meaning and concept corresponding to the technical aspects of the present invention on the basis of the principle that the inventor is allowed to appropriately define the terms for the best interpretation. Therefore, the descriptions presented here are only preferred examples for illustrative purposes and are not intended to limit the scope of the present invention, so that it should be understood that other equivalents or improvements can be obtained therefrom without departing from the spirit and scope of the present invention.

[0064] In this document, the terms "include", "including", "have", "contain" or any other similar terms are open conjunctions, which are intended to cover non-exclusive inclusions. For example, a composition or product containing multiple elements is not limited to the elements listed herein, but may also include other elements that are not explicitly listed but are generally inherent to the composition or product. In addition, unless otherwise explicitly stated, the term "or" refers to an inclusive "or" rather than an exclusive "or". For example, any of the following situations satisfies the condition "A or B": A is true (or exists) and B is false (or does not exist), A is false (or does not exist) and B is true (or exists), and A and B are both true (or exist). In addition, in this document, the interpretation of the terms "include", "including", "have", and "contain" should be considered to have been specifically disclosed and simultaneously cover closed or semi-closed conjunctions such as "consisting of" and "consisting essentially of".

[0065] In this article, all features or conditions defined in the form of numerical ranges or percentage ranges are only for brevity and convenience. Accordingly, the description of numerical ranges or percentage ranges should be deemed to have covered and specifically disclosed all possible secondary ranges and individual values ​​within the range, especially integer values. For example, the range description of "1 to 8" should be deemed to have specifically disclosed all secondary ranges such as 1 to 7, 2 to 8, 2 to 6, 3 to 6, 4 to 8, 3 to 8, etc., especially secondary ranges defined by all integer values, and should be deemed to have specifically disclosed individual values ​​such as 1, 2, 3, 4, 5, 6, 7, 8, etc. within the range. Unless otherwise specified, the above interpretation method applies to all contents of the entire present invention, regardless of whether the range is broad or not.

[0066] If the quantity or other numerical value or parameter is expressed as a range, a preferred range or a series of upper and lower limits, it should be understood that all ranges consisting of any upper limit or preferred value of the range and the lower limit or preferred value of the range have been specifically disclosed herein, regardless of whether these ranges are disclosed separately. In addition, if a numerical range is mentioned herein, unless otherwise specified, the range should include its endpoints and all integers and fractions within the range.

[0067] In this document, under the premise of achieving the purpose of the invention, numerical values ​​should be understood to have the accuracy of the number of significant digits of the numerical value. For example, the number 40.0 should be understood to cover the range from 39.50 to 40.49.

[0068] In this article, for the use of Markush groups or optional terms to describe the features or examples of the present invention, those skilled in the art should understand that all subgroups of elements in the Markush group or option list or any individual element can also be used to describe the present invention. For example, if X is described as "selected from the group consisting of X1, X2 and X3", it also means that the claim that X is X1 and the claim that X is X1 and / or X2 have been fully described. Furthermore, for the use of Markush groups or optional terms to describe the features or examples of the present invention, those skilled in the art should understand that any combination of all subgroups of elements in the Markush group or option list or individual elements can also be used to describe the present invention. Accordingly, for example, if X is described as "selected from the group consisting of X1, X2 and X3", and Y is described as "selected from the group consisting of Y1, Y2 and Y3", it means that the claim that X is X1 or X2 or X3 and Y is Y1 or Y2 or Y3 has been fully described.

[0069] definition

[0070] The compounds described herein may contain one or more asymmetric centers and may therefore exist in various isomeric forms, such as enantiomers and / or diastereomers. For example, the compounds described herein may be in the form of individual enantiomers, diastereomers, or geometric isomers, or may be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers. Isomers may be separated from a mixture by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferably, isomers may be prepared by asymmetric synthesis. See, e.g., Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, Tables of Resolving Agents and Optical Resolutions, p. 268 (ELiel, ed., Notre Dame University Press, Notre Dame, IN 1972). The present disclosure further encompasses the compounds described herein as individual isomers substantially free of other isomers, or as mixtures of various isomers.

[0071] When a range of values ​​is listed, it is intended to include every value and sub-range within that range. For example, "C 1-6 "Aims to cover C1, C2, C3, C4, C5, C6, C 1-6 , C 1-5 , C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-5 , C 2-4 , C 2-3 , C 3-6 , C 3-5 , C 3-4 , C 4-6 , C 4-5 and C 5-6 .

[0072] The term "alkyl" refers to a straight or branched chain saturated hydrocarbon group having 1 to 10 carbon atoms ("C 1-10 In some embodiments, an alkyl group has 1 to 9 carbon atoms (“C 1-9 In some embodiments, an alkyl group has 1 to 8 carbon atoms (“C 1-8 In some embodiments, an alkyl group has 1 to 7 carbon atoms (“C 1-7 In some embodiments, an alkyl group has 1 to 6 carbon atoms (“C 1-6 In some embodiments, an alkyl group has 1 to 5 carbon atoms (“C 1-5 In some embodiments, an alkyl group has 1 to 4 carbon atoms (“C 1-4 In some embodiments, an alkyl group has 1 to 3 carbon atoms (“C 1-3 In some embodiments, an alkyl group has 1 to 2 carbon atoms (“C 1-2 In some embodiments, an alkyl group has 1 carbon atom (“C1 alkyl”). In some embodiments, an alkyl group has 2 to 6 carbon atoms (“C 2-6 C 1-6Examples of alkyl groups include methyl (C1), ethyl (C2), propyl (C3) (e.g., n-propyl, isopropyl), butyl (C4) (e.g., n-butyl, tert-butyl, sec-butyl, isobutyl), pentyl (C5) (e.g., n-pentyl, 3-pentyl, neopentyl, 3-methyl-2-butyl, tert-pentyl) and hexyl (C6) (e.g., n-hexyl). Additional examples of alkyl groups include n-heptyl (C7), n-octyl (C8), and the like. Unless otherwise specified, each example of alkyl is independently unsubstituted ("unsubstituted alkyl") or substituted ("substituted alkyl") with one or more substituents (e.g., halogen, such as F). In certain embodiments, alkyl is unsubstituted C 1-10 Alkyl (e.g. unsubstituted C 1-6 In certain embodiments, the alkyl group is a substituted C 1-10 Alkyl (e.g. substituted C 1-6 Alkyl, such as -CF3).

[0073] "Alkenyl" refers to a straight or branched chain hydrocarbon radical having 2 to 20 carbon atoms, one or more carbon-carbon double bonds and no triple bonds ("C 2-20 The carbon-carbon double bond can be located at the end of the carbon long chain or at any position allowed by the chemical structure. In some embodiments, the alkenyl group has 2 to 10 carbon atoms ("C 2-10 In some embodiments, an alkenyl group has 2 to 9 carbon atoms (“C 2-9 In some embodiments, an alkenyl group has 2 to 8 carbon atoms (“C 2-8 In some embodiments, an alkenyl group has 2 to 7 carbon atoms (“C 2-7 In some embodiments, an alkenyl group has 2 to 6 carbon atoms (“C 2-6 In some embodiments, an alkenyl group has 2 to 5 carbon atoms (“C 2-5 In some embodiments, an alkenyl group has 2 to 4 carbon atoms (“C 2-4 In some embodiments, an alkenyl group has 2 to 3 carbon atoms (“C 2-3 In some embodiments, an alkenyl group has 2 carbon atoms ("C2 alkenyl"). The one or more carbon-carbon double bonds can be internal (such as in 2-butenyl) or terminal (such as in 1-butenyl). C 2-4 Examples of alkenyl groups include ethenyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), and the like. 2-6 Examples of the alkenyl group include the aforementioned C 2-4In some embodiments, alkenyl is an unsubstituted C 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1, 1-1 2-10 In certain embodiments, alkenyl is a substituted C 2-10 In alkenyl, a C=C double bond with unspecified stereochemistry (e.g., -CH=CHCH3 or ) can be an (E)- or (Z)-double bond.

[0074] "Alkynyl" refers to a straight or branched chain hydrocarbon radical having 2 to 20 carbon atoms, one or more carbon-carbon triple bonds and optionally one or more double bonds ("C 2-20 The carbon-carbon triple bond can be located at the end of the carbon long chain or at any position permitted by the chemical structure. In some embodiments, the alkynyl group has 2 to 10 carbon atoms ("C 2-10 In some embodiments, an alkynyl group has 2 to 9 carbon atoms (“C 2-9 In some embodiments, an alkynyl group has 2 to 8 carbon atoms (“C 2-8 In some embodiments, an alkynyl group has 2 to 7 carbon atoms (“C 2-7 In some embodiments, an alkynyl group has 2 to 6 carbon atoms (“C 2-6 In some embodiments, an alkynyl group has 2 to 5 carbon atoms (“C 2-5 In some embodiments, an alkynyl group has 2 to 4 carbon atoms (“C 2-4 In some embodiments, an alkynyl group has 2 to 3 carbon atoms (“C 2-3 In some embodiments, an alkynyl group has 2 carbon atoms (a "C2 alkynyl"). The one or more carbon-carbon triple bonds can be internal (such as in 2-butynyl) or terminal (such as in 1-butynyl). 2-4 Examples of alkynyl groups include, but are not limited to, ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), and the like. 2-6 Examples of the alkenyl group include the above-mentioned C 2-4Alkynyl and pentynyl (C5), hexynyl (C6), etc. Additional examples of alkynyl include heptynyl (C7), octynyl (C8), etc. Unless otherwise specified, each instance of alkynyl is independently optionally substituted, i.e., unsubstituted (“unsubstituted alkynyl”) or substituted with one or more substituents (“substituted alkynyl”). In certain embodiments, alkynyl is unsubstituted C 2-10 In certain embodiments, the alkynyl group is a substituted C 2-10 Alkynyl.

[0075] "Cycloalkyl" refers to a non-aromatic ring system having 3 to 10 ring carbon atoms ("C 3-10 In some embodiments, the carbocyclyl group has 3 to 8 ring carbon atoms ("C 3-8 In some embodiments, a carbocyclyl group has 3 to 7 ring carbon atoms (“C 3-7 In some embodiments, a carbocyclyl group has 3 to 6 ring carbon atoms (“C 3-6 In some embodiments, a carbocyclyl group has 5 to 10 ring carbon atoms (“C 5-10 Carbocyclyl”). Exemplary C 3-6 Carbocyclic groups include, but are not limited to, cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), and the like. Exemplary C 3-8 Carbocyclic groups include but are not limited to the above C 3-6 Carbocyclic groups and cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptatrienyl (C7), cyclooctyl (C8), cyclooctenyl (C8), bicyclo [2.2.1] heptyl (C7), bicyclo [2.2.2] octyl (C8), etc. Exemplary C 3-10 Carbocyclic groups include but are not limited to the above C 3-8 Carbocyclic groups and cyclononyl (C9), cyclononenyl (C9), cyclodecyl (C 10 ), cyclodecenyl (C 10 ), octahydro-1H-indenyl (C9), decahydronaphthyl (C 10 ), spiro[4.5]decyl (C 10) etc. As shown in the foregoing examples, in certain embodiments, the carbocyclyl is a monocyclic ring ("monocyclic carbocyclyl") or contains a fused ring, a bridged ring or a spirocyclic ring system, such as a bicyclic ring system ("bicyclic carbocyclyl") and can be saturated or can be partially unsaturated. "Carbocyclyl" also includes a ring system in which the carbocyclic ring as defined above is fused to one or more aryl or heteroaryl groups at a point of attachment on the carbocyclic ring, and in this case, the carbon number continues to refer to the carbon number in the carbocyclic ring system. Unless otherwise indicated, each example of a carbocyclyl group is independently optionally substituted, i.e., unsubstituted ("unsubstituted carbocyclyl") or substituted ("substituted carbocyclyl") with one or more substituents. In certain embodiments, the carbocyclyl is an unsubstituted C 3-10 In certain embodiments, a carbocyclyl group is a substituted C 3-10 Carbocyclic group.

[0076] In some embodiments, a "carbocyclyl" is a monocyclic saturated carbocyclyl ("C 3-10 In some embodiments, a cycloalkyl group has 3 to 8 ring carbon atoms (“C 3-8 In some embodiments, a cycloalkyl group has 3 to 6 ring carbon atoms (“C 3-6 In some embodiments, a cycloalkyl group has 5 to 6 ring carbon atoms (“C 5-6 In some embodiments, a cycloalkyl group has 5 to 10 ring carbon atoms (“C 5-10 Cycloalkyl”) C 5-6 Examples of cycloalkyl groups include cyclopentyl (C5) and cyclohexyl (C6). 3-6 Examples of the cycloalkyl group include the aforementioned C 5-6 Cycloalkyl as well as cyclopropyl (C3) and cyclobutyl (C4). 3-8 Examples of the cycloalkyl group include the aforementioned C 3-6 Cycloalkyl and cycloheptyl (C7) and cyclooctyl (C8). Unless otherwise specified, each instance of cycloalkyl is independently unsubstituted (“unsubstituted cycloalkyl”) or substituted (“substituted cycloalkyl”) with one or more substituents. In certain embodiments, cycloalkyl is unsubstituted C 3-10 In certain embodiments, the cycloalkyl group is a substituted C 3-10 Cycloalkyl.

[0077] "Heterocyclyl" refers to a group of a 3 to 10-membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus and silicon ("3-10-membered heterocyclyl"). In heterocyclyls containing one or more nitrogen atoms, the point of attachment may be a carbon atom or a nitrogen atom as long as the valence permits. The heterocyclyl may be a monocyclic ("monocyclic heterocyclyl") or a fused ring, bridged ring or spirocyclic system, such as a bicyclic system ("bicyclic heterocyclyl"), and may be saturated or may be partially unsaturated. The heterocyclyl bicyclic system may contain one or more heteroatoms in one or both rings. "Heterocyclyl" also includes a ring system in which the point of attachment of a heterocycle as defined above to one or more carbocyclyl groups is fused to a carbocyclyl or heterocycle, or a ring system in which the point of attachment of a heterocycle as defined above to one or more aryl or heteroaryl groups is fused to a heterocycle, and in this case, the number of ring members continues to refer to the number of ring members in the heterocyclic system. Unless otherwise specified, each instance of heterocyclyl is independently optionally substituted, i.e., unsubstituted (“unsubstituted heterocyclyl”) or substituted (“substituted heterocyclyl”) with one or more substituents. In certain embodiments, heterocyclyl is an unsubstituted 3-10 membered heterocyclyl. In certain embodiments, heterocyclyl is a substituted 3-10 membered heterocyclyl.

[0078] In some embodiments, heterocyclyl is a 5-10 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus and silicon (“5-10 membered heterocyclyl”). In some embodiments, heterocyclyl is a 5-8 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur (“5-8 membered heterocyclyl”). In some embodiments, heterocyclyl is a 5-6 membered non-aromatic ring system having ring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur (“5-6 membered heterocyclyl”). In some embodiments, 5-6 membered heterocyclyl has 1-3 ring heteroatoms selected from nitrogen, oxygen and sulfur. In some embodiments, 5-6 membered heterocyclyl has 1-2 ring heteroatoms selected from nitrogen, oxygen and sulfur. In some embodiments, the 5-6 membered heterocyclyl has 1 ring heteroatom selected from nitrogen, oxygen, and sulfur.

[0079] "Aryl" refers to a group having 6-14 ring carbon atoms and zero heteroatoms provided in the aromatic ring system, a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10 or 14 π electrons shared in the cyclic array) ("C 6-14 In some embodiments, an aryl group has 6 ring carbon atoms ("C6 aryl"; e.g., phenyl). In some embodiments, an aryl group has 10 ring carbon atoms ("C 10In some embodiments, an aryl group has 14 ring carbon atoms ("C 14 "Aryl"; e.g., anthracenyl). "Aryl" also includes ring systems in which an aryl ring as defined above is fused to one or more carbocyclic or heterocyclic groups, wherein the radical or point of attachment is on the aromatic ring, and in this case, the number of carbon atoms continues to refer to the number of carbon atoms in the aromatic ring system. Unless otherwise specified, each instance of aryl is independently optionally substituted, i.e., unsubstituted ("unsubstituted aryl") or substituted ("substituted aryl") with one or more substituents. In certain embodiments, aryl is unsubstituted C 6-14 In certain embodiments, aryl is substituted C 6-14 Aryl.

[0080] "Aralkyl" is a subset of alkyl and aryl and refers to an optionally substituted alkyl substituted by an optionally substituted aryl. In certain embodiments, the aralkyl is an optionally substituted benzyl. In certain embodiments, the aralkyl is benzyl. In certain embodiments, the aralkyl is an optionally substituted phenethyl. In certain embodiments, the aralkyl is phenethyl.

[0081] "Heteroaryl" refers to a group of a 5-10 membered monocyclic or bicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 π electrons shared in a cyclic array) having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen and sulfur ("5-10 membered heteroaryl"). In heteroaryl groups containing one or more nitrogen atoms, the point of attachment may be a carbon atom or a nitrogen atom as long as valence permits. Heteroaryl bicyclic ring systems may contain one or more heteroatoms in one or both rings. "Heteroaryl" includes ring systems in which a heteroaryl ring as defined above is fused to one or more carbocyclyl or heterocyclyl groups, wherein the point of attachment is on the heteroaryl ring, and in this case, the number of ring members continues to refer to the number of ring members in the heteroaryl ring system. "Heteroaryl" also includes ring systems in which a heteroaryl ring as defined above is fused to one or more aryl groups, wherein the point of attachment is on the aryl or heteroaryl ring, and in this case, the number of ring members refers to the number of ring members in the fused (aryl / heteroaryl) ring system. For bicyclic heteroaryls in which one ring does not contain heteroatoms (e.g., indolyl, quinolyl, carbazolyl, etc.), the point of attachment can be on either ring, i.e., the ring with heteroatoms (e.g., 2-indolyl) or the ring without heteroatoms (e.g., 5-indolyl).

[0082] "Halogen" includes a halogen atom selected from F, Cl, Br and I.

[0083] The following examples are only listed as examples of embodiments of the present invention and do not constitute any limitation to the present invention. It can be understood by those skilled in the art that modifications within the scope of the essence and concept of the present invention fall within the scope of protection of the present invention. Unless otherwise specified, the reagents and instruments used in the following examples are all commercially available products.

[0084] In addition, unless otherwise stated, the biochemical reagents used in this application are all commercial products, for example, purchased from Anage Chemical, Beijing Inokai Technology Co., Ltd. or Beijing Jinrui Meixiang Technology Co., Ltd., with analytical grade specifications and used directly without further treatment.

[0085] Example

[0086] Example 1: Preparation of caprylic acid prodrug (Compound 0836)

[0087] Step 1: Preparation of 1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-yl) (5)

[0088]

[0089] Take a 500ml three-necked flask and dry it, then add uridine (30.00g, 122.85mmol), anhydrous acetonitrile (225ml) and N-methylpyrrolidine (192.0ml, 1.843mol), slowly add TMSCl (78.0ml, 614.25mmol) under nitrogen protection and 0℃ ice bath, and stir at room temperature for 1h. Then slowly add phosphorus oxychloride (22.5ml, 245.7mmol) under 0℃ ice bath, and stir under ice bath for 20 minutes after the addition is completed. Add triazole (84.60g, 1.23mol) to the reaction solution under 0℃ ice bath, ice bath for 1 hour and room temperature for 2 hours. After the reaction is completed, pour the reaction solution into the prepared 600ml sodium chloride saturated solution to quench the reaction, extract it three times with dichloromethane (200ml), combine the organic phases, dry it over anhydrous sodium sulfate, filter it with suction, and concentrate it to obtain an orange-red oil. A mixed solvent (120 mL) of methanol and acetic acid (v / v=10:1) was added thereto, and the mixture was stirred at room temperature overnight. The mixture was filtered and dried to obtain 30.24 g of a light yellow solid with a yield of 83.4%.

[0090] Step 2: Preparation of methyl ((2R,3S,4R,5R)-3,4-dihydroxy-5-(2-oxo-4-(1H-1,2,4-triazol-1-yl)pyrimidin-1(2H)-yl)tetrahydrofuran-2-yl)isobutyrate (6)

[0091]

[0092] 1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-yl)(5)(10 g, 33.87 mmol) was dissolved in a mixed solution of 1,4-dioxane (40 ml) and N,N-dimethylformamide (20 ml), N,N-dimethylpropyleneurea was added, and then isobutyryl chloride (6.72 g,

[0093] 67.74mmol), reacted at room temperature for eight hours. The reaction solution was extracted with ethyl acetate / water six times, the organic phase was vortexed until it was no longer reduced to a solution state, extracted three times with petroleum ether: methyl tert-butyl ether (20:1), and then slurried with ethyl acetate: petroleum ether (1:10) to obtain 8.80g of an oily substance with a yield of 71.31%.

[0094] Step 3: Preparation of ((2R,3S,4R,5R)-3,4-dihydroxy-5-(4-(hydroxyamino)-2-oxopyrimidin-1(2H)-yl)tetrahydrofuran-2-yl)methyl isobutyrate (9)

[0095]

[0096] The compound ((2R, 3S, 4R, 5R)-3, 4-dihydroxy-5-(2-oxo-4-(1H-1, 2, 4-triazol-1-yl) pyrimidine-1(2H)-yl) tetrahydrofuran-2-yl) isobutyric acid methyl ester (6) (8.80 g, 24.10 mmol) obtained above was added to a 100 ml reaction bottle, and after adding isopropanol and stirring, the prepared hydroxylamine aqueous solution (36.15 mmol of hydroxylamine hydrochloride and 36.15 mmol of sodium bicarbonate were dissolved in 6 ml of water) was added, and the reaction was carried out at room temperature for three hours. After isopropanol was removed by concentration under reduced pressure, ethyl acetate / water was extracted four times, and the organic layer was spin-dried to obtain an oily substance. The above oily substance was slurried with ethyl acetate: petroleum ether (1:10) to obtain 6.82 g of the compound, with a yield of 85.89%.

[0097] Step 4: Synthesis of caprylic acid prodrug (Compound 0836)

[0098]

[0099] N4-hydroxycytidine isobutyrate (400 mg, 1.22 mmol) was added to a 50 ml reaction bottle, and dichloromethane (10 ml), octanoic acid (166 mg, 1.15 mmol) and condensing agent 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (256 mg, 1.34 mmol), 4-dimethylaminopyridine (122 mg, 0.49 mmol) were added, and stirred at room temperature for 6 hours, extracted with dichloromethane / water, and the organic phase was concentrated. The obtained residue was purified by column chromatography (DCM:MeOH (v / v) = 80:1 to 50:1) to obtain a solid product with a yield of 30.2%.

[0100] 1 H NMR (600MHz, chloroform-d) δ8.82(s,1H),7.09(d,J=8.3Hz,1H),5.79(d,J=8.3Hz,1H),5.76(d,J=4. 0Hz,1H),4.34(d,J=3.6Hz,2H),4.28(q,J=3.9Hz,1H),4.20(t,J=4.8Hz,1H),4.17(t,J=5.2H z,1H),3.18(s,2H),2.58(p,J=7.0Hz,1H),2.48(t,J=7.5Hz,2H),1.69(p,J=7.5Hz,2H),1.37 –1.25(m,8H),1.18(dd,J=6.9,1.2Hz,6H),0.88(t,J=6.8Hz,3H).MS(ESI):m / z=456.23(M+H) + .

[0101] Example 2: Preparation of DL-lipoic acid prodrug (Compound 0838)

[0102] Step 1: Preparation of 1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-yl) (5)

[0103]

[0104] Take a 500ml three-necked flask and dry it, then add uridine (30.00g, 122.85mmol), anhydrous acetonitrile (225ml) and N-methylpyrrolidine (192.0ml, 1.843mol), slowly add TMSCl (78.0ml, 614.25mmol) under nitrogen protection and 0℃ ice bath, and stir at room temperature for 1h. Then slowly add phosphorus oxychloride (22.5ml, 245.7mmol) under 0℃ ice bath, and stir under ice bath for 20 minutes after the addition is completed. Add triazole (84.60g, 1.23mol) to the reaction solution under 0℃ ice bath, ice bath for 1 hour and room temperature for 2 hours. After the reaction is completed, pour the reaction solution into the prepared 600ml sodium chloride saturated solution to quench the reaction, extract it three times with dichloromethane (200ml), combine the organic phases, dry it over anhydrous sodium sulfate, filter it with suction, and concentrate it to obtain an orange-red oil. A mixed solvent (120 mL) of methanol and acetic acid (v / v=10:1) was added thereto, and the mixture was stirred at room temperature overnight. The mixture was filtered and dried to obtain 30.24 g of a light yellow solid with a yield of 83.4%.

[0105] Step 2: Preparation of ((3aR,4R,6R,6aR)-2,2-dimethyl-6-(2-oxo-4-(1H-1,2,4-triazol-1-yl)pyrimidin-1(2H)-yl)tetrahydrofuran[3,4-d][1,3]dioxolan-4-yl)methyl isobutyrate (7)

[0106]

[0107] The compound 1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-4-(1H-1,2,4-triazol-1-yl)pyrimidin-2(1H)-yl) (5) (29.07 g, 98.46 mmol) obtained above was added to a round-bottom flask, 300 ml of anhydrous acetonitrile was added, 2,2-dimethoxypropane (24.21 ml, 196.41 mmol) was added under stirring, concentrated sulfuric acid (4.92 mmol) was added dropwise, and the reaction was carried out at room temperature for 6 h. The reaction solution was filtered to obtain a light yellow solid. The light yellow solid was added to the reaction bottle, and 350 ml of anhydrous acetonitrile was added. Triethylamine (109.50 ml, 787.68 mmol), 4-dimethylaminopyridine (3.01 g, 24.61 mmol), and isobutyric anhydride (18.00 ml, 108.31 mmol) were added under stirring, and stirred at room temperature for 1 hour. Most of the solvent was removed by rotary evaporation, and the mixture was dissolved in ethyl acetate and washed once with a saturated sodium bicarbonate aqueous solution and a saturated sodium chloride solution, and then dried by rotary drying to obtain 32.94 of a light yellow solid with a yield of 82.53%.

[0108] Step 3: Preparation of ((3aR,4R,6R,6aR)-6-(4-(hydroxyamino)-2-oxopyrimidin-1(2H)-yl)-2,2-dimethyltetrahydrofuran[3,4-d][1,3]dioxolan-4-yl)methyl isobutyrate (8).

[0109]

[0110] The compound ((3aR, 4R, 6R, 6aR)-2,2-dimethyl-6-(2-oxo-4-(1H-1,2,4-triazol-1-yl)pyrimidin-1(2H)-yl)tetrahydrofuran[3,4-d][1,3]dioxolan-4-yl)methyl isobutyrate (7) (30 g, 74.00 mmol) obtained above was added to a 500 ml reaction bottle, 300 ml of isopropanol was added and stirred, and then the prepared hydroxylamine aqueous solution (111.00 mmol of hydroxylamine hydrochloride and 111.00 mmol of sodium bicarbonate were dissolved in 60 ml of water) was added and reacted at room temperature for three hours. After concentrating under reduced pressure to remove most of the isopropanol, 50 ml of ice water was added and the mixture was beaten in an ice bath for 1 hour, and 23.58 g of white solid was obtained by suction filtration, with a yield of 85.63%.

[0111] Step 4: Preparation of ((2R,3S,4R,5R)-3,4-dihydroxy-5-(4-(hydroxyamino)-2-oxopyrimidin-1(2H)-yl)tetrahydrofuran-2-yl)methyl isobutyrate (9)

[0112]

[0113] The compound ((3aR, 4R, 6R, 6aR)-6-(4-(hydroxyamino)-2-oxopyrimidin-1(2H)-yl)-2,2-dimethyltetrahydrofuran[3,4-d][1,3]dioxolan-4-yl)methyl isobutyrate (8) (20 g, 54.15 mmol) obtained above was added to a 500 ml reaction bottle, 100 ml of 25% methanol aqueous solution was added, sulfuric acid (43.32 mmol) was slowly added dropwise under stirring, and the mixture was stirred at 70°C for 1 h. After cooling to room temperature, sodium bicarbonate was added to adjust the pH to neutral, and the mixture was filtered. The filtrate was extracted with ethyl acetate 10 times (80 ml×10), the solvent was spin-dried, EA:DCM:MeOH=15:5:2, and 20 ml was slurried for purification to obtain a total of compound 12.52 with a yield of 70.2%.

[0114] Step 5: Synthesis of DL-lipoic acid prodrug (Compound 0838)

[0115]

[0116] N4-hydroxycytidine isobutyrate (400 mg, 1.22 mmol) was added to a 50 ml reaction bottle, and dichloromethane (10 ml), thioctic acid (237 mg, 1.15 mmol) and condensing agent 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (256 mg, 1.34 mmol), 4-dimethylaminopyridine (122 mg, 0.49 mmol) were added, stirred at room temperature for 6 hours, extracted with dichloromethane / water, and the organic phase was concentrated. The residue was purified by column chromatography (DCM:MeOH (v / v) = 80:1 to 50:1) to obtain a solid product with a yield of 48.9%.

[0117] 1 H NMR (400MHz, chloroform-d) δ8.90 (s, 1H), 7.10 (d, J = 8.3Hz, 1H), 5.81–5.73 (m, 2H), 4.35 (d,J=3.5Hz,2H),4.28(q,J=3.8Hz,1H),4.18(dq,J=10.6,5.2Hz,2H),3.60–3.55 (m,1H),3.45(s,1H),3.22–3.06(m,3H),2.54–2.46(m,3H),1.95–1.91(m,1H),1 .82(s,1H),1.75–1.68(m,4H),1.19(d,J=6.8Hz,6H).MS(ESI):m / z=518.08(M+H) + .

[0118] Example 3: Preparation of Decanoic Acid Prodrug (Compound 0839)

[0119]

[0120] Compound 0839 was prepared in the same manner as in Example 1 except that octanoic acid was replaced by decanoic acid. A solid product was obtained with a yield of 21.1%.

[0121] 1H NMR (600MHz, chloroform-d) δ8.88 (s, 1H), 7.09 (d, J = 8.3Hz, 1H), 5.79 (dd, J = 8.3, 1.6Hz, 1H), 5. 76(d,J=4.1Hz,1H),4.34(d,J=3.6Hz,2H),4.27(dd,J=5.0,3.4Hz,1H),4.23–4.14(m,2 H),2.58(hept,J=7.0Hz,1H),2.48(t,J=7.6Hz,2H),1.69(p,J=7.5Hz,2H),1.37–1.23( m,12H),1.18(dd,J=6.9,1.1Hz,6H),0.88(t,J=7.0Hz,3H).MS(ESI):m / z=484.26(M+H) + .

[0122] Example 4: Preparation of octadecenoic acid prodrug (Compound 0844)

[0123]

[0124] Compound 0844 was prepared in the same manner as in Example 1 except that octanoic acid was replaced by octadecenoic acid. Finally, a solid product was obtained with a yield of 36.0%.

[0125] 1 H NMR (600MHz, chloroform-d) δ8.62(s,1H),7.10(d,J=8.4Hz,1H),5.79(d,J=8.3Hz,1H),5.73(d,J=4.0Hz,1H), 5.36–5.33(m,2H),4.34(t,J=3.2Hz,2H),4.31–4.29(m,1H),4.20(dd,J=5.5,4.0Hz,1H),4.17(t,J=5. 1Hz,1H),2.57(p,J=7.0Hz,1H),2.47(t,J=7.6Hz,2H),2.01(dd,J=7.2,3.0Hz,4H),1.70(p,J=7.5Hz,2 H),1.40–1.21(m,20H),1.18(dd,J=6.9,2.0Hz,6H),0.88(t,J=7.0Hz,3H).MS(ESI):m / z=594.37(M+H) + .

[0126] Example 5: Preparation of linoleic acid prodrug (Compound 0845)

[0127]

[0128] Compound 0845 was prepared in the same manner as in Example 1 except that linoleic acid was used instead of caprylic acid to obtain a solid product with a yield of 37.2%.

[0129] 1 H NMR (600MHz, chloroform-d) δ7.10(d,J=8.3Hz,1H),5.79(dd,J=8.3,2.0Hz,1H),5.73(d,J=3.9Hz,1H),5 .39–5.30(m,4H),4.34(dd,J=3.4,1.8Hz,2H),4.31–4.27(m,1H),4.21–4.16(m,2H),2.77(t,J=6 .9Hz,2H),2.57(p,J=7.0Hz,1H),2.47(t,J=7.6Hz,2H),2.07–2.02(m,4H),1.72–1.67(m,2H),1. 37–1.29(m,14H),1.18(dd,J=6.9,1.7Hz,6H),0.89(t,J=6.9Hz,3H).MS(ESI):m / z=592.37(M+H) + .

[0130] Example 6: Preparation of 4-methyloctanoic acid prodrug (Compound 0850)

[0131]

[0132] Compound 0850 was prepared in the same manner as in Example 1 except that octanoic acid was replaced by 4-methyloctanoic acid. Finally, a solid product was obtained with a yield of 44.5%.

[0133] 1 H NMR (600MHz, chloroform-d) δ8.83 (s, 1H), 7.09 (d, J = 8.3Hz, 1H), 5.79 (dd, J = 8.4, 1.8H z,1H),5.76(d,J=4.0Hz,1H),4.34(d,J=3.6Hz,2H),4.28–4.25(m,1H),4.20– 4.13(m,2H),2.58(p,J=6.9Hz,1H),2.51–2.43(m,2H),1.52–1.44(m,2H),1.2 9(tdd,J=13.4,9.6,6.6Hz,6H),1.18(d,J=7.1Hz,6H),0.89(t,J=6.5Hz,6H).

[0134] MS (ESI): m / z = 470.25 (M+H) + .

[0135] Example 7: Preparation of Undecanoic Acid Prodrug (Compound 0901)

[0136]

[0137] Compound 0901 was prepared in the same manner as in Example 1 except that octanoic acid was replaced by undecanoic acid. Finally, a solid product was obtained with a yield of 22.1%.

[0138] 1 H NMR(400MHz,DMSO-d6)δ7.18(d,J=8.1Hz,1H),5.76–5.71(m,2H),5.47(d,J=5.5Hz, 1H),5.29(d,J=5.3Hz,1H),4.23–4.15(m,2H),4.06–4.02(m,1H),3.98–3.92(m,2H), 2.62–2.56(m,1H),2.44(t,J=7.5Hz,2H),1.54(dd,J=10.4,3.9Hz,2H),1.25(d,J=5 .2Hz,13H),1.10(d,J=6.9Hz,5H),0.86(d,J=6.2Hz,3H).MS(ESI):m / z=498.27(M+H) + .

[0139] Example 8: Preparation of undecylenic acid prodrug (Compound 0902)

[0140]

[0141] Compound 0902 was prepared in the same manner as in Example 1 except that octanoic acid was replaced by undecylenic acid. Finally, a solid product was obtained with a yield of 40.0%.

[0142] 1 H NMR (600MHz, chloroform-d) δ8.84(s,1H),7.09(d,J=8.3Hz,1H),5.81–5.77(m,2H),5.75(d,J=4.0Hz,1 H),4.99(dt,J=17.2,1.9Hz,1H),4.93(dq,J=10.2,1.5Hz,1H),4.34(d,J=3.7Hz,2H),4.29–4. 26(m,1H),4.21–4.15(m,2H),2.60–2.56(m,1H),2.47(t,J=7.5Hz,2H),2.04–2.02(m,2H),1.6 8(d,J=7.6Hz,2H),1.32–1.27(m,10H),1.18(dd,J=7.1,1.2Hz,6H).MS(ESI):m / z=496.27(M+H)+ .

[0143] Example 9: Preparation of palmitoleic acid prodrug (Compound 0903)

[0144]

[0145] Compound 0903 was prepared in the same manner as in Example 1 except that octanoic acid was replaced by palmitoleic acid. Finally, a solid product was obtained with a yield of 40.7%.

[0146] 1 H NMR (600MHz, chloroform-d) δ8.74(s,1H),7.10(d,J=8.3Hz,1H),5.79(dd,J=8.3,1.7Hz,1H),5.74(d,J= 4.0Hz,1H),5.34(q,J=5.5Hz,2H),4.34(d,J=3.5Hz,2H),4.31–4.25(m,1H),4.23–4.13(m,2H),2 .58(dq,J=14.0,7.0Hz,1H),2.47(t,J=7.6Hz,2H),2.04–1.98(m,4H),1.69(p,J=7.5Hz,2H),1. 39–1.24(m,18H),1.18(dd,J=7.0,1.5Hz,6H),0.88(t,J=6.9Hz,3H).MS(ESI):m / z=566.35(M+H) + .

[0147] Example 10: Preparation of azelaic acid monomethyl ester prodrug (Compound 0904)

[0148]

[0149] Compound 0904 was prepared in the same manner as in Example 1 except that octanoic acid was replaced by azelaic acid monomethyl ester. Finally, a solid product was obtained with a yield of 23.1%.

[0150] 1H NMR (600 MHz, chloroform-d) δ8.79 (s, 1H), 7.09 (d, J = 8.4 Hz, 1H), 5.79 (dd, J = 8.3, 2.2 Hz, 1H), 5.77 (d, J = 4.0 Hz, 1H), 4.34 (d, J = 3.6 Hz, 2H), 4.29 (q, J = 3.8 Hz, 1H), 4.18 (dq, J = 10.1, 5.4 Hz, 2H), 3.67 (s, 4 H),2.58(p,J=7.0Hz,1H),2.48(t,J=7.5Hz,2H),2.31(t,J=7.5Hz,3H),1.69(p,J=7.5Hz,2H),1 .62(p,J=7.4Hz,3H),1.35–1.30(m,5H),1.18(dd,J=7.0,1.4Hz,6H).MS(ESI):m / z=514.22(M+H) + .

[0151] Example 11: Preparation of cyclopentanepropionic acid prodrug (Compound 0905)

[0152]

[0153] Compound 0905 was prepared in the same manner as in Example 1 except that octanoic acid was replaced by cyclopentanepropionic acid. Finally, a solid product was obtained with a yield of 27.6%.

[0154] 1 H NMR(400MHz,DMSO-d6)δ10.88(s,1H),7.11(d,J=8.2Hz,1H),5.70–5.65(m,2H),5 .40(d,J=5.6Hz,1H),5.22(d,J=5.3Hz,1H),4.17–4.09(m,2H),3.97(t,J=5.2Hz, 1H),3.91–3.85(m,2H),3.34(s,2H),2.52(td,J=6.9,1.5Hz,1H),2.39(t,J=7.8H z,2H),1.69–1.63(m,3H),1.56–1.36(m,8H),1.03(d,J=7.0Hz,6H).MS(ESI):m / z

[0155] =454.22(M+H) + .

[0156] Example 12: Preparation of 6-phenylhexanoic acid prodrug (Compound 0906)

[0157]

[0158] Compound 0906 was prepared in the same manner as in Example 1 except that octanoic acid was replaced by 6-phenylhexanoic acid. A solid product was obtained with a yield of 45.8%.

[0159] 1 H NMR (400MHz, DMSO-d6) δ7.29–7.15(m,6H),5.77–5.67(m,2H),5.47(d,J=5.5Hz,1H), 5.29(d,J=5.3Hz,1H),4.27–4.13(m,2H),4.05(q,J=5.3Hz,1H),4.01–3.90(m,2H),3 .39(s,19H),2.58(q,J=7.3,6.9Hz,3H),2.45(t,J=7.5Hz,2H),1.59(q,J=7.6Hz,4H) ,1.31(td,J=7.2,3.9Hz,2H),1.10(dd,J=7.0,3.9Hz,6H).MS(ESI):m / z=504.25(M+H) + .

[0160] Example 13: Preparation of 10-Undecynoic Acid Prodrug (Compound 0907)

[0161]

[0162] Compound 0907 was prepared in the same manner as in Example 1 except that octanoic acid was replaced by 10-undecynoic acid. Finally, a solid product was obtained with a yield of 16.0%.

[0163] 1 H NMR (600MHz, chloroform-d) δ8.77(s,1H),7.09(d,J=8.3Hz,1H),5.79(dd,J=8.3,2.0Hz,1H),5.74(d, J=3.9Hz,1H),4.35–4.34(m,2H),4.29–4.27(m,1H),4.22–4.16(m,2H),2.58(dt,J=14.0,7.0H z,1H),2.48(t,J=7.6Hz,2H),2.19–2.17(m,2H),1.94(t,J=2.6Hz,1H),1.69(d,J=7.5Hz,2H), 1.54–1.50(m,2H),1.40–1.31(m,8H),1.18(dd,J=7.1,1.4Hz,6H).MS(ESI):m / z=494.25(M+H) + .

[0164] Example 14: Preparation of 2-methylheptanoic acid prodrug (Compound 0908)

[0165]

[0166] Compound 0908 was prepared in the same manner as in Example 1 except that octanoic acid was replaced by 2-methylheptanoic acid. Finally, a solid product was obtained with a yield of 33.2%.

[0167] 1H NMR (600MHz, chloroform-d) δ8.49(s,1H),7.08(dd,J=8.2,2.2Hz,1H),5.82(d,J=8.2Hz,1H),5.75(dd, J=7.8,4.0Hz,1H),4.34(d,J=3.6Hz,2H),4.28(q,J=3.8Hz,1H),4.23–4.16(m,2H),2.59(ddt, J=24.1,14.0,7.0Hz,2H),1.48(ddt,J=13.2,10.0,6.4Hz,1H),1.33–1.26(m,6H),1.23(d,J=6 .9Hz,3H),1.18(dd,J=7.0,1.4Hz,6H),0.88(td,J=7.0,2.2Hz,3H).MS(ESI):m / z=456.23(M+H) + .

[0168] Example 15: Preparation of 4-ethyloctanoic acid prodrug (Compound 0909)

[0169]

[0170] Compound 0909 was prepared in the same manner as in Example 1 except that octanoic acid was replaced by 4-ethyloctanoic acid. A solid product was obtained with a yield of 36.0%.

[0171] 1 H NMR (600 MHz, chloroform-d) δ8.69 (s, 1H), 7.10 (d, J = 8.2 Hz, 1H), 5.79 (d, J = 8.3 Hz, 1H), 5.73 (d, J = 3.9 Hz, 1H), 4.35 (dd, J = 3.5, 2.0 Hz, 2H), 4.30–4.28 (m, 1H), 4.21–4.16 (m, 2 H),2.60–2.56(m,1H),2.46(dd,J=9.0,6.9Hz,2H),1.68–1.64(m,2H),1.32–1.25( m,9H),1.18(dd,J=6.9,1.6Hz,6H),0.91–0.86(m,6H).MS(ESI):m / z=484.28(M+H) + .

[0172] Example 16: Preparation of trans-4-butylcyclohexanecarboxylic acid prodrug (Compound 0910)

[0173]

[0174] Compound 0910 was prepared in the same manner as in Example 1 except that octanoic acid was replaced by trans-4-butylcyclohexanecarboxylic acid. Finally, a solid product was obtained with a yield of 30.6%.

[0175] 1 H NMR (600 MHz, chloroform-d) δ8.45 (s, 1H), 7.08 (d, J = 8.2 Hz, 1H), 5.81 (d, J = 8.3 Hz, 1H), 5.75 (d, J = 4.1 Hz, 1H), 4.34 (d, J = 3.6 Hz, 2H), 4.28 (q, J = 3.8 Hz, 1H), 4.19 (dt, J =

[0176] 16.1,5.3Hz,2H),2.61–2.56(m,1H),2.01–1.97(m,2H),1.85(dd,J=13.5,3.3Hz,2H),1.52(dddd,J=17.2,13.1 ,8.6,4.1Hz,2H),1.30–1.20(m,9H),1.18(dd,J=6.9,1.6Hz,6H),0.90–0.87(m,3H).MS(ESI):m / z=496.28(M+H) + .

[0177] Example 17: Preparation of cyclohexylbutyric acid prodrug (Compound 0912)

[0178]

[0179] Compound 0912 was prepared in the same manner as in Example 1 except that octanoic acid was replaced by trans-cyclohexylbutyric acid. Finally, a solid product was obtained with a yield of 17.8%.

[0180] 1H NMR (600MHz, chloroform-d) δ8.90 (s, 1H), 7.09 (d, J = 8.3Hz, 1H), 5.78 (dd, J = 12.2, 6.1Hz, 2H),4.34(d,J=3.5Hz,2H),4.26(t,J=4.1Hz,1H),4.18(dt,J=25.4,5.2Hz,2H),2 .58(p,J=7.0Hz,1H),2.45(t,J=7.6Hz,2H),1.73–1.67(m,7H),1.28–1.21(m,6H) ,1.18(d,J=7.1Hz,6H),0.88(dt,J=10.0,5.3Hz,2H).MS(ESI):m / z=482.28(M+H) + .

[0181] Example 18: Preparation of BOC-11-aminoundecanoic acid prodrug (Compound 0923)

[0182]

[0183] Compound 0923 was prepared in the same manner as in Example 1 except that octanoic acid was replaced by BOC-11-aminoundecanoic acid. Finally, a solid product was obtained with a yield of 12.6%.

[0184] 1 H NMR (400MHz, DMSO-d6) δ10.93(s,1H),6.76(s,1H),5.75(d,J=3.1Hz,1H),5.47(d,J =5.4Hz,1H),5.29(d,J=5.2Hz,1H),4.26–4.13(m,3H),4.03(t,J=5.1Hz,1H),3.98–3 .91(m,2H),2.88(d,J=6.5Hz,2H),2.60–2.56(m,1H),2.44(t,J=7.5Hz,2H),1.54(s ,2H),1.36(s,11H),1.23(s,12H),1.10(d,J=7.0Hz,6H).MS(ESI):m / z=613.36(M+H) + .

[0185] Example 19: Preparation of BOC-8-aminocaprylic acid prodrug (Compound 0924)

[0186]

[0187] Compound 0923 was prepared in the same manner as in Example 1 except that octanoic acid was replaced by BOC-8-aminooctanoic acid. Finally, a solid product was obtained with a yield of 15.4%.

[0188] 1 H NMR (400MHz, DMSO-d6) δ10.93(s,1H),7.18(d,J=8.2Hz,1H),6.78(s,1H),5.75(d,J=4.2Hz, 1H),5.47(d,J=5.5Hz,1H),5.29(d,J=5.4Hz,1H),4.26–4.14(m,3H),4.04(q,J=5.5Hz,1H),3 .98–3.91(m,2H),2.89(d,J=6.5Hz,2H),2.61–2.56(m,1H),2.44(t,J=7.5Hz,2H),1.55–1.5 1(m,2H),1.37(s,11H),1.29–1.24(m,6H),1.10(d,J=6.9Hz,6H).MS(ESI):m / z=571.31(M+H) + .

[0189] Example 20: Preparation of R-(+)-Lipoic Acid Prodrug (Compound 0926)

[0190]

[0191] Compound 0926 was prepared in the same manner as in Example 1 except that octanoic acid was replaced by R-(+)-lipoic acid. Finally, a solid product was obtained with a yield of 28.9%.

[0192] 1 H NMR (400MHz, DMSO-d6) δ10.94(s,1H),7.19(d,J=8.2Hz,1H),5.75–5.73(m,1H),5.47(d,J=5.6H z,1H),5.29(d,J=5.3Hz,1H),4.19(dtt,J=20.8,8.8,3.5Hz,3H),4.04(q,J=5.3Hz,1H),3.98–3 .91(m,2H),3.24–3.10(m,3H),2.46(q,J=7.1Hz,3H),1.88(dd,J=12.9,6.6Hz,1H),1.71–1.66( m,1H),1.62–1.54(m,4H),1.43–1.38(m,2H),1.10(d,J=7.0Hz,6H).MS(ESI):m / z=518.20(M+H) + .

[0193] Example 21: Preparation of Chenodeoxycholic Acid Prodrug (Compound 0939)

[0194]

[0195] 1. Dissolve chenodeoxycholic acid in dichloromethane, add 4-dimethylaminopyridine, add acetic anhydride under stirring, and stir at room temperature for 3 hours. The reaction solution is washed with 1mol / L HCl (3×30ml), saturated sodium bicarbonate solution, and saturated sodium chloride solution, respectively, and the organic layer is concentrated. The residue is purified by column chromatography (PE:EA (v / v) = 5:1) to obtain a white solid.

[0196] 2. Dissolve N4-hydroxycytidine isobutyrate in dichloromethane, add the above white solid (hydroxyl protective acid) and condensing agent 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 4-dimethylaminopyridine, stir at room temperature for 6 hours, extract with dichloromethane / water, spin dry, and column chromatography (DCM:MeOH (v / v) = 80:1, 50:1) to obtain a white solid product with a yield of 18.9%.

[0197] 1 H NMR (400MHz, chloroform-d) δ7.11(d,J=8.3Hz,1H),5.80(dd,J=8.3,2.0Hz,1H),5.74(d,J=3.6Hz,1H),4.58(tt,J=11.3,4. 5Hz,1H),4.35(d,J=3.5Hz,2H),4.28(dt,J=5.1,3.5Hz,1H),4.21–4.13(m,2H),3.86(q,J=3.0Hz,1H),2.58(dt,J= 14.0,7.0Hz,1H),2.51(dt,J=9.8,4.9Hz,1H),2.41(dd,J=9.4,6.4Hz,1H),2.30(d,J=12.4Hz,1H),2.01(s,3H),1. 88–1.43(m,20H),1.25–1.13(m,11H),0.96(d,J=6.3Hz,3H),0.92(s,3H),0.67(s,3H).MS(ESI):m / z=746.48(M+H) + ..

[0198] Example 22: Preparation of Ursodeoxycholic Acid Prodrug (Compound 0941)

[0199]

[0200] Compound 0941 was prepared in the same manner as in Example 21 except that chenodeoxycholic acid was replaced by ursodeoxycholic acid. Finally, a solid product was obtained with a yield of 11.3%.

[0201] 1H NMR (400 MHz, chloroform-d) δ 5.79 (dd, J = 8.2, 2.1 Hz, 1H), 5.73 (d, J = 3.6 Hz, 1H), 4.71 (dtd, J = 40.6, 10.7, 5.3 Hz, 3H), 4.36–4.34 (m, 2H), 4.29 (dd, J = 4.7, 3.3 Hz, 1H), 4.21–4.15 (m, 2H), 3.33 (s, 1H), 2.61–2.48(m,2H),2.03(s,3H),1.98(s,3H),1.77–1.35(m,20H),1.19(dd,J=7.0,1.0Hz,7H), 1.14–1.05(m,3H),0.97(s,3H),0.95(d,J=6.2Hz,3H),0.69(s,3H).MS(ESI):m / z=788.41(M+H) + .

[0202] Example 23: Lithocholic acid prodrug (Compound 0943)

[0203]

[0204] Except that chenodeoxycholic acid was replaced by lithocholic acid, compound 0941 was prepared in the same manner as in Example 21. Finally, a solid product was obtained with a yield of 12.2%.

[0205] 1 H NMR (400 MHz, chloroform-d) δ7.10 (d, J = 8.1 Hz, 1H), 5.79 (dd, J = 8.3, 2.0 Hz, 1H), 5.75 (d, J = 3.7 Hz, 1H), 4.72 (dt, J = 11.3, 6.3 Hz, 1H), 4.35 (d, J = 3.4 Hz, 2H), 4.27 (dd, J = 5.0, 3.3 Hz, 1H), 4.17 (dq, J = 10.5, 5.4Hz,3H),2.62–2.50(m,2H),2.39(td,J=9.7,4.8Hz,1H),2.03(s,3H),1.88–1.38(m,21H),1.1 9(d,J=6.9Hz,6H),1.10–1.03(m,5H),0.95–0.92(m,6H),0.65(s,3H).MS(ESI):m / z=730.43(M+H) + .

[0206] Example 24: Preparation of Deoxycholic Acid Prodrug (Compound 0945)

[0207]

[0208] Compound 0945 was prepared in the same manner as in Example 21 except that chenodeoxycholic acid was replaced by deoxycholic acid. A solid product was obtained with a yield of 13.9%.

[0209] 1 H NMR (400 MHz, CHLOROFORM-d) δ7.20 (d, J = 8.2 Hz, 1H), 5.79 (dd, J = 8.3, 1.7 Hz, 1H), 5.71 (d, J = 1.7 Hz, 1H), 4.73 (dq, J = 11.0, 5.5, 4.7 Hz, 1H), 4.43–4.36 (m, 2H), 4.25 (dt, J = 6.6, 3.2 Hz, 1H), 4.18 (d, J = 3.8 Hz, 2H), 4.00 (d, J =3.1Hz,1H),2.62–2.56(m,1H),2.51–2.31(m,3H),2.01(s,3H),1.85–1.47(m,19H),1.20(dd,J=7.0,1. 7Hz,6H),1.17–1.03(m,5H),1.01(d,J=6.0Hz,3H),0.93(s,3H),0.69(s,3H).MS(ESI):m / z=746.48(M+H) + .

[0210] Test Example 1: Screening of Anti-Vesicular Stomatitis Virus Cell Activity

[0211] Test principle: African green monkey kidney cells (Vero) are used as host cells to measure the green fluorescence signal of the recombinant vesicular stomatitis virus inserted with green fluorescent protein (GFP) to measure the inhibition efficiency of the sample.

[0212] Test materials and methods:

[0213] Vesicular stomatitis virus (VSV) was donated by Professor Zhang Weina's laboratory at the Academy of Military Medical Sciences.

[0214] Cell culture: African green monkey kidney cells (Vero) were used as host cells and cultured in MEM medium (CM50011, MACGENE) containing 10% fetal bovine serum (HQ30071-T500, Hongquan Biotechnology).

[0215] Virus strain: Vesicular stomatitis virus (containing green fluorescent protein reporter gene), cultured and passaged in African green monkey kidney cells (Vero), -80°C.

[0216] Sample processing: The sample was prepared into a stock solution with DMSO and then diluted with cell culture medium.

[0217] Positive control drug: Molnupiravir

[0218] Test method: Vero cells were inoculated into 96-well plates (2×104 cells / well) and cultured at 37°C and 5% CO2 for 12 hours. After the cells adhered to the wall, the old culture medium was replaced with the culture medium containing the drug, and 3 duplicate wells were set up, namely the solvent control group, the virus control group, and the drug administration group. In the virus control group and the drug administration group, MDCK cells were infected with vesicular stomatitis virus with a virus titer of 50 PFU and cultured at 37°C and 5% CO2. A fluorescence microplate reader was used for 24 hours to detect the GFP signal at an excitation wavelength of 488nm and an emission wavelength of 535nm. The inhibition rate calculation formula is: inhibition rate = 1-[(fluorescence intensity of the drug group-background fluorescence intensity) / (fluorescence intensity of the DMSO group-background fluorescence intensity)].

[0219] The results of the anti-VSV virus test are shown in Table 1 below.

[0220] Test Example 2: MDCK cytotoxicity test

[0221] Test method: MDCK cells (purchased from ATCC) were cultured in high-glucose DMEM medium (CM10017, MACGENE) containing 10% fetal bovine serum (10091148, Gibco). When the cell density reached about 90%, they were plated in 96-well plates and 2×10 4 cells / well, cultured at 37°C and 5% CO2 for 12h, discarded the old culture medium after the cells adhered, added 200μl of the compound of corresponding concentration to each well, set up 3 replicates, and set up a solvent control group. After 72h of culture at 37°C, discarded the supernatant, added 100μl of freshly prepared MTS (G3580, Promega) detection solution to each well, set up a background control group, placed the 96-well plate in an incubator for 1h, and measured the OD value at 490nm with an enzyme reader (Thermo Scientific). The value measured by the enzyme reader is the value obtained after automatically subtracting the background value. Cell viability (%) = OD value of drug experimental group / OD value of solvent control group × 100%.

[0222] The effects of drugs on cell viability are shown in Table 2.

[0223] Table 1: Anti-VSV efficacy test results of the example compounds

[0224]

[0225] Table 2. MDCK cell toxicity test results of the example compounds

[0226]

[0227]

[0228] Test Example 3: Anti-Staphylococcus aureus Experiment

[0229] Reagents:

[0230] MHB culture medium Beijing Aoboxing Biotechnology Co., Ltd. Batch number: 20200806

[0231] Brain Heart Infusion (BHI) Broth Beijing Aoboxing Biotechnology Co., Ltd. Batch No.: 20210813

[0232] Penicillin Potassium for Injection Sichuan Jishanzhijia Pharmaceutical Co., Ltd. Batch No.: 20190905

[0233] The reference compound ZXC-0323 is the lauroyl prodrug (Compound 19) in CN114805458B.

[0234]

[0235] Strains:

[0236] Gram-positive bacteria: Staphylococcus aureus ATCC 29213

[0237] Gram-negative bacteria: Escherichia coli ATCC 25922

[0238] Experimental methods:

[0239] The antibacterial activity of the compound was determined using the microbroth dilution method (CLSI standard, 2017). Single bacterial colonies were picked and placed in BHI broth, and the bacteria were cultured in a shaking incubator at 37°C until the logarithmic growth phase. The bacterial turbidity was adjusted to a McFarland turbidity of 0.5 using a McFarland turbidimeter, and the mixture was diluted 100 times (about 1.0×106 CFUs / mL) with MHB medium for later use. The drug to be tested was diluted with MHB medium, 100 μL was added to a 96-well U-shaped plate, and then 100 μL of the diluted test bacterial solution was added to each well. After the 96-well plate was placed at 37°C for 18 hours, the MIC result was read, and the lowest drug concentration that could inhibit bacterial growth that could be distinguished by the naked eye was the MIC value of the drug. The test results are shown in Table 3 below.

[0240] Table 3 Antimicrobial activity test results

[0241]

[0242]

[0243] Test Example 4 Beas-2B cytotoxicity experiment

[0244] Seeding cells: digest the cells, draw out the culture medium, discard it, add 3ml of normal saline to rinse, rinse the culture medium clean, add 2ml of trypsin, place in a 37℃ incubator for 3min, take it out and add 2ml of culture medium to neutralize the cells, use a 1ml pipette to blow down the attached cells, transfer them to a 15ml centrifuge tube, centrifuge (800r, 3min), discard the supernatant, add 2ml of culture medium to suspend and count: prepare a 1.5ml EP tube, add 10μl of trypan blue and cell suspension respectively, blow and suspend, draw 10μl and add it to the counting plate for counting. Assuming that each well of a 96-well plate requires 8*106cell viable cells, and 4 plates need to be seeded, 8 concentrations, and 6 rows per plate, the required cell suspension = 4*8*6*(8*106) / number of viable cells after counting. 100 μl needs to be added to each well of a 96-well plate. For 4 plates and 6 rows of 8 concentrations, the total liquid required is 4*6*8*100. Add cell suspension and culture medium in proportion. After suspension, use a dispenser to add to the 96-well plate.

[0245] Dosing: Weigh 5 mg of each drug, use a molar calculator to calculate the required DMSO volume (mass = concentration * volume * molecular weight), the concentration is 100mM (the required volume is too small, converted to 10mM), the 8 concentrations are - NT, 0μM, 5μM, 10μM, 20μM, 40μM, 80μM, 160μM. Add 500μl of culture medium to the EP tube, prepare drugs of different concentrations, add to 500μl of culture medium, mark and turn off the air, remove the culture medium from the 96-well plate, add the prepared culture medium of different concentrations in turn, 100μl to each well, and incubate in a 37℃ incubator for 24-48h.

[0246] Detection: Then extract the 96-well plate culture medium, add 100 μl of a 5:1 mixture of fresh culture medium and MTS (G3580, Promega) to each well as the detection solution, and set up a background control group in the upper right corner of each plate. Place the 96-well plate in a 37°C incubator for 1-4 hours, and measure the OD value at 490nm with an enzyme reader (Thermo Scientific). The value measured by the enzyme reader is the value obtained after automatically subtracting the background value. Cell viability (%) = (OD value of drug experimental group / OD value of solvent control group) × 100%. See the results below Figure 1 .

[0247] The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art can easily think of changes or substitutions within the technical scope disclosed by the present invention, which should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A compound derivative represented by formula I or an isotope-labeled compound thereof, or an optical isomer, geometric isomer, tautomer or isomer mixture thereof, or a pharmaceutically acceptable salt thereof: in, R1 is substituted or unsubstituted C2-C 10 Alkyl, substituted or unsubstituted C 12 Alkyl, substituted or unsubstituted C 14 Alkyl, substituted or unsubstituted C 16 -C 27 Alkyl, substituted or unsubstituted C2-C 27 Alkenyl, substituted or unsubstituted C2-C 27 Alkynyl, substituted or unsubstituted C3-C6 cycloalkyl, wherein the "substituted" means that the group contains 1 to 3 R a The substituent, R a Selected from deuterium, hydroxyl, amino, cyano, nitro, halogen, C1-C5 alkoxy, C1-C5 alkoxycarbonyl, C3-C6 cycloalkyl, -NHCOOR a1 、C6-C 10 Aryl, saturated or unsaturated three- to six-membered heterocyclic group containing 1-3 heteroatoms selected from N, O, and S, three- to six-membered heteroaryl group containing 1-3 heteroatoms selected from N, O, and S, unsubstituted or substituted with 1 to 2 R a2 Substituted steroid structure, the R a1 is selected from C1-C6 alkyl, said R a2 Selected from hydroxyl group, C2-C6 ester group; R2 is a substituted or unsubstituted C1-C6 alkyl group, wherein the "substituted" means that the group contains 1 to 3 R b replaced by the R b Selected from deuterium, hydroxyl, amino, cyano, nitro, halogen, C1-C5 alkoxy, C1-C5 alkyl substituted by halogen, and C3-C6 cycloalkyl.

2. The compound derivative represented by formula I according to claim 1 or its isotope-labeled compound, or its optical isomer, geometric isomer, tautomer or isomer mixture, or its pharmaceutically acceptable salt, characterized in that: R1 is substituted or unsubstituted C4-C 10 Alkyl, substituted or unsubstituted C 12 Alkyl, substituted or unsubstituted C 14 Alkyl, substituted or unsubstituted C 16 -C 20 Alkyl, substituted or unsubstituted C3-C 17 Alkenyl, substituted or unsubstituted C3-C 17 Alkynyl, substituted or unsubstituted C5-C6 cycloalkyl, wherein the "substituted" means that the group contains 1 or 2 R a The substituent, R a Selected from deuterium, hydroxyl, amino, cyano, nitro, halogen, C1-C3 alkoxy, C1-C3 alkoxycarbonyl, C5-C6 cycloalkyl, -NHCOOR a1 、C6-C 10 Aryl, saturated or unsaturated five- to six-membered heterocyclic group containing 1 or 2 heteroatoms selected from N, O, and S, five- to six-membered heteroaryl group containing 1 or 2 heteroatoms selected from N, O, and S, unsubstituted or with 1 to 2 R a2 Substituted steroid structure, the R a1 is selected from C1-C4 alkyl, said R a2 Selected from hydroxyl group, C2-C6 ester group.

3. The compound derivative represented by formula I according to claim 1 or its isotope-labeled compound, or its optical isomer, geometric isomer, tautomer or isomer mixture, or its pharmaceutically acceptable salt, characterized in that: More preferably, R1 is a substituted or unsubstituted C7-C 10 Alkyl, substituted or unsubstituted C 12 Alkyl, substituted or unsubstituted C 14 Alkyl, substituted or unsubstituted C 16 -C 17 Alkyl, substituted or unsubstituted C8-C 17 Alkenyl, substituted or unsubstituted C8-C 17 Alkynyl, substituted or unsubstituted C3-C6 cycloalkyl, wherein the "substituted" means that the group contains 1 or 2 R a The substituent, R a Selected from deuterium, hydroxyl, amino, cyano, nitro, halogen, methoxy, ethoxy, propoxy, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, cyclopentyl, cyclohexyl, -NHCOOR a1 , phenyl, naphthyl, a saturated or unsaturated five- to six-membered heterocyclic group containing 1 or 2 heteroatoms selected from N, O, and S, a five- to six-membered heteroaryl group containing 1 or 2 heteroatoms selected from N, O, and S, unsubstituted or replaced by 1 or 2 R a2 Substituted steroid structure, the R a1 is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, wherein R a2 Selected from hydroxyl, CH3COO-, CH3CH2COO-, CH3CH2CH2COO-; More preferably, the substituent R a wherein the residue is unsubstituted or is substituted with 1 or 2 R a2 The substituted steroid is selected from the following structures:

4. The compound derivative represented by formula I according to claim 1 or its isotope-labeled compound, or its optical isomer, geometric isomer, tautomer or isomer mixture, or its pharmaceutically acceptable salt, characterized in that: R2 is a substituted or unsubstituted C1-C4 alkyl group, wherein the "substituted" means that the group contains 1 or 2 R b replaced by the R b Selected from deuterium, hydroxyl, amino, cyano, nitro, halogen, C1-C3 alkoxy, C1-C3 alkyl substituted by halogen, C3-C6 cycloalkyl.

5. The compound derivative represented by formula I according to claim 1 or its isotope-labeled compound, or its optical isomer, geometric isomer, tautomer or isomer mixture, or its pharmaceutically acceptable salt, characterized in that: More preferably, R2 is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, n-hexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl; More preferably, R2 is isopropyl.

6. The compound derivative represented by formula I according to claim 1 or its isotope-labeled compound, or its optical isomer, geometric isomer, tautomer or isomer mixture, or its pharmaceutically acceptable salt is selected from the following structures:

7. Use of a compound derivative represented by formula I according to any one of claims 1 to 6 or an isotope-labeled compound thereof, or an optical isomer, geometric isomer, tautomer or isomer mixture thereof, or a pharmaceutically acceptable salt thereof as a prodrug of N4-hydroxycytidine.

8. A pharmaceutical composition comprising a therapeutically effective amount of a compound derivative represented by formula I according to any one of claims 1 to 6 or an isotope-labeled compound thereof, or an optical isomer, geometric isomer, tautomer or isomer mixture thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

9. Use of a compound derivative represented by formula I according to any one of claims 1 to 6 or an isotope-labeled compound thereof, or an optical isomer, geometric isomer, tautomer or isomer mixture thereof, or a pharmaceutically acceptable salt thereof in the preparation of a broad-spectrum antiviral or antibacterial drug; Preferably, the broad-spectrum antiviral or antibacterial drug is used to treat or prevent Gram-positive bacterial infection; Preferably, the Gram-positive bacteria include but are not limited to Staphylococcus, Streptococcus, Pneumococcus, Bacillus anthracis, Corynebacterium diphtheriae, and Corynebacterium tetani; Preferably, the broad-spectrum antiviral or antibacterial drug is prepared into an inhalant, a patch, a transdermal agent, an oral immediate-release preparation, a sustained-release or controlled-release preparation, a pill, a capsule, or a granule; Preferably, the inhalant includes a dry powder inhaler, a solution inhaler, or a vortex inhaler.

10. A method for treating or preventing a disease caused by a viral or bacterial infection, the method comprising administering to a subject a therapeutically effective amount of a compound derivative represented by formula I according to any one of claims 1 to 6 or an isotope-labeled compound thereof, or an optical isomer, geometric isomer, tautomer or isomer mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 8.

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