Medical use of cytosine derivatives

Abstract

The present application provides a compound (I) or a pharmaceutically acceptable salt, ester, optical isomer, tautomer, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, chelate, complex, clathrate or prodrug thereof, and a pharmaceutical composition containing the compound of the present application, and also provides the use of the compound of the present application as a double-stranded DNA virus inhibitor.

Description

Technical Field

[0001] This invention belongs to the field of antiviral drug technology based on cytosine derivatives, specifically relating to a cytosine derivative used as a double-stranded DNA virus, its preparation method, and its uses. Background Technology

[0002] Common double-stranded DNA (dsDNA) viruses that can cause human infection include smallpox virus, adenovirus, cytomegalovirus, human herpesvirus, Epstein-Barr virus (EBV), and BK virus. Most dsDNA viruses typically remain dormant in the human body, but can be reactivated when the body's immunity is weakened, leading to severe or even fatal illnesses in immunocompromised patients. For example, in hematopoietic stem cell transplantation (where multiple dsDNA viruses are often detected in the patient's plasma), the mortality rate from AdV-related infections alone can reach 50%–70%. Smallpox virus, belonging to the orthopoxvirus genus, is highly contagious and has a mortality rate as high as 30%, causing 3 to 5 million deaths in the 20th century alone, making it one of the most dangerous pathogens to humans. Monkeypox virus, also belonging to the orthopoxvirus genus, caused a monkeypox outbreak in 2022, which garnered widespread attention and was declared a Public Health Emergency of International Concern by the WHO. However, approved antiviral drugs against dsDNA viruses are very limited, and for some diseases, there are currently no marketed treatments available. Therefore, developing novel antiviral drugs for infectious diseases caused by dsDNA viruses is of great significance.

[0003] Cidofovir (CDV) Figure 1 CDV is a non-cyclic nucleoside viral DNA polymerase competitive inhibitor with broad-spectrum anti-dsDNA virus activity, exhibiting good antiviral effects against various dsDNA viruses, including cytomegalovirus, smallpox virus, monkeypox virus, adenovirus, Epstein-Barr virus, human herpesvirus, and BK virus. The U.S. Food and Drug Administration officially approved CDV for the treatment of cytomegalovirus (CMV) retinitis in AIDS patients in 1996. Due to the inherent negative charge of the phosphate group in the CDV structure under physiological pH conditions, its cell permeability is poor, resulting in low oral bioavailability and requiring injection for administration. Furthermore, as a substrate of organic anion transporter 1 (hOAT1), CDV exhibits significant nephrotoxicity, greatly limiting its therapeutic efficacy and scope of use. Structural modification of CDV using a prodrug strategy to prepare its derivatives is an effective means to address these issues. Brincidofovir (BCV) Figure 1Sildofovir (BCV) is a long fatty chain derivative of CDV. A hexadecyloxypropyl side chain is introduced onto the phosphate group of sildofovir, significantly enhancing its antiviral activity while retaining its broad spectrum of activity. It also greatly reduces nephrotoxicity while improving oral bioavailability, allowing for oral administration. The U.S. Food and Drug Administration approved BCV for marketing in 2021 for the treatment of smallpox, making it the first drug suitable for smallpox treatment in patients of all ages. The specific structures of sildofovir and brincedofovir are shown below.

[0004]

[0005] However, BCV has adverse side effects in clinical application, such as abdominal pain, diarrhea, and elevated transaminase and bilirubin levels. Therefore, developing more effective and safer anti-dsDNA virus drugs is of great significance. Summary of the Invention

[0006] To explore better-active cidofovir and improve its oral activity, the inventors conducted in-depth research. They discovered that by introducing a long hydrophobic side chain containing a disulfide bond (-SS-) onto the phosphate group in the cidofovir structure, a cidofovir derivative with a long hydrophobic side chain containing a disulfide bond was prepared. This derivative not only retained its activity but could also theoretically be used as a prodrug. Specifically, through testing its anti-dsDNA virus activity, a cidofovir derivative with stronger antiviral activity and a higher therapeutic index was discovered. Furthermore, the designed and synthesized derivative exhibited good stability in artificial gastric juice, artificial intestinal juice, and mixed human plasma, which meets the requirements for the development of oral formulations and can be used for the further development of broad-spectrum anti-dsDNA virus drugs.

[0007] More specifically, the inventors of this invention have discovered that compounds having the structure shown in formula (I), or their pharmaceutically acceptable salts, esters, optical isomers, tautomers, stereoisomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites, chelates, complexes, inclusion compounds, or prodrugs, possess good double-stranded DNA virus inhibitory activity.

[0008]

[0009] In formula (I), X is selected from C1-C20 alkylene, C1-C20 haloalkylene, C1-C20 ynylene, C1-C20 alkenylene, and C3-6 cyclic alkylene. The CH2 in X can be replaced by -O-.

[0010] Y is selected from -O-, -O-CH2-O-, -CO-, -C(=O)O-, -CONH-, -NHCO-, -NHCONH-, -NH-, -NR a -、-C(R a)2-、、-SS-、 Or any combination of them containing divalent groups, where R a Independently selected from H, substituted or unsubstituted C1-C10 alkyl, saturated or partially unsaturated C3-6 cyclic hydrocarbon, saturated or partially unsaturated 3-10 membered heterocyclic group, or C6-10 aryl group, R a CH2 in R can be replaced by -O- or -S-. a "Substitution" refers to substitution by hydroxyl, halogen, or C1-C3 alkoxy groups.

[0011] Z is selected from C1-C20 alkyl, C1-C20 haloalkyl, C1-C20 alkynyl, C1-C20 alkenyl, and C1-C20 cycloalkyl.

[0012] In a preferred embodiment of the present invention, the compound of (I) has the structure of the following formula (II).

[0013]

[0014] m is an integer from 1 to 9; n is an integer from 1 to 15.

[0015] Y 1 Divalent groups selected from -O-, -CO-, -NH-, -SS-, or any combination thereof

[0016] In a preferred embodiment of the present invention, Y 1 Preferred -SS-

[0017] In a preferred embodiment of the present invention, m is an integer from 1 to 5; n is an integer from 7 to 18.

[0018] Pharmacological experiments have shown that the compounds of the present invention exhibit inhibitory activity against vaccinia virus and adenovirus 5 in vitro under Vero and A549 cell conditions, and their therapeutic indices are close to or significantly superior to those of existing clinically used positive control compounds.

[0019] Therefore, in a preferred embodiment of the present invention, the double-stranded DNA virus is selected from any of the following viruses:

[0020] (1) Orthopoxviridae viruses: smallpox virus, monkeypox virus, cowpox virus, rabbitpox virus, mousepox virus, vaccinia virus;

[0021] (2) Adenoviridae viruses: Adenovirus 1, Adenovirus 2, Adenovirus 3, Adenovirus 4, Adenovirus 5, Adenovirus 7, Adenovirus 8, Adenovirus 14, Adenovirus 37, Adenovirus 40, Adenovirus 41;

[0022] (3) Herpesviridae viruses: Herpes simplex virus type 1, Herpes simplex virus type 2, Human cytomegalovirus, Epstein-Barr virus;

[0023] (4) Polyomaviridae virus: BK virus.

[0024] In a further preferred embodiment of the present invention, the double-stranded DNA virus is selected from any one of smallpox virus, monkeypox virus, cowpox virus, rabbitpox virus, mousepox virus, vaccinia virus, adenovirus 1, adenovirus 2, adenovirus 3, adenovirus 4, adenovirus 5, adenovirus 7, adenovirus 8, adenovirus 14, adenovirus 37, adenovirus 40, and adenovirus 41.

[0025] In a further preferred embodiment of the present invention, the double-stranded DNA virus is vaccinia virus or adenovirus 5.

[0026] The present invention also provides the use of cytosine compounds of formula (I) or their pharmaceutically acceptable salts, esters, optical isomers, tautomers, stereoisomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites, chelates, complexes, inclusion compounds or prodrugs in the preparation of medicaments for treating and alleviating diseases associated with double-stranded DNA viruses. These pharmaceutical uses are not limited to the mechanism speculations described in the present invention, and whether the mechanism speculations are correct does not limit the technical effects of the technical solutions of the present invention.

[0027] Diseases associated with double-stranded DNA viruses are selected from:

[0028] Smallpox, monkeypox, cowpox, and other infections caused by viruses of the Orthopoxviridae family, or vaccinia virus infection caused by smallpox vaccination and its complications;

[0029] Disseminated adenoviremia caused by adenoviral infections, adenovirus infection in organ transplant or hematopoietic stem cell transplant patients, adenovirus ocular infection and other diseases and their complications;

[0030] Herpesvirus infections, including oral herpes simplex, genital herpes simplex, human cytomegalovirus infection, Epstein-Barr virus infection, and their complications;

[0031] BK virus infection and its complications in the Polyomaviridae family.

[0032] In a preferred embodiment of the present invention, the drug for treating and alleviating diseases associated with double-stranded DNA viruses is an oral dosage form, which includes capsules, tablets, pills, powders, granules, emulsions, solutions, suspensions, syrups, or tinctures.

[0033] The following describes the other elements of the invention in more detail.

[0034] definition

[0035] Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as commonly understood by one of ordinary skill in the art. References to technical terms herein refer to techniques commonly understood in the art, including variations or equivalent substitutions of techniques that are obvious to one of ordinary skill in the art. While it is believed that the following terms will be well understood by one of ordinary skill in the art, the following definitions are set forth to better explain the invention.

[0036] In this invention, the terms “comprising,” “including,” “having,” “containing,” or “involving,” and their other variations herein, are inclusive or open-ended and do not exclude other elements or method steps not listed.

[0037] As used herein, the term "alkylene" means a saturated divalent hydrocarbon group, preferably a saturated divalent hydrocarbon group having 1, 2, 3, 4, 5 or 6 carbon atoms, such as methylene, ethylene, propylene or butylene.

[0038] As used herein, the term "alkyl" is defined as a linear or branched saturated aliphatic hydrocarbon. In some embodiments, the alkyl group has 1 to 12, for example, 1 to 6 carbon atoms. For example, as used herein, the term "C1-6 alkyl" refers to a linear or branched group (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, or n-hexyl) of 1 to 6 carbon atoms, optionally substituted with one or more (e.g., 1 to 3) suitable substituents such as halogens (in which case the group is referred to as "haloalkyl") (e.g., CH2F, CHF2, CF3, CCl3, C2F5, C2Cl5, CH2CF3, CH2Cl, or -CH2CH2CF3, etc.). The term “C1-4 alkyl” refers to a linear or branched aliphatic hydrocarbon chain with 1 to 4 carbon atoms (i.e., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl).

[0039] As used herein, the term "alkenyl" refers to a linear or branched monovalent hydrocarbon group containing a double bond and having 2–6 carbon atoms ("C"). 2-6 The alkenyl group is, for example, vinyl, 1-propenyl, 2-propenyl, 2-butenyl, 3-butenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 2-methyl-2-propenyl, and 4-methyl-3-pentenyl. When the compounds of the present invention contain an alkenyl group, the compounds may exist in pure E (iso-side) form, pure Z (iso-side) form, or any mixture thereof.

[0040] As used herein, the term "alkynyl" refers to a monovalent hydrocarbon group containing one or more triple bonds, preferably having 2, 3, 4, 5 or 6 carbon atoms, such as ethynyl or propynyl.

[0041] As used herein, the term "cycloalkyl" refers to a saturated monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (e.g., monocyclic, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or bicyclic, including spirocyclic, fused, or bridged systems (e.g., bicyclic [1.1.1]pentyl, bicyclic [2.2.1]heptyl, bicyclic [3.2.1]octyl, or bicyclic [5.2.0]nonyl, decahydronaphthyl, etc.)) which is optionally substituted with one or more (e.g., one to three) suitable substituents. The cycloalkyl group has 3 to 15 carbon atoms. For example, the term "C 3-6 "Cycloalkyl" refers to a saturated monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) with 3 to 6 cyclic carbon atoms, which is optionally substituted with one or more (such as 1 to 3) suitable substituents, such as methyl-substituted cyclopropyl.

[0042] As used herein, the terms “cycloalkylene group,” “cycloalkylene group,” and “hydrocarbon ring” refer to a saturated (i.e., “cycloalkylene group” and “cycloalkylene group”) or unsaturated (i.e., having one or more double and / or triple bonds within the ring) monocyclic or polycyclic hydrocarbon ring having, for example, 3 to 10 (suitably 3 to 8, more preferably 3 to 6) cyclic carbon atoms, including but not limited to (cycloalkylene group) propyl(ring), (cycloalkylene group) butyl(ring), (cycloalkylene group) pentyl(ring), (cycloalkylene group) hexyl(ring), (cycloalkylene group) heptyl(ring), (cycloalkylene group) octyl(ring), (cycloalkylene group) nonyl(ring), (cycloalkylene group) hexenyl(ring), etc.

[0043] As used herein, the terms “(aryl)aryl” and “aromatic ring” refer to all-carbon monocyclic or fused-ring polycyclic aromatic groups having a conjugated π-electron system. For example, as used herein, the term “C…” 6-10 (Asyl) aryl" and "C 6-10 "Aromatic ring" refers to an aromatic group containing 6 to 10 carbon atoms, such as ()phenylene (benzene ring) or ()naphthyl (naphthalene ring). The ()aryl and aromatic rings are optionally substituented with one or more (such as 1 to 3) suitable substituents (e.g., halogen, -OH, -CN, -NO2, C). 1-6 Alkyl groups, etc., are substituted.

[0044] As used herein, the term "aralkyl" preferably refers to an aryl or heteroaryl-substituted alkyl group, wherein the aryl, heteroaryl, and alkyl groups are as defined herein. Typically, the aryl group may have 6-14 carbon atoms, the heteroaryl group may have 5-14 ring atoms, and the alkyl group may have 1-6 carbon atoms. Exemplary aralkyl groups include, but are not limited to, benzyl, phenylethyl, phenylpropyl, and phenylbutyl.

[0045] A more specific explanation of the terminology is as follows:

[0046] "alkyl" refers to a saturated aliphatic hydrocarbon group comprising 1-20 carbon atoms, or 1-10 carbon atoms, or 1-6 carbon atoms, or 1-4 carbon atoms, or 1-3 carbon atoms, or 1-2 carbon atoms of a saturated straight-chain or branched monovalent hydrocarbon group, wherein the alkyl group may be independently and optionally substituted by one or more substituents described in this invention. Further examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, etc. The alkyl group may be optionally substituted or unsubstituted.

[0047] "Alkenyl" refers to a monovalent hydrocarbon group with 2-12 carbon atoms, or 2-8 carbon atoms, or 2-6 carbon atoms, or 2-4 carbon atoms, in which at least one C or C is sp. 2 The double bond, wherein the alkenyl group may be independently and optionally substituted by one or more substituents described in this invention, specific examples of which include, but are not limited to, vinyl, allyl, and olefinic groups. The alkenyl group may be optionally substituted or unsubstituted.

[0048] "Cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, wherein the cycloalkyl ring comprises 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, and more preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclohepttrienyl, cyclooctyl, etc.; polycyclic cycloalkyl groups include spirocyclic, fused-ring, and bridged-ring cycloalkyl groups. The cycloalkyl group may be optionally substituted or unsubstituted.

[0049] "Spirocycloalkyl" refers to a polycyclic aromatic system with 5 to 18 quintiles, two or more cyclic structures, where the monocyclic rings share a carbon atom (called a spiro atom) with each other, and containing one or more double bonds within the rings, but none of the rings has fully conjugated π electrons. Preferably, it is 6 to 14 quintiles, more preferably 7 to 10 quintiles. Based on the number of shared spiro atoms between the rings, spirocycloalkyl is classified into monospiro, bispiro, or polyspirocycloalkyl, preferably monospiro and bispirocycloalkyl, and preferably 4 / 5, 4 / 6, 5 / 5, or 5 / 6 quintiles. Non-limiting examples of "spirocycloalkyl" include, but are not limited to:

[0050]

[0051] "Fused cycloalkyl" refers to a 5- to 18-membered polycyclic aromatic group containing two or more ring structures sharing a pair of carbon atoms. One or more rings may contain one or more double bonds, but none of the rings has fully conjugated π electrons. It is preferably a 6- to 12-membered aromatic system, more preferably a 7- to 10-membered system. Depending on the number of rings, it can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic fused cycloalkyl, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic alkyl. Non-limiting examples of "fused cycloalkyl" include, but are not limited to:

[0052]

[0053] "Bridged cycloalkyl" refers to an aromatic system consisting of 5 to 18 quintiles, containing two or more cyclic structures sharing two non-directly bonded carbon atoms, and wherein one or more rings may contain one or more double bonds, but none of the rings possesses fully conjugated π electrons. Preferably, it is a 6 to 12 quintile, more preferably a 7 to 10 quintile. Depending on the number of constituent rings, it can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic bridged cycloalkyl, preferably bicyclic, tricyclic, or tetracyclic, and more preferably bicyclic or tricyclic. Non-limiting examples of "bridged cycloalkyl" include, but are not limited to:

[0054]

[0055] The cycloalkyl ring can be fused to an aryl, heteroaryl, or heterocyclic ring, wherein the ring connected to the parent structure is a cycloalkyl ring, and non-limiting examples include indanyl, tetrahydronaphthyl, benzocycloheptyl, etc.

[0056] "Aryl" refers to a carbocyclic aromatic system containing one or two rings, wherein the rings may be connected together in a fused manner. The term "aryl" includes aromatic groups such as phenyl, naphthyl, and tetrahydronaphthyl. Preferably, aryl groups are C6-C. 10Aryl, more preferably phenyl and naphthyl, most preferably phenyl. The aryl group may be substituted or unsubstituted. The "aryl" group may be fused with a heteroaryl, heterocyclic, or cycloalkyl group, wherein the aryl ring is attached to the parent structure. Non-limiting embodiments include, but are not limited to:

[0057]

[0058] "Heteroaryl" refers to an aromatic 5- to 6-membered monocyclic or 9- to 10-membered bicyclic ring, which may contain 1 to 4 atoms selected from nitrogen, oxygen, and / or sulfur. Examples of "heteroaryl" include, but are not limited to, furanyl, pyridinyl, 2-oxo-1,2-dihydropyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, thiopheneyl, isoxazolyl, oxazolyl, oxadiazolyl, imidazolyl, pyrroleyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, benzo[a]dioxacyclopentenyl, benzimidazolyl, indoleyl, isoyindolyl, 1,3-dioxo-isoindolyl, quinolinyl, indazoleyl, benzisothiazolyl, benzo[a]oxazolyl, and benzisothiazolyl. Heteroaryl groups may be optionally substituted or unsubstituted. The heteroaryl ring may be fused to an aryl, heterocyclic, or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring. Non-limiting embodiments include, but are not limited to:

[0059]

[0060] "Alkoxy" refers to an (alkyl-O-) group. Alkyl groups are defined in the relevant section of this document. C1-C6 alkoxy groups are preferred. Examples include, but are not limited to: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, etc.

[0061] "Halogenated alkyl" refers to an alkyl group having one or more halogen substituents, wherein the alkyl group has the meaning as described in this invention. Examples of halogenated alkyl groups include, but are not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, perfluoroethyl, 1,1-dichloroethyl, 1,2-dichloropropyl, etc.

[0062] "Hydroxy" refers to the -OH group.

[0063] "Halogen" refers to fluorine, chlorine, bromine and iodine, with fluorine, chlorine and bromine being preferred.

[0064] "Amino" refers to -NH2.

[0065] “Cyano” refers to -CN.

[0066] "Nitro" refers to -NO2.

[0067] "Benzyl" refers to -CH2-phenyl.

[0068] "Carboxyl group" refers to -C(O)OH.

[0069] "Acetyl" refers to -C(O)CH3 or Ac.

[0070] "Carboxylic acid ester group" refers to -C(O)O (alkyl) or (cycloalkyl), where alkyl and cycloalkyl are defined as described above.

[0071] As used herein, the term “halogenated” or “halogenated” is defined as including F, Cl, Br, or I.

[0072] The term "substitution" refers to the selective replacement of one or more (e.g., one, two, three, or four) hydrogen atoms on a specified atom by a designated group, provided that the substitution does not exceed the normal valence of the specified atom in the present case and that the substitution forms a stable compound. Combinations of substituents and / or variables are permitted only if such combinations form a stable compound.

[0073] If a substituent is described as “optionally substituted,” then the substituent may be (1) unsubstituted or (2) substituted. If the carbon of the substituent is described as being optionally substituted with one or more of the substituents in the list, then one or more hydrogens on the carbon (to the extent that any hydrogens are present) may be substituted individually and / or together with independently selected optional substituents. If the nitrogen of the substituent is described as being optionally substituted with one or more of the substituents in the list, then one or more hydrogens on the nitrogen (to the extent that any hydrogens are present) may each be substituted with independently selected optional substituents.

[0074] If a substituent is described as being “independently selected” from a group, then each substituent is selected independently of the others. Therefore, each substituent may be the same as or different from another (other) substituent. As used herein, the term “one or more” means one or more under reasonable conditions, such as two, three, four, five, or ten.

[0075] Unless otherwise specified, as used herein, the connection point of a substituent may be derived from any suitable location of the substituent.

[0076] When the bond of a substituent is such that it passes through the ring and connects two atoms, then such a substituent can be bonded to any cyclic atom in the substituted ring.

[0077] This invention also includes all pharmaceutically acceptable isotopically labeled compounds that are identical to the compounds of this invention, except that one or more atoms are replaced by atoms having the same atomic number but with an atomic mass or mass number different from the dominant atomic mass or mass number in nature. Examples of isotopes suitable for inclusion in the compounds of this invention include (but are not limited to) isotopes of hydrogen (e.g., deuterium). 2 H), tritium ( 3 H); carbon isotopes (e.g., H);11 C 13 C and 14 C); isotopes of chlorine (e.g.) 36 Cl); isotopes of fluorine (e.g., Cl); 18 F); isotopes of iodine (e.g., F); 123 I and 125 I); nitrogen isotopes (e.g.) 13 N and 15 N); isotopes of oxygen (e.g., N); 15 O、 17 O and 18 O); isotopes of phosphorus (e.g., O); phosphorus isotopes ... 32 P); and isotopes of sulfur (e.g. 35 S). Certain isotope-labeled compounds of the present invention (e.g., those doped with radioactive isotopes) can be used in drug and / or substrate tissue distribution studies (e.g., analysis). Radioactive isotope tritium (i.e. 3 H) and carbon-14 (i.e. 14 C) It is particularly suitable for this purpose due to its ease of incorporation and detection. Using positron-emitting isotopes (e.g.) 11 C 18 F, 15 O and 13 N) Substitution can be used in positron emission tomography (PET) studies to examine substrate acceptor occupancy. The isotopically labeled compounds of the present invention can be prepared by methods similar to those described in the accompanying routes and / or examples and preparations, by using a suitable isotopically labeled reagent instead of the previously used unlabeled reagent. Pharmaceutically acceptable solvates of the present invention include those in which the crystalline solvent can be isotopically substituted, for example, D2O, acetone-d6, or DMSO-d6. “Substituted” refers to one or more hydrogen atoms in a group, preferably up to five, more preferably one to three hydrogen atoms, independently substituted by the corresponding number of substituents. It goes without saying that substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without much effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom having an unsaturated (e.g., alkene) bond.

[0078] Unless otherwise specified, the terms "substitution" or "substituted" in this specification refer to the substitution of a group by one or more groups selected from the following: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic, aryl, heteroaryl, cycloalkoxy, heterocyclic alkoxy, cycloalkylthio, heterocyclic alkylthio, amino, haloalkyl, hydroxyalkyl, carboxyl, carboxylic acid ester, =O, -C(O)R b -OC(O)Rb -NR b R b -C(O)NR b R b -NR b C(O)R b -S(O)NR b R b or -S(O)2NR b R b , where R b The definition is as stated in general formula (I).

[0079] As used herein, the “effective amount” of a compound refers to an amount sufficient to inhibit viral survival. This dosage can be used as a single dose or taken according to a regimen to be effective.

[0080] As stated herein, “to improve the symptoms of a particular disease by using a particular compound or pharmaceutical composition” means any reduction, whether permanent or temporary, lasting or transient, that is attributable to or related to the use of the composition.

[0081] The definition and conventional use of stereochemistry in this invention are generally referenced in the following literature:

[0082] SP Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984), McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994. The compounds of this invention may contain asymmetric or chiral centers, and therefore exist as different stereoisomers. All stereoisomers of the compounds of this invention, including, but not limited to, diastereomers, enantiomers, transisomers, and mixtures thereof, such as racemic mixtures, constitute a part of this invention. Diastereomers can be separated into individual diastereomers based on their physicochemical differences by methods such as chromatography, crystallization, distillation, or sublimation. Enantiomers can be separated to convert a mixture of chiral isomers into a mixture of diastereomers by reacting with a suitable optically active compound (e.g., a chiral auxiliary agent, such as a chiral alcohol or Mosher's acyl chloride), separating the diastereomers and converting individual diastereomers into their corresponding pure enantiomers. The intermediates and compounds of this invention can also exist in different tautomeric forms, and all such forms are included within the scope of this invention. Many organic compounds exist in optically active forms, meaning they are capable of rotating the plane of polarized light. In describing optically active compounds, the prefixes D, L, or R, S are used to indicate the absolute configuration of the chiral center of the molecule. The prefixes d, l, or (+), (-) are used to name the symbol for the plane polarization rotation of the compound; (-) or l indicates that the compound is levorotatory, and the prefix (+) or d indicates that the compound is dextrorotatory. These stereoisomers have the same order of atomic or atomic groups connected to each other, but their stereostructures are different. Specific stereoisomers can be enantiomers, and mixtures of isomers are generally called enantiomer mixtures. A 50:50 enantiomer mixture is called a racemic mixture or racemate, which may result in a lack of stereoselectivity or stereodirection in chemical reactions. The terms "racemic mixture" and "racemate" refer to a mixture of two equimolar enantiomers that lack optical activity.

[0083] "Tautomer" or "tautomer form" refers to isomers of structures with different energies that can interconvert through a low energy barrier. For example, proton tautomers (i.e., proton-transfer tautomers) include interconversions via proton transfer, such as isomerizations of keto-enol and imine-enamine forms. Valence tautomers include interconversions involving the recombination of bonding electrons. Unless otherwise indicated, the structural formulas described in this invention include all isomer forms (e.g., enantiomers, diastereomers, and geometric isomers): for example, R, S configurations containing an asymmetric center, (Z), (E) isomers of double bonds, and (Z), (E) conformational isomers. Therefore, individual stereochemical isomers of the compounds of this invention, or mixtures of their enantiomers, diastereomers, or geometric isomers, are within the scope of this invention.

[0084] "Pharmaceutically acceptable salts" refers to salts of the compounds of this invention that are safe and effective when used in humans or animals. Salts of the compounds can be obtained by adding sufficient amounts of base or acid in a pure solution or suitable inert dissolution. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, etc., and pharmaceutically acceptable acid addition salts include inorganic acid salts and organic acid salts, including hydrochloric acid, hydrobromic acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, monohydrogen sulfate, acetic acid, maleic acid, malonic acid, succinic acid, benzoic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, and methanesulfonic acid, etc. (See Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science 66:1-19 (1977)).

[0085] Solid lines may be used in this article. solid wedge Or virtual wedge The chemical bonds of the compounds of the present invention are depicted. Solid lines are used to depict bonds to asymmetric carbon atoms to indicate all possible stereoisomers (e.g., specific enantiomers, racemic mixtures, etc.) at that carbon atom. Solid or dashed wedges are used to depict bonds to asymmetric carbon atoms to indicate the presence of the indicated stereoisomers. When present in racemic mixtures, solid and dashed wedges are used to define relative stereochemistry, not absolute stereochemistry. Unless otherwise specified, the compounds of the present invention are intended to exist as stereoisomers (including cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, geometric isomers, rotational isomers, conformational isomers, trans-blocking isomers, and mixtures thereof). The compounds of the present invention may exhibit more than one type of isomerism and may consist of mixtures thereof (e.g., racemic mixtures and diastereomer pairs).

[0086] This invention covers all possible crystalline forms or polymorphs of the compounds of this invention, which may be a single polymorph or a mixture of more than one polymorph in any proportion.

[0087] It should also be understood that certain compounds of the present invention may exist in their free form for therapeutic purposes, or, where appropriate, in their pharmaceutically acceptable derivative forms. In the present invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, solvates, N-oxides, metabolites, chelates, complexes, inclusion compounds, or prodrugs, which, upon administration to a patient in need, can directly or indirectly provide the compounds of the present invention or their metabolites or residues. Therefore, when referring to "compounds of the present invention" herein, it is also intended to cover the various derivative forms of the compounds described above.

[0088] Pharmaceutically acceptable salts of the compounds of the present invention include their acid addition salts and base addition salts, including but not limited to salts containing hydrogen bonds or coordination bonds.

[0089] Suitable acid addition salts are formed from acids that form pharmaceutically acceptable salts. Examples include acetates, adipates, aspartates, benzoates, benzenesulfonates, bicarbonates / carbonates, bisulfates / sulfates, borates, camphor sulfonates, citrates, cyclohexanesulfonates, ethanedisulfonates, ethanesulfonates, formates, fumarates, glucohepanoates, glucuronates, hexafluorophosphates, hymenates, hydrochlorides / chlorides, hydrobromates / bromines, hydroiodates / iodides, hydroxyethyl sulfonates, lactates, malates, maleates, malonates, methanesulfonates, methyl sulfates, naphthylcarbamates, 2-naphthalenesulfonates, nicotinates, nitrates, orotates, oxalates, palmitates, dihydroxynaphthyl salts, phosphates / hydrogen phosphates / dihydrogen phosphates, pyroglutamates, glycosides, stearates, succinates, tannins, tartrates, toluenesulfonates, trifluoroacetates, and xinofoate.

[0090] Suitable base addition salts are formed from bases that form pharmaceutically acceptable salts. Examples include aluminum salts, arginine salts, benzathine penicillin salts, calcium salts, choline salts, diethylamine salts, diethanolamine salts, glycine salts, lysine salts, magnesium salts, meglumine salts, ethanolamine salts, potassium salts, sodium salts, tromethamine salts, and zinc salts.

[0091] For a review of suitable salts, see Stahl and Wermuth's "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" (Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptable salts for the compounds of the present invention are known to those skilled in the art.

[0092] The compounds of the present invention can exist as solvates (preferably hydrates), wherein the compounds of the present invention contain a polar solvent, particularly, for example, water, methanol, or ethanol, as a structural element of the lattice of the compound. The amount of the polar solvent, particularly water, can be stoichiometric or non-stoichiometric.

[0093] Those skilled in the art will understand that not all nitrogen-containing heterocycles can form N-oxides because nitrogen requires available lone pairs of electrons to be oxidized into oxides; those skilled in the art will identify nitrogen-containing heterocycles that can form N-oxides. Those skilled in the art will also recognize that tertiary amines can form N-oxides.

[0094] The scope of this invention also includes metabolites of the compounds of this invention, i.e., substances formed in the body when the compounds of this invention are administered. Such products can be generated, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, etc., of the administered compound. Therefore, this invention includes metabolites of the compounds of this invention, including compounds obtained by methods that expose the compounds of this invention to mammals for a time sufficient to produce their metabolites.

[0095] This invention also covers compounds of the invention containing protecting groups. In any process of preparing the compounds of the invention, protection of sensitive or reactive groups on any relevant molecule may be necessary and / or desired, thereby forming a form of chemical protection for the compounds of the invention. This can be achieved by conventional protecting groups, for example, those described in TW Greene & P. ​​GMWuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991, which are incorporated herein by reference. Protecting groups can be removed at appropriate subsequent stages using methods known in the art.

[0096] The term “about” means within ±10% of the stated value, preferably within ±5%, and more preferably within ±2%.

[0097] Preferred compounds of the present invention

[0098] The general formula and preferred scope of the compounds of the present invention have been described. More preferably, specific examples of the compounds of the present invention may be selected from any of the following structures, but are not limited to the following compounds:

[0099]

[0100]

[0101]

[0102] Typical compounds of the present invention include, but are not limited to, the compounds mentioned above. The compounds in the present invention are named according to systematic nomenclature, or, using ChemDraw software.

[0103] General method for obtaining the compounds of the present invention

[0104] General formula synthesis method

[0105] The cytosine derivative compounds represented by general formula (I) of this invention can be obtained by known methods, such as synthesis by known organic synthesis methods. The examples described below provide exemplary synthetic routes, but those skilled in the art can also obtain them by other known methods.

[0106] Pharmaceutical compositions and treatment methods

[0107] The present invention provides a pharmaceutical composition comprising an effective amount of the compound of the present invention or a pharmaceutically acceptable salt, ester, optical isomer, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, chelate, complex, inclusion compound or prodrug, and a pharmaceutically acceptable carrier, wherein the pharmaceutical composition is preferably a solid dosage form, a semi-solid dosage form, a liquid dosage form or a gaseous dosage form.

[0108] In this invention, "pharmaceutically acceptable carrier" refers to a diluent, excipient, vehicle, or medium that is administered co-administered with a therapeutic agent and is suitable, to the extent of reasonable medical judgment, for contact with human and / or other animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications commensurate with a reasonable benefit / risk ratio.

[0109] Pharmaceutically acceptable carriers that can be used in the pharmaceutical compositions of the present invention include, but are not limited to, sterile liquids such as water and oils, including those of petroleum, animal, plant, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. Water is an exemplary carrier when the pharmaceutical composition is administered intravenously. Physiological saline and aqueous solutions of glucose and glycerol can also be used as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, maltose, chalk, silica gel, sodium stearate, glyceryl monostearate, talc, sodium chloride, skim milk powder, glycerol, propylene glycol, water, ethanol, etc. The compositions may also contain small amounts of wetting agents, emulsifiers, or pH buffers as needed. Oral formulations may contain standard carriers such as pharmaceutical-grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, etc. Examples of suitable pharmaceutically acceptable carriers are described in Remington's Pharmaceutical Sciences (1990).

[0110] The pharmaceutical compositions of the present invention can act systemically and / or locally. For this purpose, they can be administered via suitable routes, such as by injection (e.g., intravenous, intra-arterial, subcutaneous, intraperitoneal, intramuscular injection, including infusion) or transdermal administration; or by oral, sublingual, nasal, transmucosal, topical, ophthalmic formulations or by inhalation.

[0111] For these routes of administration, the pharmaceutical compositions of the present invention can be administered in suitable dosage forms.

[0112] The dosage forms include, but are not limited to, tablets, capsules, lozenges, hard candies, powders, sprays, creams, ointments, suppositories, gels, pastes, lotions, ointments, aqueous suspensions, injectable solutions, elixirs, and syrups.

[0113] The pharmaceutical compositions of this invention contain a safe and effective amount of the compound of this invention and a pharmaceutically acceptable carrier or excipient. Such carriers include (but are not limited to): one or more of saline, buffer solutions, glucose, water, glycerol, ethanol, powders, etc. The pharmaceutical formulation should be matched to the route of administration.

[0114] The pharmaceutical compositions of the present invention can be formulated into injectable forms, for example, by conventional methods using physiological saline or an aqueous solution containing glucose and other excipients. Pharmaceutical compositions such as tablets and capsules can be prepared by conventional methods. Pharmaceutical compositions such as injections, solutions, tablets, and capsules are preferably manufactured under sterile conditions. The pharmaceutical compositions of the present invention can also be formulated into powders for nebulized inhalation.

[0115] The dosage of the active ingredient is a therapeutically effective amount, for example, about 1 microgram / kg body weight to about 50 milligrams / kg body weight per day; preferably, about 5 micrograms / kg body weight to about 10 milligrams / kg body weight; more preferably, about 10 micrograms / kg body weight to about 5 milligrams / kg body weight. Furthermore, the compounds of the present invention can also be used with other therapeutic agents.

[0116] The pharmaceutical compositions of the present invention can be administered to the desired subjects (such as humans and non-human mammals) in a conventional manner. Representative administration methods include (but are not limited to): oral administration, injection, nebulized inhalation, etc.

[0117] When using the pharmaceutical composition, a safe and effective amount of the drug is administered to mammals, wherein this safe and effective amount is generally at least about 10 micrograms per kilogram of body weight, and in most cases does not exceed about 50 milligrams per kilogram of body weight. Preferably, this dose is about 10 micrograms per kilogram of body weight to about 20 milligrams per kilogram of body weight. Of course, the specific dosage should also take into account factors such as the route of administration and the patient's health condition, which are all within the scope of the skill of a skilled physician. This invention is particularly applicable to oral formulations. In artificial gastric juice, artificial intestinal juice, and plasma, the compounds of this invention can achieve the stability required for formulation. The compounds of this invention themselves, as well as their hydrolysates, enzymatic hydrolysis products in vivo, and hydrolysates all possess antiviral activity, making them ideal candidate compounds for oral formulations.

[0118] The present invention will be further illustrated below with reference to specific embodiments. The processes, conditions, reagents, experimental methods, etc., used to implement the present invention, except as specifically mentioned below, are all common knowledge and general knowledge in the art, and the present invention does not have any particular limitations. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments that do not specify specific conditions are generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.

[0119] As used herein, the term "effective amount" refers to an amount of compound that, when administered, provides some relief for one or more symptoms of the treated condition. Specifically, as used herein, an "effective amount" of a compound is an amount sufficient to inhibit double-stranded DNA viruses. As used herein, a "therapeutic effective dose" of a compound is an amount sufficient to improve or reduce symptoms in some way, stop or reverse disease progression, or inhibit double-stranded DNA viruses. This dose can be used as a single dose or as part of a regimen to be effective.

[0120] The dosing regimen can be adjusted to provide the optimal required response. For example, a single bolus injection can be administered, several fractions can be administered over time, or the dose can be proportionally reduced or increased as indicated by the urgency of the treatment situation. It should be noted that dosage values ​​can vary depending on the type and severity of the condition to be alleviated, and may include single or multiple doses. To further understand, for any given individual, the specific dosing regimen should be adjusted over time based on individual needs and the professional judgment of the person administering the composition or supervising its administration.

[0121] As used herein, “treatment” means any improvement or alteration of a patient’s condition, disorder, or disease symptoms or pathology in any way. As described herein, “improvement of symptoms of a particular disease by using a particular compound or pharmaceutical composition” means any reduction, whether permanent or temporary, lasting or transient, attributable to or related to the use of the composition.

[0122] As used herein, “individual” includes humans or non-human animals. Exemplary human individuals include individuals suffering from a disease (such as the disease described herein) (referred to as patients) or normal individuals. In this invention, “non-human animals” includes all vertebrates, such as non-mammals (e.g., birds, amphibians, reptiles) and mammals, such as non-human primates, livestock, and / or domesticated animals (e.g., sheep, dogs, cats, cows, pigs, etc.). In some embodiments, the pharmaceutical compositions of this invention may also contain one or more additional therapeutic or preventative agents. Attached Figure Description

[0123] Figure 1 For compound V-3 1 H NMR spectrum.

[0124] Figure 2 For compound V-3 13 C NMR spectrum.

[0125] Figure 3 For compound V-3 31 P NMR spectrum.

[0126] Figure 4 Here is the HRMS plot of compound V-3.

[0127] Figure 5 For compound V-4 1 H NMR spectrum.

[0128] Figure 6 For compound V-4 13 C NMR spectrum.

[0129] Figure 7 For compound V-4 31 P NMR spectrum.

[0130] Figure 8 Here is the HRMS plot of compound V-4.

[0131] Figure 9 For compound V-14 1 H NMR spectrum.

[0132] Figure 10 For compound V-14 13 C NMR spectrum.

[0133] Figure 11 For compound V-14 31 PNMR plot.

[0134] Figure 12 Here is the HRMS plot of compound V-14.

[0135] Figure 13 For compound V-17 1 H NMR spectrum.

[0136] Figure 14 For compound V-17 13 C NMR spectrum.

[0137] Figure 15 For compound V-17 31 PNMR plot.

[0138] Figure 16 This is the HRMS plot of compound V-17.

[0139] Figure 17 A diagram illustrating the antiviral activity of compounds V-3, V-4, and BCV in Vero cells.

[0140] Figure 18 A diagram illustrating the antiviral activity of compounds V-3, V-4, and BCV in A549 cells.

[0141] Figure 19 A diagram illustrating the cytotoxicity of compounds V-3, V-4, and BCV in Vero cells.

[0142] Figure 20 A diagram illustrating the cytotoxicity of compounds V-3 and V-4 in A549 cells.

[0143] Figure 21 A graph showing the stability of compounds V-3, V-4, V-14, V-17 and BCV in artificial gastric juice.

[0144] Figure 22A graph showing the stability of compounds V-3, V-4, V-14, V-17 and BCV in artificial intestinal fluid. Detailed Implementation

[0145] The method of the present invention will be described below through specific embodiments to make the technical solution of the present invention easier to understand and master, but the present invention is not limited thereto. In the following embodiments... 1 The 1H NMR spectra were obtained using a Bruker instrument (400 MHz), and chemical shifts are expressed in ppm. Tetramethylsilane internal standard (0.00 ppm) was used. 1 ¹H NMR representation: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, br = broadened, dd = doublet of doublet, dt = doublet of triplet. If the coupling constant is provided, the unit is Hz. CD₃OD: deuterated methanol. CDCl₃: deuterated chloroform. DMSO-d₆: deuterated dimethyl sulfoxide.

[0146] High-resolution mass spectrometry (HRMS) was performed using a Thermo Scientific Q Exactive high-resolution mass spectrometer with an electron spray ionization (ESI) source.

[0147] Solvent removal under reduced pressure was performed using an IKA RV-8 rotary evaporator; thin-layer chromatography was performed using Huanghai GF254 (5×20cm) silica gel plates; rapid column chromatography purification was performed using a Biotage Selekt automated rapid chromatography system, employing Biotage-Rening columns or Biotage- Silica gel column; Sample purity analysis was performed on an Agilent 1100 HPLC system equipped with a photodiode array (PDA) detector and a SunFire C18 column (4.6 mm × 150 mm × 5 μm, Waters). Thin-layer chromatography (TLC) used Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates. The silica gel plates used for TLC were 0.2 mm–0.3 mm, and the plates used for TLC separation and purification were 0.4 mm–0.5 mm. Column chromatography generally used Yantai Huanghai 200–300 mesh silica gel as the carrier.

[0148] In the following examples, unless otherwise specified, all temperatures are in Celsius; unless otherwise specified, all starting materials and reagents are commercially available or synthesized according to known methods; commercially available materials and reagents are used directly without further purification; unless otherwise specified, commercially available manufacturers include, but are not limited to, Sinopharm Group, Bailingwei Technology Co., Ltd., TCI (Shanghai) Chemical Industry Development Co., Ltd., Shanghai Bid Pharmaceutical Technology Co., Ltd., Shanghai Mairui Chemical Technology Co., Ltd., SigmaAldrich Reagents Co., Ltd., Shaoyuan Technology (Shanghai) Co., Ltd., Anaiji Reagents Co., Ltd., Jiuding Chemical Reagents Co., Ltd., etc.

[0149] Unless otherwise specified in the examples, the solution in the reaction refers to an aqueous solution. Unless otherwise specified in the examples, the reaction temperature is room temperature, between 20°C and 30°C.

[0150] The reaction process in the examples was monitored using thin-layer chromatography (TLC). The developing solvent used in the reaction, the eluent system used for column chromatography to purify the compounds, or the developing solvent system for TLC included: A: petroleum ether and ethyl acetate system; B: dichloromethane and methanol system; C: n-hexane: ethyl acetate. The volume ratio of the solvent varied depending on the polarity of the compound and could also be adjusted by adding a small amount of acidic or basic reagents, such as acetic acid or triethylamine.

[0151] Specific compound synthesis methods:

[0152] Example 1. Synthesis of target compound V-1

[0153]

[0154] 0.585 g (1.2 mmol) of starting material 1 was added to the reaction flask and dissolved in 8 mL of dichloromethane. The mixture was then magnetically stirred at room temperature. 2.5 mL of oxalyl chloride (2.0 M dichloromethane solution, 4.80 mmol) was added to the reaction flask, along with 20 μL (0.250 mmol) of DMF. After reacting at room temperature for 2 h, the solvent and excess oxalyl chloride were evaporated under reduced pressure. The residue was dissolved in 8 mL of dichloromethane under N2 protection and magnetically stirred at 0 °C. 0.250 g (1.00 mmol) of 2-(decyldithio)ethane-1-ol and 0.316 g (4.00 mmol) of pyridine were dissolved in dichloromethane and slowly added dropwise to the reaction solution. After the addition was complete, the temperature was slowly raised to room temperature, and the reaction was continued at room temperature for 3 h. Then, 100 μL of H2O was added, and the reaction was continued for 30 min before the reaction was stopped. The mixture was washed twice with 20 mL of 0.2 M dilute hydrochloric acid and once with 20 mL of 10% NaCl solution. The organic layer was collected, dried over anhydrous Na2SO4, and the solvent was evaporated under reduced pressure. The mixture was separated by silica gel column chromatography under the following elution conditions: dichloromethane: dichloromethane / dichloromethane (80 / 20; V / V) 0–60% gradient elution to give intermediate IV-1, a white solid of 0.384 g, yield 53%.

[0155] Its nuclear magnetic resonance 1 H NMR (600MHz, CD3OD / CDCl3) δ8.20(d,J=13.8Hz,1H),8.09-7.99(m,4H),7.65-7.48(m,6H),4.62(dd,J=9.6,45.6Hz,1H),4.44-4.29(m,3H),4 .15-3.98(m,4H),3.84-3.72(m,1H),2.88-2.80(m,2H),2.65-2.58(m, 2H),1.62-1.56(m,2H),1.38-1.27(m,14H),0.90(t,J=15.6Hz,3H); NMR 13 C NMR (150MHz, CD3OD / CDCl3) δ 134.07, 130.34, 129.41, 129.22, 128.90, 128.83, 39.33, 39.31, 32.60, 30.20 (d, J = 3.6Hz), 29.99, 29.87, 29.69, 29.04, 23.28, 14.01; NMR 31 PNMR (240MHz, CD3OD / CDCl3) δ 16.42; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 34 H 47 N3O8PS2[M+H] +720.2537,found720.2529,C 34 H 46 N3O8PS2Na[M+Na] +

[0156] 742.2356, found 742.2302.

[0157] 0.324 g (0.45 mmol) of intermediate b1 and 20 mL of ammonia-methanol solution (7.0 M) were added to the reaction flask. The reaction was carried out at room temperature with magnetic stirring for 48 h. The solvent was removed by vacuum distillation, and the solution was purified by silica gel column chromatography. The elution conditions were dichloromethane: dichloromethane / methanol / ammonia (4 g / 4 g / 2; V / V / V).

[0158] Elution was performed using a gradient of 20% to 80% to obtain the target product a1, a white solid weighing 0.113 g, with a yield of 48%.

[0159] Its nuclear magnetic resonance 1 H NMR(600MHz,CD3OD)δ7.81(d,J=7.8Hz,1H),5.96(d,J=7.2Hz,1H),4.16-4.10(m,3H) ,3.84(dd,J=7.8,14.4Hz,1H),3.79-3.75(m,2H),3.72-3.68(m,1H),3.64(dd,J=9.0, 12.6Hz, 1H), 3.56(dd, J = 4.2, 12.6Hz, 1H), 2.92(t, J = 6.6Hz, 2H), 2.74(t, J = 7.2Hz, 2H), 1.72-1.67(m, 2H), 1.42-1.39(m, 2H), 1.36-1.29(m, 12H), 0.92(t, J = 7.2Hz, 3H); NMR 13 C NMR (150MHz, CD3OD) δ 164.02, 154.30, 149.09, 93.73, 80.08 (d, J = 11.6Hz), 65.34 (d, J = 158.7Hz), 63.00 (d, J = 5.55Hz) 60.17, 50.03, 39.01 (d, J = 6.0Hz), 38.55, 31.67, 29.30, 29.28, 29.06, 28.98, 28.79, 28.12, 22.34, 13.05; NMR 31 PNMR (240MHz, CD3OD) δ 16.12; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 20 H 39 N3O6PS2[M+H] +512.2012,found 512.2009,C 20 H 38 N3O6PS2Na[M+Na] + 534.1832, found 534.1837.

[0160] Example 2. Synthesis of target compound V-2

[0161]

[0162] The target compound V-2 was synthesized using the same method as compound V-1, except that the starting material 2-(decyldithio)ethane-1-ol was replaced with 0.279 g (1.00 mmol) of 2-(dodecyldithio)ethane-1-ol, yielding intermediate IV-2, a white solid of 0.363 g, with a yield of 49%.

[0163] Its nuclear magnetic resonance 1 ¹H NMR (600MHz, CDCl₃) δ 8.00–7.97 (m, 4H), 7.56–7.36 (m, 6H), 4.62–3.85 (m, 9H), 2.82–2.72 (m, 2H), 2.54–2.51 (m, 2H), 1.53–1.51 (m, 2H), 1.33–1.13 (m, 18H), 0.87 (t, J = 6.6 Hz, 3H); NMR 13 C NMR (150MHz, CDCl3) δ 166.11, 133.26, 129.72, 129.57, 128.71, 128.51, 38.83, 31.93, 29.69, 29.66, 29.65, 29.58, 29.37, 29.28, 29.10, 28.55, 22.70, 14.14; 31 P NMR (240MHz, CDCl3) δ 15.62; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 36 H 51 N3O8PS2[M+H] + 748.2850, found 748.2834,

[0164] C 36 H 50 N3O8PS2Na[M+Na] + 770.2669, found 770.2652.

[0165] The intermediate IV-2 was fed in an amount of 0.337 g (0.45 mmol), yielding the target product V-2, a white solid of 0.132 g, with a yield of 53%.

[0166] Its nuclear magnetic resonance 1 H NMR (600MHz, CD3OD) δ7.83(d,J=7.2Hz,1H),5.96(d,J=7.2Hz,1H),4.17-4.10(m,3H),3.84(dd,J=7.2,13.8Hz,1H),3.79-3.75(m,2H),3.7 2-3.69(m,1H),3.64(dd,J=9.0,12.6Hz,1H),3.56(dd,J=4.2,12.0Hz,1H),2.92(t,J=7.2Hz,2H),2.74(t,J=7.2Hz,2H),1.72-1.67(m,2H),

[0167] 1.43–1.29 (m, 18H) 0.92 (t, J = 6.6 Hz, 3H); NMR 13 C NMR (150MHz, CD3OD) δ 163.75, 153.86, 149.26, 93.64, 80.04 (d, J = 11.7Hz), 65.32 (d, J = 159.0Hz), 62.99 (d, J = 5.7Hz), 60.16, 50.04, 39.01 (d, J = 6.0Hz), 38.55, 31.68, 29.39, 29.36, 29.33, 29.27, 29.08, 28.98, 28.79, 28.12, 22.34, 13.04; NMR 31 P NMR (240MHz, CD3OD) δ 16.11; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 22 H 43 N3O6PS2[M+H] + 540.2325, found 540.2323, C 22 H 42 N3O6PS2Na[M+Na] + 562.2145, found 562.2142.

[0168] Example 3. Synthesis of target compound V-3

[0169]

[0170] The target compound V-3 was synthesized using the same method as compound V-1, except that the starting material 2-(decyldithio)ethane-1-ol was replaced with 0.307 g (1.00 mmol) of 2-(tetradecyldithio)ethane-1-ol, yielding intermediate IV-3, a white solid of 0.440 g, with a yield of 57%.

[0171] Its nuclear magnetic resonance 1 ¹H NMR (600MHz, CDCl₃) δ 8.06–7.94 (m, 4H), 7.55–7.36 (m, 6H), 4.59–3.86 (m, 9H), 2.83–2.71 (m, 2H), 2.54–2.52 (m, 2H), 1.53–1.51 (m, 2H), 1.41–1.21 (m, 22H), 0.88 (t, J = 6.6 Hz, 3H); NMR 13 C NMR (150MHz, CDCl3) δ 166.11, 133.25, 129.72, 129.57, 128.72, 128.51, 63.41, 63.08, 38.84, 38.74, 31.94, 29.72, 29.70, 29.68, 29.66, 29.59, 29.40, 29.38, 29.29, 29.10, 28.56, 22.70, 14.14; 31 P NMR (240MHz, CDCl3) δ 15.98; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 38 H 55 N3O8PS2[M+H] + 776.3163,found 776.3146,C 38 H 54 N3O8PS2Na[M+Na] + 798.2982, found 798.2965.

[0172] The intermediate IV-3 was fed in an amount of 0.349 g, yielding the target product V-3, a white solid of 0.128 g, with a yield of 49%.

[0173] Its nuclear magnetic resonance 1H NMR(600MHz,CD3OD)δ7.74(d,J=7.2Hz,1H),5.90(d,J=7.2Hz,1H),4.14-4.10(m,3H) ,3.83(dd,J=7.2,13.8Hz,1H),3.76-3.73(m,2H),3.70-3.66(m,1H),3.64(dd,J=9.0, 12.6Hz, 1H), 3.54(dd, J = 4.2, 12.0Hz, 1H), 2.92(t, J = 6.6Hz, 2H), 2.74(t, J = 7.2Hz, 2H), 1.72-1.67(m, 2H), 1.43-1.40(m, 2H), 1.36-1.29(m, 20H), 0.92(t, J = 6.6Hz, 3H); NMR 13 C NMR (150MHz, CD3OD) δ 165.56, 156.46, 148.26, 93.94, 80.39 (d, J = 11.7 Hz), 65.55 (d, J = 159.0 Hz), 63.04 (d, J = 5.4 Hz), 60.25, 50.00, 39.00 (d, J = 6.2 Hz), 38.55, 31.67, 29.39, 29.38, 29.36, 29.31, 29.26, 29.07, 28.97, 28.78, 28.11, 22.33, 13.03; NMR 31 PNMR (240MHz, CD3OD) δ 16.10; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 24 H 47 N3O6PS2[M+H] + 568.2638,found 568.2636,C 24 H 46 N3O6PS2Na[M+Na] + 590.2458, found 590.2456.

[0174] Example 4. Synthesis of target compound V-4

[0175]

[0176] The target compound V-4 was synthesized using the same method as compound V-1, except that the starting material 2-(decyldithio)ethane-1-ol was replaced with 0.335 g (1.00 mmol) of 2-(hexadecyldithio)ethane-1-ol, yielding intermediate IV-4, a white solid of 0.413 g, with a yield of 51%.

[0177] Its nuclear magnetic resonance 1¹H NMR (600MHz, CDCl₃) δ 7.94–7.91 (m, 4H), 7.47–7.33 (m, 6H), 4.52–3.78 (m, 9H), 2.75–2.71 (m, 2H), 2.50–2.47 (m, 2H), 1.50–1.45 (m, 2H), 1.26–1.14 (m, 26H), 0.81 (t, J = 6.6 Hz, 3H); NMR 13 C NMR (150MHz, CDCl3) δ 166.09, 133.29, 129.73, 129.55, 128.74, 128.61, 128.53, 128.46, 127.44, 63.46, 63.06, 38.87, 31.94, 30.95, 29.73, 29.71, 29.68, 29.66, 29.59, 29.38, 29.30, 29.11, 28.56, 22.70, 14.14; 31 P NMR (240MHz, CDCl3) δ 16.17; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 40 H 59 N3O8PS2[M+H] + 804.3476, found 804.3470, C 40 H 58 N3O8PS2Na[M+Na] + 826.3295, found 826.3289.

[0178] The intermediate IV-4 was fed in an amount of 0.361 g, yielding the target product V-4, a white solid of 0.110 g, with a yield of 40%.

[0179] Its nuclear magnetic resonance 1 H NMR (600MHz, CD3OD) δ7.71(d,J=7.2Hz,1H),5.88(d,J=7.2Hz,1H),4.14-4.09(m,3H),3.83(dd,J=7.2,13.8Hz,1H),3.76-3.72(m,2H),3.6 9-3.66(m,1H),3.64(dd,J=9.0,12.6Hz,1H),3.53(dd,J=4.2,12.6Hz,1H),2.92(t,J=7.2Hz,2H),2.74(t,J=7.8Hz,2H),1.72-1.67(m,2H),

[0180] 1.43–1.40 (m, 2H), 1.36–1.29 (m, 24H), 0.92 (t, J = 6.6 Hz, 3H); NMR 13 C NMR (150MHz, CD3OD) δ 166.08, 157.19, 147.98, 94.01, 80.49 (d, J = 11.9Hz), 65.61 (d, J = 159.6Hz), 63.05 (d, J = 5.5Hz), 60.27, 50.00, 38.97 (d, J = 5.5Hz), 38.55, 31.67, 29.37, 29.35, 29.31, 29.26, 29.07, 28.97, 28.78, 28.11, 22.33, 13.03; NMR 31 P NMR (240MHz, CD3OD) δ 16.11; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 26 H 51 N3O6PS2[M+H] + 596.2951,found 596.2946,C 26 H 50 N3O6PS2Na[M+Na] + 618.2771, found 618.2767.

[0181] Example 5. Synthesis of target compound V-5

[0182]

[0183] The target compound V-5 was synthesized using the same method as compound V-1, except that the starting material 2-(decyldithio)ethane-1-ol was replaced with 0.363 g (1.00 mmol) of 2-(octadecyldithio)ethane-1-ol, yielding intermediate IV-5, a white solid of 0.458 g, with a yield of 55%.

[0184] Its nuclear magnetic resonance 1 ¹H NMR (600MHz, CDCl₃) δ 8.00–7.97 (m, 4H), 7.55–7.39 (m, 6H), 4.59–3.85 (m, 9H), 2.80–2.73 (m, 2H), 2.54–2.51 (m, 2H), 1.55–1.50 (m, 2H), 1.31–1.20 (m, 30H), 0.88 (t, J = 7.2 Hz, 3H); NMR 13C NMR (150MHz, CDCl3) δ 166.11, 133.25, 129.72, 129.58, 128.70, 128.51, 63.41, 63.03, 38.82, 31.94, 29.74, 29.73, 29.68, 29.60, 29.38, 29.31, 29.10, 28.57, 22.70, 14.14; 31 P NMR (240MHz, CDCl3)δ

[0185] 15.58; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 42 H 63 N3O8PS2[M+H] + 832.3789, found 832.3787, C 42 H 62 N3O8PS2Na[M+Na] + 854.3608, found 854.3613.

[0186] The intermediate IV-5 was fed in an amount of 0.374 g, yielding the target product V-5, a white solid of 0.135 g, with a yield of 47%.

[0187] Its nuclear magnetic resonance 1 H NMR (600MHz, CD3OD / CDCl3) δ7.69(d,J=7.2Hz,1H),5.87(d,J=7.2Hz,1H),4.13-4.08(m,3H),3.83(dd,J=7.2,13.8Hz,1H),3.76-3.71(m, 2H),3.68-3.61(m,2H),3.52(dd,J=4.2,12.6Hz,1H),2.90(t,J=6.6Hz,2H),2.72(t,J=7.2Hz,2H),1.71-1.66(m,2H),1.42-1.38(m,2H),

[0188] 1.35–1.29 (m, 28H), 0.90 (t, J = 6.6 Hz, 3H); NMR 13C NMR (150MHz, CD3OD / CDCl3) δ 169.28, 148.76, 95.32, 80.67 (d, J = 11.8Hz), 65.43 (d, J = 158.6Hz), 63.09 (d, J = 5.5Hz), 60.37, 50.00, 39.21 (d, J = 6.0Hz), 38.55, 31.67, 29.45, 29.43, 29.42, 29.40, 29.31, 29.12, 29.04, 28.86, 28.21, 22.39, 13.26; NMR 31 P NMR (240MHz, CD3OD / CDCl3) δ 16.07; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 28 H 55 N3O6PS2[M+H] + 624.3264,found 624.3262,C 28 H 54 N3O6PS2Na[M+Na] + 646.3084, found 646.3084.

[0189] Example 6. Synthesis of target compound V-6

[0190]

[0191] The target compound V-6 was synthesized using the same method as compound V-1, except that the starting material 2-(decyldithio)ethane-1-ol was replaced with 0.264 g (1.00 mmol) of 3-(decyldithio)prop-1-ol, yielding intermediate IV-6, a white solid of 0.368 g, with a yield of 50%.

[0192] Its nuclear magnetic resonance 1 H NMR (600MHz, CD3OD / CDCl3) δ8.18(d,J=10.8Hz,1H),8.04-7.87(m,4H),7.64-7.47(m,6H),4.62(dd,J=9.6,45.6Hz,1H),4.43-4.09(m,4H), 4.02-3.94(m,3H),3.78-3.64(m,1H),2.71-2.56(m,4H),1.94-1.92(m ,2H),1.66-1.58(m,2H),1.38-1.27(m,14H),0.89(t,J=6.6Hz,3H); NMR 13C NMR (150MHz, CD3OD / CDCl3) δ 134.04, 133.78, 130.55, 129.42, 129.25, 129.21, 129.12, 98.00, 64.06, 64.00, 39.17, 39.15, 35.02, 32.62, 30.93, 30.22 (d, J = 2.9Hz), 30.01, 29.90, 29.88, 29.86, 29.77, 29.74, 29.07, 29.05, 23.29, 14.06; NMR 31 P NMR (240MHz, CD3OD / CDCl3) δ 20.69, 20.08; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 35 H 49 N3O8PS2[M+H] + 734.2693, found 734.2697, C 35 H 48 N3O8PS2Na[M+Na] +

[0193] 756.2513, found 756.2516.

[0194] The intermediate IV-6 was fed in an amount of 0.330 g, yielding the target product V-6, a white solid of 0.123 g, with a yield of 50%.

[0195] Its nuclear magnetic resonance 1 H NMR (600MHz, CD3OD) δ7.77(d,J=7.2Hz,1H),5.93(d,J=7.8Hz,1H),4.12(dd,J=3.6,13.8Hz,1 H),3.99-3.96(m,2H),3.84(dd,J=7.2,13.8Hz,1H),3.77-3.73(m,2H),3.71-3.67(m,1H),3. 63 (dd, J = 9.6, 13.2 Hz, 1H), 3.55 (dd, J = 3.6, 12.0 Hz, 1H), 2.80 (t, J = 7.2 Hz, 2H), 2.71 (t, J = 7.2 Hz, 2H), 2.01–1.97 (m, 2H), 1.71–1.66 (m, 2H), 1.43–1.29 (m, 14H), 0.92 (t, J = 6.6 Hz, 3H); NMR 13C NMR (150MHz, CD3OD) δ 164.79, 155.41, 148.65, 93.82, 80.18 (d, J = 11.6 Hz), 65.30 (d, J = 158.6 Hz), 62.81 (d, J = 5.9 Hz), 60.18, 50.04, 38.22, 34.42, 31.66, 30.35 (d, J = 6.2 Hz), 29.28, 29.27, 29.05, 28.95, 28.83, 28.12, 22.33, 13.04; NMR 31 P NMR (240MHz, CD3OD) δ 16.09; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 21 H 41 N3O6PS2[M+H] + 526.2169,found 526.2165,C 21 H 40 N3O6PS2Na[M+Na] + 548.1988, found 548.1984.

[0196] Example 7. Synthesis of target compound V-7

[0197]

[0198] The target compound V-7 was synthesized using the same method as compound V-1, except that the starting material 2-(decyldithio)ethane-1-ol was replaced with 0.293 g (1.00 mmol) of 3-(dodecyldithio)prop-1-ol, yielding intermediate IV-7, a white solid of 0.383 g, with a yield of 50%.

[0199] Its nuclear magnetic resonance 1 ¹H NMR (600MHz, CDCl₃) δ 7.99–7.87 (m, 4H), 7.54–7.41 (m, 6H), 4.60–3.86 (m, 9H), 2.60–2.53 (m, 4H), 1.90–1.82 (m, 2H), 1.56–1.53 (m, 2H), 1.34–1.23 (m, 18H), 0.87 (t, J = 7.2 Hz, 3H); NMR 13 C NMR (150MHz, CDCl3) δ 166.11, 133.31, 129.72, 129.54, 128.74, 128.54, 38.81, 34.43, 31.93, 29.69, 29.66, 29.64, 29.58, 29.37, 29.29, 29.18, 28.57, 22.70, 14.14;1 P NMR (240MHz, CDCl3) δ 15.97; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 37 H 53 N3O8PS2[M+H] + 762.3006, found 762.3000,

[0200] C 37 H 52 N3O8PS2Na[M+Na] + 784.2826, found 784.2820.

[0201] The intermediate IV-7 was fed in an amount of 0.342 g, yielding the target product V-7, a white solid of 0.122 g, with a yield of 48%.

[0202] Its nuclear magnetic resonance 1 H NMR (600MHz, CD3OD) δ7.80(d,J=7.2Hz,1H),5.95(d,J=7.2Hz,1H),4.14(dd,J=3.0,13.8Hz,1 H),3.99-3.96(m,2H),3.84(dd,J=7.8,13.8Hz,1H),3.78-3.74(m,2H),3.71-3.68(m,1H),3. 62 (dd, J = 9.0, 13.8 Hz, 1H), 3.55 (dd, J = 4.2, 12.0 Hz, 1H), 2.80 (t, J = 7.2 Hz, 2H), 2.71 (t, J = 7.2 Hz, 2H), 2.01–1.97 (m, 2H), 1.71–1.66 (m, 2H), 1.42–1.29 (m, 18H), 0.92 (t, J = 6.6 Hz, 3H); NMR 13 C NMR (150MHz, CD3OD) δ 164.11, 154.39, 149.05, 93.67, 80.06 (d, J = 11.7Hz), 65.23 (d, J = 158.6Hz), 62.80 (d, J = 5.6Hz), 60.15, 50.05, 38.23, 34.42, 31.68, 30.34 (d, J = 6.2Hz), 29.38, 29.36, 29.31, 29.26, 29.07, 28.96, 28.84, 28.12, 22.34, 13.04; NMR 31 P NMR (240MHz, CD3OD) δ 16.11; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C23 H 45 N3O6PS2[M+H] + 554.2482,found 554.2475,C 23 H 44 N3O6PS2Na[M+Na] + 576.2301 found 576.2296.

[0203] Example 8. Synthesis of target compound V-8

[0204]

[0205] The target compound V-8 was synthesized using the same method as compound V-1, except that the starting material 2-(decyldithio)ethane-1-ol was replaced with 0.321 g (1.00 mmol) of 3-(tetradecyldithio)prop-1-ol, yielding intermediate IV-8, a white solid of 0.450 g, with a yield of 57%.

[0206] Its nuclear magnetic resonance 1 H NMR (600MHz, CDCl3) δ8.26-7.96(m,4H),7.59-7.39(m,6H),4.65-3.69(m,9H),2.69-2.48(m,4H),2.05-1.20(m,26H),0.88(t,J=6.6Hz,3H); Nuclear Magnetic 13 C NMR (150MHz, CDCl3) δ 166.01, 133.46, 129.74, 129.71, 128.62, 128.51, 38.89, 34.12, 31.94, 29.74, 29.72, 29.69, 29.68, 29.65, 29.62, 29.58, 29.38, 29.33, 29.30, 29.23, 29.17, 28.59, 22.71, 14.15; NMR 31 P NMR (240MHz, CDCl3) δ 20.73; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 39 H 57 N3O8PS2[M+H] + 790.3319, found 790.3305,

[0207] C 39 H 56 N3O8PS2Na[M+Na] + 812.3139, found 812.3127.

[0208] The intermediate IV-8 was fed in an amount of 0.355 g, yielding the target product V-8, a white solid of 0.149 g, with a yield of 55%.

[0209] Its nuclear magnetic resonance 1 H NMR(600MHz,CD3OD)δ7.80(d,J=7.2Hz,1H),5.95(d,J=7.2Hz,1H),4.14(dd,J=3.6,14.4Hz,1H), 3.99-3.96(m,2H),3.84(dd,J=7.8,14.4Hz,1H),3.78-3.74(m,2H),3.71-3.68(m,1H),3.62(dd,J =9.0, 12.6 Hz, 1H), 3.55 (dd, J = 4.2, 12.6 Hz, 1H), 2.80 (t, J = 7.2 Hz, 2H), 2.71 (t, J = 7.2, 2H), 2.01-1.97 (m, 2H), 1.71-1.66 (m, 2H), 1.43-1.39 (m, 2H), 1.36-1.29 (m, 20H), 0.92 (t, J = 6.6 Hz, 3H); NMR 13 C NMR (150MHz, CD3OD) δ 164.15, 154.47, 149.02, 93.69, 80.06 (d, J = 11.7 Hz), 65.20 (d, J = 158.1 Hz), 62.80 (d, J = 5.6 Hz), 60.13, 50.05, 38.21, 34.40, 31.69, 30.33 (d, J = 6.2 Hz), 29.41, 29.40, 29.38, 29.33, 29.28, 29.09, 28.97, 28.84, 28.13, 22.35, 13.06; NMR 31 P NMR (240MHz, CD3OD) δ 16.13; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 25 H 49 N3O6PS2[M+H] + 582.2795,found582.2792,C 25 H 48 N3O6PS2Na[M+Na] + 604.2614, found 604.2611.

[0210] Example 9. Synthesis of target compound V-9

[0211]

[0212] The target compound V-9 was synthesized using the same method as compound V-1, except that the starting material 2-(decyldithio)ethane-1-ol was replaced with 0.349 g (1.00 mmol) of 3-(hexadecyldithio)prop-1-ol, yielding intermediate IV-9, a white solid of 0.460 g, with a yield of 56%.

[0213] Its nuclear magnetic resonance 1 H NMR (600MHz, CDCl3) δ8.00-7.86(m,4H),7.55-7.38(m,6H),4.83-3.85(m,9H),2.64-2.51(m,4H),1.85-1.22(m,30H),0.87(t,J=7.2Hz,3H); Nuclear Magnetic 13 C NMR (150MHz, CDCl3) δ 166.12, 133.28, 129.71, 129.55, 128.72, 128.52, 63.48, 62.99, 38.78, 34.39, 31.94, 30.00, 29.73, 29.71, 29.68, 29.60, 29.38, 29.31, 29.18, 28.58, 22.71, 14.14; NMR 31 P NMR (240MHz, CDCl3) δ 15.76; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 41 H 61 N3O8PS2[M+H] + 818.3632, found 818.3605, C 41 H 60 N3O8PS2Na[M+Na] + 840.3452, found 840.3420.

[0214] The intermediate IV-9 was fed in an amount of 0.368 g, yielding the target product V-9, a white solid of 0.145 g, with a yield of 51%.

[0215] Its nuclear magnetic resonance 1H NMR(600MHz,CD3OD)δ7.72(d,J=7.2Hz,1H),5.89(d,J=7.2Hz,1H),4.10(dd,J=3.6,14.4Hz,1H), 3.99-3.94(m,2H),3.83(dd,J=7.2,13.8Hz,1H),3.76-3.71(m,2H),3.69-3.66(m,1H),3.62(dd,J =9.0, 12.6 Hz, 1H), 3.53 (dd, J = 4.2, 12.6 Hz, 1H), 2.80 (t, J = 7.2 Hz, 2H), 2.71 (t, J = 7.2, 2H), 2.01-1.97 (m, 2H), 1.72-1.67 (m, 2H), 1.43-1.40 (m, 2H), 1.36-1.29 (m, 24H), 0.92 (t, J = 6.6 Hz, 3H); NMR 13 C NMR (150MHz, CD3OD) δ 165.75, 156.72, 148.17, 93.91, 80.36 (d, J = 11.9Hz), 65.39 (d, J = 158.7Hz), 62.83 (d, J = 5.6Hz), 60.21, 50.04, 38.20, 34.40, 31.68, 30.36 (d, J = 6.3Hz), 29.39, 29.37, 29.36, 29.30, 29.26, 29.08, 28.95, 28.83, 28.12, 22.34, 13.04; NMR 31 P NMR (240MHz, CD3OD) δ 16.12; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 27 H 53 N3O6PS2[M+H] + 610.3108,found610.3099,C 27 H 52 N3O6PS2Na[M+Na] + 632.2927, found 632.2921.

[0216] Example 10. Synthesis of target compound V-10

[0217]

[0218] The target compound V-10 was synthesized using the same method as compound V-1, except that the starting material 2-(decyldithio)ethane-1-ol was replaced with 0.377 g (1.00 mmol) of 3-(octadecyldithio)prop-1-ol, yielding intermediate IV-10, a white solid of 0.433 g, with a yield of 51%.

[0219] Its nuclear magnetic resonance 1 H NMR (600MHz, CDCl3) δ7.99-7.87(m,4H),7.56-7.35(m,6H),4.60-3.72(m,9H),2.61-2.53( m,4H),1.90-1.83(m,2H),1.55-1.53(m,2H),1.41-1.23(m,30H),0.88(t,J=7.2Hz,3H); NMR 3 C NMR (150MHz, CDCl3) δ 166.11, 133.30, 129.72, 129.55, 128.71, 128.53, 38.80, 34.42, 31.94, 29.73, 29.68, 29.60, 29.38, 29.31, 29.19, 28.59, 22.70, 14.14; 31 P NMR (240MHz, CDCl3) δ 15.90; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 43 H 65 N3O8PS2[M+H] + 846.3945, found 846.3935,

[0220] C 43 H 64 N3O8PS2Na[M+Na] + 868.3765, found 868.3755.

[0221] The intermediate IV-10 was fed in an amount of 0.380 g, yielding the target product V-10, a white solid weighing 0.142 g, with a yield of 48%.

[0222] Its nuclear magnetic resonance 1H NMR(600MHz,CD3OD / CDCl3)δ7.74(d,J=7.2Hz,1H),5.91(d,J=7.2Hz,1H),4.11(dd,J=3.6,14.4Hz, 1H),3.99-3.95(m,2H),3.83(dd,J=7.8,13.8Hz,1H),3.77-3.71(m,2H),3.69-3.66(m,1H),3.62(d d, J = 9.6, 13.2 Hz, 1H), 3.53 (dd, J = 4.2, 12.6 Hz, 1H), 2.79 (t, J = 7.2 Hz, 2H), 2.70 (t, J = 7.2, 2H), 2.01-1.97 (m, 2H), 1.71-1.66 (m, 2H), 1.42-1.39 (m, 2H), 1.35-1.30 (m, 28H), 0.91 (t, J = 6.6 Hz, 3H); NMR 13 C NMR (150MHz, CD3OD / CDCl3) δ 164.90, 148.56, 93.91, 80.20 (d, J = 11.6Hz), 65.33 (d, J = 158.3Hz), 62.83 (d, J = 5.6Hz), 60.14, 50.05, 38.32, 34.45, 31.71, 30.35 (d, J = 6.0Hz), 29.43, 29.41, 29.40, 29.35, 29.31, 29.11, 29.01, 28.89, 28.20, 22.38, 13.20; NMR 31 P NMR (240MHz, CD3OD / CDCl3) δ 16.08; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 29 H 57 N3O6PS2[M+H] + 638.3421, found 638.3419,

[0223] C 29 H 56 N3O6PS2Na[M+Na] + 660.3240, found 660.3230.

[0224] Example 11. Synthesis of target compound V-11

[0225]

[0226] The target compound V-11 was synthesized using the same method as compound V-1, except that the starting material 2-(decyldithio)ethane-1-ol was replaced with 0.279 g (1.00 mmol) of 4-(decyldithio)but-1-ol, yielding intermediate IV-11, a white solid of 0.400 g, with a yield of 53%.

[0227] Its nuclear magnetic resonance 1 H NMR (600MHz, CD3OD / CDCl3) δ8.17 (d, J = 7.2Hz, 1H), 8.04-7.98 (m, 4H), 7.65-7. 46(m,6H),4.62(dd,J=12.0,45.0Hz,1H),4.39(q,J=15.6Hz,2H),4.25(dd,J=1 2.6,33.6Hz,1H),4.14-3.96(m,2H),3.91-3.89(m,2H),3.80-3.69(m,1H),2.6 5-2.59(m,4H),1.72-1.57(m,6H),1.38-1.27(m,14H),0.89(t,J=7.2Hz,3H); NMR 13 C NMR (150MHz, CD3OD / CDCl3) δ 164.28, 134.13, 134.02, 133.78, 130.57, 130.37, 129.41, 129.26, 129.21, 129.12, 128.88, 64.05, 39.22, 39.20, 38.69, 38.65, 32.61, 30.21 (d, J = 3.8Hz), 30.00, 29.89, 29.89, 29.76, 29.74, 29.03, 29.02, 26.07, 25.98, 23.29, 14.03; NMR 31 P NMR (240MHz, CD3OD / CDCl3) δ 20.89, 20.24; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 36 H 51 N3O8PS2[M+H] +

[0228] 748.2850, found 748.2835, C 36 H 50 N3O8PS2Na[M+Na] + 770.2669, found 770.2661.

[0229] The intermediate IV-11 was fed in an amount of 0.336 g, and the target product V-11 was obtained, which was a white solid of 0.141 g, with a yield of 56%.

[0230] Its nuclear magnetic resonance 1 H NMR(600MHz,CD3OD)δ7.82(d,J=7.8Hz,1H),5.96(d,J=7.2Hz,1H),4.15(dd,J=3.6,14.4Hz,1H), 3.92-3.88(m,2H),3.84(dd,J=7.8,14.4Hz,1H),3.78-3.75(m,2H),3.71-3.68(m,1H),3.62(dd,J =9.6,13.2Hz,1H), 3.56(dd,J=4.2,12.6Hz,1H), 2.74(t,J=6.6Hz,2H), 2.70(t,J=7.2,2H), 1.82-1.77(m,2H), 1.74-1.66(m,4H), 1.44-1.39(m,2H), 1.36-1.29(m,12H), 0.92(t,J=6.6Hz,3H); NMR 13 C NMR (150MHz, CD3OD) δ 163.71, 153.81, 149.31, 93.57, 79.97 (d, J = 11.6 Hz), 65.20 (d, J = 158.3 Hz), 63.96 (d, J = 5.7 Hz), 60.12, 50.05, 38.30, 37.92, 31.67, 29.50 (d, J = 6.2 Hz), 29.29, 29.27, 29.06, 28.97, 28.83, 28.10, 25.27, 22.34, 13.06; NMR 31 P NMR (240MHz, CD3OD) δ 16.06; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 22 H 43 N3O6PS2[M+H] + 540.2325, found 540.2323, C 22 H 42 N3O6PS2Na[M+Na] + 562.2145, found 562.2145.

[0231] Example 12. Synthesis of target compound V-12

[0232]

[0233] The target compound V-12 was synthesized using the same method as compound V-1, except that the starting material 2-(decyldithio)ethane-1-ol was replaced with 0.307 g (1.00 mmol) of 4-(dodecyldithio)but-1-ol, yielding intermediate IV-2, a white solid of 0.357 g, with a yield of 46%.

[0234] Its nuclear magnetic resonance 1 H NMR (600MHz, CDCl3) δ7.99-7.90(m,4H),7.53-7.40(m,6H),4.61-4.13(m,5H),3.91-3.83( m,4H),2.61-2.54(m,4H),1.60-1.57(m,4H),1.43-1.24(m,20H),0.87(t,J=7.2Hz,3H); NMR 13 C NMR (150MHz, CDCl3) δ 166.10, 133.31, 129.71, 129.54, 128.75, 128.53, 64.81, 38.86, 38.20, 31.92, 29.68, 29.65, 29.63, 29.56, 29.36, 29.29, 29.20, 28.58, 25.25, 22.70, 14.13; NMR 31 P NMR (240MHz, CDCl3) δ 16.16; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 38 H 55 N3O8PS2[M+H] + 776.3163,found 776.3152,C 38 H 54 N3O8PS2Na[M+Na] + 798.2982, found 798.2978.

[0235] The intermediate IV-12 was fed in an amount of 0.348 g, yielding the target product V-12, a white solid weighing 0.133 g, with a yield of 51%.

[0236] Its nuclear magnetic resonance 1H NMR(600MHz,CD3OD)δ7.82(d,J=7.8Hz,1H),5.95(d,J=7.2Hz,1H),4.15(dd,J=3.0,13.8Hz,1H), 3.92-3.88(m,2H),3.84(dd,J=7.8,14.4Hz,1H),3.78-3.74(m,2H),3.71-3.68(m,1H),3.62(dd,J =9.6,13.2Hz,1H), 3.56(dd,J=4.2,12.6Hz,1H), 2.74(t,J=7.2Hz,2H), 2.70(t,J=7.2,2H), 1.82-1.77(m,2H), 1.74-1.66(m,4H), 1.42-1.39(m,2H), 1.36-1.29(m,16H), 0.92(t,J=6.6Hz,3H); NMR 13 C NMR (150MHz, CD3OD) δ 163.78, 153.92, 149.26, 93.59, 79.98 (d, J = 11.4Hz), 65.20 (d, J = 158.4Hz), 63.95 (d, J = 5.6Hz), 50.05, 38.30, 37.92, 31.68, 29.50 (d, J = 6.5Hz), 29.39, 29.37, 29.32, 29.26, 29.08, 28.97, 28.83, 28.10, 25.27, 22.35, 13.06; NMR 31 P NMR (240MHz, CD3OD) δ 16.06; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 24 H 47 N3O6PS2[M+H] + 568.2638,found 568.2632,C 24 H 46 N3O6PS2Na[M+Na] + 590.2458, found 590.2454.

[0237] Example 13. Synthesis of target compound V-13

[0238]

[0239] The target compound V-13 was synthesized using the same method as compound V-1, except that the starting material 2-(decyldithio)ethane-1-ol was replaced with 0.335 g (1.00 mmol) of 4-(tetradecyldithio)but-1-ol, yielding intermediate IV-13, a white solid of 0.388 g, with a yield of 48%.

[0240] Its nuclear magnetic resonance 1 H NMR (600MHz, CDCl3) δ8.01-7.81(m,4H),7.56-7.33(m,6H),4.62-3.77(m,9H),2. 64-2.52(m,4H),1.58-1.55(m,4H),1.44-1.24(m,24H),0.88(t,J=6.6Hz,3H); NMR 13 C NMR (150MHz, CDCl3) δ 166.11, 133.28, 129.70, 129.57, 128.70, 128.52, 38.81, 38.20, 31.94, 31.83, 29.72, 29.70, 29.67, 29.65, 29.59, 29.38, 29.31, 29.20, 28.59, 25.26, 22.70, 14.14; 31 P NMR (240MHz, CDCl3) δ 15.61; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 40 H 59 N3O8PS2[M+H] + 804.3476, found 804.3474,

[0241] C 40 H 58 N3O8PS2Na[M+Na] + 826.3295, found 826.3291.

[0242] The intermediate IV-13 was fed in an amount of 0.361 g, yielding the target product V-13, a white solid of 0.131 g, with a yield of 48%.

[0243] Its nuclear magnetic resonance 1H NMR(600MHz,CD3OD)δ7.80(d,J=7.2Hz,1H),5.94(d,J=7.2Hz,1H),4.14(dd,J=3.6,13.8Hz,1H), 3.92-3.87(m,2H),3.84(dd,J=7.8,13.8Hz,1H),3.78-3.74(m,2H),3.71-3.68(m,1H),3.62(dd,J =9.6, 13.2 Hz, 1H), 3.55 (dd, J = 4.2, 12.6 Hz, 1H), 2.74 (t, J = 7.2 Hz, 2H), 2.70 (t, J = 7.2, 2H), 1.82-1.78 (m, 2H), 1.74-1.66 (m, 4H), 1.43-1.40 (m, 2H), 1.36-1.29 (m, 20H), 0.92 (t, J = 7.2 Hz, 3H); NMR 13 C NMR (150MHz, CD3OD) δ 164.15, 154.48, 149.05, 93.67, 80.05 (d, J = 11.6 Hz), 65.24 (d, J = 158.6 Hz), 63.96 (d, J = 5.9 Hz), 60.13, 50.05, 38.30, 37.92, 31.69, 29.50 (d, J = 6.5 Hz), 29.41, 29.40, 29.38, 29.32, 29.27, 29.09, 28.97, 28.84, 28.11, 25.27, 22.35, 13.06; NMR 31 P NMR (240MHz, CD3OD) δ 16.05; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 26 H 51 N3O6PS2[M+H] + 596.2951, found 596.2947,

[0244] C 26 H 50 N3O6PS2Na[M+Na] + 618.2771, found 618.2770.

[0245] Example 14. Synthesis of target compound V-14

[0246]

[0247] The target compound V-14 was synthesized using the same method as compound V-14, except that the starting material 2-(decyldithio)ethane-1-ol was replaced with 0.363 g (1.00 mmol) of 4-(hexadecyldithio)but-1-ol, yielding intermediate IV-14, a white solid of 0.492 g, with a yield of 59%.

[0248] Its nuclear magnetic resonance 1 H NMR (600MHz, CDCl3) δ8.01-8.00(m,4H),7.57-7.42(m,6H),4.62-3.85(m,9H),2. 61-2.56(m,4H),1.63-1.59(m,6H),1.35-1.26(m,26H),0.90(t,J=6.6Hz,3H); NMR 13 C NMR (150MHz, CDCl3) δ 166.10, 133.33, 132.44, 129.71, 129.53, 128.77, 128.54, 128.47, 64.80, 63.00, 38.86, 38.18, 31.94, 29.72, 29.70, 29.67, 29.65, 29.58, 29.38, 29.31, 29.20, 28.59, 25.23, 22.70, 14.14; 31 P NMR (240MHz, CDCl3) δ 16.43; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 42 H 63 N3O8PS2[M+H] + 832.3789,found

[0249] 832.3761,C 42 H 62 N3O8PS2Na[M+Na] + 854.3608, found 854.3588.

[0250] The intermediate IV-14 was fed in an amount of 0.374 g, yielding the target product V-14, a white solid of 0.195 g, with a yield of 60%.

[0251] Its nuclear magnetic resonance 1H NMR(600MHz,CD3OD)δ7.80(d,J=7.2Hz,1H),5.94(d,J=7.2Hz,1H),4.14(dd,J=3.6,14.4Hz,1H), 3.92-3.87(m,2H),3.84(dd,J=7.8,14.4Hz,1H),3.78-3.74(m,2H),3.71-3.67(m,1H),3.61(dd,J =9.6,13.2Hz,1H), 3.55(dd,J=4.2,12.6Hz,1H), 2.74(t,J=7.2Hz,2H), 2.70(t,J=7.2,2H), 1.82-1.78(m,2H), 1.74-1.66(m,4H), 1.43-1.40(m,2H), 1.36-1.31(m,24H), 0.92(t,J=6.6Hz,3H); NMR 13 C NMR (150MHz, CD3OD) δ 164.26, 154.63, 149.01, 93.68, 80.07 (d, J = 11.9 Hz), 65.24 (d, J = 157.8 Hz), 63.96 (d, J = 6.0 Hz), 60.13, 50.05, 38.29, 37.91, 31.69, 29.50 (d, J = 6.0 Hz), 29.42, 29.39, 29.38, 29.32, 29.27, 29.10, 28.98, 28.84, 28.11, 25.27, 22.36, 13.07; NMR 31 P NMR (240MHz, CD3OD) δ 16.06; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 28 H 55 N3O6PS2[M+H] + 624.3264, found 624.3257,

[0252] C 28 H 54 N3O6PS2Na[M+Na] + 646.3084, found 646.3080.

[0253] Example 15. Synthesis of target compound V-15

[0254]

[0255] The target compound V-15 was synthesized using the same method as compound V-1, except that the starting material 2-(decyldithio)ethane-1-ol was replaced with 0.279 g (1.00 mmol) of 6-(octyldithio)hex-1-ol, yielding intermediate IV-15, a white solid of 0.362 g, with a yield of 48%.

[0256] Its nuclear magnetic resonance 1 H NMR (600MHz, CDCl3) δ8.01-7.87(m,4H),7.60-7.33(m,6H),4.62-3.65(m,9H),2.67-2.52(m,4H),1.68-1.19(m,20H),0.87(t,J=6.6Hz,3H); Nuclear Magnetic 13 C NMR (150MHz, CDCl3) δ 166.13, 133.27, 133.12, 129.70, 129.60, 128.72, 128.61, 128.50, 128.36, 65.19, 63.11, 39.07, 39.04, 38.82, 31.81, 30.68, 30.20, 29.71, 29.22, 29.19, 29.06, 28.55, 28.20, 25.29, 22.65, 14.12; 31 P NMR (240MHz, CDCl3) δ 16.00; High-resolution mass spectrometry (HRMS) (ESI) - m / z: calculated for C 36 H 49 N3O8PS2[MH] - 746.2704, found 746.2729.

[0257] The intermediate IV-15 was fed in an amount of 0.337 g, yielding the target product V-15, a white solid of 0.113 g, with a yield of 45%.

[0258] Its nuclear magnetic resonance 1H NMR(600MHz,CD3OD / CDCl3)δ7.80(d,J=7.2Hz,1H),5.94(d,J=7.2Hz,1H),4.14(dd,J =4.2,12.6Hz,1H),3.90-3.82(m,3H),3.78-3.74(m,2H),3.71-3.67(m,1H),3.61(dd, J = 9.0, 12.6 Hz, 1H), 3.55 (dd, J = 4.2, 12.6 Hz, 1H), 2.71-2.68 (m, 4H), 1.73-1.66 (m, 4H), 1.65-1.60 (m, 2H), 1.46-1.40 (m, 6H), 1.36-1.31 (m, 8H), 0.93 (t, J = 6.6 Hz, 3H); NMR 13 C NMR (150MHz, CD3OD) δ 164.13, 154.44, 149.10, 93.63, 80.02 (d, J = 11.9 Hz), 65.22 (d, J = 158.4 Hz), 64.40 (d, J = 5.9 Hz), 60.12, 50.03, 38.34, 38.24, 31.59, 30.70 (d, J = 6.0 Hz), 28.94, 28.92, 28.83, 28.82, 28.09, 27.84, 25.16, 22.33, 13.05; NMR 31 PNMR (240MHz, CD3OD) δ16.03 High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 22 H 41 N3O6PS2[MH] - 538.2180, found 538.2183.

[0259] Example 16. Synthesis of compound V-16

[0260]

[0261] The target compound V-16 was synthesized using the same method as compound V-1, except that the starting material 2-(decyldithio)ethane-1-ol was replaced with 0.307 g (1.00 mmol) of 6-(decyldithio)hex-1-ol, yielding intermediate IV-16, a white solid of 0.376 g, with a yield of 48%.

[0262] Its nuclear magnetic resonance 1H NMR(600MHz,CD3OD / CDCl3)δ8.19(d,J=7.8Hz,1H),8.04-7.87(m,4H),7.65-7 .46(m,6H),6.20(d,J=7.8Hz,1H),4.65-4.56(m,1H),4.42-4.35(m,2H),4.29 -4.08(m,2H),3.99-3.93(m,1H),4.65-4.56(m,1H),3.86-3.83(m,2H),3.72- 3.68(m,1H),2.67-2.56(m,4H),1.67-1.27(m,24H),0.89(t,J=6.6Hz,3H); NMR 13 C NMR (150MHz, CD3OD / CDCl3) δ 167.24, 134.21, 130.68, 130.47, 129.62, 129.37, 128.95, 64.13, 39.50, 39.32, 32.76, 30.36 (d, J = 4.5Hz), 30.15, 30.04, 29.93, 29.82, 29.19, 28.83, 26.06, 23.45, 14.24; NMR 31 PNMR (240MHz, CD3OD / CDCl3) δ 16.67; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 38 H 55 N3O8PS2[M+H] + 776.3163,found 776.3158,C 38 H 54 N3O8PS2Na[M+Na] + 798.2982, found 798.2979.

[0263] The intermediate IV-16 was fed in an amount of 0.361 g, yielding the target product V-16, a white solid of 0.121 g, with a yield of 46%.

[0264] Its nuclear magnetic resonance 1H NMR (600MHz, CD3OD) δ7.82 (d, J=6.0Hz, 1H), 5.94 (d, J=7.8Hz, 1H), 4.16-4. 13(m,1H),3.89-3.81(m,3H),3.78-3.74(m,2H),3.70-3.68(m,1H),3.63-3. 58(m,1H),3.57-3.53(m,1H),2.72-2.68(m,4H),1.72-1.67(m,4H),1.64-1 .61(m,2H),1.45-1.39(m,6H),1.36-1.28(m,12H),0.92(t,J=5.4Hz,3H); NMR 13 C NMR (150MHz, CD3OD) δ 163.86, 154.04, 149.24, 93.58, 79.97 (d, J = 11.9 Hz), 65.19 (d, J = 158.9 Hz), 64.40 (d, J = 6.2 Hz), 60.10, 50.03, 38.34, 38.24, 31.67, 30.70 (d, J = 6.2 Hz), 29.28, 29.26, 29.06, 28.94, 28.83, 28.07, 27.84, 25.16, 22.35, 13.06; NMR 31 P NMR (240MHz, CD3OD) δ 15.97; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 24 H 47 N3O6PS2[M+H] + 568.2638, found 568.2634,

[0265] C 24 H 46 N3O6PS2Na[M+Na] + 590.2458, found 590.2459.

[0266] Example 17. Synthesis of compound V-17

[0267]

[0268] The target compound V-17 was synthesized using the same method as compound V-1, except that the starting material 2-(decyldithio)ethane-1-ol was replaced with 0.335 g (1.00 mmol) of 6-(dodecyldithio)hex-1-ol, yielding intermediate IV-17, a white solid of 0.378 g, with a yield of 47%.

[0269] Its nuclear magnetic resonance 1H NMR (600MHz, CDCl3) δ8.00-7.91(m,4H),7.55-7.36(m,6H),4.74-4.32(m,5H),3. 95-3.81(m,4H),2.66-2.54(m,4H),1.65-1.19(m,28H),0.88(t,J=6.6Hz,3H); NMR 13 C NMR (150MHz, CDCl3) δ 166.09, 133.31, 132.51, 129.70, 129.55, 128.76, 128.53, 65.29, 63.01, 39.03, 38.79, 31.92, 30.61 (d, J = 6.3Hz), 29.67, 29.64, 29.62, 29.55, 29.36, 29.28, 29.24, 29.03, 28.57, 28.15, 25.25, 22.70, 14.13; NMR 31 P NMR (240MHz, CDCl3) δ 16.46; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 40 H 59 N3O8PS2[M+

[0270] H] + 804.3476, found 804.3472, C 40 H 58 N3O8PS2Na[M+Na] + 826.3295, found826.3302.

[0271] The intermediate IV-17 was fed in an amount of 0.361 g, yielding the target product V-17, a white solid of 0.140 g, with a yield of 51%.

[0272] Its nuclear magnetic resonance 1H NMR (600MHz, CD3OD) δ7.82(d,J=7.2Hz,1H),5.95(d,J=7.8Hz,1H),4.15(dd,J=3.0, 13.8Hz,1H),3.90-3.82(m,3H),3.78-3.74(m,2H),3.71-3.68(m,1H),3.61(dd,J=9 0, 12.6 Hz, 1H), 3.55 (dd, J = 3.6, 12.0 Hz, 1H), 2.71-2.68 (m, 4H), 1.73-1.66 (m, 4H), 1.65-1.60 (m, 2H), 1.46-1.39 (m, 6H), 1.36-1.29 (m, 16H), 0.92 (t, J = 6.6 Hz, 3H); NMR 13 C NMR (150MHz, CD3OD) δ 163.85, 154.02, 149.26, 93.58, 79.97 (d, J = 11.7Hz), 65.19 (d, J = 158.3Hz), 64.40 (d, J = 5.6Hz), 60.10, 50.04, 38.35, 38.24, 31.69, 30.71 (d, J = 6.0Hz), 29.39, 29.37, 29.32, 29.25, 29.09, 28.94, 28.84, 28.82, 28.07, 27.84, 25.17, 22.36, 13.07; NMR 31 P NMR (240MHz, CD3OD) δ 16.03; High-resolution mass spectrometry (HRMS) (ESI) + m / z: calculated for C 26 H 51 N3O6PS2[M+H] + 596.2951,found 596.2949,C 26 H 50 N3O6PS2Na[M+Na] + 618.2771, found 618.2772.

[0273] Example 18. Synthesis of compound V-18

[0274]

[0275] The target compound V-18 was synthesized using the same method as compound V-1, except that the starting material 2-(decyldithio)ethane-1-ol was replaced with 0.363 g (1.00 mmol) of 6-(tetradecyldithio)hex-1-ol, yielding intermediate IV-18, a white solid of 0.437 g, with a yield of 53%.

[0276] Its nuclear magnetic resonance1 H NMR (600MHz, CDCl3) δ8.00-7.71(m,4H),7.54-7.31(m,6H),4.61-3.72(m,9H),2.62-2.49(m,4H),1.64-1.17(m,32H),0.88(t,J=7.2Hz,3H); Nuclear Magnetic 13 C NMR (150MHz, CDCl3) δ 166.12, 133.25, 133.06, 129.69, 129.61, 128.69, 128.59, 128.51, 128.40, 38.99, 38.78, 31.94, 30.68, 30.20, 29.71, 29.69, 29.67, 29.64, 29.57, 29.37, 29.30, 29.24, 29.04, 28.59, 28.23, 25.30, 22.70, 14.14; NMR 31 P NMR (240MHz, CDCl3) δ 15.10; High-resolution mass spectrometry (HRMS) (ESI) - m / z: calculated for C 42 H 61 N3O8PS2[MH] - 830.3643,found830,3664.

[0277] The intermediate IV-18 was fed in an amount of 0.374 g, yielding the target product V-18, a white solid of 0.151 g, with a yield of 52%.

[0278] Its nuclear magnetic resonance 1 H NMR (600MHz, CD3OD) δ7.81 (d, J=7.8Hz, 1H), 5.94 (d, J=7.2Hz, 1H), 4.15 (dd, J= 3.6,14.4Hz,1H),3.90-3.82(m,3H),3.78-3.74(m,2H),3.71-3.68(m,1H),3.6 0(dd,J=9.6,12.6Hz,1H),3.55(dd,J=4.2,12.6Hz,1H),2.71-2.68(m,4H),1.7 3-1.60(m,6H),1.46-1.40(m,6H),1.35-1.31(m,20H),0.92(t,J=6.6Hz,3H); NMR 13C NMR (150MHz, CD3OD) δ 163.93, 154.16, 149.21, 93.62, 79.97 (d, J = 11.7Hz), 65.18 (d, J = 158.7Hz), 64.40 (d, J = 5.9Hz), 60.10, 50.05, 38.34, 38.24, 31.70, 30.71 (d, J = 6.0Hz), 29.42, 29.40, 29.39, 29.38, 29.31, 29.25, 29.10, 28.94, 28.84, 28.83, 28.08, 27.85, 25.17, 22.36, 13.08; NMR 31 P NMR (240MHz, CD3OD) δ 16.04; High-resolution mass spectrometry (HRMS) (ESI) - m / z: calculated for C 28 H 53 N3O6PS2[MH] - 622.3119, found 622.3124.

[0279] Pharmacological activity test

[0280] Example 19. Study on the cytotoxicity and anti-dsDNA virus activity of target compounds V-1 to V-18

[0281] Target compound cytotoxicity test

[0282] A549 cells or Vero cells were selected and cytotoxicity was tested using neutral red staining.

[0283] A549 cells or Vero cells were injected at a rate of 3 × 10⁻⁶. 4Cells were seeded at a density of 100 cells / well in 96-well plates and incubated at 37°C with 5% CO2 for approximately 24 hours. When the cells reached approximately 90% confluence, the culture medium was discarded, and each well was washed twice with PBS. 200 μL / well of DMEM medium (containing 2% fetal bovine serum) containing different concentrations of the target compound or the positive control drug BCV was added to each well. Each compound was serially diluted in 2-fold increments, with 3 replicates. A blank control and a normal cell control group were also included. Cell status was observed daily. After 72 hours of incubation at 37°C with 5% CO2, the culture medium was discarded, and each well was washed twice with PBS. 100 μL / well of DMEM medium containing 40 μg / mL neutral red (containing 2% fetal bovine serum) was added to each well for staining. After incubation at 37°C with 5% CO2 for 4 hours, the culture medium was discarded, and each well was washed twice with PBS. 48% acidic ethanol aqueous solution containing 1% glacial acetic acid was added to each well. The optical density (OD) at 540 nm was measured using a spectrophotometer, and cell viability was calculated using the following formula. The half-maximal cytotoxic concentration (CC) of the compound was calculated using GraphPad Prism 8. 50 The cytotoxicity results are shown in Table 1.

[0284]

[0285] Test of the anti-dsDNA virus activity of the target compound

[0286] Antiviral activity of vaccinia virus Tian Tan strain and adenovirus 5 was tested using neutral red staining. For vaccinia virus Tian Tan strain, antiviral activity was tested in Vero cells and A549 cells, respectively; for adenovirus 5, antiviral activity was tested in A549 cells.

[0287] A549 cells or Vero cells were respectively injected at 2×10 4 1 cell / well and 3 × 10 4 Cells were seeded at a density of 100 cells / well in 96-well plates and cultured at 37°C with 5% CO2 for 24 hours. The culture medium was then discarded, and each well was washed twice with PBS. 100 mg TCID₂ was added to each well. 50Viral fluid was added to each well, along with 100 μL / well of DMEM medium (containing 2% fetal bovine serum) containing different concentrations of the target compound. Each compound was serially diluted 3-fold, with 3 replicates. A positive control group (BCV), a viral control group, and a normal cell control group were also established. Cells were cultured at 37°C with 5% CO2, and cytopathic effect (CPE) was observed daily. After 72 h (adenovirus) or 96 h (vaccinia virus), the culture medium was discarded, and each well was washed twice with PBS. 100 μL of DMEM medium (containing 2% fetal bovine serum) containing 40 μg / mL neutral red was added to each well for staining. After incubation at 37°C with 5% CO2 for 4 h, the culture medium was discarded, and each well was washed twice with PBS. 48% acidic ethanol solution containing 1% acetic acid was added to each well. The optical density (OD) at 540 nm was measured using a spectrophotometer, and cell viability was calculated using the following formula. The half-maximal effective concentration (IC50) of the compound was calculated using GraphPad Prism 8. 50 The results of the antiviral activity are shown in Table 1.

[0288]

[0289] Table 1. Cytotoxic and in vitro antiviral activities of compounds V-1 to V-18 and positive control BCV.

[0290]

[0291]

[0292] The cytotoxicity and antiviral activity results showed that the compounds exhibited stronger cytotoxicity in Vero cells than in A549 cells. Regarding antivaccinia virus activity, the compounds showed antiviral activity in both Vero and A549 cells, with some compounds showing higher antiviral activity than the positive control drug BCV. Compounds with a disulfide bond and phosphate group linked by a -C2H4- group exhibited better antiviral activity than those with -C3H6-, -C4H8-, or -C6H12- groups. For example, compounds V-3, V-8, V-13, and V-18 all have a hexadecyl carbon chain linked to a disulfide bond; compound V-3 showed 2.8-7.6 times the activity of the other compounds. Compounds V-3, V-12, and V-16 are homologues with a hydrophobic chain length of 18 carbon atoms; compound V-3 showed 9.0 times and 65.4 times the activity of these homologues, respectively. Comparing the antiviral activity and cytotoxicity data of compounds V-1 to V-5, when the disulfide bond and phosphate group share the same linking group (-C2H4-), the cytotoxicity and antiviral activity of V-1 to V4 gradually increased with the increase in the length of the tail carbon chain linked to the disulfide bond. Compounds V-5 and V-4 exhibited comparable antiviral activity and cytotoxicity, with V-3 and V-4 showing the strongest antiviral activity. The IC50 values ​​for the antiviral activity of compounds V-3 and V-4 in Vero cells were 0.0960 μM and 0.0684 μM, respectively (see appendix). Figure 17 The antiviral activity IC50 values ​​in A549 cells were 0.0790 μM and 0.0904 μM, respectively (see attached image). Figure 18 Comparing other compounds with different linking groups, compounds V-6 to V-10, V-11 to V-14, and V-15 to V18, with linking groups of -C3H6-, -C4H8-, and -C6H12- respectively, showed similar trends in antiviral activity and cytotoxicity to compounds V-1 to V5. Among these, compounds with disulfide-bonded hydrophobic side chains of 18-20 atoms exhibited strong antiviral activity. Compounds V-1, V-6, V-11, and V-15, with chain lengths of 14-16 atoms, showed 10-250 times lower antiviral activity than the positive control drug BCV. It can be seen that among the synthesized disulfide-bonded cidofovir prodrug derivatives with long aliphatic side chains, the length of the hydrophobic chain has a significant impact on their antiviral activity. Compounds V-3 and V-4, exhibiting the strongest antiviral activity, have chain lengths of 18 and 20 atoms, respectively. Compounds V-3 and V-4 showed cytotoxicity (CC) in Vero cells. 50 The values ​​are 6.530 μM and 2.578 μM, respectively (see attached image). Figure 19 ), cytotoxic CC in A549 cells 50 The values ​​were 14.73 μM and 8.689 μM, respectively (see attached image). Figure 20Compound V-3 exhibits lower cytotoxicity than V-4. Its therapeutic index (TI) is... 50 It was 2.8 times higher than the positive control drug BCV in A549 cells.

[0293] Vaccinia virus, along with monkeypox virus, smallpox virus, cowpox virus, rabbitpox virus, and mousepox virus, belongs to the genus Orthopoxvirus of the family Poxviridae. They share high genomic homology and conservation, and their main viral characteristics are similar. Vaccinia virus can be tested in biosafety level II laboratories; therefore, it is widely used as a tool for screening and evaluating anti-poxvirus drugs. Based on this, although the antiviral activity experiment in this embodiment was conducted using vaccinia virus, it can be inferred that the synthesized compound will have similar antiviral effects in other Orthopoxviruses.

[0294] To further verify the broad-spectrum antiviral effects of the compounds, their antiviral activity against adenovirus 5 was tested in A549 cells. It can be seen that the antiviral activity of the compounds against adenovirus 5 shows a similar trend to their activity against vaccinia virus, with compounds V-3 and V-4 both exhibiting better antiviral activity than the positive control drug BCV. However, compound V-14 showed a significantly lower IC50 value for its antiviral activity. 50 The concentration was 0.1337 μM, nearly twice that of BCV, and the therapeutic index was nearly three times higher. Compound V-16 exhibited similar antiviral activity to BCV, but with significantly reduced cytotoxicity, and its therapeutic index was nearly nine times that of BCV. Therefore, the structure of the hydrophobic chain can have a significant impact on antiviral activity for different viruses.

[0295] Example 22. Stability testing of target compounds V-3, V-4, V-14 and V-17

[0296] Target compounds V-3, V-4, V-14, and V-17, exhibiting good activity and a high therapeutic index, were selected. Their stability in simulated gastric fluid, simulated intestinal fluid, and human plasma was tested to assess their potential for development into oral formulations. The stability of the compounds in simulated gastric fluid and simulated intestinal fluid was determined by HPLC, and their stability in human plasma was determined by LC-MS / MS.

[0297] Stability testing of compounds in artificial gastric fluid

[0298] The analyte was prepared into a 9000 μL working solution with a concentration of 200 μM using simulated gastric fluid (pH = 2.0) and incubated at 37 °C. At time points of 0 min, 15 min, 30 min, 60 min, 90 min, and 120 min, 200 μM of the working solution was transferred to 2 mL centrifuge tubes, 600 μM of chromatographic methanol was added, and the mixture was centrifuged at 14000 rpm for 5 min at 4 °C. The supernatant was collected and analyzed by high-performance liquid chromatography (HPLC). A Waters SunFire C18 column (4.6 mm × 250 mm × 5 mm) was used. Mobile phase A was methanol containing 0.1% trifluoroacetic acid, and mobile phase B was water containing 0.1% trifluoroacetic acid. The column temperature was 35 °C, the flow rate was 1.0 mL / min, and the injection volume was 10 μL. Gradient elution: 0-6.0 min, 80%-100% A; 6.0-10.0 min, 100% A; 10.0-11.0 min, 100%-80% A; 11.0-15.0 min, 80% A. The peak area at 0 min was used as the standard. Compound stability was calculated by comparing the peak area at different time points with the peak area at 0 min. Three parallel experimental groups were set up for each compound. The stability results of different compounds in simulated gastric fluid are shown below. Figure 21 .

[0299] In simulated gastric fluid, all tested compounds exhibited good stability within 120 min. Compounds V-14 and V17 were almost unaffected during incubation. Compounds V-3 and V-4 retained 77% and 68% of their original composition, respectively, after 120 min of incubation, demonstrating good stability. Compounds V-3, V-4, V-14, and V-17 all showed stronger stability than the control drug BCV.

[0300] Stability testing of compounds in artificial intestinal fluid

[0301] The analyte was prepared into a 9000 μL working solution with a concentration of 200 μM using artificial intestinal fluid (pH = 6.8) and incubated at 37 °C. At time points of 0 min, 15 min, 30 min, 60 min, 90 min, 120 min, 240 min, and 360 min, 200 μM of the working solution was transferred to 2 mL centrifuge tubes, 600 μM of chromatographic methanol was added, and the mixture was centrifuged at 14000 rpm for 5 min at 4 °C. The supernatant was collected and analyzed by high-performance liquid chromatography (HPLC). A Waters SunFire C18 column (4.6 mm × 250 mm × 5 mm) was used. Mobile phase A was methanol containing 0.1% trifluoroacetic acid, and mobile phase B was water containing 0.1% trifluoroacetic acid. The column temperature was 35 °C, the flow rate was 1.0 mL / min, and the injection volume was 10 μL. Gradient elution: 0-6.0 min, 80%-100% A; 6.0-10.0 min, 100% A; 10.0-11.0 min, 100%-80% A; 11.0-15.0 min, 80% A. The peak area at 0 min was used as the standard. Compound stability was calculated by comparing the peak area at different time points with the peak area at 0 min. Three parallel experimental groups were set up for each compound. The stability results of different compounds in simulated gastric fluid are shown below. Figure 22 .

[0302] In artificial intestinal fluid, the test compound was almost unaffected within 360 minutes, demonstrating strong stability.

[0303] Stability testing of compounds in human plasma

[0304] The mixed human plasma was diluted 1-fold with PBS (pH=7.4) and kept at 37°C for 30 min. 5 μL of stock solutions of different compounds at a concentration of 40.0 μM was transferred to 195 μL of the mixed human plasma, mixed thoroughly, and incubated at 37°C for 0 h, 1 h, 2 h, 4 h, 6 h, 12 h, and 24 h. Then, 400 μL of chromatographic methanol containing an internal standard was added, mixed thoroughly, and centrifuged at 14000 rpm for 10 min at 4°C. The supernatant was collected and analyzed by LC-MS / MS. The ratio of the peak area of ​​the compound to the peak area of ​​the internal standard at 0 h was used as the standard to calculate the stability of the compounds. Three parallel experimental groups were set up for each compound. In the mixed human plasma, although the stability of compounds V-3, V-4, V-14, and V-17 was weaker than that of BCV, their half-lives were all greater than 2 h, fully meeting the requirements for oral administration.

[0305] Stability studies have shown that the compound exhibits good stability in simulated gastric juice, simulated intestinal juice, and mixed human plasma, making it particularly suitable for oral administration. It should be noted that this invention is not limited to the above embodiments, i.e., it does not mean that this invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvements to this invention, equivalent substitutions of the raw materials in the product of this invention, the addition of auxiliary components, and the selection of specific methods, all fall within the protection and disclosure scope of this invention.

Claims

1. The use of cytosine compounds of formula (II) or pharmaceutically acceptable salts thereof in the preparation of medicaments for the treatment and relief of diseases associated with orchiopiveidae viruses, (II) Y 1 The expression is -SS-, where m is an integer from 1 to 5; and n is an integer from 7 to 18. The viruses in the Orthopoxviridae family are selected from smallpox virus, monkeypox virus, cowpox virus, rabbitpox virus, mousepox virus, and vaccinia virus.

2. The application as described in claim 1, wherein the compound or its pharmaceutically acceptable salt is an ammonium salt with the following chemical structure: 。

Images