WO2024259934A1 - Nucleoside analogue and medical application thereof - Google Patents

Abstract

A nucleoside analog, or a pharmaceutically acceptable salt or ester thereof, for use in the preparation of a drug for treating, inhibiting or preventing a disease caused by viral infection or cell proliferation, and in particular for being capable of treating, inhibiting or preventing a monkey poxvirus infection or a disease caused by a monkey poxvirus infection in a mammal including human beings.

Description

Nucleoside analogs and their medical uses

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is based on the Chinese patent application with application number 202310741316.5 and application date June 21, 2023, and claims the priority of the Chinese patent application. The entire content of the Chinese patent application is hereby introduced into this application as a reference.

Technical Field

The present invention relates to a nucleoside analog or a pharmaceutically acceptable salt or ester thereof, and use thereof in preparing a medicine for treating, inhibiting or preventing a virus infection disease or a cell proliferation disease.

Background Art

Based on the type and expression pattern of nucleic acid carried by the virus, viruses can be divided into DNA viruses, RNA viruses, etc. The design and discovery of new antiviral drugs are generally directed against viral or cellular targets. Drugs that inhibit viral proteins may be virus-specific and more susceptible to resistance.

The 2022 monkeypox outbreak was first discovered in the UK on May 7, 2022, local time. On May 20, local time, with more than 100 confirmed and suspected cases of monkeypox in Europe. Monkeypox is a viral zoonosis that causes symptoms in humans similar to those seen in smallpox patients in the past. However, since smallpox was eradicated from the world in 1980, smallpox no longer exists, while monkeypox is still sporadic in parts of Africa. Monkeypox occurs in monkeys in the rainforests of central and western Africa, and can also infect other animals and occasionally humans. The clinical manifestations are similar to smallpox, but the condition is milder. The disease is caused by the monkeypox virus, a double-stranded DNA virus with a relatively stable structure and a low mutation rate. The virus can be transmitted from animals to humans through direct close contact, and can also be transmitted from person to person.

Since the discovery that (S)-hydroxyphosphonylmethoxypropyladenine ((S)-HPMPA) exhibits broad-spectrum antiviral effects against DNA viruses, people have conducted extensive research on the antiviral effects of phosphonic acid nucleosides. However, since the phosphonic acid group carries a negative charge, its cell permeability and oral bioavailability are low. The currently available antiviral drugs on the market are still insufficient to cope with the spread of frequent viral infections. Therefore, there is a need to develop and produce more effective broad-spectrum antiviral drugs.

Summary of the invention

The main technical problem solved by the present application is to provide a nucleoside analog compound, which can have at least one or more of the following effects:

1) Better pharmacokinetic properties in vivo;

2) Higher drug stability and solubility;

3) Lower toxicity and adverse reactions;

4) Higher transport and distribution capacity to specific sites; and

5) Better sustained-release effect and longer duration of action.

In one aspect, the present invention provides a nucleoside analogue represented by formula (I) or a pharmaceutically acceptable salt or ester thereof:

Wherein, X is selected from -OR ⁴ , or X and R ³ are combined to form a chemical bond;

^R1 is selected from H or C4-C30 carbonyl, the C4-C30 carbonyl includes substituted or unsubstituted hydrocarbon carbonyl, substituted or unsubstituted aryl carbonyl or heterocyclic carbonyl, and substituted or unsubstituted hydrocarbon oxycarbonyl, and the carbonyl has a carbon number of 4 to 30, which can be 4 to 10, 10 to 30 or 20 to 30, and specifically can be 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30;

R ² and R ⁴ are independently selected from H, Wherein R ⁵ is selected from substituted or unsubstituted C15-C30 hydrocarbon groups, and the hydrocarbon group may be a straight-chain hydrocarbon group or a branched hydrocarbon group;

^R3 is selected from: H; formyl, C4-C30 carbonyl, wherein the C4-C30 carbonyl includes substituted or unsubstituted alkylcarbonyl, substituted or unsubstituted arylcarbonyl or heterocyclic carbonyl, and substituted or unsubstituted alkoxycarbonyl, and the carbon number of the carbonyl is not more than 30; or an amino acid residue.

As a limitation, when X is -OR ⁴ , R ¹ , R ² , R ³ , and R ⁴ are not H at the same time; when X and R ³ are combined to form a chemical bond, R ¹ and R ² are not H at the same time.

Furthermore, the present application provides a compound represented by formula (II);

Furthermore, the present application provides a compound represented by formula (III):

As a limitation, when X is -OR ⁴ , R ¹ , R ³ , and R ⁴ are not H at the same time; when X and R ³ are combined to form a chemical bond, R ¹ is not H.

In some embodiments, the present invention provides compounds represented by formula (IIIa) and formula (IIIb):

Wherein R ¹ or R ³ is as defined above. In some embodiments, in formula (IIIa) and (IIIb), R ¹ and R ³ are independently selected from formyl, C1-C3 alkylcarbonyl, (CH ₃ ) ₂ CHCOOCH(CH ₃ )C(O)-, CH ₃ COOCH(CH ₃ )C(O)-; and X is H. In some preferred embodiments, in formula (IIIa) and (IIIb), R ¹ and R ³ are independently selected from C1-C3 Alkylcarbonyl and X is H.

Furthermore, the present application provides a compound represented by formula (IV):

As a limitation, when X is -OR ⁴ , R ¹ , R ³ , and R ⁴ are not H at the same time; when X and R ³ are combined to form a chemical bond, R ¹ is not H.

In some embodiments, the present invention provides a compound represented by formula (IVa) or (IVb):

Wherein R ¹ or R ³ is as defined above. In some preferred embodiments, in formula (IVa) and (IVb), R ¹ and R ³ are independently selected from formyl, C1-C3 alkylcarbonyl, (CH ₃ ) ₂ CHCOOCH(CH ₃ )C(O)-, CH ₃ COOCH(CH ₃ )C(O)-; and X is H. In some preferred embodiments, in formula (IVa) and (IVb), R ¹ and R ³ are independently selected from C1-C3 alkylcarbonyl, and X is H.

More preferably, R ³ is selected from formyl. Further, R ¹ is selected from substituted or unsubstituted hydrocarbon carbonyl, including the groups shown below:

Further, ^R1 is selected from substituted or unsubstituted arylcarbonyl or heterocyclic carbonyl, including the groups shown below:

wherein Y is selected from H, F, Cl, Br, Me, -OMe, -OEt, _-CF3 or -CN.

Further, ^R1 is selected from substituted or unsubstituted hydrocarbonoxycarbonyl groups, including the groups shown below:

Furthermore, R ⁵ includes the following groups:

Furthermore, R ³ includes the following groups:

The compounds of the present invention include the compounds shown in Table 1a and Table 1b below:

Table 1a

Table 1b

The compounds provided in this application can be used to prepare a method for treating, inhibiting or preventing viral infection or virus Drugs for diseases caused by infection. In some embodiments, viral infections include hepatitis B virus (HBV), new coronavirus (SARS-COV-2), human immunodeficiency virus (HIV), varicella zoster virus (VZV), cytomegalovirus (CMV), herpes simplex virus (HSV), BK virus, JC virus, Epstein-Barr virus (EBV), Ebola virus, polyomavirus, papillomavirus, orthopoxvirus, hepatitis C virus (HCV), respiratory syncytial virus (RSV), dengue virus, influenza virus, adenovirus, parainfluenza virus and/or rhinovirus.

Further, the above-mentioned orthopoxviruses include severe and mild smallpox viruses, monkeypox virus, cowpox virus, camelpox virus, molluscum contagiosum, sheeppox virus, aractuba virus (ARAV), BeAn58058 virus (BAV), cantagalo orthopoxvirus (CTGV), mousepox virus, elephantpox virus, vaccinia virus (VV), rabbitpox virus, raccoonpox virus, skunkpox virus, gerbilpox virus and volepox virus. Preferably, the compounds disclosed in the present application can be used to prepare drugs for treating, inhibiting or preventing diseases caused by monkeypox virus or smallpox virus infection in mammals.

The compounds provided in the present application can be used to prepare drugs for treating, inhibiting or preventing diseases caused by viral infection. Further, the diseases caused by viral infection include diseases caused by DNA virus infection, specifically, the diseases are selected from retinitis, pneumonia, cystitis, proteinopathy, etc.

The compounds provided herein can also be used to prepare drugs for treating, inhibiting or preventing cell proliferation-induced diseases. Further, the cell proliferation-induced disease is a tumor or cancer, specifically, the tumor or cancer is selected from multiple myeloma (MM), chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), solid tumors, refractory solid tumors, non-Hodgkin's lymphoma, hematological tumors, neuroblastoma, colorectal cancer, cervical cancer, lung cancer, leukemia, breast cancer, pancreatic cancer, B-cell malignancies, metastatic tumors and colon cancer.

The present application also provides a pharmaceutical composition, comprising one or more of the above compounds and at least one pharmaceutically acceptable carrier or excipient. Specifically, the pharmaceutically acceptable carrier includes one or more of creams, emulsions, gels, liposomes and nanoparticles; the pharmaceutically acceptable excipient includes one or more of binders, fillers, disintegrants, lubricants and glidants.

Furthermore, the above pharmaceutical composition is suitable for oral administration or injection administration.

The present application also provides a kit, which includes any one or more of the above-mentioned compounds or pharmaceutically acceptable salts or esters or any one or more of the pharmaceutical compositions.

The specific compounds provided in the present application, or their pharmaceutically acceptable salts or esters, or pharmaceutical compositions containing them, can effectively treat, inhibit or prevent viral infection and/or bacterial infection in mammals. Cell proliferative diseases, especially smallpox and monkeypox, and at least one or more of the following effects: changing the pharmacokinetic properties in the body, adjusting the absorption and distribution of the drug in the body, improving the stability and solubility of the drug, reducing toxicity and adverse reactions, improving the transport and distribution to specific parts, improving the sustained release effect, and prolonging the duration of action.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG1 is a graph showing the inhibitory activity-drug concentration curve of the test compound.

FIG. 2 shows a death curve diagram of a surrogate endpoint of mouse death, wherein FIG. 2a is a graph of body weight change, and FIG. 2b is a surrogate endpoint death curve.

FIG. 3 shows a graph of the actual mortality of mice, wherein FIG. 3a is a graph of weight changes, and FIG. 3b is an actual mortality curve.

DETAILED DESCRIPTION

In order to provide a clear and consistent understanding of the terms used in the specification of the present invention, some definitions are provided below. In addition, unless otherwise specified, all technical and scientific terms used in the present invention have the same meaning as commonly understood by ordinary technicians in the field to which the present invention belongs.

When used in conjunction with the term "comprising" in the claims and/or the specification, the use of the word "a" can mean "one", but it is also consistent with the meaning of "one or more", "at least one", and "one or more than one". Similarly, the word "another" can mean at least a second or many.

As used in this specification and claims, the words "comprising", "having" and synonyms are inclusive and open-ended and do not exclude additional unlisted elements or processing steps. The term "about" or "approximately" is used to indicate that the value includes the error caused by the instruments and methods used in determining the value.

The term "pharmaceutically acceptable" as used herein refers to drugs, medicines, inert ingredients, etc. described by the term, which are suitable for contact with the tissues of humans and lower animals without abnormal toxicity, incompatibility, instability, irritation, allergic reaction, etc., commensurate with a reasonable benefit/risk ratio. It preferably refers to compounds, compositions and preparations, etc. listed in the pharmacopoeia or other recognized pharmacopoeias for use in animals, more particularly in humans.

A "pharmaceutically acceptable salt" of a compound refers to a salt of the compound that is pharmaceutically acceptable. Salts of the compounds (basic, acidic or charged functional groups) may retain or improve the biological activity and properties of the parent compound as defined herein and are not biologically undesirable.

The term "ester" refers to esters derived from the compounds of the general formulae herein, including physiologically hydrolyzable esters (which can be hydrolyzed under physiological conditions to release the compounds of the present invention in free acid or alcohol form). The compounds of the present invention themselves may also be esters.

In some embodiments, the compounds disclosed herein exist as a prodrug.

The term "prodrug" or its equivalent refers to a pharmaceutical agent that is directly or indirectly converted into an active form in vitro or in vivo (see, for example, R.B.Silverman, 1992, "The Organic Chemistry of Drug Design and Drug Action," Academic Press, Chap. 8; Bundgaard, Hans; Editor. Neth. (1985), "Design of Prodrugs". 360pp. Elsevier, Amsterdam; Stella, V.; Borchardt, R.; Hageman, M.; Oliyai, R.; Maag, H.; Tilley, J. (Eds.)(2007), "Prodrugs: Challenges and Rewards, XVIII, 1470p. Springer). Prodrugs can be used to alter the biodistribution (e.g., so that the agent does not normally enter the protease reaction site) or pharmacokinetics of a particular drug. A variety of groups such as esters, ethers, phosphates/salts, etc. have been used to modify compounds to form prodrugs. When the prodrug is administered to a subject, the group is cleaved off enzymatically or non-enzymatically, reduced, oxidatively or hydrolytically, or otherwise releases the active compound. As used herein, "prodrug" includes pharmaceutically acceptable salts, or pharmaceutically acceptable solvates, and any crystalline forms of the above. Prodrugs are generally (although not necessarily) pharmaceutically inactive until they are converted to an active form.

It should be understood that the term "substituted" or "substituted" as used in the present invention includes an implicit condition that such substitution produces a stable compound (e.g., the compound cannot spontaneously undergo rearrangement, cyclization, elimination, etc.) as the valence of the substituted atom and the substituent change. As used in the present invention, the term "substituted" includes all permissible substituents of organic compounds. In a broad sense, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic organic compounds. The substituent may be one or more. The term "substituted" means when the above groups are substituted at one or more positions, the substituents include acylamino (including carbamoyl and ureido), alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, alkoxycarbonyl, carboxyl, carboxyl, aminocarbonyl, mono- and dialkylaminocarbonyl, cyano, azido, halogen, hydroxy, nitro, trifluoromethyl, thio, alkylthio, arylthio, alkylthiocarbonyl, thiocarboxylate, lower alkyl, lower alkenyl, lower alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, lower alkoxy, aryloxy, aryloxycarbonyloxy, benzyloxy, benzyl, sulfinyl, alkylsulfinyl, sulfonyl, sulfate, sulfonate, sulfonamide, phosphate, phosphonate, imino, formyl, etc. If permitted, Any of the above substituents may be further substituted, for example, by alkyl, aryl or other groups.

The term "alkylcarbonyl", "arylcarbonyl", "heterocycliccarbonyl" or "alkyloxycarbonyl" refers to -C(=O) _Ra , where _Ra is a hydrocarbon, aryl, heteroaryl, heterocyclic or alkyloxy. The term "alkyloxy" refers to _-ORb , where _Rb is a hydrocarbon. For example, the expression "C4-C30carbonyl" refers to "-C(=O)Ra" where _Ra has 3 to 29 carbon atoms, specifically, the number of carbon atoms may be 3, 9, 11, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, and _Ra may be a substituted or unsubstituted hydrocarbon, _a substituted or unsubstituted aryl, a substituted or unsubstituted three-membered ring, a four-membered ring, a five-membered ring, a substituted or unsubstituted alkyloxy.

The term "C15-C30 hydrocarbon group" means having 15 to 30 carbon atoms in the hydrocarbon structure, specifically, the number of carbon atoms can be 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, and the term "C1-C16 hydrocarbon group" means having 1 to 16 carbon atoms in the hydrocarbon structure, specifically, the number of carbon atoms can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16.

The term "aryl" or "aryl ring" used in the present invention refers to an aromatic group having "4n+2" (π) electrons in a conjugated monocyclic or polycyclic system (condensed or non-condensed), and having 6 to 14 ring atoms, wherein n is an integer from 1 to 3. The polycyclic system includes at least one aromatic ring. The aryl group can be directly connected or connected through a C1-C3 alkyl group (also referred to as an arylalkyl group or an aralkyl group). Examples of aryl groups include, but are not limited to, phenyl, benzyl, phenethyl, 1-phenylethyl, tolyl, naphthyl, biphenyl, terphenyl, indenyl, benzocyclooctenyl, benzocycloheptenyl, azulenyl, acenaphthenyl, fluorenyl, phenanthrenyl, anthracenyl, etc. The term "aryl" includes unsubstituted aryl and substituted aryl.

The term "heteroaryl" or "heteroaryl ring" as used herein refers to an aromatic group having "4n+2" (π) electrons in a conjugated monocyclic or polycyclic system (fused or non-fused), wherein n is an integer from 1 to 3 and includes one to six heteroatoms (e.g., N, O, S, P) or a group including heteroatoms (e.g., NH, NRx (Rx is alkyl, acyl, aryl, heteroaryl or cycloalkyl), _PO2 , SO, _SO2 , etc.). The polycyclic system includes at least one heteroaromatic ring. The heteroaryl group can be directly attached or attached through a C1-C3 alkyl group (also called heteroarylalkyl or heteroaralkyl). The heteroaryl group can be attached to carbon or to a heteroatom (e.g., through a nitrogen atom). Examples of heteroaryl groups include, but are not limited to, pyridyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, tetrazolyl, furanyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolidinyl, quinolyl, isoquinolyl, indolyl, isoindolyl, chromenyl, isochromenyl, benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, pyrazinyl, triazinyl, isoindolyl, pteridinyl, furanyl, benzofuranyl, benzothiazolyl, benzothiophenyl, benzothiazolyl, benzo The term "heteroaryl" includes unsubstituted heteroaryl and substituted heteroaryl. The term "C5-Cn heteroaryl", wherein n is an integer from 6 to 15, means a heteroaryl having from 5 to "n" atoms in the ring structure, including at least one heterocyclic group or atom as defined above.

In some embodiments, the present invention provides a phosphonate compound, or a pharmaceutically acceptable salt or ester thereof, for use in the preparation of a medicament for treating, inhibiting or preventing a viral infection or a cell proliferation disease, wherein the phosphonate compound is selected from but not limited to the compounds listed in Table 1.

The term "amino acid residue" refers to the major portion of an amino acid after removal of the carboxyl group.

As used herein, the term "amino acid" generally refers to an organic compound that contains both a carboxylic acid group and an amine group. The term "amino acid" includes "natural" and "unnatural" amino acids. In addition, the term "amino acid" includes O-alkylated or N-alkylated amino acids, as well as amino acids with nitrogen-, sulfur- or oxygen-containing side chains (e.g., Lys, Cys or Ser), wherein the nitrogen, sulfur or oxygen atom may or may not be acylated or alkylated. The amino acid may be an L-amino acid, a D-amino acid or a mixed L- and D-amino acid, including (but not limited to) a racemic mixture.

The term "natural amino acid" and equivalent expressions used in the present invention refer to L-amino acids that are usually found in naturally occurring proteins. Examples of natural amino acids include, but are not limited to, alanine (Ala), cysteine (Cys), aspartic acid (Asp), glutamic acid (Glu), phenylalanine (Phe), glycine (Gly), histidine (His), isoleucine (Ile), lysine (Lys), leucine (Leu), methionine (Met), asparagine (Asn), proline (Pro), glutamine (Gln), arginine (Arg), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), β-alanine (β-Ala) and γ-aminobutyric acid (GABA), etc.

The term "non-natural amino acid" used in the present invention refers to any derivative of natural amino acids, including D-amino acids and their derivatives, as well as α- and β-amino acid derivatives. It should be noted that in the present invention, some non-natural amino acids (such as hydroxyproline) may exist in certain biological tissues or specific proteins in nature. Amino acids with many different protecting groups suitable for direct application in solid phase peptide synthesis can be purchased. In addition to the twenty most common natural amino acids, the following examples of non-natural amino acids and amino acid derivatives (common abbreviations in brackets) can be used according to the present invention: 2-aminoadipic acid (Aad), 3-aminoadipic acid (β-Aad), 2-aminobutyric acid (2-Abu), α, β-dehydro-2-aminobutyric acid (8-AU), 1-aminocyclopropane-1-carboxylic acid (ACPC), aminoisobutyric acid (Aib), 3-aminoisobutyric acid (β-Aib), 2-amino-thiazoline-4-carboxylic acid, 5-amino Valeric acid (5-Ava), 6-aminohexanoic acid (6-Ahx), 2-aminoheptanoic acid (Ahe), 8-aminooctanoic acid (8-Aoc), 11-aminoundecanoic acid (11-Aun), 12-aminododecanoic acid (12-Ado), 2-aminobenzoic acid (2-Abz), 3-aminobenzoic acid (3-Abz), 4-aminobenzoic acid (4-Abz), 4-amino-3-hydroxy-6-methylheptanoic acid (Statine, Sta), aminooxyacetic acid (Aoa), 2-aminotetralin-2-carboxylic acid (ATC), 4-amino-5-cyclohexyl-3-hydroxypentanoic acid (ACHPA), p-aminophenylalanine (4-NH2-Phe), 2-aminopimelic acid (Apm), biphenylalanine (Bip), p-bromophenylalanine (BIP), 4-Br-Phe, o-chlorophenylalanine (2-Cl-Phe), m-chlorophenylalanine (3-Cl-Phe), p-chlorophenylalanine (3-Cl-Phe), m-chlorotyrosine (3-Cl-Tyr), p-benzoylphenylalanine (Bpa), tert-butylglycine (TLG), cyclohexylalanine (Cha), cyclohexylglycine (Chg), desmosine (Des), 2,2-diaminopimelic acid (Dpm), 2,3-diaminopropionic acid (Dpr), 2,4-diaminobutyric acid (Dbu), 3,4-dichlorophenylalanine (3,4-Cl2-Phe), 3,4-difluorophenylalanine (3,4-F2-Phe), 3,5-diiodotyrosine (3,5-I2-Tyr), N-ethylglycine (Et Gly), N-ethylasparagine (EtAsn), o-fluorophenylalanine (2-F-Phe), m-fluorophenylalanine (3-F-Phe), p-fluorophenylalanine (4-F-Phe), m-fluorotyrosine (3-F-Tyr), homoserine (Hse), homophenylalanine (Hfe), homotyrosine hydroxylysine (Hyl), isohydroxylysine (aHyl), 5-hydroxytryptophan (5-OH-Trp), 3- or 4-hydroxyproline (3- or 4-Hyp), p-iodophenylalanine-isotrosine (3-I-Tyr), indoline-2-carboxylic acid (Idc), isoiduromicin (Ide), isoleucine (α-Ile), isopenepic acid (Inp), N-methylisoleucine (MeLys), m-methyltyrosine (3 -Me-Tyr), N-methylvaline (MeVal), 1-naphthylalanine (1-Nal), 2-naphthylalanine (2-Nal), p-nitrophenylalanine (4-NO2-Phe), 3-nitrotyrosine (3-NO2-Tyr), norleucine (Nle), norvaline (Nva), ornithine (Orn), o-phosphotyrosine (H2PO3-Tyr), octahydroindole-2-carboxylic acid (Penicillamine), pentafluorophenylalanine (F5-Phe), phenylglycine (Phg), pipecolic acid (Pip), propargylglycine (Pra), pyroglutamic acid (PGLU), sarcosine (Sar), tetrahydroisoquinoline-3-carboxylic acid (Tic), thiazolidine-4-carboxylic acid (thioproline, Th).

The present invention provides methods for treating mammalian diseases associated with viral infection, inappropriate cell proliferation, etc. These methods specifically comprise administering to a human or other mammal in need of treatment a therapeutically effective amount of a compound of the present invention.

In some embodiments, the compounds provided herein can be used to prepare a drug that can be used to treat, inhibit or prevent viral infection or a disease caused by viral infection. In some embodiments, the compounds provided herein and their pharmaceutically acceptable salts or esters are used to prepare a drug for treating smallpox virus infection or a disease caused by smallpox virus. In some embodiments, the compounds provided herein and their pharmaceutically acceptable salts or esters are used to prepare a drug for treating monkeypox virus infection or a disease caused by monkeypox. Drugs for diseases caused by viruses.

The compounds provided herein have good effects in treating, inhibiting or preventing viral infections and diseases caused by viral infections. At the same time, the applicant has found that the compounds provided herein also have good effects in treating cancer or tumors. In some embodiments, the compounds provided herein and their pharmaceutically acceptable salts or esters can be used to prepare drugs, which can be used to treat, inhibit or prevent tumors or cancers caused by cell proliferation.

In some embodiments, the drug provided herein further comprises at least one pharmaceutically acceptable carrier or diluent. In some embodiments, the pharmaceutically acceptable carrier or diluent is selected from creams, emulsions, gels, liposomes or nanoparticles.

The compound or medicine provided by the present invention can be applied to the subject in any appropriate manner known in the art. Suitable routes of administration include, but are not limited to, oral; parenteral, such as intramuscular, intravenous, subcutaneous (such as injection or implantation), intraperitoneal, intracisternal, intraarticular, intracerebral (intracerebral parenchyma and intraventricular); nasal; vaginal; sublingual; intraocular; rectal; topical (such as transdermal); oral and inhaled. The accumulation injection method generally administered subcutaneously or intramuscularly can also be used to release the compound or medicine disclosed in the present application within a limited time period. In some embodiments, the medicine is an injectable preparation. In other embodiments, the medicine is formulated for oral administration to the subject.

The present invention also provides a kit comprising an antiviral infection compound or drug. The kit is generally in the form of a physical structure that accommodates various components, and can be used, for example, to implement the method provided herein. For example, the kit may include one or more compounds or drugs disclosed in the present invention (e.g., provided in a sterile container), which may be in the form of a pharmaceutical composition suitable for administration to a subject. The compound may be provided in a ready-to-use form (e.g., tablet or capsule) or in a form that requires, for example, reconstruction or dilution (e.g., powder) before administration. When the compound is in a form that requires the user to be reconstructed or diluted, the kit may also include a diluent (e.g., sterile water), a buffer, a pharmaceutically acceptable excipient, etc. that is packaged together with the compound or packaged separately. When a combination therapy is used, the kit may contain several therapeutic agents independently, or they may have been combined in the kit. Each component of the kit may be encapsulated in a separate container, and all the various containers may be in a single package. The kit of the present invention may be designed to appropriately maintain the conditions required for the components contained therein (e.g., refrigeration or freezing).

The medicaments or pharmaceutical compositions described herein can be prepared by any method known in the art of pharmacology. In general, such preparation methods include the steps of combining a compound described herein ("active ingredient") with a carrier and/or one or more other auxiliary ingredients, and then, if necessary and/or desired, forming and/or packaging the product into a desired single-dose unit or multiple-dose units. Bit.

Wherein the drug or pharmaceutical composition can be prepared, packaged and/or sold in batches as a single unit dose and/or as a plurality of single unit doses. As used herein, a "unit dose" is a discrete amount of a pharmaceutical composition containing a predetermined amount of an active ingredient. The amount of the active ingredient is generally equal to the dose of the active ingredient to be administered to a subject and/or a convenient fraction of such a dose, such as half or one-third of such a dose.

In order to better understand the present invention and to more clearly show how to implement the present invention, the features of the embodiments according to the present invention are now described by way of examples.

Example

The present invention will be more readily understood by reference to the following examples, which are provided to illustrate the present invention and are not to be construed as limiting the scope of the present invention in any way.

The compounds provided by the present invention can be synthesized according to the following general formula, wherein all reagents used to prepare the compounds of the present invention are commercially available or prepared according to preparation methods disclosed in the literature.

Compound A is used as a starting material, and under the action of a condensation agent 1H-benzotriazole-1-yloxytripyrrolidino hexafluorophosphate (PYBOP), it undergoes cyclization and condensation reaction with different alcohols to obtain compound B; under the action of a base such as potassium carbonate, the amino group on compound B reacts with an acyl chloride to obtain compound C, which is then hydrolyzed with a sodium hydroxide solution and acidified to obtain compound D;

The amino group on the intermediate compound B is protected by Boc anhydride to obtain compound E, which is then hydrolyzed with sodium hydroxide solution to obtain a ring-opened compound F. Compound F is condensed with an acyl chloride to obtain compound G, which is then deprotected to obtain compound H.

All R groups such as R ¹ , R ² , R ³ , R ⁴ in the general formula are as defined in the specification and claims.

Unless otherwise defined or the context clearly dictates otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention belongs. It should be understood that any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention.

Synthesis of intermediates

Synthesis of intermediate 1

Step A: Add diethoxymethanol phosphate (20g, 118.96mmol, 1.0eq.), dichloromethane (400mL) and triethylamine (14.44g, 142.75mmol, 1.2eq.) to the reaction flask. After cooling to -60°C, slowly drop trifluoromethanesulfonic anhydride (38.6g, 136.80mmol, 23.02mL, 1.15eq.) into the reaction system and stir the reaction for half an hour. Slowly warm to room temperature and stir the reaction for 1 hour. After concentrating to remove dichloromethane, add ethyl acetate (500mL), wash the organic phase with water, 1N dilute hydrochloric acid, and brine. Dry over anhydrous sodium sulfate, filter and concentrate to obtain the product diethoxymethyl trifluorophosphate. Mesylate (28 g, yield: 78.41%).

Step B: Compound a (4.9 g, 36.26 mmol, 1.0 eq.), DMF (100 mL), DBU (1.1 g, 7.25 mmol, 1.08 mL, 0.2 eq.) and (S)-2-((triphenoxy)methyl)oxirane (10.33 g, 36.26 mmol, 0.9 eq.) were added to a reaction flask. The mixture was stirred at 100°C for 6 hours, and the reaction solvent was removed by concentration. The residue was passed through a silica gel column (dichloromethane:methanol=100:0-94:6) to obtain product b (7 g, yield: 42.75%).

Step C: Add compound b (5 g, 11.07 mmol, 1.0 eq.) and tetrahydrofuran (100 mL) to the reaction flask. Cool to -60 °C, slowly add LiHMDs (1.0 M, 33.22 mL, 3.0 eq.). After stirring at -60 °C for 15 minutes, slowly add diethoxy methyl trifluoromethanesulfonate solution (8.30 g dissolved in 20 mL tetrahydrofuran, 28.79 mmol, 2.6 eq.). Warm to room temperature and stir for 16 hours. After concentrating to remove most of the reaction solvent, add ethyl acetate (300 mL), wash the organic phase with water and brine, dry with anhydrous sodium sulfate, filter and concentrate, and pass the residue through a silica gel column (dichloromethane: methanol = 100: 0-92: 8) to obtain product c (3.5 g, yield: 52.53%).

Step D: Add c (3.5 g, 5.82 mmol, 1 eq.), acetic acid (50 mL) and water (12 mL) to the reaction flask. Stir the reaction at 90°C for 3 hours, filter and concentrate, and pass the residue through a silica gel column (dichloromethane: methanol = 100: 0-90: 10) to obtain product d (1.6 g, yield: 76.54%).

Step E: Add d (0.8 g, 2.23 mmol, 1 eq.), acetonitrile (10 mL) and TMSBr (1.36 g, 8.91 mmol, 1.18 mL, 4.0 eq.) to a reaction flask, stir at room temperature for 16 hours, concentrate to remove the reaction solvent, add water to the residue, and freeze-dry to obtain product e (675 mg, yield: 99.10%).

Step F: Add e (0.2 g, 0.66 mmol, 1 eq.), DMF (10 mL), and DIPEA (1 mL) to the reaction flask. Stir the reaction at 45°C for 2 hours. Concentrate and remove the solvent to obtain a residue, add DMF (10 mL), 2-(octadecyloxy)ethane-1-ol (0.31 g, 0.99 mmol, 1.5 eq.), DIPEA (0.51 g, 3.96 mmol, 0.69 mL, 6.0 eq.) and pyBOP (1.03 g, 1.98 mmol, 3.0 eq.). Stir the reaction at 45°C for 16 hours, concentrate and remove the reaction solvent, and purify the residue by silica gel column separation (dichloromethane: methanol = 100: 0-90: 10) to obtain product f (0.22 g, yield 56.08%). ¹ H NMR (500MHz, CDCl ₃ )δppm:0.87(t,J＝6.8Hz,3H),1.24(s,30H),1.54(s,2H),3.43(dt,J＝13.6,6.7Hz,2H),3.54-3.70(m,2H),3.89(dd, J＝13.1,6.2Hz,1H),4.04-4.66(m,8H),6.02(d,J＝34.8Hz,2H),7.87(d,J＝3.0Hz,1H),8.34(d,J＝3.8Hz,1H); m/z(ESI ⁺ ):582.5(M+H).

Step G: Add f (70 mg, 0.12 mmol, 1 eq.) and sodium hydroxide aqueous solution (0.5 M, 1.2 mmol, 5.0 eq.) to the reaction flask. Stir and react for 48 hours at room temperature. The system became clear. In an ice-water bath, 1N HCl solution was slowly added dropwise to adjust the pH to about 1. A large amount of solid precipitated, which was filtered and dried in vacuo to obtain intermediate 1 (65 mg, yield 84.56%). ¹ H NMR (500MHz, CDCl ₃ )δppm:0.84(t,J＝6.8Hz,3H),1.21(s,30H),1.51(d,J＝6.7Hz,2H),3.42(t ,J＝7.0Hz,2H),3.49(d,J＝8.9Hz,1H),3.57(t,J＝4.5Hz,2H),3.70(dd,J＝21 .7,8.8Hz,2H),3.84(d,J＝8.7Hz,2H),4.07(d,J＝6.1Hz,2H),4.34(dd,J＝1 4.2,7.3Hz,1H),4.47(d,J＝11.7Hz,1H),8.15(s,1H),8.30(s,1H); m/z(ESI ⁺ ):600.6(M+H).

Synthesis of compounds

Example 1: Synthesis of Compound 1b

Step A: Add intermediate 1 (50 mg, 0.083 mmol, 1.0 eq.), DMF (3 mL) and NaH (33.35 mg, 0.833 mmol, 60% purity, 10.0 eq.) to the reaction flask. Stir at room temperature for 20 minutes. 4-nitrophenylbutyrate (34.22 mg, 0.208 mmol, 2.5 eq.) DMF (0.5 mL) solution was added dropwise to the reaction. Stir at room temperature for 3 hours, then heated to 50 ° C for 2 hours. Add ethyl acetate (25 mL) to dilute, add 50% acetic acid aqueous solution at low temperature, and then add 1N HCl to quench the reaction, ensuring that the pH of the aqueous phase is 1-2. The organic layer was washed twice with concentrated brine, dried and concentrated, and the residue was separated and purified by silica gel column (dichloromethane: methanol = 100: 0-80: 20) to obtain the product compound 1b (22 mg, yield 32.30%). ¹ H NMR (500MHz, CDCl ₃ )δppm:0.84(t,J＝6.8Hz,3H),0.91(t,J＝7.4Hz,3H),0.97(t,J＝7.3Hz,3H),1.20(d,J＝11.6Hz,30H),1.44(s,2 H),1.60(dq,J＝14.5,7.2Hz,2H),1.74(dd,J＝14.6,7.2Hz,2H),2.27(t,J＝7.3Hz,2H),2.67(t,J＝7.1Hz,2H),3 .31(t,J＝6.9Hz,2H),3.37(s,2H),3.47(t,J＝11.8Hz,1H),3.83(dd,J＝25.5,18.8Hz,4H),4.04(d,J＝7.5Hz,1H ), 4.14 (d, J = 6.6Hz, 1H), 4.29 (dd, J = 14.5, 7.4Hz, 1H), 4.58 (d, J = 13.1Hz, 1H), 8.65 (d, J = 12.2Hz, 2H); m/z (ESI ⁺ ):740.

Example 2: Synthesis of Compound 2b

Step A: Add 4-nitrophenol (2.0 g, 14.38 mmol, 1 eq.) and Py (4 mL) to the reaction flask, add acetic anhydride (2.20 g, 21.57 mmol, 1.5 eq.) dropwise under ice bath, stir and react at room temperature for 16 hours. TLC monitors the consumption of the reaction raw materials, and 1 mol/L The reaction system was adjusted to a weakly acidic state with an aqueous solution of hydrochloric acid, and water (80 mL) and dichloromethane were added for extraction and washing (30 mL*2). The organic layer was washed once with saturated saline (60 mL), the organic layer was separated, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated and purified by a silica gel column (dichloromethane) to obtain the product 4-nitrophenyl acetate (2.2 g, yield 84.47%). Intermediate 1 (100 mg, 166.74 μmol, 1 eq.) and DMF (2 mL) were added to the reaction flask, and NaH (53.36 mg, 1.33 mmol, 60% purity, 8 eq.) was added under ice bath, and the reaction was stirred at 0°C for 20 min, and then a solution of 4-nitrophenyl acetate (120.82 mg, 666.96 μmol, 4 eq.) in DMF (1 mL) was added dropwise, and the reaction was stirred at 50°C for 2 hours after the addition. LC-MS monitoring confirmed that the reaction raw materials were consumed. The reaction system was adjusted to pH 2 with 1 mol/L hydrochloric acid aqueous solution under ice-water bath conditions. Water (50 mL) was added and washed with ethyl acetate (20 mL*4). The organic layer was washed once with saturated brine (60 mL). The organic layer was separated, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated and purified using a preparative plate to obtain compound 2b (42 mg, yield 46.27%). ¹ H NMR(500MHz,MeOD)δ0.91-0.96(m,3H),1.31(s,30H),1.51(s,2H),2.03(s,2H),2.40(s,2H),3.42(s,2H),3.55(s,2H),3 .64-3.73(m,1H),3.84-3.99(m,2H),4.09-4.21(m,3H),4.51-4.58(m,1H),4.62-4.69(m,2H),8.61-8.80(m,2H); m/z(ESI ⁺ ):684.7(M+H).

Example 3: Synthesis of Compound 3b

Step A: 2-(octadecyloxy)ethyl hydrogen (((S)-1-(6-amino-9H-purin-9-yl)-3-hydroxypropyl-2-yl)oxy)methyl)phosphonate (30 mg, 0.05 mmol, 1 eq.) and formic acid (4 mL) were added to a reaction flask. The mixture was stirred at 60°C for 16 hours, and the formic acid was removed by concentration. Methanol (5 mL) was added to the residual crude product, and the mixture was stirred for 30 minutes. The solid was filtered to obtain a solid. After the solid was dispersed in water, it was lyophilized to obtain compound 3b (20 mg, yield 59.88%). ¹ H NMR (500MHz, CD ₃ OD)δppm:0.88(d,J＝6.9Hz,3H),1.27(s,30H),1.51(s,2H),3.42(d,J＝6.6Hz,2H),3.55(s,2H),3.72(s,1H),3.82-3.90(m,1H),3.97(s,2H), 4.13(d,J＝11.4Hz,2H),4.31(d,J＝7.7Hz,1H),4.45(d,J＝8.1Hz,1H),4.60(d,J＝13.5Hz,1H),8.12(s,1H),8.30(s,1H),8.45(s,1H); m/z(ESI ⁺ ):628.39(M+H).

Example 4: Synthesis of Compound 4b

Step A: In a reaction flask, intermediate 1 (80 mg, 133.39 μmol, 1 eq.), DIPEA (68.96 mg, 533.56 μmol, 4 eq.) and DMF (2 mL) were added, and then 1-(((4-nitrobenzene) was added dropwise. oxy)carbonyl)oxy)ethyl isobutyrate (79.30 mg, 266.78 μmol, 2 eq.) in DMF (1 mL), after the addition, the reaction was stirred at 80°C for 6 hours. LC-MS monitoring showed that the reaction raw materials were basically consumed, and the reaction system was adjusted to pH 2 with 1 mol/L hydrochloric acid aqueous solution under ice-water bath conditions, water (50 mL) was added, and ethyl acetate was extracted and washed (20 mL*4), and the organic layer was washed once with saturated brine (60 mL), the organic layer was separated, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by reverse phase preparative column (acetonitrile, water, ammonium acetate system), and lyophilized to obtain compound 4b (20 mg, yield 18.83%).

¹ H NMR(500MHz,MeOD)δ0.93(t,J＝6.7Hz,3H),1.16-1.21(m,6H),1.30(d,J＝10.7Hz,30H),1.50-1.58(m,5H ),2.60(dt,J＝13.8,6.9Hz,1H),3.47(t,J＝6.3Hz,2H),3.60(d,J＝3.5Hz,2H),3.77-3.85(m,1H),3.88 -3. 95(m,1H),4.03(s,2H),4.09-4.18(m,1H),4.22(s,1H),4.36(ddd,J＝28.4,11.5,4.2Hz,1H),4.47(dd, J= 14.5,5.6Hz,1H),4.61(d,J＝14.7Hz,1H),6.70-6.75(m,1H),8.29(d,J＝6.4Hz,1H),8.46(s,1H); m/ z(ESI ⁺ ):758.5(M+H).

Example 5: Synthesis of Compound 5b

Step A: Intermediate 1 (100 mg, 166.74 μmol, 1 eq.), DIPEA (86.20 mg, 666.96 μmol, 4 eq.) and DMF (4 mL) were added to a reaction flask, and then a solution of ethyl 1-(((4-nitrophenoxy)carbonyl)oxy)acetate (67.33 mg, 250.11 μmol, 1.5 eq.) in DMF (1 mL) was added dropwise. The mixture was stirred at 80°C for 5 hours. LC-MS monitoring showed that the reaction raw materials were basically consumed. The reaction system was adjusted to pH 2 with 1 mol/L hydrochloric acid aqueous solution under ice-water bath conditions. Water (50 mL) was added, and ethyl acetate was extracted and washed (20 mL*4). The organic layer was washed once with saturated brine (60 mL). The organic layer was separated, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by reverse ^phase preparative column (acetonitrile, water, ammonium acetate system) and freeze-dried to obtain compound 5b (40 mg, yield 31.08%). NMR(500MHz,MeOD)δ0.93(t,J＝6.6Hz,3H),1.32(s,30H),1.48-1.56(m,5H),2.10(s,3 H),3.45(t,J＝6.0Hz,2H),3.55-3.61(m,2H),3.74(q,J＝11.2Hz,1H),3.86-3.93(m,1H) ,3.96-4.04(m,2H),4.13-4.19(m,2H),4.26-4.39(m,1H),4.45-4.51(m,1H),4.56-4. 63(m,1H),6.71(q,J＝5.1Hz,1H),8.28(d,J＝4.8Hz,1H),8.44(d,J＝3.0Hz,1H); m/z(ESI ⁺ ):730.5(M+H).

Example 6: Synthesis of Compound 6b

Step A: Add intermediate 1 (200 mg, 0.34 mmol, 1 eq.) and formic acid (10 mL) to the reaction flask. Stir the reaction at 60°C for 16 hours, concentrate to remove formic acid, add methanol (5 mL) and acetonitrile (5 mL) to the residual crude product, stir for 10 minutes, filter to obtain a solid, disperse the solid in methanol (15 mL), continue stirring for 10 minutes, and filter to obtain a solid. After the solid is dispersed in water, freeze-dried to obtain compound 6b (150 mg, yield 69.87%). ¹ H NMR (500 MHz, CD ₃ OD)δppm:0.89(t,J＝6.7Hz,3H),1.27(s,26H),1.51(d,J＝6.8Hz,2H),1.82(p,J＝6.4Hz,2H) ,3.39(t,J＝6.6Hz,2H),3.48(t,J＝6.3Hz,2H),3.67(dd,J＝13.1,8.9Hz,1H),3.85(dd,J＝13 .2,8.9Hz,1H),3.92(q,J＝6.4Hz,2H),4.09-4.21(m,2H),4.33(d,J＝7.0Hz,1H),4.46(dd,J ＝14.6,6.3Hz,1H),4.59(d,J＝14.6Hz,1H),8.13(s,1H),8.31(s,1H),8.42(s,1H); m/z(ESI ⁺ ):614.72(M+H).

Comparative Example 1 (Synthesis of ODE-(S)-HPMPA)

Step A: Add diethoxymethanol phosphate (20 g, 118.96 mmol, 1.0 eq.), dichloromethane (400 mL) and triethylamine (14.44 g, 142.75 mmol, 1.2 eq.) to the reaction flask. After cooling to -60 °C, slowly drop trifluoromethanesulfonic anhydride (38.6 g, 136.80 mmol, 23.02 mL, 1.15 eq.) into the reaction system and stir for half an hour. Slowly warm to room temperature and stir for 1 hour. After concentrating to remove dichloromethane, add ethyl acetate (500 mL), wash the organic phase with water, 1N dilute hydrochloric acid, and brine. Dry over anhydrous sodium sulfate, filter and concentrate to obtain the product diethoxymethyl trifluoromethanesulfonate (28 g, yield: 78.41%).

Step B: Add 9H-purine-6-amine (4.9 g, 36.26 mmol, 1.0 eq.) and DMF (100 mL), DBU (1.1 g, 7.25 mmol, 1.08 mL, 0.2 eq.) and (S)-2-((triphenoxy)methyl)oxirane (10.33 g, 36.26 mmol, 0.9 eq.) were stirred at 100°C for 6 hours, the reaction solvent was removed by concentration, and the residue was passed through a silica gel column (dichloromethane: methanol = 100: 0-94: 6) to obtain the product 1-1 (7 g, yield: 42.75%).

Step C: Add 1-1 (5 g, 11.07 mmol, 1.0 eq.) and tetrahydrofuran (100 mL) to the reaction flask. Cool to -60 °C, slowly add LiHMDs (1.0 M, 33.22 mL, 3.0 eq.). After stirring at -60 °C for 15 minutes, slowly add diethoxy methyl trifluoromethanesulfonate solution (8.30 g dissolved in 20 mL tetrahydrofuran, 28.79 mmol, 2.6 eq.). Warm to room temperature and stir for 16 hours. After concentrating to remove most of the reaction solvent, add ethyl acetate (300 mL), wash the organic phase with water and brine, dry over anhydrous sodium sulfate, filter and concentrate, and pass the residue through a silica gel column (dichloromethane: methanol = 100: 0-92: 8) to obtain the product 1-2 (3.5 g, yield: 52.53%).

Step D: Add 1-2 (3.5 g, 5.82 mmol, 1 eq.), acetic acid (50 mL) and water (12 mL) to a reaction flask. Stir the reaction at 90°C for 3 hours, filter and concentrate, and pass the residue through a silica gel column (dichloromethane: methanol = 100: 0-90: 10) to obtain product 1-3 (1.6 g, yield: 76.54%).

Step E: 1-3 (0.8 g, 2.23 mmol, 1 eq.), acetonitrile (10 mL) and TMSBr (1.36 g, 8.91 mmol, 1.18 mL, 4.0 eq.) were added to a reaction flask, stirred at room temperature for 16 hours, concentrated to remove the reaction solvent, added water to the residue, and freeze-dried to obtain product 1-4 (675 mg, yield: 99.10%).

Step F: 1-4 (0.2 g, 0.66 mmol, 1 eq.), DMF (10 mL), and DIPEA (1 mL) were added to the reaction flask. The reaction was stirred at 45°C for 2 hours. The solvent was removed by concentration to obtain a residue, and DMF (10 mL), 2-(octadecyloxy)ethane-1-ol (0.31 g, 0.99 mmol, 1.5 eq.), DIPEA (0.51 g, 3.96 mmol, 0.69 mL, 6.0 eq.) and pyBOP (1.03 g, 1.98 mmol, 3.0 eq.) were added. The reaction was stirred at 45°C for 16 hours, and the reaction solvent was removed by concentration. The residue was separated and purified by silica gel column (dichloromethane: methanol = 100: 0-90: 10) to obtain 1-5 (0.22 g, yield 56.08%). ¹ H NMR (500MHz, CDCl ₃ )δppm:0.87(t,J＝6.8Hz,3H),1.24(s,30H),1.54(s,2H),3.43(dt,J＝13.6,6.7Hz,2H),3.54-3.70(m,2H),3.89(dd, J＝13.1,6.2Hz,1H),4.04-4.66(m,8H),6.02(d,J＝34.8Hz,2H),7.87(d,J＝3.0Hz,1H),8.34(d,J＝3.8Hz,1H); m/z(ESI ⁺ ):582.5(M+H).

Step G: Add 1-5 (70 mg, 0.12 mmol, 1 eq.) and sodium hydroxide aqueous solution (0.5 M, 1.2 mmol, 5.0 eq.) to the reaction flask. Stir the reaction at room temperature for 48 hours. The reaction system becomes clear. In an ice-water bath, slowly add 1N HCl solution dropwise to adjust the pH to about 1. A large amount of solid precipitates, which is filtered and dried in vacuo to obtain Comparative Example 1 (65 mg, yield 84.56%). ¹ H NMR (500 MHz, CDCl ₃ )δppm:0.84(t,J＝6.8Hz,3H),1.21(s,30H),1.51(d,J＝6.7Hz,2H),3.42(t ,J＝7.0Hz,2H),3.49(d,J＝8.9Hz,1H),3.57(t,J＝4.5Hz,2H),3.70(dd,J＝21 .7,8.8Hz,2H),3.84(d,J＝8.7Hz,2H),4.07(d,J＝6.1Hz,2H),4.34(dd,J＝1 4.2,7.3Hz,1H),4.47(d,J＝11.7Hz,1H),8.15(s,1H),8.30(s,1H); m/z(ESI ⁺ ):600.6(M+H).

Comparative Example 2:

Steps A-E are the same as those of Comparative Example 1

Step F: Add 2-1 (0.675 g, 2.23 mmol, 1 eq.), DMF (25 mL), and DIPEA (3 mL) to the reaction flask. Stir the reaction at 45°C for 2 hours. Concentrate and remove the solvent to obtain a residue, add DMF (25 mL), 3-(hexadecyloxy)propanol (1 g, 3.34 mmol, 1.5 eq.), DIPEA (1.73 g, 13.36 mmol, 2.33 mL, 6.0 eq.) and pyBOP (3.48 g, 6.68 mmol, 3.0 eq.). Stir the reaction at 45°C for 4 hours, concentrate and remove the reaction solvent, and purify the residue by silica gel column separation (dichloromethane: methanol = 100: 0-90: 10) to obtain the product 2-2 (0.6 g, yield 47.48%).

Step G: Add 2-2 (150 mg, 0.26 mmol, 1 eq.) and sodium hydroxide aqueous solution (0.5 M, 2.64 mmol, 5.0 eq.) to the reaction flask. Stir the reaction at room temperature for 4 hours. The reaction system becomes clear. In an ice-water bath, slowly dropwise add 1N HCl solution to adjust the pH to about 1. A large amount of solid precipitates, which is filtered and dried in vacuo to obtain the product comparative example 2 (80 mg, yield 51.29%). ¹ H NMR (500 MHz, CD ₃ OD)δppm:0.92(t,J＝5.9Hz,3H),1.31(s,26H),1.55(d,J＝5.9Hz,2H),1.80-1.93( m,2H),3.41(dd,J＝17.7,12.4Hz,2H),3.52(t,J＝5.5Hz,2H),3.59(d,J＝12.3Hz,1H ),3.70(t,J＝10.9Hz,2H),3.84(dd,J＝28.7,17.4Hz,2H),3.93-3.99(m,2H),4.47( dd,J＝14.1,7.0Hz,1H),4.58(d,J＝14.4Hz,1H),8.34(s,1H),8.41(s,1H); m/z(ESI ⁺ ):586.5(M+H).

Effect Example

1. Biological assay

General approach to pharmacokinetic studies:

The test compound is dissolved in PBS (pH=8) at a concentration determined by the desired dose and dosing volume for the particular animal to which the compound is administered. The animal is administered a measured volume of dosing solution orally. Blood samples are collected at specific time points (e.g., 0.5, 1, 2, 4, 8, 24, 32, and 48 hours) after administration of the test compound. The blood samples are converted to plasma samples using standard techniques. After LC-MS/MS analysis, the concentration of the test compound and the parent drug in the comparative example in plasma was obtained.

1.1. Pharmacokinetic study of the compounds of the present invention in mice

After the compound and the comparative example were administered by single intravenous injection/oral administration to fasted ICR male mice, blood samples were collected at 0.083h (IV), 0.25h (IV), 0.5h, 1h, 2h, 4h, 8h, 24h, 32h and 48h after administration. Plasma was separated by centrifugation (3200rpm, 10min, 4°C) and frozen (-80°C) until used for analysis. The concentration of the compound in mouse plasma was determined by UPLC-MS/MS. Plasma was distributed to a suitable tube containing internal standard and precipitant, and the tube was violently shaken for 1 minute to achieve deproteinization, followed by centrifugation at 12000rmp for 5 minutes. After the supernatant was transferred to a 96-well plate for dilution, the mixture was shaken and centrifuged again (4100rpm, 5min, 4°C), and quantitative analysis was performed by LC-MS/MS. DAS 3.2.8 software was used to calculate pharmacokinetic parameters such as AUC _0-t , AUC _0-∞ , Cmax, tmax, t1/2, MRT, CL and Vd. Absolute bioavailability was calculated as follows: F = [AUC (ig) × dose (iv)] / [AUC (iv) × dose (ig)] × 100%.

Compounds 2b and 3b and the compound of Comparative Example 1 were orally administered to ICR male mice at an equimolar amount of 42.7 mg/kg. The experimental method was as described above. The obtained pharmacokinetic parameters are shown in Table 2.

Table 2

Compound 6b and the compound of comparative example 2 were orally administered to ICR male mice at an equimolar amount of 41.7 mg/kg. The experimental method was as described above. The obtained pharmacokinetic parameters are shown in Table 3.

Table 3

2. Efficacy test

2.1. In vitro efficacy test

African green monkey kidney cells (Vero cells, ATCC CRL-1587) were inoculated into 96-well plates in a medium containing MEM (Gibco Biosystems, Inc.) containing 10% fetal bovine serum, 100 U/mL penicillin, and 100 μg/mL streptomycin. Cat#41500034), and Vaccinia virus, Tiantan strain, with an infection dose of 0.05 PFU/cell was added. After incubation at 37°C in a 5% CO ₂ incubator for 2 hours, 10 series of half-diluted drugs BCV (Brincidofovir), BCV prodrug 15b, compound 3b or compound 6b (10μM, 5μM, 2.5μM, 1.25μM, 0.63μM, 0.32μM, 0.16μM, 0.08μM, 0.04μM, 0.02μM) were added, with 3 replicate wells for each concentration. The cells were cultured continuously for 3 days, and the remaining cells were fixed with 4% neutral paraformaldehyde and stained with 0.1% crystal violet to count the number of viral plaques. Vero cells not infected with the virus were used as controls. Virus inhibition activity (%) = (number of virus plaques in the drug group/number of virus plaques in the control group) × 100.

The results of the in vitro test are shown in FIG1 . The inhibitory activities of BCV and BCV prodrug 15b are similar, while the inhibitory activities of the present compound 3b and compound 6b are similar, about 10-30 times that of BCV, with EC50 values of 0.068 μM and 0.165 μM, respectively.

2.2 In vivo efficacy test

60 male Balb/c nude mice were randomly divided into 5 groups, namely blank control group, BCV administration group, BCV prodrug 15b administration group, compound 3b administration group and compound 6b administration group. The experimental mice were inoculated with vaccinia virus on D1 of the experiment, and then gavaged with the corresponding drugs on D2, D4 and D6 of the experiment (the blank control group was only given the same volume of solvent), and the three administration doses were 20 mg/kg, 5 mg/kg and 5 mg/kg respectively. The weight changes of the animals before and after administration were recorded during the experiment to evaluate the protective effect of the test substance on the animals, and blood and tissues were taken for standby.

The weight of each group of mice during the experiment was recorded, and the weight of mice on D0 was used as the benchmark, and the weight loss of mice> 30% of the initial weight was used as the death surrogate endpoint. The results are shown in Figures 2a and 2b. It can be clearly observed that the mice in the blank control group gradually reached the surrogate endpoint from D3, a large number of mice reached the surrogate endpoint on D6 and D7, and all mice reached the surrogate endpoint on D8; the BCV administration group failed to improve the situation of mice reaching the surrogate endpoint, and all mice in this group also reached the surrogate endpoint on D8; while the situation of the compound 3b and compound 6b administration groups was significantly improved, among which compound 3b had the best effect, and only one mouse reached the surrogate endpoint on D14.

The weight of each group of mice during the experiment was recorded, and the weight of mice on D0 was used as the benchmark to obtain the weight change graph and the actual death curve, as shown in Figure 3a and Figure 3b. It can be clearly observed from the weight change graph that the weight of mice in the blank control group decreased significantly, and on D14 it was only 70%-80% of the weight on D0. The weight of mice in the BCV administration group decreased significantly at the beginning, and recovered in the later period, and on D14 it was close to the level of D0; the weight of mice in the compound 3b and compound 6b administration groups was always higher than that in the BCV administration group, among which the lowest point of the weight of mice in the compound 3b administration group was 80% of D0, and on D14 it reached 110% of D0, which was similar to the blank control group. The control group had significant differences. From the actual death curve, it can be observed that only 2 mice survived in the blank control group on D14, 10 mice survived in the BCV administration group on D14, all mice survived in the compound 3b administration group on D14, and 11 mice survived in the compound 6b administration group on D14.

Whether it is weight change, alternative endpoint death curve or actual death curve, the therapeutic effects of compound 3b and compound 6b are better than BCV. They can delay the virus-induced weight loss in mice, and even restore the weight growth of mice, and can also reduce the death of mice caused by the virus.

Although the present invention has been described in detail with reference to the embodiments of the present invention, these embodiments are provided to illustrate rather than limit the present invention. Other embodiments that can be obtained according to the principles of the present invention all belong to the scope defined by the claims of the present invention.

Claims (21)

A compound represented by formula (I) or a pharmaceutically acceptable salt or ester thereof:
Wherein, X is selected from -OR ⁴ , or X and R ³ are combined to form a chemical bond; ^R1 is selected from H or C4-C30 carbonyl, wherein the C4-C30 carbonyl includes substituted or unsubstituted hydrocarbylcarbonyl, substituted or unsubstituted arylcarbonyl or heterocyclic carbonyl, and substituted or unsubstituted hydrocarbyloxycarbonyl, and the carbon number of the carbonyl is not more than 30; R ² and R ⁴ are independently selected from H, Wherein R ⁵ is selected from substituted or unsubstituted C15-C30 hydrocarbon groups, and the hydrocarbon group may be a straight-chain hydrocarbon group or a branched hydrocarbon group; ^R3 is selected from H; formyl, C4-C30 carbonyl, wherein the C4-C30 carbonyl includes substituted or unsubstituted alkylcarbonyl, substituted or unsubstituted arylcarbonyl or heterocyclic carbonyl, and substituted or unsubstituted alkoxycarbonyl, and the carbonyl has a carbon number of not more than 30, or an amino acid residue; As a limitation, when X is -OR ⁴ , R ¹ , R ² , R ³ , and R ⁴ are not H at the same time; when X and R ³ are combined to form a chemical bond, R ¹ and R ² are not H at the same time. The compound according to claim 1, wherein the compound is a compound represented by formula (III);
As a limitation, when X is -OR ⁴ , the R ¹ , R ³ , and R ⁴ are not H at the same time; when X and R ³ When they form a chemical bond, the ^R1 is not H. The compound according to claim 2, wherein the compound is a compound represented by formula (IIIa);
The compound according to claim 1, wherein the compound is a compound represented by formula (IV);
As a limitation, when X is -OR ⁴ , R ¹ , R ³ , and R ⁴ are not H at the same time; when X and R ³ are combined to form a chemical bond, R ¹ is not H. The compound according to claim 4, wherein the compound is a compound represented by formula (IVa);
The compound according to any one of claims 1 to 5, wherein R ¹ is a substituted or unsubstituted hydrocarbon carbonyl group, and the substituted or unsubstituted hydrocarbon carbonyl group includes the following groups:
The compound according to any one of claims 1 to 6, wherein R ¹ is a substituted or unsubstituted arylcarbonyl or heterocyclic carbonyl group, and the substituted or unsubstituted arylcarbonyl or heterocyclic carbonyl group includes the following groups:

wherein Y is selected from H, F, Cl, Br, Me, -OMe, -OEt, _-CF3 or -CN. The compound according to any one of claims 1 to 6, wherein the R ¹ is a substituted or unsubstituted alkoxycarbonyl group, and the substituted or unsubstituted alkoxycarbonyl group includes the groups shown below:


The compound according to any one of claims 1 to 6, wherein the R ⁵ comprises the following groups:
The compound according to any of claims 1 to 6, wherein the R ³ comprises the following groups:

Preferably, ^R3 is selected from A compound selected from the following and a pharmaceutically acceptable salt thereof:

A pharmaceutical composition comprising at least one compound according to any one of claims 1 to 11, and at least one pharmaceutically acceptable carrier or excipient. The pharmaceutical composition according to claim 12, wherein the pharmaceutically acceptable carrier comprises one or more of a cream, an emulsion, a gel, a liposome and nanoparticles; and the pharmaceutically acceptable excipient comprises one or more of a binder, a filler, a disintegrant, a lubricant and a glidant. The pharmaceutical composition according to claim 12 or 13, wherein the pharmaceutical composition is suitable for oral administration or injection administration. Use of the compound according to any one of claims 1 to 11 or the pharmaceutical composition according to any one of claims 12 to 14 in the preparation of a medicament for treating, inhibiting or preventing viral infection or a disease caused by viral infection. The use according to claim 15, wherein the viral infection comprises infection caused by hepatitis B virus (HBV), new coronavirus (SARS-COV-2), human immunodeficiency virus (HIV), varicella zoster virus (VZV), cytomegalovirus (CMV), herpes simplex virus (HSV), BK virus, JC virus, Epstein-Barr virus (EBV), Ebola virus, polyomavirus, papillomavirus, orthopoxvirus, hepatitis C virus (HCV), respiratory syncytial virus (RSV), dengue virus, influenza virus, adenovirus, parainfluenza virus and/or rhinovirus. The use according to claim 16, wherein the orthopoxvirus includes severe and mild smallpox viruses, monkeypox virus, cowpox virus, camelpox virus, molluscum contagiosum, sheeppox virus, aractuba virus, BeAn 58058 virus, cantagalo orthopoxvirus, mousepox virus, elephantpox virus, vaccinia virus (VV), rabbitpox virus, raccoonpox virus, skunkpox virus, gerbilpox virus and volepox virus. The use according to claim 16, wherein the viral infection includes infection caused by smallpox virus and monkeypox virus. Use of the compound according to any one of claims 1 to 11 or the pharmaceutical composition according to any one of claims 12 to 14 in the preparation of a medicament for treating, inhibiting or preventing diseases caused by cell proliferation. The use according to claim 19, wherein the cell proliferation-induced disease includes a tumor, cancer or other viral infection disease, and the tumor or cancer is selected from multiple myeloma (MM), chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), solid tumors, refractory solid tumors, non-Hodgkin's lymphoma, hematological tumors, neuroblastoma, colorectal cancer, cervical cancer, lung cancer, leukemia, breast cancer, pancreatic cancer, B-cell malignancies, tumors, metastatic tumors, and colon cancer; the disease caused by viral infection is selected from diseases caused by DNA virus infection, including retinitis, pneumonia, cystitis, proteinopathy, etc. A kit comprising the compound or pharmaceutically acceptable salt or ester according to any one of claims 1 to 11 or the pharmaceutical composition according to any one of claims 12 to 14.