CN115960148A

CN115960148A - Novel cytidine derivative, pharmaceutical composition and application thereof

Info

Publication number: CN115960148A
Application number: CN202211039144.9A
Authority: CN
Inventors: 张哲峰; 孟月垒
Original assignee: Nanjing Zhihe Medical Technology Co ltd
Current assignee: Nanjing Zhihe Medical Technology Co ltd
Priority date: 2021-08-27
Filing date: 2022-08-29
Publication date: 2023-04-14
Anticipated expiration: 2042-08-29
Also published as: CN117886870A; CN115960148B; WO2023025319A1

Abstract

The invention discloses a novel cytidine derivative, a pharmaceutical composition and application thereof, wherein the cytidine derivative is shown as a formula (I) ₀ ) Shown; the compound can be used for preparing medicine for resisting virus infection.

Description

Novel cytidine derivative, pharmaceutical composition and application thereof

Technical Field

The invention relates to but not limited to the technical field of medicinal chemistry, in particular to a novel cytidine derivative, a medicinal composition and application thereof.

Background

Influenza is an acute respiratory infectious disease caused by infection with influenza virus. Is a particularly important disease in high-risk people such as infants, old people and the like, and has high pneumonia complication rate in the old people.

Since influenza viruses have high variability, development of anti-influenza virus drugs has been made far from priority due to concerns about emergence of drug-resistant strains or side effects, prevalence of pathogenic or lethal novel influenza viruses, and the like, and thus development of antiviral drugs of novel structures is still desired in the art.

Disclosure of Invention

The present inventors have developed a novel cytidine derivative having an antiviral action and low cytotoxicity.

In one aspect of the invention there is provided a compound as (I) ₀ ) Novel cytidine derivatives, tautomers, stereoisomers, isotopic derivatives, and pharmaceutically acceptable salts thereof, as shown:

formula (I) ₀ ) In the step (1), the first step,

R ₁ and R ₂ Each independently selected from hydrogen,

Or, R ₁ And R ₂ An acetal or ketal with the oxygen adjacent thereto;

wherein n is ₁ Selected from 0, 1, 2 or 3;

n ₂ selected from 1,2 or 3;

R _a and R _b Each independently selected from hydroxy, the following groups substituted or unsubstituted with one or more groups A: C1-C8 alkyl, C1-C8 alkoxy, C2-C8 alkenyl, C3-C8 cycloalkyl, C6-C18 aryl, aryloxy, arylalkyl, alkylaryl;

R _c and R _d Each independently selected from hydrogen, C1-C8 alkyl substituted or unsubstituted with one or more groups A;

R ₃ and R ₄ The same or different, are each independently selected from hydrogen or

R ₃ And R ₄ Not all may be hydrogen;

wherein n is _a Selected from 0, 1, 2, 3,4, or 5;

n _b selected from 1, 2, 3,4, or 5;

n ₃ selected from 0, 1, 2, 3,4, or 5;

n ₄ selected from 0, 1, 2, 3, or 4;

R ₅ and R ₆ Identical or different, independently selected from hydrogen, C1-C8 alkyl substituted or unsubstituted by one or more groups A; or R ₅ 、R ₆ To which carbon is attached to form a cycloalkyl group;

R ₇ is hydrogen, halogen, amino, C1-C8 alkyl substituted or unsubstituted by one or more groups A;

z is selected from

Wherein n is ₅ Each independently is 0, 1, 2, 3,4, or 5;

R ₈ selected from H, hydroxy, nitro, halogen, the following groups substituted or unsubstituted with one or more groups a: amino, C1-C8 alkyl, C6-C18 aryl, C1-C8 alkoxy, aminoalkyl, C1-C8 alkylaryl, arylcarbonyl, C1-C8 alkylcarbonyloxy;

R _e and R _f Each independently selected from hydrogen, the following groups substituted or unsubstituted with one or more groups A: C1-C8 alkyl, C3-C8 cycloalkyl, heterocycloalkyl, C6-C18 aryl, heteroaryl, non-aromatic heterocyclyl;

R _X1 、R _X2 、R _X3 、R _X4 、R _X5 、R _X6 、R _X7 and R _X8 Each independently selected from hydrogen, deuterium;

the group A is: hydroxyl, carboxyl, amino, halogen, cyano, aldehyde group, nitro, trifluoromethyl, C3-C8 cycloalkyl, C1-C8 alkoxy and chlorobenzoyl.

In some embodiments, the invention provides a compound of formula (I) ₀ -1) novel cytidine derivatives, tautomers, stereoisomers, isotopic derivatives, and pharmaceutically acceptable salts thereof:

formula (I) ₀ The substituents in (1) are as defined above.

In some embodiments, the invention provides a compound of formula (I) ₀ -2) novel cytidine derivatives, tautomers, stereoisomers, isotopic derivatives, and pharmaceutically acceptable salts thereof:

formula (I) ₀ The substituents in-2) are as defined above.

In some embodiments, the invention provides a compound of formula (I) ₀ -3) novel cytidine derivatives, tautomers, stereoisomers, isotopic derivatives, and pharmaceutically acceptable salts thereof:

formula (I) ₀ The substituents in-3) are as defined above. In some embodiments, the invention provides a compound of formula (I) ₀ -4) and pharmaceutically acceptable salts thereof:

formula (I) ₀ The substituents in-4) are as defined above.

In one aspect of the present invention there is provided a compound of formula (I) ₀ -1) novel cytidine derivatives, tautomers, stereoisomers, and pharmaceutically acceptable salts thereof:

formula (I) ₀ In the step (1) of (a) to (b),

R ₁ and R ₂ Are the same or different and are each independently selected from hydrogen,

Or, R ₁ And R ₂ Together with the oxygen adjacent thereto to form an acetal or ketal;

wherein n is ₁ And n ₂ Selected from 0, 1, 2 or 3;

R _a and R _b Each independently selected from the group consisting of hydroxy, alkyl substituted or unsubstituted by group A, alkoxy substituted or unsubstituted by group A, alkenyl substituted or unsubstituted by group A, cycloalkyl substituted or unsubstituted by group A, aryl substituted or unsubstituted by group A, aryloxy substituted or unsubstituted by group A, arylalkyl substituted or unsubstituted by group A, alkylaryl substituted or unsubstituted by group A;

R _c and R _d Each independently selected from hydrogen, C1-C8 alkyl substituted or unsubstituted by group A;

R ₃ and R ₄ The same or different, are each independently selected from hydrogen and

R ₃ and R ₄ Not all hydrogen;

wherein n is _a 、n _b And n ₃ Each independently selected from 0, or 1, or 2, or 3, or 4, or 5;

n ₄ selected from 0, 1, 2, 3 or 4;

R ₅ and R ₆ Identical or different, independently selected from hydrogen, C1-C8 alkyl substituted or unsubstituted by a radical A; or R ₅ 、R ₆ C to which it is attached forms a cycloalkyl group;

R ₇ selected from hydrogen, halogen, amino, C1-C8 alkyl substituted or unsubstituted by a group A;

z is selected from

Wherein n is ₅ Each independently is 0, or 1, or 2, or 3, or 4, or 5;

R ₈ selected from the group consisting of H, hydroxy, halogen, amino substituted or unsubstituted by a group A, C1-C8 alkyl substituted or unsubstituted by a group A, aryl substituted or unsubstituted by a group A, C1-C8 alkyloxy substituted or unsubstituted by a group A, aminoalkyl substituted or unsubstituted by a group A, C1-C8 alkylaryl substituted or unsubstituted by a group A, arylcarbonyl substituted or unsubstituted by a group A, C1-C8 alkylcarbonyloxy substituted or unsubstituted by a group A;

In some embodiments, the invention provides a compound of formula (I) ₀ -1-1) novel cytidine derivatives, tautomers, stereoisomers, and isomersSite derivatives and pharmaceutically acceptable salts thereof:

formula (I) ₀ The substituents in (1-1) are as defined above.

In some embodiments, the invention provides a compound of formula (I) ₀ -1-2) and pharmaceutically acceptable salts thereof:

formula (I) ₀ The substituents in (1) to (2) are as defined above.

In some embodiments, the invention provides a compound of formula (I) ₀ -1-3) of cytidine derivatives, tautomers, stereoisomers, isotopic derivatives, and pharmaceutically acceptable salts thereof:

/>

formula (I) ₀ The substituents in (1) to (3) are as defined above.

formula (I) ₀ In the step (1) of (a) to (b),

R ₁ and R ₂ Each independently selected from hydrogen,

wherein n is ₁ And n ₂ Selected from 0, 1, 2 or 3;

R _c and R _d Each independently selected from hydrogen, C1-C8 alkyl substituted or unsubstituted with a group A;

R ₃ and R ₄ The same or different, are each independently selected from hydrogen, or

R ₃ And R ₄ Not both may be hydrogen;

wherein n is _a And n _b Each independently selected from 1, or 2, or 3, or 4, or 5;

n ₃ selected from 0, or 1, or 2, or 3, or 4, or 5;

n ₄ selected from 0, 1, 2, 3 or 4;

R ₅ and R ₆ Identical or different, independently selected from hydrogen, C1-C8 alkyl substituted or unsubstituted by a radical A; or R ₅ 、R ₆ To which carbon is attached to form a cycloalkyl group;

R ₇ is hydrogen, halogen, amino, C1-C8 alkyl which is substituted or unsubstituted by a radical A;

z is selected from

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

n ₅ are independent of each otherIs 0, or 1, or 2, or 3, or 4, or 5;

R _e and R _f Each independently selected from hydrogen, the following groups substituted or unsubstituted with one or more groups a: C1-C8 alkyl, C3-C8 cycloalkyl, heterocycloalkyl, C6-C18 aryl, heteroaryl, non-aromatic heterocyclyl; wherein R is _e And R _f Cannot be simultaneously hydrogen;

In some embodiments, the invention provides a compound of formula (I) ₀ -1-1) and pharmaceutically acceptable salts thereof:

formula (I) ₀ The substituents in (1-1) are as defined above.

formula (I) ₀ The substituents in (1) to (2) are as defined above.

In some embodiments, the invention provides a compound of formula (I) ₀ -1-3) and pharmaceutically acceptable salts thereof:

formula (I) ₀ The substituents in (1) to (3) are as defined above.

In one aspect of the invention there is provided a compound as (I) ₀ -5) novel cytidine derivatives, tautomers, stereoisomers, and pharmaceutically acceptable salts thereof:

formula (I) ₀ In (1) of (5) above,

R ₁ and R ₂ Each independently selected from the group consisting of hydroxy,

In particular, when R _x6 And R _x7 When both are hydrogen or deuterium, R ₁ 、R ₂ Independently selected from C1-8 alkoxy, R ₁ And R ₂ May form an acetal or ketal with the carbon atom to which they are attached;

wherein n is as defined above ₁ And n ₂ Each independently selected from 0, 1, 2 or 3;

R _a and R _b Each independently selected from hydroxy, the following groups substituted or unsubstituted with one or more groups A: alkyl, alkoxy, alkenyl, cycloalkyl, aryl, aryloxy, arylalkyl, alkylaryl;

R ₃ and R ₄ Are the same or different and are each independently selected from hydrogen, or

R ₃ And R ₄ Not both may be hydrogen;

n ₄ selected from 0, 1, 2, 3 or 4;

z is selected from

n ₅ each independently is 0, or 1, or 2, or 3, or 4, or 5;

R ₈ selected from H, hydroxy, halogen, the following groups substituted or unsubstituted with one or more groups a: amino, C1-C8 alkyl, aryl, alkyloxy, aminoalkyl substituted or unsubstituted with one or more groups A, C1-C8 alkylaryl, arylcarbonyl, C1-C8 alkylcarbonyloxy;

R _X1 、R _X2 、R _X3 、R _X4 、R _X5 、R _X6 、R _X7 and R _X8 Each independently selected from hydrogen, deuterium, in particular, R _X1 、R _X2 、R _X3 、R _X4 、R _X5 、R _X6 、R _X7 And R _X8 Not both hydrogen.

In some embodiments, the above formula (I) ₀ ) And/or (I) ₀ -2)-(I ₀ In-4), R _X1 、R _X2 、R _X3 、 R _X4 Each independently selected from hydrogen, deuterium;

in some embodiments, the above formula (I) ₀ In (1) R _X1 、R _X2 、R _X3 、R _X4 、R _X5 、R _X6 、 R _X7 And R _X8 Are all hydrogen.

In some embodiments, the above formula (I) ₀ -2)-(I ₀ In (4), R _X5 、R _X6 、R _X7 And R _X8 Are all hydrogen.

In some embodiments, the above formula (I) ₀ )-(I ₀ In-4), R ₁ And R ₂ Are all hydrogen;

in some embodiments, the above formula (I) ₀ )-(I ₀ In-1), R ₁ Is hydrogen, R ₂ Is selected from

Wherein n is ₁ Selected from 0, 1, 2 or 3;

in some more specific embodiments, n ₁ Is selected from 0 or 1;

n ₂ selected from 1, 2 or 3;

in some more specific embodiments, n ₂ Is 1;

R _a and R _b Each independently selected from hydroxy, the following groups substituted or unsubstituted with one or more groups A: C1-C8 alkyl, C1-C8 alkoxy, aryloxy, alkylaryl;

in some more specific embodiments, R is as defined above _a And R _b Each independently preferably selected from hydroxy, substituted by one or more groups A orThe following unsubstituted groups: C1-C8 alkyl, C1-C8 alkyloxy;

in some embodiments, the above formula (I) ₀ )-(I ₀ In-1), R ₂ Is hydrogen, R ₁ Is selected from

Wherein n is as defined above ₁ 、n ₂ 、R _a And R _b Respectively as defined above;

in some embodiments, the above formula (I) ₀ )-(I ₀ In-1), R ₁ And R ₂ Are not both hydrogen and are each independently selected from

Wherein n is as defined above ₁ 、n ₂ 、R _a And R _b Respectively as defined above.

In some embodiments, the above formula (I) ₀ )-(I ₀ In (2), R ₄ Is hydrogen, R ₃ Is composed of

In some embodiments, n is _a Selected from 0, 1, 2, or 3;

in some more specific embodiments, n _a Is 0;

in some more specific embodiments, n _a Is 1;

in some embodiments, n is _b Selected from 0, 1, 2, or 3;

in some more specific embodiments, n _b Is 1;

in some embodiments, n is ₃ Selected from 0, 1, 2, 3;

in some more specific embodiments, n ₃ Is 0;

in some more specific embodiments, n ₃ Is 2;

in some more specific embodiments, n ₃ Is 3;

in some embodiments, n is ₄ Selected from 0, 1, 2;

in some more specific embodiments, n ₄ Is 0;

in some more specific embodiments, n ₄ Is 2;

in some embodiments, the above R _e Is hydrogen, R _f The following groups, substituted or unsubstituted with one or more groups a: C1-C8 alkyl, C3-C8 cycloalkyl, heterocycloalkyl, C6-C18 aryl, heteroaryl, non-aromatic heterocyclyl;

in some more specific embodiments, R is as defined above _e Is hydrogen, R _f Is methyl;

in some embodiments, the above R _e And R _f C1-C8 alkyl which is unsubstituted or substituted by a group A;

in some more specific embodiments, R is as defined above _e And R _f Are each C1-C8 alkyl;

in some embodiments, the above R ₅ And R ₆ Identical or different, independently selected from hydrogen, C1-C8 alkyl substituted or unsubstituted by one or more groups A; or R ₅ 、R ₆ C to which it is attached forms a cycloalkyl group;

in some more specific embodiments, R is as defined above ₅ And R ₆ Are each C1-C4 alkyl; preferably, R ₅ And R ₆ A homomethyl group;

in some embodiments, the above R ₅ Is methyl, R ₆ Selected from C1-C4 alkyl, unsubstituted or substituted by one or more groups A; preferably, R ₅ Is methyl, R ₆ Is ethyl;

in some embodiments, the above R ₇ Is hydrogen, halogen, amino, C1-C8 alkyl substituted or unsubstituted by one or more groups A;

in some more specific embodiments, R is as defined above ₇ Selected from hydrogen, or methyl;

in some embodiments, the aforementioned Z is

In some embodiments, n is as defined above ₅ Selected from 0, 1, 2, 3,4, or 5;

in some more specific embodiments, n is as defined above ₅ Preferably 0, 1, or 2;

in some embodiments, the above R ₈ Selected from hydrogen, hydroxy, nitro, halogen, the following groups substituted or unsubstituted with one or more groups a: amino, C1-C8 alkyl, C6-C18 aryl, C1-C8 alkoxy, C1-C8 alkylaryl, C1-C8 alkylcarbonyloxy;

in some more specific embodiments, R is as defined above ₈ Preferably selected from hydrogen, nitro, chloro, bromo, the following groups substituted or unsubstituted with one or more groups a: amino, C1-C3 alkyl, C6-C18 aryl, C1-C3 alkoxy, C1-C3 alkylcarbonyloxy;

in some more specific embodiments of the present invention, Z is selected from 4-chlorobenzoyl, hydrogen, chlorine, 4-chlorobenzyl,

In some embodiments, the above formula (I) ₀ )-(I ₀ -1) and/or (I) ₀ In-3), R ₃ And R ₄ Are all made of

Wherein, Z and R ₅ 、R ₆ 、R ₇ 、R _e 、R _f 、n _a 、 n _b 、n ₃ And n ₄ As defined above.

In some embodiments, the above formula (I) ₀ )-(I ₀ -1) and/or (I) ₀ In-4), R ₃ Is hydrogen, R ₄ Is composed of

In some embodiments, the novel cytidine derivatives, tautomers, stereoisomers, isotopic derivatives, and pharmaceutically acceptable salts thereof, provided herein above, are selected from the group consisting of the following compounds:

/>

/>

/>

/>

/>

in another aspect, the present invention provides, in some embodiments, pharmaceutical compositions comprising the above-described novel cytidine derivatives, tautomers, stereoisomers, isotopic derivatives, and pharmaceutically acceptable salts thereof.

In some embodiments, the invention discloses a pharmaceutical composition, which comprises the compound, isomer or pharmaceutically acceptable salt thereof as an active ingredient or a main active ingredient, and a pharmaceutically acceptable carrier.

In yet another aspect, the present invention provides in some embodiments the above-described novel cytidine derivatives, tautomers, stereoisomers, isotopic derivatives, and pharmaceutically acceptable salts thereof, that are useful for the treatment and/or prevention of diseases associated with antiviral disorders.

In some embodiments, the present invention provides the use of the above pharmaceutical composition for the preparation of an antiviral medicament; wherein, the viruses include but are not limited to: arenaviridae, filoviridae, and coronaviridae, and the like, including, but not limited to, adenovirus, rhinovirus, influenza virus, lassa virus, respiratory syncytial virus, severe acute respiratory syndrome virus, parainfluenza virus, coronavirus, and the like.

In some embodiments, the present invention provides the use of the above pharmaceutical composition for the preparation of an antiviral medicament; among these, the influenza viruses and coronaviruses include, but are not limited to: influenza A virus, influenza B virus, SARS virus, MERS virus, and COVID-19 virus.

In some embodiments, the novel cytidine derivatives of the present invention may be formulated as pharmaceutical compositions for administration to a patient in accordance with a variety of suitably selected modes of administration, including systemically, e.g., orally or parenterally, intravenously, intramuscularly, transdermally, or subcutaneously, etc.

Compared with the compound A disclosed in CN111372592A, the compound disclosed by the invention has better stability.

Compared with the compound A disclosed in CN111372592A, the compound disclosed by the invention has better anti-influenza virus activity, lower cytotoxicity and higher selection index.

Compared with the compound A, the compound disclosed by the invention has better anti-new coronavirus activity.

The bioavailability of the compound disclosed by the invention is about 1.4 times that of the compound A, and the compound has a better safety range.

Compared with the compound A and the compound C which are used together, the compound disclosed by the invention has more excellent treatment effect and prevention effect on the aspect of resisting influenza viruses, and the two metabolic components have a certain synergistic effect after the compound enters the body through the analysis of the excellent treatment effect and the excellent prevention effect. The compound can be used as an antiviral drug with a novel structure.

The structure of compound a is as follows:

compound C has the structure:

defining:

the following terms and phrases used herein are intended to have the following meanings unless otherwise indicated. A particular term or phrase, unless specifically defined, should not be considered as indefinite or unclear, but rather construed according to ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding commodity or its active ingredient.

Certain compounds of the invention may exist in unsolvated forms as well as solvated forms, such as hydrated, ethanolic forms. In general, the solvated forms are equivalent to unsolvated forms and are intended to be encompassed within the scope of the present invention.

The term "pharmaceutically acceptable" is intended to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salts" refers to salts of the compounds of the present invention, prepared from the compounds of the present invention found to have particular substituents, with relatively nontoxic acids or bases. When compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of a base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include aluminum, sodium, potassium, calcium, manganese, iron, ammonium, organic ammonia or magnesium salts or similar salts. When compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include salts with inorganic acids including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, and the like; and salts of organic acids including such acids as acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-toluenesulfonic, citric, tartaric, methanesulfonic, and the like; also included are salts of amino acids such as arginine and the like, and salts of organic acids such as glucuronic acid and the like. Certain specific compounds of the invention contain both basic and acidic functionalities and can thus be converted to any base or acid addition salt.

The term "alkyl" denotes saturated aliphatic groups, including straight and branched chain groups, alkyl groups may be substituted or unsubstituted. When substituted alkyl, the substituent is preferably one or more, more preferably 1 to 3, most preferably 1 or 2 substituents.

The term "alkenyl" denotes aliphatic radicals containing unsaturated carbon-carbon double bonds, including straight and branched radicals, alkyl radicals which may be substituted or unsubstituted. The carbon-carbon double bond may be one or more.

The term "cycloalkyl" denotes a monocyclic or fused ring of all carbons (a "fused" ring meaning that each ring in the system shares an adjacent pair of carbon atoms with other rings in the system) group in which one or more rings do not have a fully linked pi-electron system, examples of cycloalkyl (without limitation) being cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, adamantane, cyclohexadiene, cycloheptane, and cycloheptatriene. Cycloalkyl groups may be substituted and unsubstituted.

The term "aryl" denotes an all-carbon monocyclic or fused polycyclic group of 1 to 12 carbon atoms with a completely conjugated pi-electron system. Non-limiting examples of aryl groups are phenyl, naphthyl and anthracenyl. The aryl group may be substituted or unsubstituted. When substituted, the substituent is preferably one or more, more preferably one, two or three, and still more preferably one or two.

The term "arylalkyl" refers to an aryl-substituted alkyl group.

The term "heteroaryl" denotes a multi-atom monocyclic or fused ring group containing one, two, three or four ring heteroatoms selected from N, O or S, the remaining ring atoms being C, and additionally having a completely conjugated pi-electron system. Non-limiting examples of unsubstituted heteroaryl groups are pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyrimidine, quinoline, isoquinoline, purine, tetrazole, triazine and carbazole.

The term "alkoxy" denotes a group wherein an alkyl group, which may be straight chain, branched or cyclic, is attached to the oxygen.

The term "hydroxy" denotes an-OH group.

The term "amino" denotes-NH ₂ A group.

The term "carboxyl" denotes the-COOH group.

The term "halogen" denotes fluorine, chlorine, bromine or iodine.

The term "pharmaceutically acceptable carrier" refers to any formulation or carrier medium capable of delivering an effective amount of an active agent of the present invention, without interfering with the biological activity of the active agent, and without toxic side effects to the host or patient, and representative carriers include water, oils, vegetables and minerals, cream bases, lotion bases, ointment bases, and the like. These include suspending agents, viscosity enhancers, skin penetration enhancers, and the like.

The term "stereoisomers" refers to compounds that have the same chemical constitution, but differ in the arrangement of atoms or groups in space.

Reference herein to a numerical range, such as "C1-C8," means that the group may contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, and the like, up to and including 8 carbon atoms.

Drawings

FIG. 1 shows the results of an anti-influenza virus (H1N 1) therapeutic administration test.

FIG. 2 shows the results of a prophylactic administration test against influenza virus (H1N 1).

Detailed Description

A number of exemplary methods of preparing the compounds of the present invention are provided in the examples below. The present invention is described in detail below by way of examples, but is not meant to be limited to any of the disadvantages of the present invention. Having described the invention in detail and having disclosed specific embodiments thereof, it will be apparent to one skilled in the art that various changes and modifications can be made therein without departing from the spirit and scope thereof. Certain compounds of the invention can be used as intermediates for the preparation of other compounds of the invention, all of which are structurally determined by liquid chromatography or nuclear magnetic resonance.

The starting materials in the examples of the present application were all purchased commercially.

Example 1: synthesis of Compound PY-01

The reaction formula is as follows:

the preparation method comprises the following steps:

step 1: preparation of Compound PY-0102:

tetrahydrofuran (300 ml), PY-01-SM2 (31.8g, 100mmol) and 4-dimethylaminopyridine (DMAP, 18.33g, 150momol) are sequentially added into a reaction bottle, 1-ethyl- (3-dimethylaminopropyl) carbonyldiimine (EDC, 18.63g, 120mmol) and a compound PY-01-SM1 (28.43 g,100 mmol) are added after dissolution, the temperature is raised to 70 ℃, stirring reaction is carried out, TLC monitoring reaction is finished, the system is cooled and evaporated to dryness, 200ml of ethyl acetate and 100ml of water are added, an organic phase is separated out, water is washed twice, dried and evaporated to dryness under reduced pressure, and the remainder is purified by a column to obtain a product PY-0102 (44.34 g) with the yield of 75.8%. ESI-MS (+): m/z =585.16.

And 2, step: preparation of Compound PY-0101:

adding a compound PY-0102 (44.00g, 75.2mmol), N-dimethylformamide (250 ml) and DIPEA (19.44g, 150.4mmol) into a reaction bottle, dissolving, adding PyBroP (38.55 g, 82.7 mmol), stirring the system at room temperature for 30min, adding hydroxylamine hydrochloride (62.71g, 90.24mmol), reacting at 40-50 ℃ for 4-6 h, detecting by TLC (thin layer chromatography), cooling, adding water, extracting with ethyl acetate for 3 times, combining organic phases, washing twice with water, evaporating to dryness under reduced pressure, recrystallizing the residue with methyl tert-butyl ether/N-heptane to obtain a product PY-0101 (39.00 g), wherein the yield is 86.4%. ESI-MS (+): m/z =600.17.

And step 3: preparation of Compound PY-01:

PY-0101 (38.00g, 63.3 mmol) and formic acid (500 ml) were added to the reaction flask, and the system was reacted at room temperature for 20 hours. After the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was recrystallized from isopropyl alcohol/methyl t-butyl ether to give PY-01 (29.21) in 82.4% yield. ESI-MS (+): m/z =560.2.1H NMR (DM SO-D6, 500MHz) < delta > 10.02 (s, 1H), 9.49 (s, 1H), 7.72-7.75 (m, 4H), 7.63-7.64 (d, 2H), 6.94-6.96 (d, 2H), 6.78-6.80 (d, 1H), 5.71-5.72 (d, 1H), 5.46-5.48 (d, 1H), 5.32-5.33 (d, 1H), 5.20-5.21 (d, 1H), 4.28-4.36 (m, 2H), 3.97 (s, 1H), 3.89-3.92 (m, 1H), 3.78-3.79 (d, 1H), 1.6-1.66 (d, 6H).

Example 2: synthesis of Compound PY-02

The reaction formula is as follows:

the preparation method comprises the following steps:

step 1: preparation of Compound PY-0201

Reference example 1 preparation of step 1, substituting the compound PY-0101 for the compound PY-01-SM1, produced the compound PY-0201 (13.21 g) in 73.5% yield. ESI-MS (+): m/z =900.22.

And 2, step: preparation of Compound PY-02

Reference example 1, step 3, was carried out in the presence of PY-0201 in place of PY-0101 to prepare PY-02 (10.14 g) in 83.2% yield. ESI-MS (+): m/z =860.19.

Example 3: synthesis of Compound PY-03

The reaction formula is as follows:

the preparation method comprises the following steps:

step 1: preparation of compound PY-0303

Under nitrogen protection, compound PY-03-SM3 (25.0 g,102.0mmo 1) and 500ml of dichloromethane were added to a three-necked flask. The resulting solution was cooled to 0 deg.C, and DMAP (1.3g, 10.6 mmol) and imidazole (27.9 g,409.0 mmol. O.1) were added in that order. TBSCl (61.7 g,40.0 mmol) was added over 10 minutes and the resulting mixture was warmed to ambient temperature and stirred for 18 hours. 300mL of water was added to the system, stirred at room temperature for 2 hours, the layers were separated, the aqueous layer was extracted 3 times with dichloromethane, the combined organic layers were washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and the residue was purified by column chromatography to give PY-0303 (43.4 g) in 72.5% yield. ESI-MS (+): m/z =587.33.

Step 2: preparation of compound PY-0302

A1L round-bottomed flask was charged with compound PY-0303 (28.0 g,47.7mo 1) and dichloromethane (700 mL). The solution was cooled to 0 ℃ using an ice bath; DMAP (0.583 g, 4.77mmo1) and N, N-diisopropylethylamine (30.9g, 239mmo1) were added successively. 2,4, 6-Triisopropylphenyl-1-sulfonyl chloride (28.9 g, 95mmol) was slowly added to the flask, and after the addition was complete, the flask was warmed to ambient temperature and stirred for 18 hours. The system was cooled to 0 deg.C and N, N diisopropylethylamine (24.6 g, 19lmol) was added dropwise, followed immediately by solid hydroxylamine hydrochloride (13.26g, 191mmol). The mixture was warmed to room temperature and stirred for 3 hours. The reaction was quenched with water (200 mL) and the resulting layer was separated. The aqueous layer was extracted with dichloromethane (200 mL) and the combined organics were washed with brine, dried over sodium sulfate and concentrated under reduced pressure to give the residue which was purified on a column to give the compound PY-0302 (20.0 g) in 69.5% yield. ESI-MS (+): m/z =602.34.

And step 3: preparation of Compound PY-0301

Referring to preparation of step 1 of example 1, compound PY-0301 (18.6 g) was prepared in 76.2% yield by replacing compound PY-01-SM1 with compound PY-0302. ESI-MS (+): m/z =902.40.

And 4, step 4: preparation of Compound PY-03

Compound PY-0301 (17.5g, 19.4 mol) and tetrahydrofuran (50 mL) were added to a three neck flask followed by triethylamine trihydrofluoride (3.1g, 19.4mmo 1) and the mixture stirred at ambient temperature for 18 hours. The mixture was concentrated under reduced pressure and the residue was dissolved in a minimum amount of MeOH, the solution was slowly added to an erlenmeyer flask containing rapidly stirred dichloromethane (50 mL), and the mixture was stirred at room temperature for 15 minutes. Filtration and recrystallization from dichloromethane and petroleum ether gave the title compound PY-03 (7.65 g) in 70.5% yield. ESI-MS (+): m/z =560.25.1H NMR (DMS O-D6, 400MHz) < delta > 11.34-11.35 (s, 1H), 10.79-10.80 (s, 1H), 7.70-7.74 (m, 4H), 7.61-7.63 (d, 2H), 7.48-7.51 (d, 1H), 6.94-6.96 (d, 2H), 5.72-5.78 (m, 2H), 5.32-5.34 (d, 1H), 5.02-5.08 (d, 2H), 3.96-4.04 (m, 2H), 3.83-3.84 (m, 1H), 3.55 (s, 2H), 1.76 (s, 3H), 1.69 (s, 3H).

Example 4: synthesis of Compound ZJT1

The reaction formula is as follows:

the preparation method comprises the following steps:

step 1: preparation of compound ZJT1-03

Adding 500ml of methylamine solution with the mass fraction of 80%, anisole (36.4 g, 0.34mol) and cuprous chloride (35.6 g, 0.36mol) into a three-necked bottle, controlling the solution temperature to 35 ℃, dropwise adding p-bromobenzamide (101.9 g, 0.35 mol), raising the solution temperature to 45 ℃ after the addition, maintaining the stirring speed for reaction for 7 hours, reducing the solution temperature to 30 ℃, pouring the reaction solution into sodium bisulfite solution with the mass fraction of 30%, maintaining the solution temperature at 0-5 ℃, separating an organic layer, extracting a water layer for 6 times by using methylamine solution, merging the organic layers, and distilling out methylamine to obtain a compound ZJT1-03 for later use.

Step 2: preparation of compound ZJT1-02

Adding 300m1 of hexane into the spare product of the compound ZJT1-03, heating to 60 ℃, adding cuprous chloride (59.4g and 0.6 mol), carrying out reflux reaction for 2 hours, distilling out the hexane, reducing the temperature of the solution to 5 ℃, adding 200ml of potassium bicarbonate solution with the mass fraction of 60%, stirring for 2 hours, filtering, and washing with potassium sulfate solution to obtain the compound ZJT1-02 (79.2 g), wherein the yield is 84.1%. ESI-MS (+): m/z =276.98.

And step 3: preparation of compound ZJT1

Acetone (120g, 2.07mol) and a compound ZJT1-02 (59.6g, 0.21mol) are added into a three-necked flask and stirred for about 10 minutes, sodium hydroxide (60.0 g,1.5 mol) is added, chloroform (45 ml) is dropwise added under stirring at the temperature of 20-30 ℃, after the dropwise addition of the chloroform is finished, the temperature is kept at the temperature of 20-30 ℃ for 1.5 hours, and then the temperature is increased to reflux for 3.5 hours. The organic solvent was distilled off under reduced pressure, 100ml of an appropriate amount of water and 130ml of toluene were added to the residue, 36% hydrochloric acid (about 60 g) was added dropwise, the feed liquid was acidified to p =3.5 to 4.5, then 100ml was added, the system was cooled to room temperature, stirred for 2 hours, filtered, and dried to obtain compound ZJT1 (66.5 g), yield 87.2%. ESI-MS (-): m/z =361.01.

Example 5: synthesis of compound ZJT2

The reaction formula is as follows:

the preparation method comprises the following steps:

referring to the procedure for the steps of example 4, the starting material was replaced with p-nitrobenzamide for bromobenzamide to give compound ZJT2 (55.2 g) in 85.8% yield. ESI-MS (-): m/z =328.09.

Example 6: synthesis of compound ZJT3

The reaction formula is as follows:

the preparation method comprises the following steps:

referring to the procedure of the steps of example 4, the starting material was replaced with p-aminobenzamide for p-bromobenzamide to give compound ZJT3 (51.9 g) in 86.1% yield. ESI-MS (-): m/z =298.12.

Example 7: synthesis of Compound ZJT4

The reaction formula is as follows:

the preparation method comprises the following steps:

referring to the procedure for the steps of example 4, the starting material was replaced with 3-chlorobenzamide for bromobenzamide to give compound ZJT4 (57.6 g) in 82.9% yield. ESI-MS (-): m/z =317.07.

Example 8: synthesis of Compound ZJT5

The reaction formula is as follows:

the preparation method comprises the following steps:

referring to the procedure for the steps of example 4, the starting material was replaced with 3, 4-dichlorobenzamide to give ZJT5 (46.1 g) in 79.6% yield. ESI-MS (-): m/z =351.03.

Example 9: synthesis of compound ZJT6

The reaction formula is as follows:

the preparation method comprises the following steps:

referring to the procedure for the steps of example 4, the starting material was replaced with p-methoxybenzamide for p-bromobenzamide to give compound ZJT6 (31.3 g) in 87.1% yield. ESI-MS (-): m/z =313.12.

Example 10: synthesis of compound ZJT7

The reaction formula is as follows:

the preparation method comprises the following steps:

referring to the procedure for the steps of example 4, the starting material was replaced with 3,4, 5-trimethoxybenzamide to give ZJT7 (32.6 g) in 84.3% yield. ESI-MS (-): m/z =373.14.

Example 11: synthesis of compound PY-ZJT8

The reaction formula is as follows:

the preparation method comprises the following steps:

referring to the procedure for each step of example 4, the starting material was replaced with 3- (trifluoromethyl) benzamide to give compound ZJT8 (36.8 g) in 81.1% yield. ESI-MS (-): m/z =351.09.

Example 12: synthesis of compound PY-ZJT9

The reaction formula is as follows:

/>

the preparation method comprises the following steps:

the compound PY-01-SM2 (21.5g, 67.5mmol), water (300 mL), methylene chloride (300 mL), tetrabutylammonium hydrogensulfate (2.3g, 6.75mmol) and sodium hydrogencarbonate (22.7g, 270mmol) were charged into a three-necked flask. Chloromethyl chlorosulfonate (13.5g, 81.7 mmol) was added dropwise at room temperature, and the reaction was stirred at room temperature overnight. The organic and aqueous layers were separated, and the aqueous layer was extracted with dichloromethane (300 mL). The combined organic phases were dried, concentrated and purified by column chromatography (0 to 5% ethyl acetate/hexanes gradient elution) to give ZJT9 (18.7 g) in 75.4% yield.

Example 13: synthesis of Compound PY-04

The reaction formula is as follows:

the preparation method comprises the following steps:

referring to the procedure of each step of example 1, the starting material was replaced with the compound ZJT1 instead of the compound PY-01-SM2 to obtain the compound PY-04 (10.8 g) in a total yield of 51.1%. ESI-MS (+): m/z =604.09.

Example 14: synthesis of Compound PY-05

The reaction formula is as follows:

the preparation method comprises the following steps:

referring to the procedure of each step of example 2, the starting material was replaced with the compound ZJT1 instead of the compound PY-01-SM2 to obtain the compound PY-05 (12.3 g) in a total yield of 57.6%. ESI-MS (+): m/z =948.09.

Example 15: synthesis of Compound PY-06

The reaction formula is as follows:

the preparation method comprises the following steps:

referring to the procedure of each step of example 1, the compound PY-01-SM2 was replaced with the compound ZJT2 as a starting material to obtain the compound PY-06 (14.1 g) in an overall yield of 46.9%. ESI-MS (+): m/z =571.16.

Example 16: synthesis of Compound PY-07

The reaction formula is as follows:

the preparation method comprises the following steps:

referring to the procedure of each step of example 1, compound PY-07 (13.6 g) was obtained in an overall yield of 48.5% by replacing compound PY-01-SM2 with compound ZJT 3. ESI-MS (+): m/z =541.19.

Example 17: synthesis of Compound PY-08

The reaction formula is as follows:

the preparation method comprises the following steps:

step 1: preparation of Compound PY-0804:

PY-01-SM2 (1.91g, 6 mmol) was added to dichloromethane (20 mL) under nitrogen, cooled to 0 deg.C, thionyl chloride (0.86g, 7.2mmol) was added slowly, and after the addition was complete, the reaction was stirred at room temperature for 2.0h. Concentrating the system to dryness, removing the residual thionyl chloride by toluene (10 mL multiplied by 3) to obtain a compound PY-0804 as the residue, and keeping the compound PY-0804 untreated for later use;

and 2, step: preparation of Compound PY-0803:

dichloromethane (25 mL) was added to the residue obtained in step 1 (compound PY-0804,6 mmol), and the above mixture was slowly added to a solution of aluminum trichloride (0.48g, 3.6 mmol) in dichloromethane (20 mL), and the mixture was stirred at room temperature for 20 minutes. The system was cooled to 0 ℃ and acetaldehyde (0.27g, 6 mmoles) was added dropwise over 10 minutes. The reaction mixture was stirred at room temperature for 1 hour and the system was gradually added to a vigorously stirred ice-water slurry. Extraction was carried out 3 times with dichloromethane, the organic phases were combined, washed 2 times with ice water, the organic phase was separated, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to give the compound PY-0803 (1.11 g) with a yield of 48.7%. ESI-MS (+): m/z =381.06.

And 3, step 3: preparation of Compound PY-0802:

to a reaction flask were added PY-01-SM1 (0.56g, 2.0 mmol), triethylamine (0.31 g,3 mmol) and PY-0803 (0.92g, 2.4 mmol), and N, N-dimethylformamide (50 mL) was added, and the system was heated to 50 ℃ and stirred for 24 hours. Cooled to room temperature, added with water and stirred, filtered to obtain a crude product, which is purified by silica gel column to obtain the compound PY-0802 (0.96 g) with yield 76.4%, ESI-MS (+): m/z =628.20.

And 4, step 4: preparation of Compound PY-0801:

adding PY-0802 (0.47g, 0.75mmol), N-dimethylformamide (25 ml) and DIPEA (0.20g, 1.50mmol) into a reaction bottle, dissolving, adding PyBroP (0.39g, 0.83mmol), stirring the system at room temperature for 30min, adding hydroxylamine hydrochloride (0.63g, 0.90mmol), reacting at 40-50 ℃ for 4-6 h, detecting by TLC, cooling, adding water, extracting with ethyl acetate for 3 times, combining organic phases, washing with water twice, evaporating to dryness under reduced pressure, and purifying residues by a column to obtain a product compound PY-0801 (0.36 g), wherein the yield is 74.5%. ESI-MS (+): m/z =644.19.

And 5: preparation of Compound PY-08:

the compound PY-0801 (0.36g, 0.56mmol) and formic acid (10 mL) were added to a reaction flask, and the system was reacted at room temperature for 20 hours. After the reaction was completed, concentration was performed under reduced pressure, and the residue was purified by column chromatography to obtain compound PY-08 (0.23 g) in 68.7% yield and ESI-MS (+): m/z =604.16.1H NMR (DMSO-D6, 400 MHz) < delta > 11.32-11.33 (s, 1H), 10.77-10.78 (s, 1H), 7.71-7.76 (m, 4H), 7.59-7.61 (D, 2H), 7.45-7.49 (D, 1H), 6.91-6.94 (D, 2H), 6.61 (m, 4H), 5.70-5.75 (m, 2H), 5.29-5.31 (D, 1H), 5.01-5.06 (D, 2H), 3.94-4.03 (m, 2H), 3.81-3.83 (m, 1H), 3.53 (s, 2H), 1.72 (D, 3H), 1.70 (s, 6H).

Example 18: synthesis of Compound PY-09

The reaction formula is as follows:

the preparation method comprises the following steps:

referring to the procedures of step 3 and step 4 of example 3, the compound PY-01-SM2 was replaced with the compound ZJT5 to give the compound PY-09 (9.3 g) in a total yield of 50.1%. ESI-MS (+): m/z =592.12.

Example 19: synthesis of Compound PY-10

The reaction formula is as follows:

the preparation method comprises the following steps:

referring to the procedures of step 3 and step 4 of example 3, the compound PY-01-SM2 was replaced with the compound ZJT6 to give the compound PY-10 (19.4 g) in a total yield of 55.3%. ESI-MS (+): m/z =554.21.

Example 20: synthesis of Compound PY-11

The reaction formula is as follows:

the preparation method comprises the following steps:

referring to the procedures of step 1 and step 2 in example 1, compound PY-1102 was prepared by replacing compound PY-01-SM2 with compound ZJT 7;

referring to the procedures of example 2, PY-11 (5.7 g) was prepared in 35.3% total yield by replacing PY-01-SM2 with ZJT7 and PY-0101 with PY-1102, respectively. ESI-MS (+): m/z =972.33.

Example 21: synthesis of Compound PY-24

The reaction formula is as follows:

the preparation method comprises the following steps:

referring to the procedure of each step of example 1, the compound PY-01-SM2 was replaced with the compound ZJT8 as a starting material to give the compound PY-24 (5.1 g) in an overall yield of 45.9%. ESI-MS (+): m/z =594.16.

Example 22: synthesis of Compound PY-26

The reaction formula is as follows:

the preparation method comprises the following steps:

step 1: synthesis of Compound PY-2602

Referring to the procedure of step 3 in example 1, PY-2602 (6.1 g) was prepared in 78.3% yield. ESI-MS (+): m/z =545.12.

Step 2: compound PY-2601

Compound PY-2602 (5.6 g, 10.24mol) and ethyl acetate (120 mL) were added to a three-necked flask. Triethylamine (5.2g, 51.12mol) and DMAP (0.063g, 5.12mmol) were added to the above system with stirring. The temperature was reduced to below 10 ℃ and trifluoroacetic anhydride (6.45g, 30.72mmol) was slowly added to the system over 5 minutes. The system exothermed during the addition. After the addition, the system was stirred at room temperature for 1h, and the reaction was completed by TLC. The reaction was quenched with 40mL of water and stirred at room temperature for 20 minutes. The layers were separated and the organics were washed with water (2X 50 mL), saturated aqueous bicarbonate (50 mL X2), water (50 mL), brine (50 mL X2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give compound PY-2601 as the residue. Used in the next step without further purification. ESI-MS (+): m/z =737.09.

And step 3: preparation of Compound PY-26

Referring to the procedure of step 2 in example 1, PY-26 (2.2 g) was prepared in 83.6% yield. ESI-MS (+): m/z =754.12.

Example 23: synthesis of Compound PY-27

The reaction formula is as follows:

the preparation method comprises the following steps:

step 1: synthesis of Compound PY-2702

The compound PY-01-SM1 (14.2g, 50.0 mmol), triethylamine (75.9 g, 75.0 mmol) and ZJT9 (36.7g, 100.0 mmol) were charged into a reaction flask, 500ml of N, N-dimethylformamide was added, and the system was heated to 50 ℃ and stirred for 24 hours. The temperature is reduced to room temperature, water is added for stirring, the crude product is obtained after filtration, and the crude product is purified by silica gel column to obtain the compound PY-2702 (17.2 g), the yield is 55.8 percent, and ESI-MS (+): m/z =615.17.

And 2, step: compound PY-2701

Referring to the procedure of step 2 in example 1, the compound PY-2701 (10.1 g) was prepared in 82.3% yield. ESI-MS (+): m/z =630.18.

And 3, step 3: preparation of Compound PY-27

Referring to the procedure of step 3 in example 1, PY-27 (5.3 g) was prepared in 79.6% yield. ESI-MS (+): m/z =590.15.

Example 24: synthesis of Compound PY-28

The reaction formula is as follows:

the preparation method comprises the following steps:

step 1: synthesis of Compound PY-2801

Referring to the procedure of step 1 of example 23, the compound PY-2801 (12.2 g) was prepared in 51.4% yield and ESI-MS (+): m/z =932.41.

Step 2: compound PY-28

Referring to the procedure of step 4 in example 3, PY-28 (5.2 g) was prepared in 68.3% yield. ESI-MS (+): m/z =590.15.

Example 25: synthesis of Compound PY-34

The reaction formula is as follows:

the preparation method comprises the following steps:

step 1: preparation of compound PY-3404:

PY-01-SM2 (1.91g, 6 mmol) was added to dichloromethane (20 mL) under nitrogen, cooled to 0 deg.C, thionyl chloride (0.86g, 7.2mmol) was added slowly, and after the addition was complete, the reaction was stirred at room temperature for 2.0h. Concentrating the system to dryness, removing the residual thionyl chloride with toluene (10 mL. Times.3) to obtain a compound PY-3404 as the residue, and keeping the compound PY-3404 as the spare material without treatment;

step 2: preparation of compound PY-3403:

methylene chloride (25 mL) was added to the residue obtained in step 1 (the compound PY-3404,6 mmol), and the above mixture was slowly added to a solution of aluminum trichloride (0.48g, 3.6 mmol) in methylene chloride (20 mL), and the mixture was stirred at room temperature for 20 minutes. The system was cooled to 0 ℃ and acetaldehyde (0.27g, 6 mmoles) was added dropwise over 10 minutes. The reaction mixture was stirred at room temperature for 1 hour, and the system was gradually added to a vigorously stirred ice-water slurry. Extraction was performed 3 times with dichloromethane, the organic phases were combined, washed 2 times with ice water, the organic phase was separated, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to give the compound PY-3403 (1.11 g) with a yield of 48.7%. ESI-MS (+): m/z =381.06.

And 3, step 3: preparation of compound PY-3402:

to a reaction flask were added PY-01-SM1 (0.56g, 2.0 mmol), triethylamine (0.31 g,3 mmol) and PY-3403 (0.92g, 2.4 mmol), and N, N-dimethylformamide (50 mL) was added, and the system was heated to 50 ℃ and stirred for 24 hours. Cooled to room temperature, added with water and stirred, filtered to obtain a crude product, which is purified by silica gel column to obtain the compound PY-3402 (0.96 g) with yield of 76.4%, ESI-MS (+): m/z =628.20.

And 4, step 4: preparation of compound PY-3401:

adding a compound PY-3402 (0.47g, 0.75mmol), N-dimethylformamide (25 ml) and DIPEA (0.20g, 1.50mmol) into a reaction bottle, dissolving, adding a tripyrrolidinyl phosphonium bromide hexafluorophosphate (PyBroP, 0.39g, 0.83mmol), stirring the system at room temperature for 30min, adding hydroxylamine hydrochloride (0.63g, 0.90mmol), reacting at 40-50 ℃ for 4-6 h, detecting by TLC, cooling, adding water, extracting for 3 times with ethyl acetate, combining organic phases, washing twice with water, evaporating to dryness under reduced pressure, and purifying the residue by a column to obtain a product compound PY-3401 (0.36 g), wherein the yield is 74.5%. ESI-MS (+): m/z =644.19.

And 5: preparation of Compound PY-34:

the compound PY-3401 (0.36g, 0.56mmol) and formic acid (10 mL) were charged into a reaction flask, and the system was reacted at room temperature for 20 hours. After the reaction was completed, concentration was performed under reduced pressure, and the residue was purified by column chromatography to obtain the compound PY-34 (0.23 g) in 68.7% yield, ESI-MS (+): m/z =604.16.1H NMR (DMSO-D6, 400 MHz) < delta > 11.32-11.33 (s, 1H), 10.77-10.78 (s, 1H), 7.71-7.76 (m, 4H), 7.59-7.61 (D, 2H), 7.45-7.49 (D, 1H), 6.91-6.94 (D, 2H), 6.61 (m, 4H), 5.70-5.75 (m, 2H), 5.29-5.31 (D, 1H), 5.01-5.06 (D, 2H), 3.94-4.03 (m, 2H), 3.81-3.83 (m, 1H), 3.53 (s, 2H), 1.72 (D, 3H), 1.70 (s, 6H).

Example 26: synthesis of Compound PY-35

The reaction formula is as follows:

the preparation method comprises the following steps:

step 1: preparation of Compound PY-3501

Referring to the preparation of step 3 of example 25, the compound PY-3501 (1.26 g) was prepared in 69.5% yield by replacing the compound PY-01-SM1 with the compound PY-3401. ESI-MS (+): m/z =988.27.

Step 2: preparation of Compound PY-35

Reference example 25 preparation of step 5, substituting compound PY-3501 for compound PY-3401, compound PY-35 (0.19 g) was prepared in a yield of 63.2%. ESI-MS (+): m/z =948.24.

Example 27: synthesis of Compound PY-36

The reaction formula is as follows:

the preparation method comprises the following steps:

step 1: preparation of Compound PY-3603

Under nitrogen protection, the compound PY-03-SM3 (12.5 g,51.0 mmol) and dichloromethane (250 mL) were added to a three-necked flask. The resulting solution was cooled to 0 ℃ and 4-dimethylaminopyridine (DMAP, 0.70g,5.7 mmol) and imidazole (14.0g, 205.6 mmol) were added in this order. Tert-butyldimethylsilyl chloride (TBSC 1, 30.9g,205.0 mmol) was added over 10 minutes and the resulting mixture was warmed to ambient temperature and stirred for 18 hours. Water was added to the system, stirred at room temperature for 2 hours, the layers were separated, the aqueous phase was extracted 3 times with dichloromethane, the combined organic layers were washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and the residue was purified by column chromatography to give the compound PY-3603 (21.1 g) in 70.6% yield. ESI-MS (+): m/z =587.33.

Step 2: preparation of compound PY-3602

Compound PY-3603 (14.0 g, 23.9mmol) and dichloromethane (350 mL) were added to a three-necked flask. The solution was cooled to 0 ℃ using an ice bath. DMAP (0.29g, 0.24mmol) and N, N-diisopropylethylamine (15.5g, 120mmol) were added successively. 2,4,6-Triisopropylbenzene-1-sulfonyl chloride (14.5 g,47.5 mmol) was slowly added to the flask, and after the addition was complete, the flask was warmed to ambient temperature and stirred for 18 hours. The system was cooled to 0 deg.C, N diisopropylethylamine (12.3g, 95.5mmol) was added dropwise, and then solid hydroxylamine hydrochloride (6.63g, 95.5mmol) was added immediately. The mixture was warmed to room temperature and stirred for 3 hours. The reaction was quenched with water and the resulting layer was separated. The aqueous layer was extracted with dichloromethane and the combined organics were washed with brine, dried over sodium sulfate, concentrated under reduced pressure and the resulting residue was purified on a column to give compound PY-3602 (9.41 g) in 65.4% yield. ESI-MS (+): m/z =602.34.

And 3, step 3: preparation of Compound PY-3601

Reference example 25 preparation of step 3, substitution of PY-3602 for PY-01-SM1 gave PY-3601 (10.1 g) in 71.3% yield. ESI-MS (+): m/z =946.42.

And 4, step 4: preparation of Compound PY-36

In a three-necked flask was charged the compound PY-0301 (9.5g, 1.0 mmol) and tetrahydrofuran (100 mL) followed by triethylamine trihydrofluoride (1.61g, 1.0 mmol), and the mixture was stirred at ambient temperature for 18 hours. The mixture was concentrated under reduced pressure and the residue was dissolved in a minimum amount of methanol, the solution was slowly added to dichloromethane containing a rapid stirrer, and the mixture was stirred at room temperature for 15 minutes. Filtration and recrystallization from dichloromethane and petroleum ether gave the title compound PY-36 (4.05 g) in 67.1% yield. ESI-MS (+): m/z =604.16.

Example 28: synthesis of Compound ZJT10

The reaction formula is as follows:

the preparation method comprises the following steps:

step 1: preparation of Compound ZJT 10-02:

under the protection of nitrogen, adding pyrimidine-5-D (11.31g, 0.1mol), BSA (40.7g, 0.2mol) and anhydrous acetonitrile (250 mL) into a reaction bottle, heating to 80 ℃, and stirring for reacting for 30min; cooling to room temperature, adding tetraacetyl ribose (63.3 g, 0.2mol), dripping TMSOTf (44.5 g, 0.2mol), heating to 80 ℃, reacting for 2 hours after the addition is finished, cooling to room temperature, and distilling out acetonitrile under reduced pressure; adding ethyl acetate into the residue, washing with saturated sodium bicarbonate water solution for 2 times, separating organic phase, washing with saturated saline solution for 2 times, drying with anhydrous sodium sulfate, filtering, concentrating under reduced pressure, and performing column chromatography to obtain ZJT10-02 (31.6 g) with yield of 85.2%. ESI-MS (+): m/z =372.13.

And 2, step: preparation of Compound ZJT 10-01:

compound ZJT10-02 (31.0 g,0.084 mol) and 7M ammonia/methanol solution (500mL, 3.5 mol) were added to a reaction flask, stirred at room temperature for 36 hours, the solvent was distilled off, and column chromatography was performed to give compound ZJT10-01 (15.5 g), 75.2% yield. ESI-MS (+): m/z =246.10.

And step 3: preparation of compound ZJT10

Dissolving compound ZJT10-01 (15.0g, 0.061mol) in acetone (300 mL), slowly adding 2, 2-dimethoxypropane (31.8g, 0.305mol), stirring for 10 minutes, slowly adding concentrated sulfuric acid (2.0g, 0.020mol), and stirring for 30 minutes; saturated sodium bicarbonate was added to the system to quench, the solvent was distilled off under reduced pressure, and the residue was subjected to column chromatography to give ZJT10 (11.5 g), yield 66.1%. ESI-MS (+): m/z =286.20.

Example 29: synthesis of compound ZJT11

The reaction formula is as follows:

the preparation method comprises the following steps:

referring to the procedure of example 1, substituting uracil-2D for pyrimidine-5-D as the starting material, compound ZJT11 (10.2 g) was obtained in an overall yield of 40.1%. ESI-MS (+): m/z =287.22.

Example 30: synthesis of compound ZJT12

The reaction formula is as follows:

the preparation method comprises the following steps:

step 1: preparation of compound ZJT12-01

PY-01-SM1 (6.3 g,22.0 mmol) and methylene chloride (200 mL) were added to the reaction flask under nitrogen at room temperature. Pyridinium dichromate (16.6 g, 44.1mmol), acetic anhydride (22.5g, 220mmol) and tert-butanol (16.3 g, 220mmol) were then added to the system in this order with stirring. The above system was stirred at room temperature for 24 hours, washed with water, the aqueous phase was separated, extracted with dichloromethane 2 times, the organic phases were combined, washed with saturated brine 2 times, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was subjected to column chromatography to give compound ZJT12-01 (5.5 g), with a yield of 70.5%. ESI-MS (+): m/z =355.15.

Step 2: preparation of compound ZJT12

Under nitrogen protection, the compound ZJT12-01 (5.4 g, 15.2mmol) was added to ethanol-D1 (200 mL) at room temperature, and sodium borodeuteride-D4 (2.5g, 60.8mmol) was added in one portion with stirring. After the addition was complete, the mixture was stirred at room temperature for 1 hour, heated to 55 ℃ for 7 hours, and then stirred at room temperature overnight. Cooling the system to 0 deg.C, quenching with acetic acid-D1, evaporating to dryness under reduced pressure to obtain residue, and performing column chromatography to obtain compound ZJT12 (3.0 g) with yield of 68.2%. ESI-MS (+): m/z =287.15

Example 31: synthesis of Compound PY-49

The reaction formula is as follows:

the preparation method comprises the following steps:

step 1: preparation of Compound PY-4902

Tetrahydrofuran (30 mL), PY-01-SM2 (3.18g, 10mmol) and 4-dimethylaminopyridine (DMAP, 1.83g, 15mmol) are sequentially added into a reaction bottle, 1-ethyl- (3-dimethylaminopropyl) carbonyl diimine (EDC, 1.86g, 12mmol) and ZJT10 (2.85g, 10mmol) are added after dissolution, the temperature is increased to 70 ℃, stirring reaction is carried out, TLC monitoring reaction is finished, the temperature of a system is reduced, evaporation is carried out, ethyl acetate and water are added into residues, an organic phase is separated out, washing is carried out twice, drying and reduced pressure evaporation are carried out, and the residues are purified by a column to obtain a product PY-4902 (4.2 g) with the yield of 71.7%. ESI-MS (+): m/z =586.20.

And 2, step: preparation of Compound PY-4901

Adding PY-4902 (4.1g, 7.0mmol), N-dimethylformamide (25 mL) and N, N-diisopropylethylamine (DIPEA, 1.81g, 14.0mmol) into a reaction bottle, dissolving, adding tripyrrolidinyl phosphonium bromide hexafluorophosphate (PyBroP, 3.59g, 7.7mmol), stirring the system at room temperature for 30min, adding hydroxylamine hydrochloride (0.59g, 8.4mmol), reacting at 40-50 ℃ for 4-6 h, detecting by TLC, cooling, adding water, extracting with ethyl acetate for 3 times, combining organic phases, washing twice with water, evaporating to dryness under reduced pressure, and obtaining PY-4901 (2.93 g) as a column chromatography product with the yield of 69.6%. ESI-MS (+): m/z =601.17.

Step 6: preparation of Compound PY-49

PY-4901 (2.9g, 4.83mmol) and formic acid (50 mL) were added to the reaction flask, and the system was reacted at room temperature for 20 hours. After the reaction was completed, concentration was performed under reduced pressure, and the residue was recrystallized from isopropanol/methyl t-butyl ether to give the compound PY-49 (2.17 g) in 80.1% yield, ESI-MS (+): m/z =561.14.1H NMR (DM SO-D6, 500MHz) < delta > 10.05 (s, 1H), 9.44 (s, 1H), 7.74-7.77 (m, 4H), 7.65-7.66 (d, 2H), 6.96-6.98 (d, 2H), 6.79-6.81 (d, 1H), 5.73-5.75 (d, 1H), 5.33-5.34 (d, 1H), 5.22-5.23 (d, 1H), 4.29-4.37 (m, 2H), 3.98 (s, 1H), 3.91-3.95 (m, 1H), 3.79-3.80 (d, 1H), 1.65-1.67 (d, 6H).

Example 32: preparation of Compound PY-50

The reaction formula is as follows:

/>

the preparation method comprises the following steps:

referring to the procedure of example 31, substituting ZJT12 for ZJT10 as the starting material gave PY-50 (1.21) in 42.2% overall yield. ESI-MS (+): m/z =562.18.

Example 33: preparation of Compound PY-51

The reaction formula is as follows:

the preparation method comprises the following steps:

referring to the procedure of example 31, substituting ZJT12 for ZJT10 as the starting material gave PY-51 (1.52) in an overall yield of 38.5%. ESI-MS (+): m/z =562.20.

Example 34: preparation of Compound PY-52

The reaction formula is as follows:

the preparation method comprises the following steps:

step 1: preparation of Compound PY-5203

Adding a compound ZJT10-01 (2.5g, 10.2mmol) and dichloromethane (50 mL) into a reaction bottle under the protection of nitrogen, and cooling to 0 ℃; DMAP (0.13g, 1.02mmol) and imidazole (2.8g, 40.9mmol) were then added in that order. Tert-butyldimethylsilyl chloride (TBSCl, 6.17g,4.0 mmol) was added over 10 minutes and the resulting mixture was warmed to ambient temperature and stirred for 18 hours. Water (30 mL) was added to the system, stirred at room temperature for 2 hours, separated, the aqueous phase extracted 3 times with dichloromethane, the combined organic layers washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and the residue was purified on a column to give the compound PY-5203 (4.14 g) in 69.0% yield. ESI-MS (+): m/z =588.33.

Step 2: preparation of Compound PY-5202

The compound PY-5203 (4.00g, 6.8mmol) and dichloromethane (100 mL) were charged to a reaction flask, and after cooling to 0 ℃ with stirring, DMAP (0.083 and g,0.68 mmol) and N, N-diisopropylethylamine (4.41g, 34.1mmol) were added successively to the above system, and 2,4, 6-triisopropylbenzene-1-sulfonyl chloride (4.13g, 13.6 mmol) was slowly added to the flask with stirring, and after completion of the addition, the mixture was stirred at room temperature for 18 hours. The system was again cooled to 0 deg.C, N-diisopropylethylamine (3.51g, 27.2 mmol) was added dropwise, followed immediately by solid hydroxylamine hydrochloride (1.9g, 27.3mmol). The mixture was warmed to room temperature and stirred for 3 hours. The reaction was quenched with water, separated, the aqueous layer was extracted 2 times with dichloromethane, the organic phases were combined, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure to give the compound PY-5202 (2.70 g) in 65.9% yield. ESI-MS (+): m/z =603.34.

And 3, step 3: preparation of Compound PY-5201

Reference example 31 preparation of step 1 substituting the compound PY-5202 for the compound ZJT10 gave the compound PY-5201 (2.22 g) in 66.5% yield. ESI-MS (+): m/z =903.43.

And 4, step 4: preparation of Compound PY-52

The compound PY-5201 (1.75g, 1.94mol) and tetrahydrofuran (20 mL) were added to a three-necked flask followed by triethylamine trihydrofluoride (0.31g, 1.94mmo 1) and the mixture was stirred at ambient temperature for 18 hours. The mixture was concentrated under reduced pressure, and column chromatography was performed on the residue to give PY-52 (0.53 g) in 48.7% yield. ESI-MS (+): m/z =561.16.

Example 35: preparation of Compound PY-54

The reaction formula is as follows:

the preparation method comprises the following steps:

referring to the procedure of example 34, substituting ZJT11-01 for ZJT10-01 as the starting material, compound PY-54 (0.46) was obtained in 16.2% overall yield. ESI-MS (+): m/z =562.20.

Example 36: preparation of Compound PY-55

The reaction formula is as follows:

the preparation method comprises the following steps:

referring to the procedures of step 1 and step 3 of example 31, starting from the compound PY-4901, compound PY-55 (0.25 g) was prepared in an overall yield of 50.2%. ESI-MS (+): m/z =861.22.

Example 37: preparation of Compound PY-56

The reaction formula is as follows:

the preparation method comprises the following steps:

step 1: synthesis of Compound PY-5604

PY-01-SM2 (3.82g, 12mmol) was added to dichloromethane (50 mL) under nitrogen, cooled to 0 deg.C, thionyl chloride (1.72g, 14.4 mmol) was added slowly, and after the addition was complete, the reaction was stirred at room temperature for 2.0h. Concentrating the system to dryness, removing the rest thionyl chloride with toluene for 3 times, and collecting the rest as compound PY-5604 without treatment;

and 2, step: synthesis of Compound PY-5603

Dichloromethane (50 mL) was added to the residue obtained in step 1 (Compound PY-5604, 12 mmol), and the above mixture was slowly added to a solution of aluminum trichloride (0.96g, 7.2 mmol) in dichloromethane (40 mL), and the mixture was stirred at room temperature for 20 minutes. The system was cooled to 0 ℃ and acetaldehyde (0.54g, 12mmol) was added dropwise over 10 minutes. The reaction mixture was stirred at room temperature for 1 hour, and the system was gradually added to a vigorously stirred ice-water slurry. Extraction with dichloromethane was carried out 3 times, the organic phases were combined, washed with ice water 2 times, the organic phase was separated, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to give the compound PY-5603 (2.07 g) in 45.2% yield. ESI-MS (+): m/z =381.06.

Step 3-step 5: synthesis of Compound PY-56

Referring to the procedures of the respective steps of example 31, PY-01-SM2 was replaced with PY-5603 as a starting material to obtain the compound PY-56 (1.12) in a total yield of 35.3% in three steps. ESI-MS (+): m/z =605.20.

The compounds of the following examples were synthesized in the same manner as in the above examples, using commercially available compounds or intermediate compounds appropriately synthesized from commercially available compounds.

/>

/>

/>

/>

Example 38: stability test

The compounds PY-01, PY-02, PY-03, PY-12, PY-15, PY-18, PY-21, PY-34, PY-49 and compound A were each prepared by mixing a sample with a 0.5% sodium carboxymethylcellulose solution and a 1.0% methylcellulose solution, placing the above samples (sealed at the mouth) in a 25.0 ℃ stability box with a humidity of 37% and keeping out of the sun, and measuring the stability by HPLC (with a DAD detector) on days 0, 3 and 7. The results are shown in Table I.

Table-stability test results

The results show that the series of compounds of the invention have better stability than the reference compound A.

Example 39: in vitro anti-influenza virus activity and cytotoxicity assays

Toxicity assay of test compounds on MDCK:

taking MDCK cells in logarithmic growth phase, inoculating them in 24-well cell culture plates, 37 ℃,5% CO ₂ The culture box is cultured for 24h, then the medicines with different dilution concentrations act for 2h, then the plate is washed for 3 times by Hanks liquid, 1ml of DMEM maintaining liquid containing 2% newborn bovine serum is added into each hole after being dried to maintain the growth, the temperature is 37 ℃, and the content of CO is 5% ₂ Cultured in an incubator. The degree of cytopathic effect (CPE) was observed over 7 days, and the half-toxic concentration of the sample (TC) to the cells was calculated by the Reed-Muench method ₅₀ )。

Determination of the anti-influenza Virus Activity of test Compounds:

MDCK cells in logarithmic growth phase were seeded in 24-well plates at approximately cell content per well (1X 10) ⁴ B), 37 ℃,5% CO ₂ Culturing in an incubator. 24H later the cells were infected with virus (H1N 1, A/WSN/33), washed 2 times with Hanks solution, treated with drug diluent, washed 3 times with Hanks solution, and finally maintained at 37 ℃ in DMEM containing 2% newborn bovine serum, and 5% CO ₂ Culturing in an incubator. The degree of cytopathic effect (CPE) was observed under an inverted microscope at 1, 2, 3,4,5, 6 and 7 days after the culture, and the half Inhibitory Concentration (IC) against virus of the cells was calculated for each sample by the Reed-Muench method ₅₀ ) Then, a Selection Index (SI) is calculated, SI = TC ₅₀ /IC ₅₀ . The results are shown in Table II.

TABLE II in vitro anti-influenza Virus Activity data

The experimental results show that the tested compound sample of the invention has better inhibitory activity to influenza virus (H1N 1), lower cytotoxicity and higher selection index compared with the disclosed compound A.

Example 40: in vitro anti-novel coronavirus activity (EC) ₅₀ ) Measurement of

Vero E6 cells were seeded in the microplate at a certain density and 5% CO ₂ And cultured overnight in an incubator at 37 ℃. The next day, compound diluted by multiple fold (8 concentration points, triple wells) and SARS-CoV-2 virus (B.1.1.7 (Alpha)) were added. Cell controls (cells, no compound treatment or viral infection) and viral controls (cells infected with virus, no compound treatment) were set. The cells were cultured in an incubator for 3 or 4 days.

The antiviral activity of the compound was represented by the inhibition rate (%) of the compound at various concentrations against the cytopathic effect caused by viruses. Compounds were calculated as EC using nonlinear fit analysis of inhibition of compounds using GraphPad Prism ₅₀ . The results are shown in Table three.

Data on the activity of the three-Table in vitro anti-novel coronavirus

Sample numbering	EC ₅₀ (μmol)
		Compound A	1.62
PY-01	0.35
		PY-02	0.58
PY-03	0.66
		PY-12	0.79
PY-15	0.57
		PY-18	0.66
PY-21	0.77
		PY-34	0.82
PY-49	0.33

The test results show that the tested compound sample of the invention has better inhibitory activity against SARS-CoV-2 virus (B.1.1.7 (Alpha)) compared with the disclosed compound A.

Example 41: pharmacokinetic profiling of Compound PY-01 in rats

12 SD rats, male, 180-220 g. The breeding conditions are as follows: 20-26 ℃, humidity: 40-70%, light illumination: dark =12h:12h; rats were acclimatized for 3 days during which time drinking water was consumed ad libitum. The groups were randomly divided into 4 groups of 3 animals each. 2 test samples (PY-01 and compound A) were administered by single equimolar oral gavage and equimolar tail vein injection, respectively. Fasting is started for 16-17h at 5 pm before administration, and animals are fed 4h after administration, and water is not forbidden in the whole process.

Blood is taken 0.25h, 0.5h, 1h, 2h, 3h, 4h, 8h and 24h after single oral gavage administration or 0.083h, 0.25h, 0.5h, 1h, 2h, 4h, 8h and 24h before single intravenous injection administration.

At each blood collection time point, about 300 μ L of venous blood is taken from rat eyeballs, added into a centrifugal tube with heparin sodium precooled by ice water, placed in an ice bath, kept stand, centrifuged (4000rpm, 10min), subpackaged by taking 50 μ L as unit volume, and placed in a sterile EP tube for storage at minus 80 ℃ for later use. The concentration of compound B in plasma was determined as soon as possible. The drug concentration in plasma was determined by HPLC (High Performance Liquid Chromatography) and LC/MS, and the bioavailability of the drug in rats was calculated by calculating the area under the concentration-time curve (AUC) in plasma using a non-linear least squares program.

Absolute bioavailability: f = AUClast-po/AUClast-iv × 100%. The results are shown in Table four.

The structure of compound B is as follows:

TABLE four pharmacokinetic results

The result of SD rat pharmacokinetics experiments shows that the compound PY-01 has longer half-life, larger exposure in vivo after oral administration and bioavailability which is nearly 1.4 times that of the compound A. The Cmax of compound B after administration of compound PY-01 was lower than compound a, indicating a better safety margin for the compounds of the present invention.

Example 42: in vivo anti-influenza virus therapeutic dosing test

C57BL/6J mice 30, 6-8 weeks old, SPF rating, female, randomly divided into 6 groups numbered as group 1, group 2, group 3, group 4, group 5 and group 6, respectively, with 5 per group. After 24H of nasal drip inoculation with a semi-lethal dose of 100pfu of influenza virus (H1N 1, A/WSN/33), group 1 (vehicle group) was gavaged with a 0.5% sodium carboxymethylcellulose solution; group 2 was gavaged with control drugs (equimolar, 17.33mg/kg compound A and 19.01mg/kg compound C, combination); groups 3-5 were gavaged with test drugs (29.46 mg/kg PY-01, 29.46mg/kg PY-03, 29.46mg/kg PY-49); group 6 administration of 29.46mg/kg of PY-01 was started 48h after the nasal drip. The administration in each group was equimolar twice daily, with groups 1-5 administered or vehicle for 4 days and group 6 administered for 3 days. Animals were sacrificed on day 5 post virus inoculation, lung tissue samples were harvested and virus titer was determined after homogenization. The results are shown in FIG. 1.

The results show that when the compound PY-01, the compound PY-03 and the compound PY-49 are applied 24H after the infection of the virus, the compound PY-01, the compound PY-03 and the compound PY-49 all show excellent anti-influenza virus (H1N 1) effects and show stronger anti-influenza virus (H1N 1) effects than the combined administration. The compound PY-01 can also obviously reduce the titer of the virus 48 hours after the virus is infected.

Example 43: in vivo anti-influenza virus (H1N 1) prophylactic dosing test

BALB/c mice 30, 6-8 weeks old, SPF rating, female, randomly divided into 6 groups numbered as group 1, group 2, group 3, group 4, group 5 and group 6, respectively, with 5 mice per group. Each group was started on day-1, and 100PFU of influenza virus (H1N 1, A/Puerto Rico/8/1934) was nasally inoculated on day 0, and 2 hours after the day of administration of the virus. Wherein, the group 1 (solvent group) is administered with 0.5% sodium carboxymethylcellulose solution by intragastric administration; group 2 was gavaged with control drugs (17.33 mg/kg of Compound A and 19.01mg/kg of Compound C, in combination); groups 3-5 were gavaged with different doses of test drugs (high, medium, and low dose groups of PY-01: 88.37mg/kg, 29.46mg/kg, 9.82 mg/kg), respectively; group 6 high doses (88.37 mg/kg) of the test drug PY-01 were administered intragastrically. Each group was given equimolar once daily, group 1-5 or vehicle 5 days, group 62 days. Animals were sacrificed on day 4 post virus inoculation, lung tissue samples were harvested and virus titer was determined after homogenization. The results are shown in FIG. 2.

The results show that: compared with the vehicle group, each group of the experiment shows lower virus titer; the PY-01 high dose group and the PY-01 medium dose group had lower virus titers compared to the combination group on the same days of administration; group 6 (high dose of PY-01 was not administered after virus inoculation 2 days before virus inoculation) had slightly higher virus titers than group 3 (high dose of PY-01 was administered 2 days before virus inoculation and 3 days after virus inoculation), but still lower than the other groups. This demonstrates that the compounds of the present invention have significant prophylactic effects against influenza virus.

Although the present invention has been described with respect to the preferred embodiments, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

Claims

1. A compound of formula (I) ₀ ) Novel cytidine derivatives, tautomers, stereoisomers, isotopic derivatives, and pharmaceutically acceptable salts thereof, as shown:

formula (I) ₀ ) In (1),

R ₁ and R ₂ Each independently selected from hydrogen,

Or, R ₁ And R ₂ Together with the oxygen to which they are attached form an acetal or ketal;

wherein n is ₁ Selected from 0, 1, 2 or 3;

n ₂ selected from 1, 2 or 3;

R ₃ And R ₄ Not all may be hydrogen;

wherein n is _a Selected from 0, 1, 2, 3,4, or 5;

n _b selected from 1, 2, 3,4, or 5;

n ₃ selected from 0, 1, 2, 3,4, or 5;

n ₄ selected from 0, 1, 2, 3, or 4;

z is selected from

Wherein n is ₅ Each independently is 0, 1, 2, 3,4, or 5;

2. The novel cytidine derivatives, tautomers, stereoisomers, isotopic derivatives and pharmaceutically acceptable salts thereof according to claim 1, having the structure of formula (I) ₀ -1)：

Formula (I) ₀ The substituents in (1) are as defined in claim 1 for formula (I) ₀ ) As defined.

3. The novel cytidine derivatives, tautomers, stereoisomers, isotopic derivatives and pharmaceutically acceptable salts thereof according to claim 1, having the structure of formula (I) ₀ -2)：

Formula (I) ₀ The substituents in (2) are as defined in claim 1(I ₀ ) As defined.

4. The novel cytidine derivatives, tautomers, stereoisomers, isotopic derivatives and pharmaceutically acceptable salts thereof according to claim 1, having the structure of formula (I) ₀ -3)：

/>

Formula (I) ₀ The substituents in (3) are as defined in claim 1 for formula (I) ₀ ) As defined.

5. The novel cytidine derivatives, tautomers, stereoisomers, isotopic derivatives, and pharmaceutically acceptable salts thereof, according to claim 1, having the structure of formula (I) ₀ -4)：

Formula (I) ₀ The substituents in (4) are as defined in claim 1 for formula (I) ₀ ) As defined.

6. Novel cytidine derivatives, tautomers, stereoisomers, isotopic derivatives, and pharmaceutically acceptable salts thereof, as claimed in claims 1-5, wherein the compounds include, but are not limited to, the following:

/>

/>

/>

/>

/>

/>

7. a pharmaceutical composition comprising the novel cytidine derivative, tautomer, stereoisomer, isotopic derivative, or pharmaceutically acceptable salt thereof according to any one of claims 1 to 6.

8. Use of a novel cytidine derivative, tautomer, stereoisomer, isotopic derivative according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 7 for the preparation of an antiviral medicament.

9. The use of claim 8, wherein the virus includes, but is not limited to: arenaviridae, filoviridae, and coronaviridae, and the like, including, but not limited to, adenovirus, rhinovirus, influenza virus, lassa virus, respiratory syncytial virus, severe acute respiratory syndrome virus, parainfluenza virus, coronavirus, and the like.

10. The use of claim 9, wherein the influenza and coronavirus include but are not limited to: influenza A virus, influenza B virus, SARS virus, MERS virus, and COVID-19 virus.