CN116462728B - Purine nucleoside derivative for resisting virus infection, pharmaceutical composition and application thereof - Google Patents

Purine nucleoside derivative for resisting virus infection, pharmaceutical composition and application thereof Download PDF

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CN116462728B
CN116462728B CN202211615605.2A CN202211615605A CN116462728B CN 116462728 B CN116462728 B CN 116462728B CN 202211615605 A CN202211615605 A CN 202211615605A CN 116462728 B CN116462728 B CN 116462728B
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CN116462728A (en
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张哲峰
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Nanjing Zhihe Medical Technology Co ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
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    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
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    • C07—ORGANIC CHEMISTRY
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    • C07J9/005—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane containing a carboxylic function directly attached or attached by a chain containing only carbon atoms to the cyclopenta[a]hydrophenanthrene skeleton
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    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
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    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

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Abstract

The invention discloses an antiviral purine nucleoside derivative, a pharmaceutical composition and application thereof, wherein the antiviral purine nucleoside derivative is shown as a formula (I) 0 ) Shown; the compound can be used for preparing antiviral drugs.

Description

Purine nucleoside derivative for resisting virus infection, pharmaceutical composition and application thereof
Technical Field
The invention relates to the technical field of pharmaceutical chemistry, in particular to a purine nucleoside derivative for resisting viral infection, a pharmaceutical composition and application thereof.
Background
Since the outbreak of coronavirus disease-19 (covd-19, also known as SARS-CoV-2), it has evolved into a pandemic that has a significant impact on the public medical system and global economy. By 3 months of 2021, over 1.1 hundred million new coronavirus-19 cases and over 250 ten thousand deaths have been reported.
Coronaviruses belong to the single positive strand RNA virus, and the family of coronaviruses mainly includes the novel coronavirus (SARS-CoV-2), the SARS coronavirus (SARS-CoV), the middle east respiratory syndrome coronavirus (MERS-CoV), HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU. Coronaviruses often cause respiratory and intestinal diseases, neurological symptoms and myocarditis.
In view of the severity, high variability and pandemic of SARS-CoV-2 infection, and because of the problems of emergence or side effects on resistant strains, although approved new coronavirus-19 therapeutic vaccines and drugs bring our hope for coping with pandemic, the development of various specific inhibitors with different mechanisms of action to treat new coronaviruses is urgent in order to reduce the incidence, severity and mortality of SARS-CoV-2 infection. Thus, there remains a need in the art for the development of antiviral drugs of novel structure.
Disclosure of Invention
The inventor develops a purine nucleoside derivative for resisting virus infection, and the purine nucleoside derivative can be used for preparing antiviral medicines.
One aspect of the present invention provides a method as described in (I 0 ) Purine nucleoside derivatives, tautomers, stereoisomers, solvates and pharmaceutically acceptable salts thereof, which are shown to be resistant to viral infection:
formula (I) 0 ) In,
R X1 and R is X2 Each independently selected from hydrogen, hydroxy, C1-C8 alkyl, and halogen;
R X3 selected from hydrogen, or azido;
R X4 selected from hydrogen, or cyano;
R 1a and R is 1b Independently selected from hydrogen, -OR 5 C1-C8 alkyl substituted or unsubstituted by the group A; wherein,
r is as described above 5 Selected from hydrogen, Wherein,
above n 1 Selected from 0, 1, 2 or 3, when n 1 When 0, the carbonyl is directly connected with oxygen to form an ester bond;
n 2 selected from 1, 2 or 3;
R a selected from the group consisting ofOr the following groups, substituted or unsubstituted with one or more groups a: C1-C8 alkyl, C1-C8 alkoxy, C3-C8 cycloalkyl, C3-C8 cycloalkoxy, C2-C8 alkenyl, C6-C18 aryl, C6-C18 aryloxy; wherein,
R b selected from hydrogen or the following groups substituted or unsubstituted with one or more groups a: C1-C8 alkyl, C1-C8 alkoxy, C3-C8 cycloalkyl, C3-C8 cycloalkoxy, C2-C8 alkenyl, C6-C18 aryl, C6-C18 aryloxy;
R c1 and R is c2 Each independently selected from hydrogen, C1-C8 alkyl substituted or unsubstituted with one or more groups A;
R 2 selected from hydrogen or amino;
R 3 and R is 4 Independently selected from hydrogen,
Wherein R is 3 And R is 4 Cannot be hydrogen at the same time;
r is as described above a 、R c1 、R c2 、n 1 And n 2 As defined above;
n 3 and n 4 Each independently selected from 0, 1, 2, 3 or 4;
R 6a and R is 6b Each independently selected from hydrogen, C1-C8 alkyl substituted or unsubstituted with group A; or R is 6a And R is 6b Cycloalkyl groups attached to the carbon;
R 7 selected from hydrogen, halogen, amino, C1-C8 alkyl substituted or unsubstituted with group A;
z is selected from hydrogen, hydroxy, halogen,Wherein,
above n 5 Selected from 0, 1, 2, 3 or 4;
R 8 selected from H, hydroxy, halogen, the following substituted or unsubstituted with one or more groups a: amino, C1-C8 alkyl, C6-C18 aryl, C1-C8 alkyloxy, amino C1-C8 alkyl, C1-C8 alkylaryl, C6-C18 arylcarbonyl, C1-C8 alkylcarbonyloxy;
x is selected from O, S, or NH;
R d1 and R is d2 Each independently selected from hydrogen, halogen, the following substituted or unsubstituted with one or more groups a: C1-C8 alkyl, C1-C8 alkoxy, C1-C8 alkylamino, or R d1 And R is d2 The carbon to which it is attached forms a 3-7 membered ring, the atom or group on this 3-7 membered ring being optionally substituted with one or more groups A;
R e selected from the following groups, substituted or unsubstituted with one or more groups a: C1-C8 alkyl, C1-C8 alkoxy, C1-C8 alkylamino;
R f1 and R is f2 Each independently selected from hydrogen, halogen or C1-C8 alkyl substituted or unsubstituted with one or more groups A;
R g 、R h each independently selected from the following groups substituted or unsubstituted with one or more groups a: C1-C8 alkyl, C1-C8 alkoxy, or R g 、R h And the carbon to which it is attached to form a 3-7 membered ring, the atom or group on this 3-7 membered ring being optionally substituted with one or more groups A;
R i1 And R is i2 Are independently selected from hydrogen, cyano, and,Or substituted or unsubstituted by one or more groups AThe following groups: C1-C8 alkyl, C1-C8 alkoxy, C2-C8 alkenyl, C2-C8 alkynyl; wherein Ra is as defined above;
the group A is: C1-C8 alkyl, C1-C8 alkoxy, Amino, mercapto C1-C8 alkyl, halogen, cyano, aldehyde, nitro, trifluoromethyl, chlorobenzoyl, C3-C8 cycloalkyl, C3-C8 cycloalkoxy, C6-C18 aryl, C6-C18 aryloxy; wherein R is a 、R b As defined above; m is selected from 1 or 2.
In some embodiments, the invention provides a purine nucleoside derivative, tautomer, stereoisomer, solvate, and pharmaceutically acceptable salt thereof, having an antiviral infection as shown in formula (I):
the definition of the substituent in the formula (I) is as shown in the formula (I) 0 ) Said method.
In some embodiments, the invention provides a purine nucleoside derivative, tautomer, stereoisomer, solvate, and pharmaceutically acceptable salt thereof, having an antiviral infection as shown in formula (II):
the definition of the substituent in the formula (II) is as shown in the formula (I) 0 ) Said method.
In some embodiments, the invention provides a purine nucleoside derivative, tautomer, stereoisomer, solvate, and pharmaceutically acceptable salt thereof, having an antiviral infection of formula (iii):
The definition of the substituent in the formula (III) is as shown in the formula (I) 0 ) Said method.
In some embodiments, the invention provides a purine nucleoside derivative, tautomer, stereoisomer, solvate, and pharmaceutically acceptable salt thereof having a multiple antiviral infection of formula (IV):
the definition of the substituent in the formula (IV) is as shown in the formula (I) 0 ) Said method.
In some embodiments, the above formula (I 0 ) Wherein R is X1 And R is X2 Each independently selected from hydrogen, C1-C8 alkyl, and halogen; preferably, R X1 And R is X2 Each independently selected from hydrogen, hydroxy, methyl, fluorine atoms.
In some embodiments, the above formula (I 0 ) Wherein R is X3 Is hydrogen, or azido;
in some embodiments, the above formula (I 0 ) Wherein R is X4 Hydrogen, or cyano;
in some embodiments, the above formula (I 0 ) In- (IV), R 1a And R is 2b Independently selected from hydrogen, C1-C8 alkyl substituted or unsubstituted with group A;
in some more specific embodiments, the above formula (I 0 ) In- (IV), R 1a And R is 2b Independently selected from hydrogen and C1-C6 alkyl.
In some embodiments, the above formula (I 0 ) In- (IV), R 2 Is hydrogen;
in some embodiments, the above formula (I 0 ) In- (IV), R 2 Is amino.
In some embodiments, the above formula (I 0 ) In (I), R 3 Selected from hydrogen,
Wherein when R is 3 When hydrogen, R 4 Is not hydrogen;
in some embodiments, the above formula (I 0 ) In- (II), R 3 Is thatWherein,
in some embodiments, the above formula (I 0 ) - (II) in which n is as defined above 1 Selected from 0, 1 or 2;
in some more specific embodiments, the above formula (I 0 ) - (II) in which n is as defined above 1 Is 0;
in some more specific embodiments, the above formula (I 0 ) - (II) in which n is as defined above 1 1.
In some embodiments, the above formula (I 0 ) - (II) in which n is as defined above 2 1 is shown in the specification;
in some embodiments, the above formula (I 0 ) - (II) in which n is as defined above 2 2.
In some embodiments, the above formula (I 0 ) - (II) in which R is as defined above a Selected from the group consisting ofOr the following groups, substituted or unsubstituted with one or more groups a: C1-C8 alkyl, C2-C8 alkenyl, C3-C8 cycloalkyl, C6-C18 aryl;
in some more specific embodiments, the above formula (I 0 ) - (II) in which R is as defined above a Selected from the group consisting ofWherein,
in some more specific embodiments, the above formula (I 0 ) - (II) in which R is as defined above b Selected from hydrogen or the following groups substituted or unsubstituted with one or more groups a: C1-C8 alkyl, C3-C8 cycloalkyl, C2-C8 alkenyl, C6-C18 aryl;
in some more specific embodiments, the above formula (I 0 ) - (II) in which R is as defined above a Selected from the following groups, substituted or unsubstituted with one or more groups a: C1-C8 alkyl, C3-C8 cycloalkyl, C6-C18 aryl.
In some embodiments, the above formula (I 0 ) - (II) in which R is as defined above c1 And R is c2 Are all hydrogen;
in some embodiments, the above formula (I 0 ) - (II) in which R is as defined above c1 And R is c2 Wherein one hydrogen and the other is C1-C8 alkyl substituted or unsubstituted with one or more groups A;
in some embodiments, the above formula (I 0 ) - (II) in which R is as defined above c1 And R is c2 Are each C1-C8-alkyl substituted or unsubstituted by one or more groups A.
In some embodiments, the above formula (I 0 ) In (I) and/or (III), R 3 Selected from the group consisting ofWherein,
in some embodiments, the above formula (I 0 ) - (I) and/or (III) in which n is as defined above 1 、n 2 、R c1 And R is c2 Respectively as defined above.
In some embodiments, the above formula (I 0 ) - (I) and/or (III) in which n is as defined above 3 Selected from 0, 1, 2, 3;
in some more specific embodiments, the above formula (I 0 ) - (I) and/or (III) in which n is as defined above 3 Is 0;
in some more specific embodiments, the above formula (I 0 ) - (I) and/or (III) in which n is as defined above 3 Is 2;
in some more specific embodimentsWherein the above formula (I) 0 ) - (I) and/or (III) in which n is as defined above 3 3.
In some embodiments, the above formula (I 0 ) - (I) and/or (III) in which n is as defined above 4 Selected from 0, 1 or 2;
in some more specific embodiments, the above formula (I 0 ) - (I) and/or (III) in which n is as defined above 4 Is 0;
in some more specific embodiments, the above formula (I 0 ) - (I) and/or (III) in which n is as defined above 4 1 is shown in the specification;
in some more specific embodiments, the above formula (I 0 ) - (I) and/or (III) in which n is as defined above 4 2.
In some embodiments, the above formula (I 0 ) - (I) and/or (III), R is as defined above 6a And R is 6b Are all hydrogen;
in some embodiments, the above formula (I 0 ) - (I) and/or (III), R is as defined above 6a And R is 6b Wherein one hydrogen and the other is C1-C8 alkyl substituted or unsubstituted with one or more groups A, preferably C1-C4 alkyl;
in some embodiments, the above formula (I 0 ) - (I) and/or (III), R is as defined above 6a And R is 6b Are each C1-C8-alkyl substituted or unsubstituted by one or more groups A.
In some embodiments, the above formula (I 0 ) - (I) and/or (III), R is as defined above 6a And R is 6b The C attached thereto is a 3-7 membered cycloalkyl group, the atom or group on this 3-7 membered ring may be substituted or unsubstituted with one or more groups A.
In some embodiments, the above formula (I 0 ) - (I) and/or (III), R is as defined above 7 Is hydrogen; or C1-C8 alkyl substituted or unsubstituted by the group A.
In some embodiments, the above formula (I 0 ) - (I) and/or (III) wherein Z is selected from the group consisting of hydrogen, F, cl, br,Wherein,
in some embodiments, the above formula (I 0 ) - (I) and/or (III), R is as defined above 8 Selected from hydrogen, hydroxy or the following groups substituted or unsubstituted with one or more groups a: amino, C1-C8 alkyl, C6-C18 aryl, C1-C8 alkyloxy, C1-C8 alkylaryl.
In some embodiments, the above formula (I 0 ) - (I) and/or (IV), R 3 Selected from the group consisting ofWherein,
in some embodiments, the above formula (I 0 ) - (I) and/or (IV), n is as defined above 1 、n 2 、R c1 And R is c2 Respectively as defined above.
In some embodiments, the above formula (I 0 ) In (I) and/or (IV), X is selected from O;
in some embodiments, the above formula (I 0 ) In (I) and/or (IV), X is selected from S;
in some embodiments, the above formula (I 0 ) In (I) and/or (IV), X is selected from NH.
In some embodiments, the above formula (I 0 ) - (I) and/or (IV), R as described above d1 And R is d2 Each independently selected from hydrogen, halogen, or the following substituted or unsubstituted with one or more groups a: C1-C8 alkyl, C1-C8 alkylamino, or R d1 And R is d2 The carbon connected with the catalyst forms a 3-7 membered ring;
in some more specific embodiments, the above formula (I 0 ) - (I) and/or (IV), R as described above d1 And R is d2 Each independently selected from hydrogen, F, cl or Br.
In some embodiments, the above formula (I 0 ) - (I) and/or (IV), R as described above f1 Is hydrogen, R f2 Is halogen or C1-C8 alkyl substituted or unsubstituted by one or more groups A;
in some embodiments, the above formula (I 0 ) - (I) and/or (IV), R as described above f1 Is halogen orC1-C8 alkyl optionally substituted by one or more groups A, R f2 Is hydrogen;
in some embodiments, the above formula (I 0 ) - (I) and/or (IV), R as described above f1 And R is f2 Are all hydrogen.
In some embodiments, the above formula (I 0 ) - (I) and/or (IV), R as described above g And R is h Each independently selected from the following groups: C1-C8 alkyl, C1-C8 alkoxy, or R g 、R h And the carbon to which it is attached is linked to form a 3-7 membered ring, the atom or group on this ring being optionally substituted with one or more groups A;
in some more specific embodiments, the above formula (I 0 ) - (I) and/or (IV), R as described above g 、R h And the carbon to which it is attached is linked to form a 3-7 membered ring, the atom or group on this ring being optionally substituted with one or more groups A;
In some more specific embodiments, the above formula (I 0 ) - (I) and/or (IV), R as described above g 、R h And the carbon to which it is attached, is linked to a 3-7 membered ring, the atoms on this 3-7 membered ring being substituted by the following principles or groups: C1-C8 alkyl, C1-C8 alkoxy, halogen, cyano, nitro, trifluoromethyl, C3-C8 cycloalkyl, C3-C8 cycloalkoxy, C6-C18 aryl, C6-C18 aryloxy.
In some embodiments, the above formula (I 0 ) - (I) and/or (IV), R as described above i1 And R is i2 Are independently selected from hydrogen, cyano, and,Or the following groups, substituted or unsubstituted with one or more groups a: C1-C8 alkyl, C1-C8 alkoxy;
in some more specific embodiments, the above formula (I 0 ) - (I) and/or (IV), R as described above i1 And R is i2 Are respectively and independently selected fromCyano groups or covered by one or moreThe group A is a substituted or unsubstituted C1-C8 alkyl group.
In some embodiments, the above formula (I 0 ) In- (IV), R 4 Selected from hydrogen,
And is provided that when R 4 When hydrogen, R 3 Is not hydrogen;
r is as described above a 、R c1 、R c2 、R d1 、R d2 、R e 、R f1 、R f2 、R g 、R h 、R i1 、R i2 、R 6a 、R 6b 、R 7 、Z、n 1 、n 2 、n 3 And n 4 As defined above.
The group A is: C1-C8 alkyl, C1-C8 alkoxy, Amino, mercapto C1-C8 alkyl, halogen, cyano, aldehyde, nitro, trifluoromethyl, chlorobenzoyl, C3-C8 cycloalkyl, C3-C8 cycloalkoxy, C6-C18 aryl, C6-C18 aryloxy; wherein R is a 、R b As defined above; m is selected from 1 or 2.
In some embodiments, the invention provides the above purine nucleoside derivatives against viral infection selected from the following compounds:
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in another aspect, in some embodiments, the invention provides pharmaceutical compositions comprising purine nucleoside derivatives, tautomers, stereoisomers, solvates, and pharmaceutically acceptable salts thereof, described above, that are resistant to viral infections.
In some embodiments, the invention discloses a pharmaceutical composition, which is composed of the compound, isomer or pharmaceutically acceptable salt thereof of the invention as an active ingredient or a main active ingredient, and a pharmaceutically acceptable carrier.
In some embodiments, the present invention provides that the above pharmaceutical compositions are useful for the treatment and 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 drug.
In yet another aspect, the present invention provides a pharmaceutical composition comprising the above for use in the treatment of any viral-caused disease in a human or animal; such viruses include, but are not limited to: arenaviridae, filoviridae, and coronaviridae, and the like, including, but not limited to, adenovirus, rhinovirus, hepatitis a virus, hepatitis c virus, pneumo virus b, pneumo virus c, HIV virus, polio virus, measles virus, ebola virus, coxsackie virus, west nile virus, smallpox virus, yellow fever virus, dengue virus, influenza virus, lassa virus, respiratory syncytial virus, severe acute respiratory syndrome virus, parainfluenza virus, coronavirus, and the like.
Further, the influenza viruses include, but are not limited to, influenza a virus, influenza b virus, and the like;
further, the above coronaviruses include, but are not limited to, SARS virus, MERS virus, covd-19 virus, etc.;
in some embodiments, the purine nucleoside derivatives of the present invention may be formulated as pharmaceutical compositions for administration to a patient in a variety of suitably selected modes of administration, including systemic, e.g., oral, inhaled or parenteral, by intravenous, intramuscular, transdermal or subcutaneous, and the like.
In some embodiments, the purine nucleoside derivatives of the present invention may be formulated as pharmaceutical compositions including, but not limited to, tablets, capsules or solutions for oral administration, or solutions, sprays, emulsions, ointments, emulsions or gels for transdermal administration.
The compound disclosed by the invention has a spectrum antiviral effect, has better activity in the aspects of resisting influenza virus and coronavirus compared with the compound A, and has lower cardiotoxicity;
the compound disclosed by the invention can more effectively protect mice from death caused by lethal dose infection in a K18-ACE2 transgenic mouse infection model infected by new coronavirus than compound A, and unexpectedly shows remarkable effect in terms of disease course shortening.
The compound disclosed by the invention shows good effect of preventing new coronavirus infection in the research of Omicron mutant infection, and shows a dose-dependent trend under a set dose.
The compounds of the invention can be used as antiviral drugs with novel structures.
The structure of compound a is as follows:
definition:
the following terms and phrases used herein are intended to have the following meanings unless otherwise indicated. A particular term or phrase, unless otherwise specifically defined, should not be construed as being ambiguous or otherwise clear, but rather should be construed in a generic sense. When trade names are presented herein, it is intended to refer to their corresponding commercial products or active ingredients thereof.
Certain compounds of the invention may exist in unsolvated forms or solvated forms such as, for example, hydrated, ethanolic forms. In general, solvated forms, which are equivalent to unsolvated forms, are intended to be encompassed within the scope of the present invention.
The term "substituted" means that one, two or three or more hydrogen atoms thereon are independently substituted with a substituent, which may be the same or different when substituted with more than one substituent.
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 salt" refers to salts of the compounds of the present invention prepared from the compounds of the present invention which have the specified substituents found herein with relatively non-toxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts may be obtained by contacting neutral forms of such compounds with a sufficient amount of a base in pure 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 the compounds of the present invention contain relatively basic functional groups, the acid addition salts may be obtained by contacting the neutral form of such compounds with a sufficient amount of an acid in pure solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts 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 organic acid salts including acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; also included are salts of amino acids (e.g., arginine, etc.), and salts of organic acids such as glucuronic acid. Certain specific compounds of the invention contain basic and acidic functionalities that can be converted to either base or acid addition salts.
The term "alkyl" means a saturated aliphatic radical, including straight and branched chain groups, alkyl groups which 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" means an aliphatic radical having an unsaturated carbon-carbon double bond, and includes straight and branched chain alkyl groups which may be substituted or unsubstituted. The carbon-carbon double bond may be one or more.
The term "cycloalkyl" means a single or fused ring of all carbons ("fused" ring means that each ring in the system shares an adjacent pair of carbon atoms with the other rings in the system), wherein one or more of the rings does not have a fully attached pi-electron system, examples of cycloalkyl (without limitation) are cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, adamantane, cyclohexadiene, cycloheptane, and cycloheptatriene. Cycloalkyl groups may be substituted and unsubstituted.
The term "aryl" means an all-carbon monocyclic or fused multicyclic group of 1 to 12 carbon atoms having a fully conjugated pi-electron system. Non-limiting examples of aryl groups are phenyl, naphthyl and anthracenyl. Aryl groups may be substituted or unsubstituted. When substituted, the substituents are preferably one or more, more preferably one, two or three, and even more preferably one or two.
The term "arylalkyl" denotes an alkyl group substituted with an aryl group.
The term "heteroaryl" means a multi-atom monocyclic or fused ring radical containing one, two, three or four ring heteroatoms selected from N, O or S, the remaining ring atoms being C, additionally having a fully 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" refers to a group in which an alkyl group is attached to oxygen, where the alkyl group may be straight chain, branched or cyclic alkyl.
The term "hydroxy" denotes an-OH group.
The term "amino" means-NH 2 A group.
The term "halogen" means fluorine, chlorine, bromine or iodine.
The term "pharmaceutically acceptable carrier" refers to any formulation or carrier medium representative of a carrier capable of delivering an effective amount of the active agents of the present invention, which does not interfere with the biological activity of the active agents and which does not have toxic or side effects to the host or patient, including water, oils, vegetables and minerals, cream bases, lotion bases, ointment bases, and the like. Such matrices include suspending agents, viscosity enhancers, transdermal enhancers, and the like.
The term "stereoisomer" refers to a compound that has the same chemical constitution but differs in the arrangement of atoms or groups in space.
By "solvate" is meant that certain compounds may exist in unsolvated forms or solvated forms, including hydrated forms. In general, solvated forms, which are equivalent to unsolvated forms, are intended to be encompassed within the scope of the present application.
The numerical ranges stated herein, e.g., "C1-C8", means that the group can contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 8 carbon atoms.
The compounds of the present application may exist in specific geometric or stereoisomeric forms. The present application contemplates all such compounds, including cis and trans isomers, (-) -and (+) -pairs of enantiomers, (R) -and (S) -enantiomers, diastereomers, (D) -isomers, (L) -isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the application. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of the present application.
Description of the drawings:
FIG. 1 is a graph showing the change in body weight of mice in example 17.
FIG. 2 shows a schematic representation of mouse lung tissue virus titers in example 18.
Detailed Description
A number of exemplary methods for preparing the compounds of the present application are provided in the examples below. The present application is described in detail below by way of examples, but is not meant to be limiting in any way. The present application has been described in detail herein, and specific embodiments thereof are also disclosed, it will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the application without departing from the spirit and scope of the application. Certain compounds of the application can be used as intermediates for preparing other compounds of the application, all of which have structures determined by MS.
The starting materials, solubles, catalysts in the examples of the present application were all purchased commercially.
Example 1: synthesis of Compound ZJT1
The reaction formula:
the preparation method comprises the following steps:
step 1: preparation of Compound ZJT 1-03:
under the protection of nitrogen, the compound ZJT1-SM1 (37.2 g,100 mmol) is added into anhydrous tetrahydrofuran (370 mL), cooled to-25 to-35 ℃, and an anhydrous tetrahydrofuran solution (35 mL) dissolved with lithium tri-tert-butyl aluminum hydride (25.4 g,100 mmol) is added dropwise at the temperature; after the addition was complete, the reaction was carried out for 2h and tlc detection was complete. Heating to-15 ℃, adding saturated ammonium chloride aqueous solution into the system, stirring and quenching the reaction, filtering by diatomite, concentrating the filtrate under reduced pressure to remove tetrahydrofuran, adding ethyl acetate into the residue to extract twice, combining organic phases, drying by anhydrous sodium sulfate, filtering, concentrating, and directly carrying out the next step without further treatment on the residue, wherein the residue is the compound ZJT 1-03.
Step 2: preparation of Compound ZJT 1-02:
the residue was taken up in methylene chloride (280 mL), nitrogen-protected, cooled to about-25℃and triphenylphosphine (28.9 g,110 mmol) was added, and the mixture was stirred for 20 minutes after the addition was completed, and carbon tetrabromide (CBr) 4 39.8g,120 mmol), and slowly heating the system to about 0 ℃ after the addition is completed; after the TLC monitoring, the reaction was completed, the filtrate system was concentrated to dryness, the residue was recrystallized from petroleum ether/n-hexane, and the obtained solid was purified by a silica gel column and eluted to obtain the compound ZJT1-02 (18.6 g), the total yield of the two steps was 42.5%. ESI-MS (+): m/z= 437.03.
Step 3: preparation of Compound ZJT 1-01:
2-amino-6-chloropurine (8.5 g,50 mmol) was dissolved in t-butanol (120 mL) under argon and potassium t-butoxide (5.6 g,50 mmol) was added. Stirring at room temperature for 1 hour, slowly adding anhydrous acetonitrile solution (25 mL) dissolved with the compound ZJT1-02 (18.6 g,42.5 mmol) to the system, heating the system to 65 ℃, reacting for 20 hours, cooling to room temperature after TLC detection, quenching the reaction with saturated ammonium chloride solution (100 mL), extracting with ethyl acetate (100 mL×3), merging organic phases, drying with anhydrous sodium sulfate, filtering, concentrating, and performing column chromatography to obtain the compound ZJT1-01 (9.8 g), wherein the yield is 43.8%. ESI-MS (+): m/z= 526.17.
Step 4: preparation of compound ZJT 1:
compound ZJT1-01 (5.26 g,10 mmol) is dissolved in absolute ethanol (100 mL) under argon, and 33% anhydrous solution of monomethylamine (1.4 g,15 mmol) is added and the system is heated to 80deg.C for 24 hours. After the completion of the reaction, the mixture was concentrated to dryness, and column chromatography gave compound ZJT1 (2.14 g) in a yield of 68.6%. ESI-MS (+): m/z= 313.13.
Example 2: synthesis of Compound ZJT2
The reaction formula:
the preparation method comprises the following steps:
step 1: preparation of Compound ZJT 2-07:
ZJT2-SM1 (27.2 g,0.1 mol) was added to 300mL of methylene chloride under argon protection, triethylamine (30.4 g,0.3 mol) was added, isobutyl chloroformate (13.7 g,0.1 mol) was slowly added under ice bath, the reaction was continued at room temperature with stirring for 3 hours, methanol solution of ammonia gas (2.0M, 50 mL) was added under ice bath, the reaction was continued at room temperature for 5 hours, concentration was performed under reduced pressure, the resulting residue was added to DMF (500 mL), 2,4, 6-collidine (36.4 g,0.3 mol) and cyanuric chloride (55.3 g,0.3 mol) were slowly added under ice bath, reaction was performed at room temperature for 20 hours, DMF was distilled off under reduced pressure, the residue was added to water, extraction was performed 3 times with ethyl acetate, ethyl acetate was combined, saturated brine was washed, dried over anhydrous sodium sulfate, concentration was performed, and the residue was recrystallized from ethanol/water to obtain the compound ZJT2-07 (13.2 g, 52.1%) ESI-MS: 254.14 (+).
Step 2: compound ZJT2-06 preparation:
ZJT2-07 (2.5 g,10.0 mmol) was added to 10mL of ethyl acetate under argon, 5mL of ethyl acetate hydrochloride solution was added, and the mixture was stirred at room temperature for 1.5 hours, and the mixture was neutralized to basicity with triethylamine to obtain a stock solution of compound ZJT2-06 ethyl acetate.
Step 3: preparation of Compound ZJT 2-05:
ZJT2-SM2 (11.1 g,0.1 mol) was added to water (1000 mL), sodium persulfate (71.4 g,0.3 mol) and silver nitrate (1.7 g,0.01 mol) were added, heated to reflux for 8 hours, cooled, concentrated, the remainder was added to ethanol (200 mL), stirred for 2 hours, filtered, the filtrate was concentrated, and the remainder was purified by column chromatography to give compound ZJT2-05 (5.93 g, 54.3%), ESI-MS (+): m/z=110.10.
Step 4: compound ZJT2-04 preparation:
ZJT2-05 (5.5 g,50 mmol) was added to toluene (100 mL) under nitrogen protection, rhodium dicarbonyl acetylacetonate (5.2 g,20 mmol) and toluene solution of N-Bn-Yanphos ligand (1.2 g) were added to 20mL, benzoic acid (2.4 g,20 mmol) was added, the reaction solution was transferred to an autoclave, carbon monoxide (3.0 bar) and hydrogen (3.0 bar) were introduced, the pressure reflux reaction was maintained for 48 hours, cooled to room temperature, magnesium monoperoxyphthalate hexahydrate (40.4 g,0.1 mol) in methanol (50 mL) and phosphoric acid buffer solution (pH=6.5, 50 mL) were added under ice bath, the reaction was continued for 2 hours under heating, cooled, the organic phase was separated, concentrated, and the compound ZJT2-04 (1.5 g, 22.0%) was separated by column chromatography.
Step 5: preparation of Compound ZJT 2-03:
ZJT2-04 (1.4 g,10.0 mol) was added to methanol (20 mL), water (2 mL) and sodium hydroxide (0.8 g,20.0 mol) were added sequentially, heated to reflux for 2 hours, pH was adjusted to 5-6 with hydrochloric acid (0.1M), concentrated, and the liquid phase was separated to give compound ZJT2-03 (0.38 g, 24.5%), ESI-MS (-):m/z= 154.09.
Step 6: compound ZJT2-02 preparation:
ZJT2-03 (1.5 g,10.0 mmol) was added to tetrahydrofuran (50 mL), 1-hydroxybenzotriazole (HOBt, 1.6g,12.0 mmol), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDCI, 2.3g,12.0 mmol), triethylamine (3.0 g,30.0 mmol), stirred at room temperature for 2 hours, the above-mentioned ZJT2-06 ethyl acetate stock solution was added, the reaction was continued at room temperature for 4 hours, the resulting residue was added to water, extracted twice with ethyl acetate, the organic phase was separated, washed twice with saturated brine, the organic phase was dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to give the compound ZJT2-02 (1.13 g, 38.9%), ESI-MS (+): m/z= 291.17.
Step 7: preparation of Compound ZJT 2-01:
Boc-L-tert-leucine (1.2 g,5.0 mmol) was added to 40mL tetrahydrofuran, HOBt (0.81 g,6.0 mmol) EDCI (1.15 g,6.0 mmol), triethylamine (1.5 g,15.0 mmol), a solution of ZJT2-02 (1.45 g,5.0 mmol) in tetrahydrofuran (20 mL) was added and the reaction was continued at room temperature for 3 hours, concentrated, added with water and ethyl acetate and shaken, the organic phase was separated, 1M solution of ethyl acetate hydrochloride was added, stirring was continued for 1 hour, the system was made alkaline with triethylamine twice under ice bath, the organic phase was dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to give the compound ZJT2-01 (0.65 g, 32.4%) ESI-MS (+): M/z= 404.26.
Step 8: compound ZJT2 preparation:
2, 2-difluoro-2-hydroxyacetic acid (0.6 g,5.0 mmol) was added to tetrahydrofuran (40 mL), HOBt (0.81 g,6.0 mmol), EDCI (1.15 g,6.0 mmol), triethylamine (1.5 g,15.0 mmol), a solution of ZJT2-01 (2.0 g,5.0 mmol) in tetrahydrofuran (20 mL) was added and the reaction was continued at room temperature for 3 hours, the mixture was concentrated, the remainder was shaken with water and ethyl acetate, the organic phase was washed twice, dried over anhydrous sodium sulfate, concentrated, and column chromatographed to give compound ZJT2 (0.78 g, 31.2%), ESI-MS (+): m/z= 498.24.
Example 3: synthesis of Compound VIZ98-01
The reaction formula:
the preparation method comprises the following steps:
tetrahydrofuran (30 mL), a compound ZJT1 (3.12 g,10 mmol) and isobutyric acid-2, 2-trifluoroethyl ester (1.7 g,10 mmol) are sequentially added into a reaction bottle, a potassium dihydrogen phosphate sodium hydroxide buffer solution (pH-6.8, 30 mL) is added, after shaking and dissolving, an esterifying enzyme protease N (1.0 g) is added, and the temperature is raised to 35-40 ℃ for shaking and reacting for 36-40 hours. After the reaction, the mixture was cooled to room temperature, filtered, extracted 3 times with ethyl acetate, the organic phases were combined, washed 2 times with water, concentrated, and the residue was purified by column to give VIZ-01 (0.83 g) in 21.7% yield. ESI-MS (+): m/z= 383.18.
Example 4: synthesis of Compound VIZ98-02
The reaction formula:
the preparation method comprises the following steps:
tetrahydrofuran (30 ml), a compound ZJT1 (3.12 g,10 mmol) and isobutyric acid (1.76 g,20 mmol) are sequentially added into a reaction bottle, 4-dimethylaminopyridine (DMAP, 2.44g,20 mmol) is added after dissolution, EDCI (3.82 g,20 mmol) is added, the temperature is raised to 70 ℃, the reaction is stirred, TLC monitors the reaction completion, the system is cooled, the concentration is carried out, the remainder is added into ethyl acetate and water, an organic phase is separated, washed twice again, dried, evaporated to dryness under reduced pressure, and the remainder is purified by a column to obtain a product VIZ-02 (2.24 g), and the yield is 49.5%. ESI-MS (+): m/z= 453.22.
Example 5: synthesis of Compound VIZ98-03
The reaction formula:
the preparation method comprises the following steps:
step 1: synthesis of compound VIZ98-0301
Isobutyric acid (26.4 g,0.3 mol), water (300 mL), methylene chloride (300 mL), tetrabutylammonium bisulfate (10.2 g,0.03 mol) and sodium bicarbonate (100.g, 1.2 mol) were charged into a three-necked flask. Chloromethyl chlorosulfonate (59.4 g,0.36 mol) 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 to give compound VIZ-0301 (29.3 g) in 71.5% yield. ESI-MS (+): m/z= 137.03.
Step 2: synthesis of Compound VIZ98-03
To the reaction flask was added compound ZJT1 (15.6 g,50.0 mmol), triethylamine (75.9 g,75.0 mmol) and VIZ-0301 (6.8 g,50.0 mmol), 100ml of N, N-dimethylformamide was added, and the system was heated to 50℃and stirred for 24 hours. Cooling to room temperature, adding water, stirring, filtering to obtain crude product, purifying with silica gel column to obtain compound VIZ-03 (4.7 g), yield 22.8%, ESI-MS (+): m/z= 413.19.
Example 6: synthesis of Compound VIZ98-04
The reaction formula:
the preparation method comprises the following steps:
to the reaction flask was added compound ZJT1 (7.8 g,25.0 mmol), triethylamine (63.3 g,62.5 mmol) and VIZ-0301 (6.8 g,50.0 mmol), 100ml of N, N-dimethylformamide was added, and the system was heated to 50℃and stirred for 24 hours. Cooling to room temperature, adding water, stirring, filtering to obtain crude product, purifying with silica gel column to obtain compound VIZ-04 (6.1 g), yield 47.6%, ESI-MS (+): m/z= 513.24.
Example 7: synthesis of Compound VIZ98-21
The reaction formula:
the preparation method comprises the following steps:
triphosgene (1.2 g,4.0 mmol) was added to anhydrous tetrahydrofuran (50 mL), triethylamine (1.6 g,16.0 mmol) was added, a tetrahydrofuran solution (10 mL) of ZJT2 (2.0 g,4.0 mmol) was slowly added under ice-bath, stirring was performed at room temperature for 2 hours, a tetrahydrofuran solution (10 mL) of ZJT1 (1.25 g,4.0 mmol) was slowly added under ice-bath, reflux was performed for 3 hours, cooling, concentration was performed, ice-water quenching was performed, stirring was performed for 2 hours, and filtration was performed to obtain a solid column chromatography separated to obtain compound VIZ-21 (0.62 g, 18.6%) ESI-MS (+): m/z= 836.36.
Example 8: synthesis of Compounds VIZ98-22
The reaction formula:
the preparation method comprises the following steps:
triphosgene (0.3 g,1.0 mmol) was added to anhydrous tetrahydrofuran (30 mL), triethylamine (0.8 g,8.0 mmol) was added, a tetrahydrofuran solution (10 mL) of ZJT2 (0.5 g,1.0 mmol) was slowly added under ice-bath, stirring was performed at room temperature for 2 hours, a tetrahydrofuran solution (5 mL) of ZJT1 (0.16 g,0.5 mmol) was slowly added under ice-bath, reflux was performed for 3 hours, cooling, concentration was performed, ice-water quenching was performed, stirring was performed for 2 hours, and filtration was performed to obtain a solid column chromatography separated to obtain a compound VIZ-22 (0.32 g, 47.1%) ESI-MS (+): m/z= 1359.58.
Example 9: synthesis of Compounds VIZ98-26
The reaction formula:
the preparation method comprises the following steps:
tetrahydrofuran (100 ml), a compound ZJT1 (3.12 g,10 mmol) and DMAP (1.34 g,11 mmol) are sequentially added into a reaction bottle, EDCI (2.11 g,11 mmol) and VIZ-26-SM (3.2 g,10 mmol) are sequentially added after dissolution, the temperature is raised to 70 ℃, the reaction is stirred, TLC monitors the reaction completion, the system is cooled, evaporated to dryness, ethyl acetate and water are added, an organic phase is separated, washed twice again, dried, evaporated to dryness under reduced pressure, and the residue is purified by a column to obtain a product VIZ-26 (0.75 g), and the yield is 12.2%. ESI-MS (+): m/z= 613.19.
Example 10: synthesis of Compounds VIZ98-27
The reaction formula:
the preparation method comprises the following steps:
tetrahydrofuran (100 ml), a compound ZJT1 (3.12 g,10 mmol) and DMAP (2.57 g,21 mmol) are sequentially added into a reaction bottle, EDCI (4.02 g,21 mmol) and VIZ-26-SM (6.4 g,20 mmol) are sequentially added after dissolution, the temperature is raised to 70 ℃, the reaction is stirred, TLC monitors the reaction completion, the system is cooled, evaporated to dryness, ethyl acetate and water are added, an organic phase is separated, washed twice again, dried, evaporated to dryness under reduced pressure, and the residue is purified by a column to obtain a product VIZ-27 (3.28 g), and the yield is 35.9%. ESI-MS (+): m/z= 913.25.
Example 11: synthesis of Compounds VIZ98-28
The reaction formula:
the preparation method comprises the following steps:
step 1: preparation of Compounds VIZ98-2801
The compound VIZ-26-SM (9.6 g,30 mmol), water (100 mL), methylene chloride (100 mL), tetrabutylammonium bisulfate (1.02 g,3 mmol) and sodium bicarbonate (10.1 g,120 mmol) were added to a three-necked flask. Chloromethyl chlorosulfonate (5.95 g,36 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. The organic phases were combined, dried, concentrated and purified by column to give compound VIZ-2801 (7.92 g) in 71.9% yield. ESI-MS (+): m/z= 367.04.
Step 2: preparation of Compounds VIZ98-28
To the reaction flask were added compound ZJT1 (3.12 g,10 mmol), potassium t-butoxide (1.68 g,15 mmol) and VIZ-2801 (3.67 g,10 mmol), 100ml of N, N-dimethylformamide was added, and the system was heated to 50℃and stirred for 24 hours. Cooling to room temperature, adding water, stirring, filtering to obtain crude product, and purifying with silica gel column to obtain compound VIZ-28 (1.82 g), yield 28.3%, ESI-MS (+): m/z= 643.20.
Example 12: synthesis of Compounds VIZ98-29
The reaction formula:
the preparation method comprises the following steps:
to the reaction flask were added compound ZJT1 (1.56 g,5 mmol), potassium tert-butoxide (1.35 g,12 mmol) and VIZ-2801 (3.67 g,10 mmol), 50ml of N, N-dimethylformamide was added, and the system was heated to 50℃and stirred for 24 hours. Cooling to room temperature, adding water, stirring, filtering to obtain crude product, and purifying with silica gel column to obtain compound VIZ-29 (3.35 g), yield 68.8%, ESI-MS (+): m/z= 973.27.
Example 13: synthesis of Compound VIZ98-40
The reaction formula:
the preparation method comprises the following steps:
step 1-step 4: preparation of Compounds VIZ98-4008
Referring to the procedure of example 1, the use of VIZ98-40-SM and the replacement of ZJT1-SM1 gave compounds VIZ98-4008 (3.66 g), in a four-step yield of 13.2%. ESI-MS (+): m/z= 299.14.
Step 5: preparation of Compounds VIZ98-4007
To the reaction solution were added triphenylphosphine (4.30 g,16.4 mmol) and elemental iodine (3.92 g,15.4 mmol) in this order, respectively, dissolved compound VIZ to 4008 (3.5 g,11.73 mmol) in a mixed solvent of acetonitrile (100 mL) and pyridine (5 mL). Stirring at room temperature for 18h, and thin layer detection showed the starting material to be substantially completely reacted. Water (50 mL) was added, the organic solvent was evaporated under reduced pressure, and dichloromethane was used: isopropyl alcohol (3:1) extraction (70 mL. Times.5). Drying over anhydrous sodium sulfate, and column chromatography gave compound VIZ-4007 (2.08 g) in 43.4% yield. ESI-MS (+): m/z= 409.04.
Step 6: preparation of Compounds VIZ98-4006
Compounds VIZ-4007 (2.00 g,4.90 mmol) (1.65 g,4.04 mmol) were dissolved in 0.4N sodium methoxide methanol solution and reacted at 65℃for 4 hours, a thin layer showed the disappearance of starting material (dichloromethane: methanol=15:1. Times.). Cooled to room temperature, pH was adjusted to neutrality with acetic acid, concentrated under reduced pressure, and the residue was subjected to column chromatography to give compound VIZ-4006 (1.14 g) in 83.2% yield. ESI-MS (+): m/z= 281.15.
Step 7: preparation of Compounds VIZ98-4005
Sodium azide (0.66 g,10.17 mmol) was dissolved in 10mL anhydrous tetrahydrofuran (30 mL), cooled to 0deg.C, and iodine chloride (1.1 g,6.78 mmol) was added and stirred for 10 min. A solution of compounds VIZ-4006 (0.95 g,3.39 mmol) in anhydrous tetrahydrofuran (15 mL) was added dropwise to the reaction solution, and the mixture was reacted at 0℃for 5 hours. Ethyl acetate extraction drying, concentration and column chromatography separation to obtain the compound VIZ 98.98-4005 (1.03 g) with a yield of 67.7%. ESI-MS (+): m/z= 450.11.
Step 8: preparation of Compounds VIZ98-4004
Compounds VIZ-4005 (0.90 g,2.11 mmol) were dissolved in anhydrous dichloromethane (40 mL), N-diisopropylethylamine (1.10 g,8.44 mmol) and t-butyldimethylsilyl triflate (TBSOTf, 1.95g,7.38 mmol) were added to an ice water bath and stirred at room temperature overnight. Dichloromethane (200 mL) was added thereto, followed by washing with saturated sodium bicarbonate, saturated ammonium chloride and saturated brine (80 mL). Drying over anhydrous sodium sulfate, concentrating and column chromatography to obtain compound VIZ-4004 (1.03 g), yield 86.6%. ESI-MS (+): m/z= 564.15.
Step 9: preparation of Compounds VIZ98-4003
Compound VIZ-4004 (1.0 g,1.77 mmol) was dissolved in dichloromethane (30 mL), water (10 mL) and dipotassium hydrogen phosphate (0.49 g,3.54 mmol), tetrabutylammonium hydrogen sulfate (0.66 g,1.95 mmol) and m-chlorobenzoic acid (0.31 g,2.00 mmol) were added, cooled to 0deg.C, and m-chloroperoxybenzoic acid (0.92 g,5.33 mmol) was added and stirred overnight at room temperature, and liquid detection showed a raw material product ratio of approximately 3:7, adding 30mL of ethyl acetate, washing with saturated sodium sulfite (20 mL), drying the organic phase with anhydrous sodium sulfate, and concentrating to obtain a crude compound VIZ-4003, and directly using the crude compound for the next step without treatment.
Step 10: preparation of Compounds VIZ98-4002
The crude product of the compound VIZ-4003 obtained in the last step is dissolved in ammonia methanol solution (6N, 50 mL), stirred at room temperature for 15h, the reaction is complete, the solvent is evaporated, and the compound VIZ-4002 (0.61 g) is obtained by column chromatography, and the total yield of the two steps of the step 9 and the step 10 is 76.0%. ESI-MS (+): m/z= 454.28.
Step 11: preparation of Compounds VIZ98-4001
Tetrahydrofuran (300 ml), compound VIZ-3802 (0.55 g,1.21 mmol) and 4-dimethylaminopyridine (DMAP, 0.18g,1.47 mmol) are sequentially added into a reaction bottle, EDCI (0.28 g,1.46 mmol) and VIZ98-26-SM (0.46 g,1.44 mmol) are sequentially added after dissolution, the temperature is raised to 70 ℃, stirring reaction is carried out, TLC monitoring reaction is completed, the system is cooled down, evaporated to dryness, ethyl acetate and water are added, an organic phase is separated, water is washed twice again, dried, evaporated to dryness under reduced pressure, and residues are purified by a column to obtain a product VIZ-4001 (0.43 g), and the yield is 47.1%. ESI-MS (+): m/z= 754.28.
Step 12: preparation of Compounds VIZ98-40
Compounds VIZ-4001 (0.36 g,0.48 mmol) were dissolved in anhydrous tetrahydrofuran (10 mL), and a solution of hydrogen fluoride in pyridine (65%, 8 mL) was added at 0deg.C and stirred overnight at room temperature. Saturated sodium bicarbonate was added, ethyl acetate was extracted 30mL X4 times, the organic phases were combined, and the column chromatography was concentrated over anhydrous sodium sulfate to give compound VIZ-40 (0.22 g) in 71.7% yield. ESI-MS (+): m/z= 640.21.
Example 14: synthesis of Compounds VIZ98-46
The reaction formula:
the preparation method comprises the following steps:
compound VIZ-46 (0.24 g) was produced in 16.3% yield by the procedure of example 9. ESI-MS (+): m/z= 687.45.
Example 15: synthesis of Compounds VIZ98-47
The reaction formula:
the preparation method comprises the following steps:
with reference to the procedure of step 11 and step 12 in example 13, compounds VIZ-47 (0.63 g) were produced in a total yield of 36.1%. ESI-MS (+): m/z= 714.44.
The following examples were synthesized in the same manner as in the above examples, using commercially available compounds or intermediate compounds appropriately synthesized from the commercially available compounds.
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Example 16: anti-influenza virus Activity assay
Virus strain: influenza A/FM/1/47 (H1N 1) and A/Han Fang/359/95 (H3N 2).
And (3) cells: the canine kidney cell line MDCK, derived from ATCC.
Test compound and positive control compound configuration: compounds VIZ-98-01, VIZ-05, VIZ98-07, VIZ-98-21, VIZ98-26, VIZ98-30, VIZ98-32, VIZ-34, VIZ98-36, VIZ98-38, VIZ-40, VIZ-98-42, VIZ98-44, VIZ98-46 and Compound A (positive control compound) were diluted with DMEM medium to 8 different concentrations, respectively.
The testing method comprises the following steps:
(1) Will be 5X 10 4 Mu.l of a canine kidney cell (MDCK) suspension per ml was inoculated into 96-well plates at 37℃and 5% CO 2 Culturing in incubator, and respectively applying cell half-toxicity concentration (CC 50 ) Experiments and cytopathic Effect half-effective concentration (EC 50 )；
(2) Inoculating MDCK cells for 24 hr, removing culture medium, adding 8 kinds of gradient diluted maintaining solutions containing test compound and positive control compound, simultaneously arranging cell control holes, arranging 3 parallel holes at each concentration, and culturing for 48 hr to obtain cell half-toxicity concentration (CC 50 ) Measuring;
(3) The MDCK cells were inoculated for 24 hours, the medium was discarded, 100. Mu.l of the medium containing the influenza virus (MOI=0.04) was added to infect the cells, the virus-containing medium was adsorbed for 2 hours, 8 kinds of test compound-containing and positive control compound-containing maintenance solutions were added in a gradient dilution, cell control wells and virus control wells were simultaneously provided, 3 parallel wells were provided for each concentration, and the culture was continued for 48 hours to give a cytopathic effect half-effective concentration (EC 50 ) Measuring;
(4) 100 μl CellTiter-Glo fluorescent cell activity detection reagent is added to each well of 96-well plates cultured for 48h in (2) and (3), and the specific operation method is that the cell Titer-Glo fluorescent cell activity detection test is carried out according to the specific operation method Kit (Promega Co.) instructions. Calculation of the cytotoxicity concentration half-times (CC) using graphpad5.0 software 50 ) And cytopathic effect half-effective concentration (EC 50 ) And calculates a selection index (si=cc 50 /EC 50 ). The results are shown in Table 1.
TABLE 1 determination of anti-influenza Virus Activity
From the above results, the compounds disclosed by the invention have good activity on influenza A/FM/1/47 (H1N 1) and A/Han Fangji/359/95 (H3N 2) and are more excellent than the activity of positive control compounds, wherein the selection indexes of the compounds VIZ-26 on A/FM/1/47 (H1N 1) and A/Han Fangji/359/95 (H3N 2) are even 156 times and 126 times that of the positive control compounds. Therefore, the compounds disclosed in the present invention can be used for preparing medicines for preventing/treating diseases induced by influenza virus infection. The superior anti-influenza virus activity, as well as the higher selectivity, suggests that the disclosed compounds may be administered in smaller clinical doses with lower side effects.
Example 17: determination of anti-coronavirus Activity
Test compounds and positive control compounds: VIZ98-01, VIZ-05, VIZ98-07, VIZ-21, VIZ98-26, VIZ98-30, VIZ98-32, VIZ98-34, VIZ98-36, VIZ98-38, VIZ98-40, VIZ98-42, VIZ98-44, VIZ98-46 and compound a (positive control compound).
Method 1: cell Activity assay
(1) 5X 10 per well 4 The Vero E6 cells were seeded in 96-well plates at 37℃with 5% CO 2 Culturing overnight in an incubator for experiment;
(2) Incubation of drug with cells: the test compound and positive control compound were diluted with DMEM-2% fetal bovine serum medium. Three-fold ratio for diluting the drug, 8 drug gradients, 3 duplicate wells per concentration were set; DMSO is used as a control group, the control group is diluted by a culture medium containing total volume DMEM-2% fetal bovine serum, and the DMSO with the same volume as the drug is given; after removal of the cell supernatant, 100. Mu.L of diluted compound was added to a 1.2 48-well plate, the control group was added with an equivalent volume of diluted DMSO, and incubated at 37℃for 1h.
(3) SARS-CoV-2 infected cells: mu.L of SARS-CoV-2,HongKong strain (Isolate Hong Kong/VM 20001061/2020) virus dilution (MOI=0.01VeroE6) was added to each well of the 48 well plate and incubation was continued for lh at 37℃and the infectious supernatant was removed and washed once with 200. Mu.LPBS (phosphate buffer). Adding 200 mu L of culture medium containing the drug with corresponding concentration into the hole again, continuously culturing for 24 hours, and collecting 150 mu L of supernatant to be detected;
(4) Viral RNA extraction was performed using the kit Takara MiniBEST Viral RNA/DNA Extraction Kit: virus lysis, to 150 u L cell culture supernatant added to 50 u L PBS solution to make the total volume of 200 u L. 200. Mu.L of BufferVGB buffer, 20. Mu.L of protease K and 1. Mu.L of Carrier RNA are added in sequence, mixed by shaking, and placed in a water bath at 56 ℃ for 10min for complete lysis. Then 200 mu L of absolute ethyl alcohol is added, and the mixture is stirred and mixed uniformly. Then passing through a column, washing and eluting to obtain viral RNA;
(5) Reverse transcription of viral RNA: methods of operation see TakaraPrimeScript TM RT reagent Kit with gDNA Eraser kit. Removing DNA in the eluent, and performing reverse transcription reaction for standby;
(6) The virus copy number is obtained by a standard curve method: reference kit Takara TBPremix Ex Taq TM II, using RBD plasmid with known copy number as standard, specific primer targeting RBD, and calculating copy number of each sample. Compound-treated group inhibition rates were obtained with blank DMSO group copy numbers as reference. According to the inhibition rate of the compound treatment groups with different concentrations, using prism6.0.1 software to fit an inhibition rate curve, and calculating half effective concentration EC 50 . The results are shown in Table 2.
Method 2: cytotoxicity test
(1) Obtaining Vero E6 cells (Changsha Ai Bi vitamin technology Co., ltd.) in logarithmic growth phase, and adjusting cell density to 5×10 4 Each of the holes respectively to100. Mu.L/well was inoculated in 96-well plates and incubated overnight;
(2) The test compound and the positive control compound are diluted by 8 concentration gradients by adopting the DMEM culture medium multiple ratio containing 2% of total volume of fetal bovine serum before administration, and each hole of diluted compound is added into Vero E6 cells in a 96-well plate respectively, and the final volume is 200 mu L. Setting 3 compound holes, wherein the DMSO solvent treatment group is blank control;
(3) After 48h incubation in the incubator, the compound-containing medium was discarded, and the cytotoxicity of the test compound was measured by CCK8 kit (Shanghai Shake Co., ltd.) and the half-toxicity concentration CC50 against Vero E6 cells was determined. The results are shown in Table 2.
Selection index (si=cc 50 /EC 50 )。
TABLE 2 determination of anti-coronavirus Activity
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From the above results, it was found that all test compounds had stronger activity against SARS-CoV-2 virus and higher selectivity than the positive control compound (Compound A). Wherein the anti-SARS-CoV-2 virus activity of compounds VIZ-01, VIZ-05, VIZ-98-26, VIZ-98-40 and VIZ-98-44 is more than 7 times that of compound A, and the anti-SARS-CoV-2 virus selectivity index of compounds VIZ-01, VIZ98-05 and VIZ-98-26 is 9 times, 11 times and 22 times that of compound A, respectively. Thus, the disclosed compounds can be used for the preparation of preventive/therapeutic drugs for symptoms/diseases induced by coronavirus infection. The higher anti-coronavirus activity and higher selectivity of the disclosed compounds indicate that the disclosed compounds treat diseases of coronaviridae infection, and have smaller administration dosage and lower side effect.
Example 18: hERG assay
In vitro hERG potassium ion inhibition experiments were performed on test compounds VIZ-01, VIZ-05, VIZ98-07, VIZ98-21, VIZ98-26, VIZ98-30, VIZ98-32, VIZ98-34, VIZ98-36, VIZ98-38, VIZ98-40, VIZ98-42, VIZ98-44, VIZ98-46 and positive control compound (compound A) to examine the potential cardiotoxicity of the presently disclosed compounds.
Cell preparation:
(1) CHO-hERG cells (Shanghai Huiyang Biotech Co., ltd.) were cultured at 175cm 2 In a culture flask, after the cell density grows to 60-80%, removing the culture solution, washing the culture solution once with 7mLPBS, and then adding 3mL of a detachin for digestion (Beijing Hua Taixin biomedical technology Co., ltd.);
(2) After digestion is completed, 7mL of culture solution is added for neutralization, then centrifugation is carried out, supernatant is sucked away, and 5mL of culture solution is added for resuspension, so that the cell density is ensured to be 2-5 multiplied by 10 6 /mL。
Electrophysiological recording procedure:
the single cell high impedance sealing and whole cell mode formation process is all completed automatically by Qpatch instrument, after obtaining whole cell record mode, the cell is clamped at-80 millivolts, before a depolarization stimulus of +40 millivolts for 5 seconds is given, a pre-voltage of-50 millivolts is given for 50 milliseconds, then repolarization is carried out to-50 millivolts for 5 seconds, and then the cell returns to-80 millivolts. The voltage stimulus was applied every 15 seconds, the extracellular fluid was applied for 1 minute after 2 minutes of recording, and after the generated current was stabilized, the extracellular fluid (NaCl: 145mmol/L, KCl:4mmol/L, caCl) containing the test compound and the positive control compound (the concentrations of the test compound and the positive control compound were 15. Mu. Mol/L) was applied 2 2.0mmol/L, mgCl2-6H2O:1mmol/L, glucose: 10mmol/L, HEPES:10mmol/L, pH 7.4.4), was applied to cells for 1 min at room temperature. At least 3 cells (n.gtoreq.3) were tested per compound.
Data analysis: experimental data were analyzed using XLFit software. The structure is shown in Table 3.
Results of cardiac hEGFR potassium current detection of the compounds of Table 3 at 15. Mu. Mol/L
Numbering of compounds Inhibition ratio (%)
VIZ98-01 7.2
VIZ98-05 7.5
VIZ98-07 7.4
VIZ98-21 8.5
VIZ98-26 6.1
VIZ98-30 8.4
VIZ98-32 7.6
VIZ98-34 8.4
VIZ98-36 9.1
VIZ98-38 8.1
VIZ98-40 7.9
VIZ98-42 8.9
VIZ98-44 7.4
VIZ98-46 7.6
Compound A 25.9
The results show that the inhibition rate of the compound disclosed by the invention on IKr is obviously smaller than that of the compound A at 15 mu mol/L, and the inhibition rate is obviously reduced compared with that of the compound A. Indicating that all test compounds were less cardiotoxic than compound a. Demonstrating that the disclosed compounds greatly reduce cardiotoxicity.
Example 19: mice survival and weight protection test
Definition of experimental days: the day of virus inoculation was defined as day 0 of the experiment, day 1 after the day, and so on.
Experimental animals and groupings: male K18-ACE2 transgenic mice (B6. Cg-Tg (K18-ACE 2) 2Primn/J mice) of a specific pathogen-free class, 40, 6-8 weeks old, purchased from Jackson laboratories, USA, were equally divided into 5 groups, 8 per group, each vehicle-infected group, VIZ-26 low dose group (44.5 mpk), VIZ-26 medium dose group (89.5 mpk), VIZ98-26 high dose group (178.5 mpk) and compound A group (143 mpk);
virus inoculation: on day 0, all mice were anesthetized by inhalation of isoflurane and vaccinated by nasal drip, and each of the 5 groups was vaccinated with SARS-CoV-2,HongKong strain (Isolate Hong Kong/VM 20001061/2020) at an inoculum size of 5,000p.f.u., at an inoculum size of 50. Mu.L.
Administration: mice were treated with compound or vehicle, and 5 groups were given 2 times daily, each for 5 days, starting on day 1, with continuous gavage.
Health monitoring: during the experiment, mice were observed daily and their body weight, survival status and clinical symptoms were recorded.
Humanity endpoint: any mice that had lost more than 20% of their body weight (based on day 0 body weight) during the experiment, or/and had a clinical symptom score of 3 points or more, or exhibited an dying state, will be euthanized according to IACUC protocol, and scored as dead animals in the results.
Clinical symptoms scoring, namely, observing the symptoms of infection of animals, recording the symptoms of erectile hair, bow back, somnolence/inactivity and heavy respiration, and recording 1 score of any symptoms. The sum of the symptom scores is the clinical symptom score.
Mice were analyzed for weight change using Two-way ANOVA and survival analysis was performed using Log-rank (Mantel-Cox).
The 14-day survival rate of each group of mice after virus inoculation is shown in Table 4, and the weight change is shown in FIG. 1.
Table 4 14 day survival of mice
Group of Number of experimental mice (Only) Number of surviving mice (Only) Survival (%)
Vehicle-infected group 8 0 0
VIZ98-26 low dose group 8 4 50
VIZ98-26 medium dose group 8 8 100
VIZ98-26 high dose group 8 8 100
Group A of compounds 8 4 50
Mice died or euthanized by the humane endpoint on days 5-6 of vehicle-infected group with a final survival of 0%; the VIZ-26 low dose group had 4 mice dying or euthanized by the humane end-point at days 6-8 with a final survival rate of 50%; the final survival rate in the VIZ-26 medium dose group and VIZ-26 high dose group was 100%; this shows that the compound disclosed by the invention can obviously improve the survival condition of animals compared with the vehicle-infected group, wherein the improvement effect of the compound in the low-dose group is equivalent to that of the compound in the group A, and the improvement effect of the compound in the medium-high-dose group is most obvious.
The mice weight change of fig. 1 shows that the maximum weight loss of animals at low, medium and high doses of test compounds VIZ-26, respectively, -9.3%, -8.7% and-8.9% was effectively protected compared to vehicle-infected group (maximum weight loss of 15.2), and also more effectively protected compared to compound a group (maximum weight loss of 10.5%), and that each dose group VIZ-26 exhibited better effects in both weight protection and course shortening in mice.
Example 20: virus titre test
Definition of experimental days: the day of virus inoculation was defined as day 0 of the experiment, day 1 after the day, and so on.
Experimental animals and groupings: male K18-ACE2 transgenic mice (B6. Cg-Tg (K18-ACE 2) 2 Prim/J mice) of a specific pathogen-free class, 6-8 weeks old, purchased from Jackson laboratories, USA, were equally divided into 5 groups according to body weight, each group of 5, vehicle-infected group, VIZ-26 low dose group (31.5 mpk), VIZ-26 medium dose group (62.5 mpk), VIZ98-26 high dose group 1 (125 mpk) and VIZ98-26 high dose group 2 (125 mpk), wherein VIZ98-26 high dose group 1 and VIZ98-26 high dose group 2 were dosed at different times;
virus inoculation: on day 0, all mice were anesthetized by inhalation of isoflurane and inoculated with virus by nasal drip, and each of the 5 groups was inoculated with SARS-CoV-2,Omicron variant (Isolate hCoV-19/USA/MD-HP 20874/2021) at an inoculum size of 100,000p.f.u., and an inoculum size of 50. Mu.L.
Administration: vehicle-infected groups of mice, VIZ98-26 low dose group (31.5 mpk), VIZ98-26 medium dose group (62.5 mpk), VIZ98-26 high dose group 1 (125 mpk) were continuously administered 5 days from day-1 (10 total), VIZ98-26 high dose group 2 (125 mpk) was continuously administered 2 days from day-1 (only first dose on day 0, 3 total), 2 times daily, and the administration mode was intragastric administration.
Sample collection: lung tissue: the sample tube was weighed into 1mL EMEM medium (Shanghai pure Biotechnology Co., ltd.) containing 1% FBS before and after the sample tube was placed into lung tissue, and the weight of the lung tissue sample was calculated, and the sample was stored in a-80℃refrigerator.
Sample detection: plaque assay detects new coronavirus titers in lung tissue: a) Resuspension of cultured Calu 3 cells (Wohan Shang En Biotechnology Co., ltd.) with MEM medium (Shanghai pure Biotechnology Co., ltd.) containing 10% FBS and spreading in 6-well plate with cell density of 2.5X10 5 Per mL, the amount of cells per well was 3mL. b) After thawing the collected samples, grinding with a tissue refiner, and centrifuging the homogenate to obtain a supernatant for detection. c) Lung in 96-well deep-well plateThe tissue homogenate supernatant stock was serially diluted 10-fold with MEM medium containing 1% fbs for a total of 5 dilutions. d) The above sample dilutions were added to 6-well plates with plated cells, each well having an inoculation volume of 0.2mL, and the 6 th well was 1% fbs MEM medium control. e) Placing the inoculated cell plates in an incubator for incubation for 1h, and shaking and mixing every 15 minutes during incubation. f) After incubation, cell culture medium was removed from 6-well plates, and 1:1 ratio of 1% agar-containing and 20% FBS-containing MEM medium solution was added to a 6-well plate. Placing the agar after solidification in CO 2 The culture was continued in the incubator for 3 days. g) Gently removing the agarose-containing medium, fixing the cells with 95% ethanol for 10 min, washing with PBS, re-fixing with 10% formalin solution containing 1% crystal violet, staining for 15 min, washing with PBS for 3 times, and drying. h) The number of plaques in the 6-well plate stained as described above was counted visually and the virus titer in the sample was calculated as Log 10 (number of plaques per gram of lung tissue sample) =log 10 (number of plaques per well × dilution/0.2/lung tissue weight).
The lung tissue virus titers between groups were analyzed using One-wayANOVA. The results are shown in FIG. 2.
FIG. 2 shows that in the novel coronavirus Omicron mutant infection model, the compounds disclosed herein were given prophylactically at doses of 31.5, 62.5, 125mpk (day-1 to day 3) and 125mpk (day-1 to day 0) at mouse lung tissue virus titers of 4.10,3.52,3.35 and 3.87Log (pfu/glung), respectively. Compared with the solvent group, the virus titer is reduced by 1.20,1.78,1.95 and 1.43Log (pfu/glung), respectively, which shows that the test compound VIZ-26 can obviously inhibit the replication of the virus in the lung tissue of mice after being subjected to preventive administration, and the good effect of preventing the infection of the novel coronavirus is shown. Among them, the high dose (125 mpk) group was administered for 5 consecutive days (total 10 times) to have an inhibitory effect on viruses superior to the 2-day (total 3 times) regimen.
While the invention has been described with reference to the preferred embodiments, it is not intended to limit the invention thereto, and it is to be understood that other modifications and improvements may be made by those skilled in the art without departing from the spirit and scope of the invention, which is therefore defined by the appended claims.

Claims (9)

1. A kind of liquid crystal display (I) 0 ) The purine nucleoside derivatives and pharmaceutically acceptable salts thereof are shown to be resistant to viral infection:
formula (I) 0 ) In,
R X1 and R is X2 Each independently selected from hydrogen, hydroxy, C1-C8 alkyl, and halogen;
R X3 selected from hydrogen, or azido;
R X4 selected from hydrogen, or cyano;
R 1a is hydrogen;
R 1b is C1-C8 alkyl;
R 2 selected from amino groups;
R 3 selected from the group consisting of
R 4 Selected from hydrogen;
n 1 selected from 0, in which case the carbonyl group is directly linked to oxygen to form an ester linkage;
R a selected from the following groups: C1-C8 alkyl, C1-C8 alkoxy;
n 3 and n 4 Each independently selected from 0, 1, 2, 3 or 4;
R 6a and R is 6b Each independently selected from hydrogen, C1-C8 alkyl;
R 7 selected from hydrogen, C1-C8 alkyl;
z is selected from hydrogen, halogen,Wherein,
above n 5 Selected from 0, 1, 2, 3 or 4;
R 8 selected from the following groups, substituted or unsubstituted with one or more groups a: amino, C6-C18 aryl;
x is selected from O;
R d1 and R is d2 Each independently selected from hydrogen, halogen;
R e selected from C1-C8 alkyl;
R f1 And R is f2 Are all hydrogen;
R g 、R h and the carbon to which it is attached to form a 3-7 membered ring, the atom or group on this 3-7 membered ring being optionally substituted by one or more C1-C8 alkyl groups;
R i1 and R is i2 Independently selected from cyano or quiltSubstituted C1-C8 alkyl; wherein m is selected from 1 or 2;
the group A is: halogen, chlorobenzoyl.
2. The antiviral purine nucleoside derivative of claim 1, and pharmaceutically acceptable salts thereof, having the structure of formula (I):
wherein the substituents in formula (I) are as defined in claim 1 formula (I) 0 ) As defined.
3. The antiviral purine nucleoside derivative of claim 1, having the structure of formula (II):
wherein in the formula (II)The definition of the substituents is as in claim 1, formula (I) 0 ) As defined.
4. The antiviral purine nucleoside derivative of claim 1, having the structure of formula (iii):
wherein the substituents in formula (III) are as defined in claim 1 of formula (I) 0 ) As defined.
5. The antiviral purine nucleoside derivative of claim 1, having the structure of formula (IV):
wherein the substituents in formula (IV) are as defined in claim 1 of formula (I 0 ) As defined.
6. The antiviral purine nucleoside derivative of claim 1, wherein the compound is selected from the group consisting of:
7. a pharmaceutical composition comprising the purine nucleoside derivative and pharmaceutically acceptable salt thereof of any one of claims 1 to 6 which is resistant to viral infection.
8. Use of a purine nucleoside derivative or a pharmaceutically acceptable salt thereof against an antiviral infection as defined in any one of claims 1 to 6, or a pharmaceutical composition as defined in claim 7, for the manufacture of a medicament against influenza a virus and covd-19 virus.
9. The use according to claim 8, wherein the medicament is formulated as an injectable, respiratory, dermal, mucosal, luminal or oral dosage form.
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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112574269A (en) * 2019-09-27 2021-03-30 中国科学院上海药物研究所 Purine nucleoside compound and application thereof
CN112778310A (en) * 2020-04-20 2021-05-11 中国科学院上海药物研究所 Application of nucleoside analogue or combination preparation containing nucleoside analogue in resisting virus
CN113292567A (en) * 2020-04-08 2021-08-24 迪斯凯威（深圳）新药发展有限公司 Nucleotide derivative and pharmaceutical composition and application thereof
WO2021188959A1 (en) * 2020-03-20 2021-09-23 Gilead Sciences, Inc. Prodrugs of 4'-c-substituted-2-halo-2'-deoxyadenosine nucleosides and methods of making and using the same
WO2021216722A1 (en) * 2020-04-23 2021-10-28 Atea Pharmaceuticals, Inc. Stereoselective process of manufacture of purine phosphoramidates

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112574269A (en) * 2019-09-27 2021-03-30 中国科学院上海药物研究所 Purine nucleoside compound and application thereof
WO2021188959A1 (en) * 2020-03-20 2021-09-23 Gilead Sciences, Inc. Prodrugs of 4'-c-substituted-2-halo-2'-deoxyadenosine nucleosides and methods of making and using the same
CN113292567A (en) * 2020-04-08 2021-08-24 迪斯凯威（深圳）新药发展有限公司 Nucleotide derivative and pharmaceutical composition and application thereof
CN112778310A (en) * 2020-04-20 2021-05-11 中国科学院上海药物研究所 Application of nucleoside analogue or combination preparation containing nucleoside analogue in resisting virus
WO2021216722A1 (en) * 2020-04-23 2021-10-28 Atea Pharmaceuticals, Inc. Stereoselective process of manufacture of purine phosphoramidates

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Adenosine analogs as inhibitors of tyrosyl-tRNA synthetase: Design, synthesis and antibacterial evaluation;Wei Wei等;《Bioorg. Med. Chem.》;第23卷;第6602–6611页 *
Identification of 8‑Aminoadenosine Derivatives as a New Class ofHuman Concentrative Nucleoside Transporter 2 Inhibitors;Kazuya Tatani等;《ACS Med.Chem.Lett.》;第6卷;第244−248页 *

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