WO2015197006A1 - Substituted amino acid thioester compound, and composition and application thereof - Google Patents

Abstract

Provided are a substituted amino acid thioester compound represented by general formula (A), and a composition and application thereof.

Description

Substituted amino acid thioester compound, composition thereof and application thereof Technical field

The present invention relates to a substituted amino acid thioester compound, a composition thereof and use thereof, and in particular, the present invention relates to a substituted amino acid thioester compound represented by the formula (I), a stereoisomer thereof or Pharmaceutically acceptable salts and pharmaceutical compositions containing them and use in the manufacture of a medicament for the treatment of viral infectious diseases.

Background technique

Hepatitis B is one of the world's diseases, it is caused by hepatitis B virus. A third of the world's population is infected with hepatitis B virus to some extent, including 350 million chronic carriers. In some Asian and African countries, hepatitis B has become a pandemic, especially in China. Hepatitis B virus can cause acute and chronic infections. Acute infections are usually accompanied by inflammation of the liver, vomiting, jaundice, and very few deaths. Chronic infections may induce cirrhosis and liver cancer. Although hepatitis B virus infection can be prevented by vaccines at present, there is no effective method for treating chronic hepatitis B disease.

Hepatitis B virus is a hemorrhagic DNA (DNA) virus with a circular partially double-stranded DNA genome. The shorter one chain has 1700 to 2800 nucleotides, the longer one chain has 3020 to 3320 nucleotides, and this long chain encodes the viral DNA polymerase. The genome of the hepatitis B virus encodes four known genes - C, X, P and S. Gene C encodes a nuclear protein (HBcAg), gene S encodes a surface antigen (HBsAg), and gene P encodes a DNA polymerase. The function of the protein encoded by gene X is unclear, but it is thought to be related to the occurrence of liver cancer because it activates A gene that induces cell proliferation and inactivates growth regulators.

The life cycle of hepatitis B virus is complex, enters cells through unknown receptors and endocytosis, and its genome is transferred to the nucleus by the host protein chaperones. In the nucleus, hepatitis B virus converts part of the double-stranded DNA into intact double-stranded DNA by the DNA polymerase of the host cell, and changes the morphology to a circular DNA (cccDNA) that is bound by a covalent bond. cccDNA was used as a template to transcribe four viral mRNAs. These four transcripts are used as templates to be transported into the cytoplasm and translated into viral membrane proteins, nuclear proteins and DNA polymerases. The longest mRNA (3.5 kb, longer than the viral genome) replicates as a template a new genomic copy, a transcriptional nucleocapsid protein and a viral DNA polymerase. At the same time, this 3.5 kb long RNA will be reverse transcribed from the antisense strand of hepatitis B virus DNA, followed by completion of the viral sense strand. Double-stranded DNA will be exported as a new sub-virus or returned to the nucleus to form a new cccDNA.

The synthesis of hepatitis B virus RNA and DNA depends on the hepatitis B virus DNA polymerase, and hepatitis B virus DNA polymerase is essential for viral replication. The polymerase has four domains: the beginning of replication of hepatitis B virus and the nucleocapsid The assembly is important for terminal proteins, spacer proteins, reverse transcriptase, and the RNaseH domain used to degrade pre-genomic RNA templates. Despite this, the lack of proofreading results in a high mutation rate of hepatitis B virus DNA polymerase.

The use of DNA polymerase inhibitors as anti-HBV drugs has become an attractive option. A particular viral polymerase inhibitor belongs to the family of nucleoside analogs. Treatment for patients with chronic hepatitis B has been improved by oral administration of anti-HBV nucleoside analog drugs. In serum, nucleoside analogs can rapidly reduce HBV DNA to unpredictable levels, and the mechanism of action is clear: nucleoside analogs competitively inhibit the activity of viral DNA polymerase. At the same time, nucleoside analogs showed good tolerance and fewer adverse reactions than interferon IFN-α. To date, five nucleoside analogues, hepatitis B virus DNA polymerase inhibitors, have been marketed as drugs for the treatment of chronic hepatitis B in the United States and Europe, including: lamivudine, and fosfovir dipivoxil. Entecavir, telbivudine and tenofovir disoproxil fumarate, as well as several other drugs at the stage of research. At the same time, long-term antiviral therapy may cause viral resistance and selectivity due to virus residues in the liver and mutations caused by viral polymerases, including mutations in the viral polymerase amino acid. This puts requirements on the development of new antiviral drugs.

Tenofovir, chemical name [(1R)-2-(6-aminofluoren-9-yl)-1-methyl-ethoxy]methyl phosphate (PMPA), is a nucleoside An acid reverse transcriptase inhibitor has anti-HBV and HIV; however, it has disadvantages such as having a phosphoric acid group, having a large polarity, a poor biofilm penetrating ability, and poor bioavailability in a living body. To overcome this disadvantage, a phosphonate or phosphonamide prodrug form can be made. Viread (Tinofovir dipivoxil fumarate), a drug developed by Gilead in 2002, is a prodrug of PMPA, and the prodrug form of the phosphonate is greatly improved in bioavailability. Viread plays an important role in the treatment of HIV and HBV. The transformation of the form of tenofovir prodrugs has become a hot topic of research.

European Patent No. EP206459 describes a structure comprising tenofovir for 9- (methoxy phosphate alkyl) adenine derivatives, and their use for antiviral drugs, in which R ¹ selected from hydrogen, methyl, hydroxymethyl, R ^{2 is} selected from substituted or unsubstituted ethylene, methylene, propylene, and the like. The detailed description in this patent is not considered to be part of the present invention, and its structural formula is as follows:

EP 481214 describes a novel oral phosphate nucleoside analog prodrug comprising adefovir dipivoxil, and its antiviral medical use, in particular anti-RNA, DNA virus, can also be used for the treatment of tumors, etc., wherein B is selected from嘌呤, cytosine, uracil, thymine, ornithril, etc., R ^{3 is} selected from substituted or unsubstituted C ₁ -C ₂₀ alkyl, R ¹ , R ^{2 are} independently selected from substituted or unsubstituted amino, OR ⁴ , R ⁴ is selected from _{CH 2 C (O) N (} R 5) 2, CH 2 C (O) OR 5, CH2OC (O) R 5, CH (R 5) OC (O) R 5, CH 2 C ( R ⁵ ) ₂ CH ₂ OH or CH ₂ OR ⁵ , R ^{5 is} selected from C ₄ -C ₂₀ alkyl, aryl or aryl-alkyl groups which are unsubstituted or substituted by hydroxy, oxo, nitro, halogen, R1 R2 can be looped. The specific description in this patent is not considered to be part of the present invention and its structure is as follows:

WO0208241 describes compositions comprising a derivative of adenine for disoproxil structure, in which R ¹ selected from hydrogen, methyl. The detailed description in this patent is not considered to be part of the present invention, and its structural formula is as follows:

WO02057288 describes substituted amino acid thioesters and their use in antiviral drugs, wherein Q is selected from the group consisting of purines or pyrimidines, R ⁴ and R ^{5 are} independently selected from hydrogen, alkyl, aryl, etc., R ¹ , R ² , R ³ , R ⁷ , and R ^{8 are} independently selected from the group consisting of a hydroxyl group, a halogen, a hydrogen, an amino group, an alkyl group, an alkoxy group, and an alkylamino group. The detailed description in this patent is not considered to be part of the present invention, and its structural formula is as follows:

CN200410024276.X describes 9-((phosphate)methoxyalkyl)adenine derivatives and their use in antiviral drugs, wherein R ¹ and R ^{2 are} independently selected from hydrogen or substituted biphenylmethyl . The detailed description in this patent is not considered to be part of the present invention, and its structural formula is as follows:

CN200710041280.0 describes substituted amino acid thioester compounds and their use in antiviral drugs, wherein R ^{1 is} selected from the group consisting of hydrogen, halogen, amino, cyclopropylamino, methoxy, ethoxy, etc., R ^{2 is} selected From hydrogen or an amino group, R ^{5 is} selected from methyl or hydrogen, and R ³ and R ^{4 are} independently selected from (substituted aminocarbonyloxy)alkyl. The detailed description in this patent is not considered to be part of the present invention, and its structural formula is as follows:

CN200410088840.4 describes substituted amino acid thioester compounds and their use in antiviral drugs, wherein R is hydrogen or methyl, R ^{2 is} selected from hydrogen or camphoryl, and R ^{1 is} selected from 3-8 carbons. A cycloalkyl group, an unsaturated chain hydrocarbon group of 3-8 carbons, an unsaturated cyclic alkyl group of 3-8 carbons or an aromatic hydrocarbon of 6-10 carbons. The detailed description in this patent is not considered to be part of the present invention, and its structural formula is as follows:

WO2011069322 describes substituted amino compounds and thioesters useful for the treatment and prevention of diseases associated with viral infections medical uses, in which R ¹ is selected from hydrogen or methyl, R ² is selected from -R ³ or -OR ^3, R ³ It is selected from a C _1-8 alkyl group and a C _3-8 cycloalkyl group. The detailed description in this patent is not considered to be part of the present invention, and its structural formula is as follows:

The invention designs a compound represented by the general formula (I) on the basis of tenofovir disoproxil to provide a novel structure, better medicine, safer, less toxic and side effects, good solubility or high bioavailability. Substituted amino acid thioesters, stereoisomers or pharmaceutically acceptable salts thereof, for use in the treatment of viral infectious diseases, including infectious diseases caused by hepatitis B virus and HIV virus.

Summary of the invention

The present invention provides a compound of the formula (A), a stereoisomer thereof, a pharmaceutically acceptable salt or a eutectic, wherein:

A is selected from a 6 to 10 membered aromatic ring or a 6 to 10 membered heteroaryl ring, and the heteroaryl group contains 1 to 4 hetero atoms selected from N, O, and S, and the aromatic ring or heteroaryl ring is Further selected from 0 to 5 selected from H, F, Cl, Br, I, CN, amino, hydroxy, carboxy, C _1-4 alkyl, trifluoromethyl, C _1-4 alkoxy or -C ( Substituted by a substituent of OC _1-4 alkyl;

B is

E is selected from -CH ₂ CH(CH ₃ )OCH ₂ - or -CH ₂ CH ₂ OCH ₂ -;

R ^{N is} selected from H or C _1-4 alkyl;

R ¹ and R ² are each independently selected from the group consisting of H, C _1-6 alkyl or a side chain of a natural or pharmaceutically acceptable amino acid, and if the side chain contains a carboxyl group, the carboxyl group may be optionally esterified with an alkyl group or an aryl group;

Alternatively, R ¹ , R ² may form a C _3-6 cycloalkyl group together with the carbon atom to which they are attached;

R ^{3 is} selected from H, C _1-6 alkyl, 6 to 10 membered aromatic ring or 6 to 10 membered heteroaryl ring, and the heteroaryl group contains 1 to 4 hetero atoms selected from N, O, and S, The alkyl, aromatic or heteroaryl ring is optionally further selected from 0 to 5 selected from the group consisting of H, F, Cl, Br, I, CN, amino, hydroxy, carboxy, C _1-4 alkyl or C _1- Substituted by a ₄ -alkoxy substituent.

According to a preferred embodiment of the present invention, a thioester compound of the amino acid represented by the formula (A), a stereoisomer or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the compounds of the formula (I), wherein:

A is selected from phenyl or naphthyl, and the phenyl or naphthyl group is further optionally further selected from 0 to 5 selected from the group consisting of H, F, Cl, Br, I, CN, amino, hydroxy, carboxy, C _1-4 alkane. Substituted by a substituent of a C _1-4 alkoxy group;

B is

E is selected from -CH ₂ CH(CH ₃ )OCH ₂ - or -CH ₂ CH ₂ OCH ₂ -;

R ^{N is} selected from H or C _1-4 alkyl;

R ² is a side chain of a natural or pharmaceutically acceptable amino acid, and if the side chain contains a carboxyl group, the carboxyl group may be optionally esterified with an alkyl group or an aryl group;

R ³ is a C _1-6 alkyl group.

A preferred embodiment of the present invention, a thioester compound of the amino acid of the formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein A is selected from phenyl or naphthyl, preferably phenyl; The phenyl or naphthyl group is optionally further substituted with from 0 to 5 substituents selected from the group consisting of H, F, Cl, Br, I, CN, amino, hydroxy, carboxy, methyl, ethyl, methoxy or ethoxy. The substitution is further preferably substituted with 0 to 5 substituents selected from H, F, Cl, Br, CN, amino or methoxy.

An alternative of the present invention, an amino acid thioester compound of the formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein R ² is a side chain of a natural or pharmaceutically acceptable amino acid, The amino acids thereof are preferably glycine, alanine, leucine, isoleucine, tyrosine, valine, phenylalanine, methionine, tryptophan, serine, glutamine, threonine. , cysteine, histidine, asparagine, tyrosine, aspartic acid, glutamic acid, naphthyl acid or arginine, further preferably glycine, alanine, leucine, phenylalanine Acid, asparagine or arginine, more preferably glycine, alanine or phenylalanine.

An alternative of the present invention, an amino acid thioester compound of the formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein R ³ is a C _1-6 alkyl group, preferably a C _1-4 The alkyl group is further preferably a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group or an isobutyl group.

According to a preferred embodiment of the present invention, there is provided a stereoisomer or a pharmaceutically acceptable salt of the amino acid thioester compound of the formula (I), wherein the compound is selected from the group consisting of the compound of the formula (II), wherein:

R ^{2 is} selected from the side chain of glycine, alanine, leucine, phenylalanine, asparagine or arginine, preferably a side chain of glycine, alanine or phenylalanine.

E is selected from -CH ₂ CH(CH ₃ )OCH ₂ -;

R ³ is a C _1-4 alkyl group.

According to a preferred embodiment of the present invention, there is provided a stereoisomer or a pharmaceutically acceptable salt of the amino acid thioester compound of the formula (I), wherein the compound is selected from the group consisting of the compound of the formula (III), wherein:

R ^{3 is} selected from the group consisting of methyl, ethyl, propyl, isopropyl, butyl or isobutyl.

According to a preferred embodiment of the present invention, a thioester compound of the amino acid represented by the formula (III), a stereoisomer or a pharmaceutically acceptable salt thereof, wherein

R ^{2 is} selected from the side chain of glycine, alanine, leucine, phenylalanine, asparagine or arginine, preferably a side chain of glycine, alanine or phenylalanine, further preferably alanine Side chain

R ³ is C _1-4 alkyl, preferably methyl, ethyl, propyl, isopropyl, butyl or isobutyl.

According to a preferred embodiment of the present invention, there is provided a stereoisomer or a pharmaceutically acceptable salt of the amino acid thioester compound of the formula (A), wherein the compound is selected from the compounds of the formula (IV):

A is selected from a 6 to 10 membered aromatic ring or a 6 to 10 membered heteroaryl ring, and the heteroaryl group contains 1 to 4 hetero atoms selected from N, O, and S, and the aromatic ring or heteroaryl ring is Further selected from 0 to 5 selected from H, F, Cl, Br, I, CN, amino, hydroxy, carboxy, C _1-4 alkyl, trifluoromethyl, C _1-4 alkoxy or -C ( Substituted by a substituent of OC _1-4 alkyl;

B is

E is selected from -CH ₂ CH(CH ₃ )OCH ₂ - or -CH ₂ CH ₂ OCH ₂ -;

R ¹ and R ² are each independently selected from a C _1-6 alkyl group,

Alternatively, R ¹ , R ² together with the carbon atom to which they are attached form a C _3-6 cycloalkyl group;

R ^{3 is} selected from H, C _1-6 alkyl, 6 to 10 membered aromatic ring or 6 to 10 membered heteroaryl ring, and the heteroaryl group contains 1 to 4 hetero atoms selected from N, O, and S, The aromatic or heteroaryl ring is optionally further selected from 0 to 5 selected from the group consisting of H, F, Cl, Br, I, CN, amino, hydroxy, carboxy, C _1-4 alkyl or C _1-4 alkoxy Substituted by a substituent.

A preferred embodiment of the invention, a compound of the formula (I), a stereoisomer thereof, a pharmaceutically acceptable salt or a eutectic, wherein:

A is selected from a substituted or unsubstituted phenyl, pyridyl or naphthyl group, and when substituted, optionally further 1 to 3 are selected from the group consisting of H, F, Cl, Br, I, CN, methoxy, methyl Substituted by a substituent of a trifluoromethyl or ethoxycarbonyl group;

B is

E is selected from -CH ₂ CH(CH ₃ )OCH ₂ -;

R ¹ and R ² are each independently selected from a methyl group and an ethyl group.

Alternatively, R ¹ , R ² together with the carbon atom to which they are attached form a cyclopropyl group;

R ^{3 is} selected from methyl, ethyl, isopropyl, monofluoromethyl or difluoromethyl.

A preferred embodiment of the invention, a compound of the formula (A), a stereoisomer thereof, a pharmaceutically acceptable salt or a eutectic, wherein:

A is selected from phenyl;

R ^{N is} selected from H;

R ¹ and R ² are each independently selected from methyl or ethyl;

R ^{3 is} selected from methyl, ethyl or isopropyl.

According to a preferred embodiment of the present invention, the amino acid thioester compound represented by the formula (I) is a stereoisomer or a pharmaceutically acceptable salt thereof, wherein the compound is:

According to some preferred embodiments of the invention, the compound is a compound obtained by resolution of compound 1:

The separation conditions were: instrument, MGII preparative SFC; column, ChiralPak AS-H, 250×30 mm I.D.; mobile phase, A is CO ₂ and B is Methanol; gradient, B 40%; flow rate, 40 mL/min; back pressure , 100 bar; column temperature 38 ° C; wavelength, 220 nm; period, 5.5 min;

Obtaining two compounds with short retention times and long retention times under the above separation conditions;

The compound having a short retention time is Compound 1-1 retention time of 2.21 ± 0.5 min;

The compound in which the retention time is long is the compound 1-2 retention time of 3.82 ± 0.5 min.

According to further preferred embodiments of the invention, the compound is a compound obtained by resolution of compound 2:

The separation conditions were: instrument: Thar 200prepararive SFC, column: ChiralPak AS-10u, 300×50 mm I.D., mobile phase: A is CO ₂ and B is ethanol, gradient: B 45%, flow rate: 200 mL/min, back Pressure: 100 bar, column temperature: 38 ° C, wavelength: 220 nm, cycle: ~ 15 min;

Obtaining two compounds with short retention times and long retention times under the above separation conditions;

The compound having a short retention time is a compound 2-1 retention time of 2.32 ± 0.5 min;

The compound in which the retention time is long is the compound 2-2 retention time of 3.98 ± 0.5 min.

According to further preferred embodiments of the invention, the compound is a compound obtained by resolution of compound 5:

The separation conditions were: instrument: Thar 200prepararive SFC, column: ChiralCel OD-10u, 300×50 mm I.D, mobile phase: A is CO ₂ and B is ethanol, gradient: B 25%, flow rate: 200 mL/min, back pressure 100 bar, column temperature: 38 ° C, wavelength: 220 nm, period: ~ 5 min;

Obtaining two compounds with short retention times and long retention times under the above separation conditions;

The compound having a short retention time is Compound 5-1 retention time of 3.38±0.5 min;

The compound in which the retention time is long is the retention time of the compound 5-2 of 3.77 ± 0.5 min.

According to further preferred embodiments of the invention, the compound is a compound obtained by resolution of compound 6:

The separation conditions were: instrument: MGII preparative SFC-1, column: ChiralCel OD-5u, 250×30 mm I.D. Mobile phase: A is CO2 and B is isopropanol, gradient: 30%, flow rate: 60 mL/min, Back pressure: 100 bar, column temperature: 38 ° C, wavelength: 220 nm, period: ~ 4 min;

Obtaining two compounds with short retention times and long retention times under the above separation conditions;

The compound having a short retention time is a compound 6-1 retention time of 1.93 ± 0.5 min;

The compound in which the retention time is long is the retention time of the compound 6-2 of 2.87 ± 0.5 min.

According to still further preferred embodiments of the invention, the compound is a compound obtained by resolution of compound 7:

The separation conditions were: instrument: waters SFC, column: Chiralpak AS-3 (4.6 × 100 mm), mobile phase: A is methanol and B is CO ₂ , gradient: B 10-40%, flow rate: 2 mL/min, back pressure : 2000 psi, column temperature: 35 ° C, wavelength: 260 nm; period: ~ 6 min;

Obtaining two compounds with short retention times and long retention times under the above separation conditions;

The compound having a short retention time is a compound 7-1 retention time of 1.62 ± 0.5 min;

The compound in which the retention time is long is the retention time of the compound 7-2 of 2.52 ± 0.5 min.

The present invention also provides a compound of the formula (A), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein the salt is a fumarate.

The invention also provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of the invention, and a stereoisomer or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or Shape agent.

Further, the present invention provides the use of the compound of the present invention, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for treating a viral infectious disease.

A preferred embodiment of the invention, wherein the viral infectious disease comprises an infectious disease caused by hepatitis B virus, hepatitis C virus and HIV virus.

Further, the present invention provides a method of treating a viral infectious disease, wherein the method comprises administering a compound of the present invention, a stereoisomer thereof, or a pharmaceutically acceptable salt or eutectic thereof or the Pharmaceutical composition.

A preferred embodiment of the invention, wherein the viral infectious disease comprises an infectious disease caused by hepatitis B virus, hepatitis C virus and HIV virus.

Terms used in the specification and claims have the following meanings unless stated to the contrary.

The present invention relates to the substitution of a plurality of substituents, which may be the same or different.

The present invention relates to the inclusion of a plurality of heteroatoms, each of which may be the same or different.

The elements carbon, hydrogen, oxygen, sulfur, nitrogen or halogen involved in the groups and compounds of the present invention include their isotopic conditions, and the elements, carbon, hydrogen and oxygen involved in the groups and compounds of the present invention. Sulfur or nitrogen is optionally further replaced by one or more of their corresponding isotopes, wherein the carbon isotopes include ¹² C, ¹³ C, and ¹⁴ C, and the hydrogen isotopes include helium (H), helium (D, also known as heavy hydrogen ), 氚 (T, also known as super heavy hydrogen), oxygen isotopes include ¹⁶ O, ¹⁷ O and ¹⁸ O, sulfur isotopes include ³² S, ³³ S, ³⁴ S and ³⁶ S, nitrogen isotopes including ¹⁴ N and ¹⁵ N The fluorine isotope ¹⁹ F, the chlorine isotope includes ³⁵ Cl and ³⁷ Cl, and the bromine isotopes include ⁷⁹ Br and ⁸¹ Br.

The term "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight chain and branched chain groups of 1 to 20 carbon atoms. Preference is given to alkyl groups having 1 to 10 carbon atoms, non-limiting examples including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, positive a mercapto group, and various branched isomers thereof; more preferred are lower alkyl groups having 1 to 4 carbon atoms, and non-limiting examples include methyl, ethyl, propyl, isopropyl, and Butyl, isobutyl or tert-butyl. The alkyl group may be substituted or unsubstituted, and when substituted, the substituent is preferably from 1 to 5, independently selected from the group consisting of H, F, Cl, Br, I, =O, alkyl, alkenyl, alkynyl. , alkoxy, alkylthio, alkylamino, fluorenyl, hydroxy, nitro, cyano, amino, alkyl acylamino, cycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy A cycloalkylalkyl group, a hydroxyalkyl group, a carboxylic acid, a carboxylic acid ester or a heterocycloalkyl group.

"Amino" means -NH _2, may be substituted or unsubstituted. When substituted, the substituent is preferably 1 to 3, independently selected from alkyl, alkenyl, alkynyl, alkoxy, Thio, hydroxy, amino, alkylamino, alkyl acylamino, heterocycloalkyl, cycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, hydroxyalkyl, carboxylic acid or Carboxylic acid ester.

"Aryl" means a substituted or unsubstituted 6 to 14 membered all carbon monocyclic or fused polycyclic group having a polycyclic group of a conjugated π-electron system, preferably a 6 to 10 membered aromatic ring, Non-limiting examples include phenyl or naphthyl; the aryl group may be fused to a heteroaryl, heterocyclyl or cycloalkyl group, and the moiety attached to the parent structure is an aryl group, non-limiting examples of which include benzo Furan, benzocyclopentyl or benzothiazole. When substituted, the substituent is preferably from 1 to 5, and the substituent is independently selected from the group consisting of H, F, Cl, Br, I, =O, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, Alkylamino, fluorenyl, hydroxy, nitro, cyano, amino, alkyl acylamino, cycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylindolyl, hydroxyalkane a carboxylic acid, a carboxylic acid ester or a heterocycloalkyl fluorenyl group.

"Natural or pharmaceutically acceptable amino acids": The basic skeleton of a protein molecule is an amino acid sequence, and there are 20 basic amino acids constituting a protein. These 20 basic amino acids are the basis for biological late modification of proteins, and in addition, based on these basic amino acids. The organism also synthesizes amino acid types derived from hydroxyproline, hydroxylysine, etc. These biosynthesized amino acids are collectively referred to as "natural amino acids"; artificially synthesized are "unnatural amino acids". "Pharmaceutically acceptable amino acid" refers to a pharmaceutically acceptable natural or unnatural amino acid.

"Pharmaceutically acceptable salt" means a pharmaceutically acceptable salt of a non-toxic acid or base, including salts of inorganic acids and bases, organic acids and bases.

"Crystal" refers to a combination of an active pharmaceutical ingredient (API) and a cocrystal former (CCF) under the action of hydrogen bonds or other non-covalent bonds, of which API and CCF The pure state is solid at room temperature and there is a fixed stoichiometric ratio between the components. Eutectic is a multi-component crystal that contains both a binary eutectic formed between two neutral solids and a multi-component eutectic formed by a neutral solid with a salt or solvate. The "eutectic former" includes, but is not limited to, various pharmaceutically acceptable acid, base, nonionic compounds.

"Stereoisomer" refers to isomers resulting from the spatial arrangement of atoms in a molecule, including cis and trans isomers, enantiomers and conformational isomers.

"Pharmaceutical composition" means one or more of the compounds described herein or a physiologically/pharmaceutically acceptable salt or mixture of prodrugs thereof with other chemical components, such as physiological/pharmaceutically acceptable carriers and excipient. The purpose of the pharmaceutical composition is to facilitate the administration of the compound to the organism.

"Prodrug" means a compound of the invention which can be converted to biological activity under physiological conditions or by solvolysis. Prodrugs of the invention are prepared by modifying functional groups in the compound which can be removed by conventional procedures or in vivo to provide the parent compound. A prodrug includes a compound formed by attaching a hydroxyl group, an amino group or a thiol group to any group in the compound of the present invention. When a prodrug of the compound of the present invention is administered to a mammalian individual, the prodrug is cleaved to form a free hydroxyl group, respectively. , free amino or free sulfhydryl. Examples of prodrugs include However, it is not limited to a compound formed by a hydroxyl group or an amino functional group in the compound of the present invention and formic acid, acetic acid or benzoic acid.

"Optional," "optional," or "optionally" means that the event or environment described subsequently can, but need not, occur, including where the event or environment occurs or does not occur. For example, "aryl substituted with an alkyl group" means that an alkyl group may, but need not, be present, and the description includes the case where the aryl group is substituted with an alkyl group and the case where the aryl group is not substituted with an alkyl group.

"Substituted or unsubstituted" refers to a situation in which a group may be substituted or unsubstituted, and if it is not indicated in the present invention that a group may be substituted, it means that the group is unsubstituted.

“As a choice” means that the scheme after “as a choice” is a side-by-side relationship with the scheme before “as a choice” rather than a further selection in the previous scheme.

"Substitution" refers to the case where one or more hydrogen atoms in a group are substituted by another group, and if the group is substituted by a hydrogen atom, the group formed is the same as the group substituted by a hydrogen atom. Where the group is substituted, for example, amino, C _1-4 alkyl, C _1-4 alkoxy, C _3-6 carbocyclic, 3 to 6 membered heterocyclic ring, optionally further from 0 to 4 selected from H, Substituted by a substituent of F, Cl, Br, I, hydroxy, cyano, amino, C _1-4 alkyl or C _1-4 alkoxy, the groups formed include, but are not limited to, methyl, chloromethyl, Trichloromethyl, hydroxymethyl, -CH ₂ OCH ₃ , -CH ₂ SH, -CH ₂ CH ₂ CN, -CH ₂ NH ₂ , -NHOH, -NHCH ₃ , -OCH ₂ Cl, -OCH ₂ OCH ₂ CH ₃ , -OCH ₂ CH ₂ NH ₂ , -OCH ₂ CH ₂ SH, -OCH ₂ CH ₂ OH, 1-hydroxycyclopropyl, 2-hydroxycyclopropyl, 2-aminocyclopropyl, 4-methyl Furanyl, 2-hydroxyphenyl, 4-aminophenyl or phenyl.

Specific synthesis method of the invention

method one:

The compound (A-1) is reacted with a thiol under an ester condensing agent to obtain a compound (A-2) including, but not limited to, dicyclohexylcarbodiimide, N,N-carbonyldiimidazole, N,N. '-disuccinimidyl carbonate, 1-p-methylbenzenesulfonylimidazole, 4,5-dicyanoimidazole, preferably N,N-carbonyldiimidazole;

Deprotection of compound (A-2) to give compound (A-3); said deprotection is carried out using conventional amino protecting group deprotection methods including, but not limited to, deprotection under acidic conditions, such as the use of trifluoroacetic acid ;

Compound (A-3) and compound (A-4) are reacted under basic conditions to obtain compound (A);

Compound (A-1) can be purchased or prepared by reference to CN201080036406.5 literature;

Compound (A-4) is prepared with reference to EP0206459B1, CN01813161.1 or WO2013052094;

Wherein: R ^{a is} selected from an amino protecting group, wherein the amino protecting group includes, but is not limited to, tert-butoxycarbonyl, benzyloxycarbonyl, fluorenylmethoxycarbonyl, allyloxycarbonyl, trichloroethoxycarbonyl, three Methylsilylethoxycarbonyl, methoxycarbonyl, ethoxycarbonyl, 2-biphenyl-2-propoxycarbonyl, tert-butoxy, phthaloyl, p-toluenesulfonyl, o-nitrophenylsulfonyl , p-nitrophenylsulfonyl, pivaloyl, formyl, trifluoroacetyl, benzoyl, benzyl, trityl, p-methoxybenzyl or 2,4-dimethoxybenzyl, Preferred is tert-butoxycarbonyl;

The definitions of A, B, E, R ^N , R ² and R ^{3 are} in accordance with the definitions of formula (A).

Method Two:

Compound (IA) is reacted with a thiol under an ester condensing agent to give compound (IB), including but not limited to dicyclohexylcarbodiimide, N,N-carbonyldiimidazole, N,N'-diamidium Imidyl carbonate, 1-p-methylbenzenesulfonylimidazole, 4,5-dicyanoimidazole, preferably N,N-carbonyldiimidazole;

Compound (I-B) deprotects the amino group to give compound (I-C); the deprotection is carried out using conventional amino protecting group deprotection methods including, but not limited to, deprotection under acidic conditions, such as using trifluoroacetic acid;

Compound (I-C) and compound (A-4) are reacted under basic conditions to obtain compound (I);

Compound (I-A) can be purchased or prepared by reference to CN201080036406.5 literature;

Compound (A-4) is prepared with reference to EP0206459B1, CN01813161.1 or WO2013052094;

Wherein: R ^{a is} selected from an amino protecting group, wherein the amino protecting group includes, but is not limited to, tert-butoxycarbonyl, benzyloxycarbonyl, fluorenylmethoxycarbonyl, allyloxycarbonyl, trichloroethoxycarbonyl, three Methylsilylethoxycarbonyl, methoxycarbonyl, ethoxycarbonyl, 2-biphenyl-2-propoxycarbonyl, tert-butoxy, phthaloyl, p-toluenesulfonyl, o-nitrophenylsulfonyl , p-nitrophenylsulfonyl, pivaloyl, formyl, trifluoroacetyl, benzoyl, benzyl, trityl, p-methoxybenzyl or 2,4-dimethoxybenzyl, Preferred is tert-butoxycarbonyl;

The definitions of A, B, E, R ^N , R ² and R ^{3 are} in accordance with the definitions of formula (I).

Method three:

Compound (A-4) can be purchased or prepared by the method of WO2014088923 or WO2012154698.

Compound (IV-B) can be purchased or prepared by methods such as WO2012075456, WO2011014973 or WO2012084794;

Compound (IV-C) can be prepared by referring to the methods of the present invention, WO0208241 or WO2013052094;

Compound (IV) is obtained by transesterification of compound (IV-C) with a thiol;

R ^{4 is} selected from H or C _1-6 alkyl;

The definitions of A, B, E, R ¹ , R ² and R ³ are consistent with the definitions in formula (IV).

detailed description

The technical solutions of the present invention are described in detail below with reference to the accompanying drawings and embodiments, but the scope of protection of the present invention includes but is not limited thereto.

The structure of the compound is determined by nuclear magnetic resonance (NMR) or (and) mass spectrometry (MS). The NMR shift (δ) is given in units of 10 ^-6 (ppm). NMR was measured using a (Bruker Avance III 400 and Bruker Avance 300) NMR instrument and the solvent was deuterated dimethyl sulfoxide (DMSO-d ₆ ), deuterated chloroform (CDCl ₃ ), deuterated methanol (CD ₃ OD). The internal standard is tetramethylsilane (TMS).

The measurement of MS was carried out (Agilent 6120B (ESI) and Agilent 6120B (APCI)).

The HPLC was measured using an Agilent 1260 DAD high pressure liquid chromatograph (Zorbax SB-C18100 x 4.6 mm).

Thin layer chromatography silica gel plate uses Yantai Yellow Sea HSGF254 or Qingdao GF254 silica gel plate. The specification of silica gel plate used for thin layer chromatography (TLC) is 0.15mm~0.20mm. The specification for thin layer chromatography separation and purification is 0.4mm. ~0.5mm.

Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as a carrier.

The known starting materials of the present invention may be synthesized by or according to methods known in the art, or may be purchased from Titan Technology, Anheji Chemical, Shanghai Demer, Chengdu Kelon Chemical, Suiyuan Chemical Technology, and Belling Technology. And other companies.

The nitrogen atmosphere means that the reaction flask is connected to a nitrogen balloon having a volume of about 1 L.

The hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon of about 1 L volume.

The hydrogenation reaction is usually evacuated, charged with hydrogen, and operated three times.

Unless otherwise stated in the examples, the reaction was carried out under a nitrogen atmosphere.

Unless otherwise stated in the examples, the solution means an aqueous solution.

There is no particular description in the examples, and the reaction temperature is room temperature.

The room temperature is an optimum reaction temperature of 20 ° C to 30 ° C.

Example 1

(2S)-2-[[(1R)-2-(6-Aminofluoren-9-yl)-1-methylethoxy]methyl]phenoxyphosphoramidopropionic acid thioisopropyl ester ( Compound 1)

S-isopropyl(2S)-2-[[[(1R)-2-(6-aminopurin-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]propanethioate

First step: (S)-2-(tert-butoxycarbonyl)aminopropionic acid thioisopropyl ester (1B)

(S)-S-isopropyl 2-((tert-butoxycarbonyl)amino)propanethioate

N-tert-butoxycarbonyl-L-alanine (1A) (5 g, 26.4 mmol) was dissolved in tetrahydrofuran (40 mL), and N,N'-carbonyldiimidazole (CDI) (4.7 g, 29.1 mmol) was added. Stir at room temperature for 2 hours. Thioisopropanol (6.2 g, 79.3 mmol) was added and allowed to react at room temperature overnight. 4 mol/L of sodium hydroxide solution (30 mL) was added, and the mixture was extracted with dichloromethane (50 mL×4). (v/v) = 1:0 to 9:1), the title compound (S)-2-(tert-butoxycarbonyl)aminopropionic acid thioisopropyl ester (1B), light yellow liquid (4 g, yield The rate is 61%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 3.61 (m, 1H), 2.37 - 2.16 (m, 1H), 1.46 (s, 9H), 1.36 (d, 3H), 1.30 (d, 6H).

The second step: (S)-2-aminopropionic acid thioisopropyl ester trifluoroacetate (1C)

(S)-S-isopropyl 2-aminopropanethioate triflouroacetate

(S)-2-(tert-Butoxycarbonyl)aminopropionic acid thioisopropyl ester (1B) (4 g, 16.2 mmol) was dissolved in dichloromethane (10 mL), trifluoroacetic acid (10 mL) Stir for 4 hours. Concentration under reduced pressure to dryness afforded crude (S)-2- phenyl propyl isopropyl ester trifluoroacetate (1C) (4 g).

The third step: [[(1R)-2-(6-aminofluoren-9-yl)-1-methylethoxy]methyl]phenoxyphosphoric acid (1E)

[(1R)-2-(6-aminopurin-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphinic acid

[[(1R)-2-(6-Aminofluoren-9-yl)-1-methylethoxy]methyl]phosphoric acid (ie PMPA) (1D) (5 g, 17.4 mmol) was added under a nitrogen atmosphere In a three-necked flask, acetonitrile (40 mL), triethylamine (3.5 g, 34.8 mmol), 4-dimethylaminopyridine (ie DMAP) (2.1 g, 17.4 mmol) and triphenyl phosphite (8.1 g, 26.1) were added. After the addition was completed, the mixture was heated to an internal temperature of 80 ° C for two days. The reaction mixture was concentrated under reduced pressure to dryness to ethyl acetate. ethyl acetate (10mL) and water (15mL) were added to the residue, and the aqueous layer was extracted with ethyl acetate (10mL×2). Adjust the pH to 3 with hydrochloric acid, stir for 10 minutes at room temperature, adjust the pH to 2 with concentrated hydrochloric acid, cool to 10 ° C with ice water, stir for two hours, then let stand overnight, filter, wash the filter cake with water (10 mL), collect the filter cake, and dry the title Compound [[(1R)-2-(6-aminofluoren-9-yl)-1-methylethoxy]methyl]phenoxyphosphoric acid (1E), (3.5 g, yield 56%).

^{1 H NMR (400MHz, DMSO)} δ8.16 (s, 1H), 8.14 (s, 1H), 7.55 (s, 2H), 7.32-7.25 (m, 2H), 7.09 (m, 3H), 4.30 (dd , 1H), 4.19 (dd, 1H), 3.97 (m, 1H), 3.87 - 3.69 (m, 2H), 1.05 (d, 3H).

³¹ P NMR (400 MHz, DMSO) δ 16.66.

Fourth step: [[(1R)-2-(6-aminofluoren-9-yl)-1-methylethoxy]methyl]phenoxyphosphoryl chloride (1F)

9-[(2R)-2-[[chloro(phenoxy)phosphoryl]methoxy]propyl]purin-6-amine

[[(1R)-2-(6-Aminofluoren-9-yl)-1-methylethoxy]methyl]phenoxyphosphinic acid (1E) (2 g, 5.5 mmol) was suspended in acetonitrile (20 mL) Among them, thionyl chloride (2.6 g, 22.0 mmol) was added and heated to an internal temperature of 85 ° C for 4 hours, and the reaction liquid was concentrated under reduced pressure to give a crude material.

The fifth step: (2S)-2-[[(1R)-2-(6-aminofluoren-9-yl)-1-methylethoxy]methyl]phenoxyphosphoramidopropionic acid thio Isopropyl ester (Compound 1)

S-isopropyl(2S)-2-[[[(1R)-2-(6-aminopurin-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]propanethioate

(S)-2-Aminopropionic acid thioisopropyl ester trifluoroacetate (1C) (4 g, 16.2 mmol) was dissolved in dry dichloromethane (20 mL). Triethylamine (5 mL, 35.8 mmol) was added dropwise at -50 ° C for 10 minutes, and [[(1R)-2-(6-aminopurin-9-yl)-1-methylethoxy]methyl was added dropwise. A suspension of phenoxyphosphoryl chloride (1F) (2.1 g, 5.5 mmol) in dichloromethane (20 mL) was obtained. Water (20 mL) was added to the mixture and the mixture was evaporated. 4 mol / L hydrochloric acid was adjusted to pH 2, and the aqueous layer was extracted with ethyl acetate (20 mL). The aqueous layer was taken, and dichloromethane (50 mL) was added, and the mixture was adjusted to pH 8 The mixture was separated, and the aqueous layer was evaporated, mjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj 2-(6-Aminofluoren-9-yl)-1-methylethoxy]methyl]phenoxyphosphoramidopropionic acid thioisopropyl ester (Compound 1) (300 mg, yield 11%) .

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.33 (d, 1H), 7.98 (s, 1H), 7.32 (t, 1H), 7.23 (t, 1H), 7.14 (m,, 2H), 7.00 (d) , 1H), 5.82 (d, 2H), 4.41 (ddd, 1H), 4.22 - 3.90 (m, 5H), 3.73 - 3.36 (m, 3H), 1.32 - 1.17 (m, 12H).

³¹ P NMR (162 MHz, CDCl ₃ ) δ 23.21., 22.00.

LC-MS M/Z (ESI):495.

Resolution of Compound 1

Separation analysis method: instrument, Thar analytical SFC; column, ChiralPak AS-H, 250 × 4.6 mm; mobile phase, A is CO ₂ and B is Methanol (0.05% DEA); gradient, B 40%; flow rate, 2.4 mL/ Min; back pressure, 100 bar; column temperature, 35 ° C; wavelength, 220 nm.

Preparation separation method: instrument, MGII preparative SFC; column, ChiralPak AS-H, 250×30 mm I.D.; mobile phase, A is CO ₂ and B is Methanol; gradient, B 40%; flow rate, 40 mL/min; back pressure, 100 bar; column temperature 38 ° C; wavelength, 220 nm; period, 5.5 min.

Sample preparation: (2S)-2-[[(1R)-2-(6-aminofluoren-9-yl)-1-methylethoxy]methyl]phenoxyphosphoramidopropionic acid thioderivative Propyl ester (Compound 1) (300 mg) was dissolved in methanol to prepare a solution having a sample concentration of 10 mg/mL, and 3 mL/cap each was injected, and two optical isomer compounds 1-1 were obtained after separation (retention time: 2.21 min). , 106 mg, white solid, ee% = 100%), Compound 1-2 (retention time: 3.82 min, 109 mg, white solid, ee% = 100%).

Compound 1-1

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.31 (s, 1H), 8.1 (s, 1H), 7.32 (t, 2H), 7.21 - 7.11 (m, 3H), 6.04 (s, 2H), 4.47 ( Dd,1H), 4.21–4.13(m,1H), 4.13–4.06(m,1H), 4.06–3.96(m,2H),3.69(dd,1H),3.54–3.39(m,2H),1.28– 1.17 (m, 12H).

³¹ P NMR (162 MHz, CDCl ₃ ) δ 23.15.

LC-MS M/Z (ESI):495.

Compound 1-2

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.35 (s, 1H), 7.97 (s, 1H), 7.25 - 7.17 (m, 2H), 7.13 - 7.05 (m, 1H), 7.03 - 6.95 (m, 2H) ), 5.90 (s, 2H), 4.34 (dd, 1H), 4.16 - 4.03 (m, 2H), 3.99 - 3.89 (m, 2H), 3.84 (t, 1H), 3.76 - 3.52 (m, 2H), 1.33–1.20 (m, 12H).

³¹ P NMR (162 MHz, CDCl ₃ ) δ 22.12.

LC-MS M/Z (ESI):495.

Example 2

(2S)-2-[[(1R)-2-(6-Amino嘌呤-9-yl)-1-methylethoxy]methyl]phenoxyphosphoramidopropionic acid thioethyl ester (compound 2)

S-ethyl(2S)-2-[[[(1R)-2-(6-aminopurin-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]propanethioate

First step: (S)-2-(tert-butoxycarbonylamino)propionic acid thioethyl ester (2B)

S-ethyl(S)-2-(tert-butoxycarbonylamino)propanethioate

N-tert-butoxycarbonyl-L-alanine (2A) (50,264 mmol) was dissolved in tetrahydrofuran (400 mL), and N,N'-carbonyldiimidazole (CDI) (47 g, 291 mmol) was added and stirred at room temperature 1 Ethyl thiol (18 g, 291 mmol) was added at rt overnight. Water (100 mL) was added to the reaction mixture, and the mixture was evaporated. EtOAcjjjjjjjjjjjj The mixture was concentrated under reduced pressure. EtOAc (EtOAc:EtOAc:EtOAc 2B), white solid (46 g, yield 74.6%).

_{^{1 H NMR (400MHz, CDCl 3}} ) δ4.97 (s, 1H), 4.37 (d, 1H), 2.88 (m, 2H), 1.45 (s, 9H), 1.37 (d, 3H), 1.25 (t, 3H).

The second step (2S)-2-aminopropionic acid thioethyl ester trifluoroacetate (2C)

S-isopropyl(2S)-2-aminopropanethioate triflouroacetate

(S)-2-(tert-Butoxycarbonylamino)propionic acid thioethyl ester (1B) (23 g, 98.57 mmol) was dissolved in dichloromethane (20 mL), trifluoroacetic acid (20 mL) The reaction was carried out for 2 hours, and the residue was evaporated to dryness mjjjjjjjj

The third step (2S)-2-[[(1R)-2-(6-aminofluoren-9-yl)-1-methylethoxy]methyl]phenoxyphosphoramidopropionic acid thioethyl Ester (Compound 2)

S-ethyl(2S)-2-[[[(1R)-2-(6-aminopurin-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]propanethioate

(2S)-2-Aminopropionic acid thioethyl ester (2C) (25g, 98.57mmol) was dissolved in dry dichloromethane (150mL), under nitrogen, dry ice-ethanol cooled to -50 ° C, add three Ethylamine (36.6 g, 361.6 mmol), [[(1R)-2-(6-aminofluoren-9-yl)-1-methylethoxy]methyl]phenoxyphosphoryl chloride (1F) 23 g, 60.24 mmol), naturally heated to room temperature for 1 hour. After completion of the reaction, water (50 mL) was added, and the mixture was evaporated. mjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj The residue was purified by EtOAcjjjjjjjjjjjjj Thioethyl 2-(6-aminoindol-9-yl)-1-methylethoxy]methyl]phenoxyphosphoramidopropionate (Compound 2) (8 g, yield 27.7%).

_{^{1 H NMR (400MHz, CDCl 3}} ) δ8.33 (d, 1H), 7.97 (d, 1H), 7.35-7.27 (m, 1H), 7.24-7.06 (m, 3H), 7.00 (d, 1H), 6.15(d, 2H), 4.50–4.29 (m, 1H), 4.22–4.05 (m, 2H), 4.05–3.89 (m, 2H), 3.75–3.61 (m, 1H), 2.84 (m, 1H), 2.80–2.64 (m, 1H), 1.29–1.13 (m, 9H).

³¹ P NMR (162 MHz, CDCl ₃ ) δ 23.38, 22.22.

LC-MS M/Z (ESI): 479.3.

Compound 2 resolution

Separation and analysis method: Instrument: Thar analytical SFC, column: ChiralPak AS-H, 250 × 4.6 mm, mobile phase: A is CO ₂ and B is methanol (0.05% DEA), gradient: B 40%, flow rate: 2.4 mL / Min, back pressure: 100 bar, column temperature: 35 ° C, wavelength: 220 nm,

Preparation Separation method: Instrument: Thar 200prepararive SFC, column: ChiralPak AS-10u, 300 × 50 mm I.D., mobile phase: A is CO ₂ and B is ethanol, gradient: B 45%, flow rate: 200 mL/min, back pressure : 100 bar, column temperature: 38 ° C, wavelength: 220 nm, period: ~ 15 min,

Sample preparation: Compound 2 was dissolved in ethanol to prepare a solution having a sample concentration of 60 mg/ml. Injection: 16 ml per needle, two optical isomer compounds 2-1 were obtained after separation (retention time: 2.32 min, 2.38 g, White solid, ee% = 100%), Compound 2-2 (retention time: 3.98 min, 2.24 g, white solid, ee% = 100%).

Compound 2-1

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.30 (s, 1H), 7.97 (s, 1H), 7.34 - 7.28 (m, 2H), 7.19 - 7.13 (m, 3H), 6.08 (s, 2H), 4.46 (dd, 1H), 4.21–3.95 (m, 5H), 3.74–3.60 (m, 2H), 2.84–2.65 (m, 2H), 1.25 (d, 3H), 1.20 (d, 3H), 1.16 ( t, 3H).

³¹ P NMR (162 MHz, CDCl ₃ ) δ 23.37.

LC-MS M/Z (ESI): 47:21.

Compound 2-2

_{^{1 H NMR (400MHz, CDCl 3}} ) δ8.35 (s, 1H), 7.97 (s, 1H), 7.20 (m, 2H), 7.09 (m, 1H), 7.03-6.96 (m, 2H), 6.10 ( s, 2H), 4.37–4.23 (m, 2H), 4.11 (m, 2H), 4.01–3.87 (m, 2H), 3.66 (dd, 1H), 2.84 (m, 2H), 1.31–1.19 (m, 9H).

³¹ P NMR (162 MHz, CDCl ₃ ) δ 22.20.

LC-MS M/Z (ESI): 47:21.

Example 3 Fumarate of Compound 1-1

Compound 1-1 (2 g, 4.06 mmol) was dissolved in ethyl acetate (26 mL), and then evaporated. The mixture was cooled to room temperature and stirred to crystallize for 3 hours, filtered, and the filter cake was washed with ethyl acetate (20 mL) and dichloromethane (20 mL). 1.9 g, yield 84.4%).

^{1 H NMR (400MHz, DMSO)} δ13.11 (s, 2H), 8.13 (d, 2H), 7.34 (t, 2H), 7.19-7.11 (m, 5H), 6.63 (s, 2H), 5.77 (dd , 1H), 4.26 (ddd, 2H), 4.16 - 3.76 (m, 4H), 3.36 (m 1H), 1.26 - 1.14 (m, 6H), 1.07 (dd, 6H).

³¹ P NMR (162 MHz, DMSO) δ 24.25.

LC-MS M/Z (ESI):495.

Example 4 Fumarate of Compound 1-2

Compound 1-2 (2 g, 4.06 mmol) was dissolved in ethyl acetate (18 mL), and then evaporated. After cooling to room temperature, the mixture was stirred for 3 hours, filtered, and the filter cake was washed successively with ethyl acetate (20 mL) and dichloromethane (20 mL), and the filter cake was dried under reduced pressure to give compound 1-2. g, yield 66.7%).

^{1 H NMR (400MHz, DMSO)} δ13.11 (s, 2H), 8.15 (s, 1H), 8.10 (s, 1H), 7.29 (t, 2H), 7.21 (s, 2H), 7.14 (t, 1H ), 7.05 (d, 2H), 6.64 (s, 2H), 5.94 (t, 1H), 4.28 (dd, 1H), 4.14 (dd, 1H), 3.99 - 3.83 (m, 3H), 3.76 (dd, 1H), 3.37 (m, 1H), 1.20 (dd, 6H), 1.12 (d, 3H), 1.08 (d, 3H).

³¹ P NMR (162 MHz, DMSO) δ 23.84.

LC-MS M/Z (ESI):495.

Example 5

2-[[[(1R)-2-(6-aminofluoren-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]-2-methylpropanoic acid Thioisopropyl ester (compound 5)

S-isopropyl 2-[[[(1R)-2-(6-aminopurin-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]-2-methyl-propanethioate

First step: 2-[[[(1R)-2-(6-aminofluoren-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]-2- Methyl-propionic acid ethyl ester (5B)

Ethyl 2-[[[(1R)-2-(6-aminopurin-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]-2-methyl-propanoate

Ethyl 2-amino-2-methylpropanoate (18 g, 0.138 mol) was dissolved in dichloromethane (200 mL), diisopropylamine (11.49 g, 0.114 mol) Add 9-[(2R)-2-[[chloro(phenoxy)phosphoryl]methoxy]propyl]indole-6-ammonia (16 g, 41.99 mmol), and react at room temperature for 2 hours to react The solution was washed with 20 mL of water and washed with saturated sodium hydrogen sulfate (20 mL×2). 50:1) to give the title compound 2-[[[(1R)-2-(6-aminopurin-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino] Ethyl 2-methyl-propionate (5B), red solid (5 g, yield 25.0%).

LCMS m/z = 477.1 [M + 1].

Second step: 2-[[[(1R)-2-(6-aminofluoren-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]-2- Thiopropyl methacrylate (Compound 5)

S-isopropyl 2-[[[(1R)-2-(6-aminopurin-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]-2-methyl-propanethioate

Trimethylaluminum (2 mol/L) (25.2 mL, 50 mmol, 2 mol/L) was dissolved in dichloromethane (50 mL) and diisopropylthiol (3.8 g, 50 mmol) Stir for 30 minutes in an ice bath and add 2-[[[(1R)-2-(6-aminoindol-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl] Amino]-2-methyl-propionic acid ethyl ester (5B) (3g, 6.3mmol), reacted at room temperature for 4 days, adding a saturated solution of ammonium chloride (50mL) to the reaction solution, filtering, liquid separation, water phase two The organic phase was extracted with chloromethane (50 mL×3), EtOAc (EtOAc) -9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]-2-methylpropionic acid thioisopropyl ester (Compound 5), red solid 3.1 g, Rate is 100%).

LCMS m/z = 507.0 [M + 1].

Resolution of compound 5

Separation and analysis method: Instrument: Agilent analytical SFC, column: ChiralCel OD-3, 150 × 4.6 mm, mobile phase: A is CO ₂ and B is ethanol (0.05% DEA), gradient: B 5-40%, flow rate: 2.4 mL / Min, back pressure: 100 bar, column temperature: 35 ° C, wavelength: 220 nm,

Preparation Separation method: Instrument: Thar 200prepararive SFC, column: ChiralCel OD-10u, 300×50 mm I.D, mobile phase: A is CO ₂ and B is ethanol, gradient: B 25%, flow rate: 200 mL/min, back pressure 100 bar , column temperature: 38 ° C, wavelength: 220 nm, period: ~ 5 min,

Sample preparation: Compound 5 was dissolved in ethanol and dichloromethane to prepare a solution having a sample concentration of 40 mg/ml. Injection: 3 ml per needle, two optical isomer compounds 5-1 were obtained after separation (retention time: 3.38 min) , 0.88 g, white solid, ee% = 100%), Compound 5-2 (retention time: 3.77 min, 1.36 g, white solid, ee% = 100%).

Compound 5-1

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.34 (s, 1H), 7.95 (s, 1H), 7.24 - 7.22 (m, 2H), 7.13 - 7.08 (m, 1H), 7.01 - 6.98 (m, 2H) ), 5.69 (s, 2H), 4.36 (dd, 1H), 4.19 - 4.13 (m, 1H), 4.04 - 3.85 (m, 3H), 3.73 - 3.52 (m, 2H), 1.54 (s, 3H), 1.49 (s, 3H), 1.31 (d, 3H), 1.29 (d, 3H), 1.26 (d, 3H).

³¹ P NMR (162 MHz, CDCl ₃ ) δ 22.56.

LCMS m/z = 507.0 [M + 1];

Compound 5-2

_{^{1 H NMR (400MHz, CDCl 3}} ) δ8.34 (s, 1H), 7.98 (s, 1H), 7.33-7.31 (m, 2H), 7.19-7.13 (m, 3H), 5.74 (s, 2H), 4.41 (dd, 1H), 4.19–4.14 (m, 1H), 4.07–3.85 (m, 2H), 3.82–3.65 (m, 2H), 3.57–3.46 (m, 1H), 1.47 (s, 3H), 1.40 (s, 3H), 1.28 (d, 3H), 1.26 (d, 3H), 1.22 (d, 3H).

³¹ P NMR (162 MHz, CDCl ₃ ) δ 22.36.

LCMS m/z = 507.0 [M + 1].

Example 6

2-[[[(1R)-2-(6-Amino-7H-indol-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]-2-yl Thioethyl propyl propionate (compound 6)

S-ethyl 2-[[[(1R)-2-(6-amino-7H-purin-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]-2-methyl-propanethioate

Trimethylaluminum (67 ml, 134.4 mmol, 2 mol/L) was dissolved in dichloromethane (100 mL), ethanethiol (8.33 g, 10 mmol) was added at 0 ° C, and reacted for 15 minutes, and 2-[[[(1R) was added. )-2-(6-Amino-7H-indol-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]-2-methyl-propionic acid ethyl ester ( 5B) (8.0 g, 16.8 mmol), allowed to react at room temperature overnight. After being added to a saturated aqueous solution of ammonium chloride, the mixture was evaporated. EtOAcjjjjjjjjjjjjjj (6-Amino-7H-indol-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]-2-methylpropionic acid thioethyl ester (Compound 6) , yellow oil (4.47 g, yield 54.4%).

LCMS m/z = 493.1 [M + 1].

Resolution of compound 6

Separation and analysis method: Instrument: Thar analytical SFC, column: ChiralCel OD-H, 250 × 4.6 mm, 5 um, mobile phase: A is CO ₂ and B is isopropanol (0.05% DEA), gradient: B 5 to 40% , flow rate: 2.4mL / min, back pressure: 100bar, column temperature: 35 ° C, wavelength: 220nm,

Preparation separation method: Instrument: MGII preparative SFC-1, column: ChiralCel OD-5u, 250×30 mm I.D. Mobile phase: A is CO2 and B is isopropanol, gradient: 30%, flow rate: 60 mL/min, back Pressure: 100 bar, column temperature: 38 ° C, wavelength: 220 nm, period: ~ 4 min,

Sample preparation: Compound 6 was dissolved in methanol, injection: 2.1 ml per needle, and two optical isomer compounds 6-1 were obtained after separation (retention time: 1.93 min, 1.45 g, white solid, ee% = 100%) Compound 6-2 (retention time: 2.87 min, 3.04 g, white solid, ee% = 100%).

Compound 6-1: ¹ H NMR (400MHz, CDCl ₃ ) δ 8.33 (s, 1H), 7.97 (s, 1H), 7.26 - 7.22 (m, 2H), 7.12 - 7.11 (m, 1H), 7.00 - 6.98 (m, 2H), 5.83 (s, 2H), 4.36 (dd, 1H), 4.16 (dd, 1H), 3.97–3.92 (m, 2H), 3.85 (d, 1H), 3.70–3.65 (m, 1H) ), 2.80 (q, 2H), 1.54 (s, 3H), 1.50 (s, 3H), 1.27 - 1.22 (m, 6H).

³¹ P NMR (162 MHz, CDCl ₃ ) δ 22.63.

LCMS m/z = 493.0 [M + 1];

Compound 6-2: ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.33 (s, 1H), 7.96 (s, 1H), 7.35 - 7.28 (m, 2H), 7.21 - 7.09 (m, 3H), 6.09 (s , 2H), 4.41 (dd, 1H), 4.16 (dd, 1H), 4.04–3.89 (m, 2H), 3.74–3.63 (m, 2H), 2.80–2.77 (m, 2H), 1.47 (s, 3H) ), 1.40 (s, 3H), 1.23 - 1.18 (m, 6H).

³¹ P NMR (162 MHz, CDCl ₃ ) δ 22.46.

LCMS m/z = 493.0 [M + 1].

Example 7

1-[[[(1R)-2-(6-aminofluoren-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]-cyclopropylcarboxylic acid thio Isopropyl ester (Compound 7)

S-isopropyl 1-[[[(1R)-2-(6-aminopurin-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]-cyclopropanecarbothioate

First step: 1-[[[(1R)-2-(6-aminofluoren-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]-cyclopropane Ethyl urethane (7B)

Ethyl 1-[[[(1R)-2-(6-aminopurin-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]-cyclopropanecarboxylate

Ethyl 1-aminocyclopropylcarboxylate (5.0 g, 43.9 mmol) was dissolved in dichloromethane (200 mL) and diisopropylamine (3.5 g, 35.56 mmol) was added under nitrogen. 5.0g, 13.1mmol), the reaction was heated to room temperature for 2 hours, the reaction solution was quenched with 20 mL of water, and the organic phase was washed with saturated sodium hydrogen sulfate (20 mL×2). The residue was purified by EtOAc EtOAcjjjjjjjjj

LCMS m/z = 475.1 [M + 1].

Second step: 1-[[[(1R)-2-(6-aminofluoren-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]-cyclopropane Thiopropyl methacrylate (compound 7)

S-isopropyl 1-[[[(1R)-2-(6-aminopurin-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphoryl]amino]-cyclopropanecarbothioate

Trimethylaluminum (2 mol/L) (33.7 ml, 67.5 mmol) was dissolved in 50 mL of dichloromethane, and diisopropyl mercaptan (5.13 g, 67.5 mmol) was added under ice-cooling under ice, and then ice bath Stir for 30 minutes, add 7B (4.0g, 8.4mmol), warm to room temperature for 4 days, add 50mL of saturated aqueous solution of saturated ammonium chloride to quench the reaction, separate the liquid, and extract with dichloromethane (50mL × 3 ), the organic phase is combined, the organic phase is washed with water (50 mL × 2), anhydrous sulfur The sodium salt was dried and concentrated to give the title compound <RTIgt;

Resolution of compound 7

Preparation separation method: Instrument: waters SFC, column: Chiralpak AS-3 (4.6 × 100mm), mobile phase: A is methanol and B is CO ₂ , gradient: B 10-40%, flow rate: 2mL / min, back pressure: 2000 psi, column temperature: 35 ° C, wavelength: 260 nm; period: ~ 6 min;

Sample preparation: Compound 7 was dissolved in methanol, injection: 2 ml per needle, two optical isomer compounds 7-1 were obtained after separation (retention time: 1.62 min, 30 mg, white solid, ee% = 100%), compound 7-2 (retention time: 2.52 min, 60 mg, white solid, ee% = 100%).

Compound 7-1: ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.32 (s, 1H), 7.96 (s, 1H), 7.31 - 7.29 (m, 2H), 7.18-7.13 (m, 3H), 5.94 (s , 2H), 4.44 (dd, 1H), 4.19 (dd, 1H), 4.13–3.94 (m, 3H), 3.84–3.78 (m, 1H), 3.46–3.39 (m, 1H), 1.56–1.47 (m) , 1H), 1.40 - 1.35 (m, 1H), 1.26 (d, 3H), 1.23 - 1.20 (m, 6H), 1.14 - 1.06 (m, 1H), 1.03 - 0.94 (m, 1H).

³¹ P NMR (162 MHz, CDCl ₃ ) δ 23.42.

LCMS m/z = 505.0 [M + 1];

Compound 7-2: ¹ H NMR (400MHz, CDCl ₃ ) δ 8.34 (s, 1H), 7.92 (s, 1H), 7.30 - 7.22 (m, 2H), 7.15 - 7.12 (m, 1H), 7.01–6.99 (m, 2H), 5.70 (s, 2H), 4.41 (dd, 1H), 4.27 - 4.08 (m, 3H), 4.05 - 3.96 (m, 1H), 3.77 (dd, 1H), 3.53 - 3.46 (m , 1H), 1.55–1.44 (m, 2H), 1.30–1.22 (m, 9H), 1.17–1.08 (m, 2H).

³¹ P NMR (162 MHz, CDCl ₃ ) δ 22.53.

LCMS m/z = 505.0 [M + 1].

Test case

Test Example 1. Screening for anti-hepatitis B virus activity

Compounds were tested for anti-HBV activity using HepG 2.2.15 cells. The materials and instruments used were as follows: HepG2.2.15 cells, RPMI 1640 medium, fetal bovine serum, 96-well plates, DMSO, QIAamp 96 DNA Blood Kit, Cell-titer blue, microplate reader, Applied Biosystems 7900 real-time PCR system.

Each compound was dissolved in DMSO at 20 mM and stored at -20 ° C, and a 20 mM stock solution of each compound was diluted with a DMSO 3 fold gradient for a total of 9 concentrations. It was further diluted 200-fold with RPMI 1640 medium containing 2.0% FBS. The highest test final concentration of the compound was 100M. Experimental Procedure The compounds were tested for anti-HBV activity by qPCR method and the EC ₅₀ (half effective inhibitory concentration) was calculated by following the QIAamp 96 DNA Blood Kit (QIAGEN 51161) instructions. Analysis of data and calculation of percent inhibition: The percentage of inhibition was calculated using the following formula: inhibition rate (%) = (total amount of HBV in the DMSO control group - total amount of HBV in the test sample group) / total amount of HBV in the DMSO control group × 100. Finally, EC ₅₀ values _were calculated using GraphPad Prism software compound.

The cytotoxicity of the compounds was determined by the Cell-titer blue method and the CC ₅₀ (to 50% cytotoxic concentration) was calculated. Analyze data and calculate relative cell viability: Calculate the percentage of cell viability using the following formula: cell viability (%) = (fluorescence value of the test sample - background fluorescence value) / (fluorescence value of the DMSO control group - background fluorescence value) × 100. Finally, compound CC ₅₀ values _are calculated using GraphPad Prism software. The results are shown in the following table:

Table 1: EC ₅₀ values and CC ₅₀ values for each compound

Serial number	Compound number	EC 50 (nM)	CC 50 (μM)
				1	1-1	1.96	>100
2	1-2	1.42	>100
				3	2-1	2.46	>100
4	2-2	3.12	>100
				5	5-1	1.90	>100
6	5-2	5.78	>100
				7	GS-7340	2.54	>100

Conclusion: The test compounds all showed good anti-HBV activity, comparable anti-HBV activity compared to the control, and no cytotoxicity over the range of concentrations tested.

Test Example 2 Rat Tissue Distribution

Male SD rats (purchased from Vitalius, license number SCXK (Beijing) 2012-0001), weighing 200-220 g. One day before the experiment, the animals were fasting and could not help but water. On the day of the experiment, 36 rats were intragastrically administered with 3 test compounds at a dose of 15 mg/kg (based on the original drug PMPA). Blood and tissue samples were collected at 0.5 h, 2 h, 6 h and 24 h after administration, respectively. Blood was collected from the eyelids (heparin anticoagulation), 3000 g, centrifuged at 4 ° C for 10 min, and plasma was collected. At the same time, the liver and kidney were collected, and the total weight was weighed, and then 50 mg of each tissue was taken and stored at -80 °C.

A standard solution of the test compound is prepared and added to the blank plasma, liver and kidney homogenate. The concentration of the standard is 10000 ng/ml, 5000 ng/ml, 2500 ng/ml, 1000 ng/ml, 250 ng/ml, 50 ng/ml, 25 ng/ml, 10 ng/ml, 5 ng/ml, 2 ng/ml. 30 μl of each concentration solution was added, 200 μl of acetonitrile containing internal standard was added, shaken at 2500 rpm for 2 min, then centrifuged at 13,000 rpm for 10 min at 4 ° C, and the supernatant was taken to prepare a standard curve of the test compound in blank plasma, liver and kidney homogenate.

Take 30 μl of plasma sample, add 200 μl of acetonitrile containing internal standard, shake at 2500 rpm for 2 min, then centrifuge at 13000 rpm for 10 min at 4 ° C, take the supernatant, and measure the concentration of PMPA and test compound (ng/ml) in the test compound group by HPLC-MS/MS. The tissue sample was added with 0.5 ml of physiological saline per 50 mg and homogenized. After homogenization, 30 μl of homogenate was added, 200 μl of acetonitrile containing internal standard was added, shaken at 2500 rpm for 2 min, then centrifuged at 13,000 rpm for 10 min at 4 ° C, and the supernatant was taken. The PMPA concentration in the test compound group was measured by HPLC-MS/MS (ng/ml). , ng/g). The test results are shown in Table 2 and Table 3.

Table 2: Rat tissue distribution results

Table 3: Peak results of rat tissue distribution

Conclusion: Compared with the control, the PMPA has higher liver exposure and higher liver to kidney concentration ratio, suggesting better liver targeting and lower renal metabolic concentration, which can reduce nephrotoxicity.

Test 3 whole blood stability

The whole blood of ICR mice, SD rats, Beagle dogs, cynomolgus monkeys and healthy people used in this experiment was collected freshly before the experiment (male and female). The test compound will be co-incubated with various genus whole blood, the incubation system is 400 μL, and the final concentration is 1 μM. At different time points (0, 5, 15, 30, 60 min), 40 μL of the incubated whole blood sample was taken and added to 200 μL of acetonitrile containing the internal standard. After the protein was precipitated, the supernatant was centrifuged, and the test compound in the supernatant was analyzed by the LC-MS/MS method, and the sample was parallelized in two portions.

The analyte/internal standard peak area ratio (Aanalyte/AIS) will be derived from the instrument and the remaining percentage (%Control) will be calculated from the ratio of non-zero time point samples to Aanalyte/AIS in the zero time sample. Ln (%Control) was plotted against incubation time and fitted linearly. The results are shown in Table 4.

Table 4: Whole blood metabolic stability

Conclusion: In human whole blood, the Average T _1/2 value of the compound 1-1, 1-2 of the present invention is more than 30 times that of the control compound, and its stability is obviously superior to the control compound, so the exposure of PMPA in human plasma is observed. The lower amount significantly reduces the toxic side effects of the compounds of the invention due to the metabolism of PMPA in plasma.

Claims (13)

1. a compound of the formula (A), a stereoisomer thereof, a pharmaceutically acceptable salt or a eutectic, wherein:

   

   A is selected from a 6 to 10 membered aromatic ring or a 6 to 10 membered heteroaryl ring, and the heteroaryl group contains 1 to 4 hetero atoms selected from N, O, and S, and the aromatic ring or heteroaryl ring is Further selected from 0 to 5 selected from H, F, Cl, Br, I, CN, amino, hydroxy, carboxy, C _1-4 alkyl, trifluoromethyl, C _1-4 alkoxy or -C ( Substituted by a substituent of OC _1-4 alkyl;

   B is

   

   E is selected from -CH ₂ CH(CH ₃ )OCH ₂ - or -CH ₂ CH ₂ OCH ₂ -;

   R ^{N is} selected from H or C _1-4 alkyl;

   R ¹ and R ² are each independently selected from the group consisting of H, C _1-6 alkyl or a side chain of a natural or pharmaceutically acceptable amino acid, and if the side chain contains a carboxyl group, the carboxyl group may be optionally esterified with an alkyl group or an aryl group;

   Alternatively, R ¹ , R ² may form a C _3-6 cycloalkyl group together with the carbon atom to which they are attached;

   R ^{3 is} selected from H, C _1-6 alkyl, 6 to 10 membered aromatic ring or 6 to 10 membered heteroaryl ring, and the heteroaryl group contains 1 to 4 hetero atoms selected from N, O, and S, The alkyl, aromatic or heteroaryl ring is optionally further selected from 0 to 5 selected from the group consisting of H, F, Cl, Br, I, CN, amino, hydroxy, carboxy, C _1-4 alkyl or C _1- Substituted by a ₄ -alkoxy substituent.
2. The compound according to claim 1, a stereoisomer thereof, a pharmaceutically acceptable salt or a eutectic, wherein the compound is selected from the compounds of the formula (I):

   

   A is selected from phenyl or naphthyl, and the phenyl or naphthyl group is further optionally further selected from 0 to 5 selected from the group consisting of H, F, Cl, Br, I, CN, amino, hydroxy, carboxy, C _1-4 alkane. Substituted by a substituent of a C _1-4 alkoxy group;

   R ² is a side chain of a natural or pharmaceutically acceptable amino acid, and if the side chain contains a carboxyl group, the carboxyl group may be optionally esterified with an alkyl group or an aryl group;

   R ³ is a C _1-6 alkyl group.
3. The compound according to claim 2, a stereoisomer thereof, a pharmaceutically acceptable salt or a cocrystal thereof, wherein the compound is selected from the group consisting of compounds represented by the formula (II), wherein

   

   E is selected from -CH ₂ CH(CH ₃ )OCH ₂ -;

   R ^{2 is} selected from the side chain of glycine, alanine or phenylalanine;

   R ³ is a C _1-4 alkyl group.
4. The compound according to claim 3, a stereoisomer thereof, a pharmaceutically acceptable salt or a eutectic, wherein the compound is selected from the group consisting of compounds of the formula (III), wherein:

   

   R ^{3 is} selected from the group consisting of methyl, ethyl, propyl, isopropyl, butyl or isobutyl.
5. The compound according to claim 1, a stereoisomer thereof, a pharmaceutically acceptable salt or a eutectic selected from the group consisting of compounds of the formula (IV), wherein:

   

   R ¹ and R ² are each independently selected from a C _1-6 alkyl group, or R ¹ and R ² together with the carbon atom to which they are attached form a C _3-6 cycloalkyl group.
6. A compound according to claim 5, a stereoisomer thereof, a pharmaceutically acceptable salt or a eutectic, wherein:

   A is selected from a substituted or unsubstituted phenyl, pyridyl or naphthyl group, and when substituted, optionally further 1 to 3 are selected from the group consisting of H, F, Cl, Br, I, CN, methoxy, methyl Substituted by a substituent of a trifluoromethyl or ethoxycarbonyl group;

   E is selected from -CH ₂ CH(CH ₃ )OCH ₂ -;

   R ¹ and R ² are each independently selected from methyl or ethyl, or R ¹ , R ² together with the carbon atom to which they are attached form a cyclopropyl group;

   R ^{3 is} selected from methyl, ethyl, isopropyl, monofluoromethyl or difluoromethyl.
7. A compound according to claim 1 which is a stereoisomer or a pharmaceutically acceptable salt or eutectic, wherein the compound is:

   
8. The compound according to any one of claims 1 to 7, which is a stereoisomer or a pharmaceutically acceptable salt or eutectic, wherein the salt is a fumarate.
9. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 8 and a stereoisomer thereof or a pharmaceutically acceptable salt or co-crystal thereof, and a pharmaceutically acceptable Accepted carrier or excipient.
10. The use of the compound according to any one of claims 1 to 8, a stereoisomer thereof, or a pharmaceutically acceptable salt or co-crystal thereof, for the preparation of a medicament for the treatment of a viral infectious disease.
11. The use according to claim 10, wherein the viral infectious disease comprises an infectious disease caused by hepatitis B virus, hepatitis C virus and HIV virus.
12. A method of treating a viral infectious disease, wherein the method comprises administering a compound according to any one of claims 1 to 8, a stereoisomer thereof, or a pharmaceutically acceptable salt or cocrystal thereof or a claim thereof 9. The pharmaceutical composition of 9.
13. The method according to claim 12, wherein the viral infectious disease comprises an infectious disease caused by hepatitis B virus, hepatitis C virus, and HIV virus.