TWI616453B - Substituted amino acid thioester compounds, materials and uses thereof - Google Patents

Abstract

The present invention relates to a substituted amino acid thioester compound represented by the formula (A), a composition thereof and use thereof, and the definition of each substituent in the formula (A) is the same as defined in the specification.

Description

Substituted amino acid thioester compounds, compositions and applications thereof

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 a pharmaceutically acceptable Acceptable salts and pharmaceutical compositions containing them and use in the manufacture of a medicament for the treatment of viral infectious diseases.

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 nuclear protein (HBcAg), gene S encodes surface antigen (HBsAg), gene P encodes DNA polymerase, and gene X The function of the encoded protein is unclear, but it is thought to be involved in the development of liver cancer because it activates genes that induce cell proliferation and deactivates 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: a terminal protein important for the initiation of hepatitis B virus replication and assembly of the nucleocapsid, a spacer protein, a reverse transcriptase, and an RNaseH domain for degrading the pre-genomic RNA template. 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 is a 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. So far, there are five nucleoside analogues of hepatitis B virus DNA polymerase inhibitors for the treatment of chronic hepatitis B. Drugs, marketed in the US and Europe, including: lamivudine, tenofovir dipivoxil, entecavir, telbivudine and tenofovir disoproxil Salt (tenofovir disoproxil fumarate), and several other drugs are in the research and development stage. 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-aminopurine-9-yl)-1-methyl-ethoxy]methyl phosphate (PMPA), is a nucleotide class A reverse transcriptase inhibitor having 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 phosphoniumamine 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 the group consisting of CH ₂ C(O)N(R ⁵ ) ₂ , CH ₂ C(O)OR ⁵ , CH ₂ OC(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 camphorinyl, and R ^{1 is} selected from 3-8 carbons. A cycloalkyl group, a 3-8 carbon unsaturated chain hydrocarbon group, a 3-8 carbon unsaturated cycloalkyl group or a 6-10 carbon aromatic hydrocarbon. 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 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.

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, the aromatic ring or heteroaryl ring 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 ( =O) 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 H, C a side chain of _1-6 alkyl or a natural or pharmaceutically acceptable amino acid, 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 be attached to the carbon to which it is attached The atoms together form a C _3-6 cycloalkyl group; R ^{3 is} selected from H, C _1-6 alkyl, 6 to 10 membered aromatic rings or 6 to 10 membered heteroaryl rings, said heteroaryl containing 1 to 4 a hetero atom selected from N, O, S, wherein the alkyl, aromatic or heteroaryl ring is further further selected from 0 to 5 selected from the group consisting of H, F, Cl, Br, I, CN, amino, hydroxy, carboxy Substituted with a C _1-4 alkyl group or a C _1-4 alkoxy 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 (A), wherein the compound is selected from the group consisting of the compound 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 If the side chain contains a carboxyl group, the carboxyl group may be optionally esterified with an alkyl group or an aryl group; and R ³ is a C _1-6 alkyl group.

According to a preferred embodiment of the present invention, an amino acid thioester compound represented by the formula (I), a stereoisomer or a pharmaceutically acceptable salt thereof, wherein A is selected from phenyl or naphthyl, preferably phenyl; said phenyl or naphthyl optionally further from 0 to 5 selected from H, F Substituted with a substituent of Cl, Br, I, CN, amino, hydroxy, carboxy, methyl, ethyl, methoxy or ethoxy, further preferably from 0 to 5 selected from H, F, Cl, Substituted by a substituent of Br, CN, amino or methoxy.

Preferably, the 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, wherein The amino acid is preferably glycine, alanine, leucine, isoleucine, tyrosine, valine, phenylalanine, methionine, tryptophan, serine, glutamine, threonine Acid, cysteine, histidine, aspartame, tyrosine, aspartic acid, glutamic acid, naphthyl acid or arginine, further preferably glycine, alanine, leucine , phenylalanine, aspartame or arginine, more preferably glycine, alanine or phenylalanine.

Preferably, the amino acid thioester compound of the formula (I), a stereoisomer or a pharmaceutically acceptable salt thereof, wherein R ³ is a C _1-6 alkyl group, preferably C _{1- 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, aspartame 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.

A preferred embodiment of the present invention, a thioester compound of the amino acid of the formula (III), a stereoisomer or a pharmaceutically acceptable salt thereof, wherein R ^{2 is} selected from the group consisting of glycine, alanine, leucine and styrene a side chain of a glycine, aspartame or arginine, preferably a side chain of glycine, alanine or phenylalanine, further preferably a side chain of alanine; R ³ is a C _1-4 The alkyl group is 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, 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 (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, the aromatic ring or heteroaryl ring 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 ( =O) 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 C _1-6 alkyl, and alternatively, R ¹ , R ² and The attached carbon atoms together form a C _3-6 cycloalkyl group; R ^{3 is} selected from H, C _1-6 alkyl, 6 to 10 membered aromatic rings or 6 to 10 membered heteroaryl rings, said heteroaryl containing 1 to 4 hetero atoms selected from N, O, S, optionally further 0 to 5 selected from H, F, Cl, Br, I, CN, amino, hydroxyl, Substituted by a substituent of a carboxyl group, a C _1-4 alkyl group or a C _1-4 alkoxy group.

A preferred embodiment of the invention, a compound of the formula (I), a stereoisomer thereof, a pharmaceutically acceptable salt or a cocrystal thereof, wherein: A is selected from substituted or unsubstituted phenyl, pyridyl or naphthalene a base, when substituted, optionally further substituted with from 1 to 3 substituents selected from the group consisting of H, F, Cl, Br, I, CN, methoxy, methyl, trifluoromethyl or ethoxycarbonyl ; B is E is selected from -CH ₂ CH(CH ₃ )OCH ₂ -; R ¹ and R ² are each independently selected from methyl, ethyl, and, as an alternative, 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.

Preferred embodiment of the present invention, a compound of the formula (A), a stereoisomer, a pharmaceutically acceptable salt or co-crystal, wherein: A is selected from phenyl; R ^N is selected from H; R ¹ and R ^{2 is} independently selected from methyl or ethyl; R ^{3 is} selected from methyl, ethyl or isopropyl.

In a preferred embodiment of the 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, MG II 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; two compounds with short retention time and long retention time under the above separation conditions; compounds with short retention time are compound 1-1 retention time It was 2.21 ± 0.5 min; the compound with a long retention time was Compound 1-2 with a retention time of 3.82 ± 0.5 min.

According to another preferred embodiment of the invention, the compound is a compound obtained by isolation of compound 2:

The separation conditions were: instrument: Thar 200 prepararive 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; two compounds with short retention time and long retention time under the above separation conditions; compounds with short retention time are reserved for compound 2-1 The time was 2.32 ± 0.5 min; the compound with a long retention time was Compound 2-2 with a retention time of 3.98 ± 0.5 min.

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

The separation conditions were: instrument: Thar 200 prepararive 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; under the above separation conditions, two compounds with short retention time and long retention time are obtained; wherein the compound with short retention time is compound 5-1 retention time is 3.38 ± 0.5 min; wherein the compound with a long retention time is Compound 5-2 with a retention time of 3.77 ± 0.5 min.

According to still further preferred embodiments of the invention, the compound is a compound obtained by isolation 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; two compounds with short retention time and long retention time under the above separation conditions; compounds with short retention time are reserved for compound 6-1 Time is 1.93±0.5min; 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 another preferred embodiment of the invention, the compound is a compound obtained by isolation 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; two compounds with short retention time and long retention time under the above separation conditions; wherein the compound with short retention time is compound 7-1 retention time 1.62 ± 0.5 min; wherein the compound with a long retention time is Compound 7-2 with a retention time 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 form Agent.

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

In a preferred embodiment of the invention, 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.

In a preferred embodiment of the invention, 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.

In the present invention, when substituted by a plurality of substituents, the respective substituents 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 of the groups and compounds of the present invention are carbon, hydrogen, oxygen, sulfur, nitrogen or halogen, including their isotopic conditions, and the elements and carbons, hydrogen and oxygen of 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. An alkyl group preferably having 1 to 10 carbon atoms, and non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl , n-decyl, and various branched isomers thereof; more preferably lower alkyl having 1 to 4 carbon atoms, non-limiting examples include methyl, B Base, propyl, isopropyl, n-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 H, F, Cl, Br, I, =O, alkyl, alkenyl, alkyne. Alkyl, alkoxy, alkylthio, alkylamino, fluorenyl, hydroxy, nitro, cyano, amino, alkyl decylamino, cycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocyclic Alkoxy, cycloalkylindenyl, hydroxyalkyl, carboxylic acid, carboxylic acid ester or heterocycloalkyl fluorenyl.

"Amino" means -NH ₂ and may be substituted or unsubstituted. When substituted, the substituent is preferably from 1 to 3, independently selected from alkyl, alkenyl, alkynyl, alkoxy, Alkylthio, hydroxy, amino, alkylamino, alkylnonylamino, heterocycloalkyl, cycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, hydroxyalkyl, carboxy 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 thereof include phenyl or naphthyl; the aryl group may be fused to a heteroaryl group, a heterocyclic group or a cycloalkyl group, and the moiety attached to the parent structure is an aryl group, non-limiting examples of which include Benzofuran, benzocyclopentanyl 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, hydroxy, nitro, cyano, amino, alkyl decylamino, cycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkyl fluorenyl, A hydroxyalkyl group, 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" means pharmaceutically acceptable Natural or unnatural amino acids.

"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. A eutectic is a multi-component crystal that contains both a two-membered eutectic formed between two neutral solids and a multi-membered 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 different 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 a 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, but are not limited to, compounds formed by the hydroxyl or amino functional groups of the compounds of the invention with 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. example For example, "the aryl group is optionally substituted by an alkyl group" means that the 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

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. '-disuccinimide carbonate, 1-p-methylbenzenesulfonimidazole, 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, a third butoxycarbonyl group, a benzyloxycarbonyl group, a fluorenylmethoxycarbonyl group, an allyloxycarbonyl group, a trichloroethoxycarbonyl group, Trimethyldecyl ethoxycarbonyl, methoxycarbonyl, ethoxycarbonyl, 2-biphenyl-2-propoxycarbonyl, tert-butoxy, phthalic acid, p-toluenesulfonyl, adjacent Benzosulfonyl, p-nitrophenylsulfonyl, trimethylethenyl, formyl, trifluoroethenyl, benzhydryl, benzyl, trityl, p-methoxybenzyl Or 2,4-dimethoxybenzyl, preferably a third butoxycarbonyl;

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

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醯iminocarbonate, 1-p-methylbenzenesulfonimidazole, 4,5-dicyanoimidazole, preferably: N,N-carbonyldiimidazole; Compound (IB) is deprotected to give a compound ( IC); the deprotection is a conventional amino protecting group deprotection method including, but not limited to, deprotection under acidic conditions, such as using trifluoroacetic acid; compound (IC) and compound (A-4) in basic conditions The next reaction gives the compound (I); the compound (IA) can be purchased or prepared by reference to the document CN201080036406.5; the 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, a third butoxycarbonyl group, a benzyloxycarbonyl group, a fluorenylmethoxycarbonyl group, an allyloxycarbonyl group, a trichloroethoxycarbonyl group, a trimethyldecylethoxycarbonyl group, a methoxy group. Carbonyl, ethoxycarbonyl, 2-biphenyl-2-propoxycarbonyl, tert-butoxy, phthalic acid, pair Phenylsulfonyl, o-nitrophenylsulfonyl, p-nitrophenylsulfonyl, trimethylethenyl, decyl, trifluoroethenyl, benzhydryl, benzyl, trityl , p-methoxybenzyl or 2,4-dimethoxybenzyl, preferably a third butoxycarbonyl; A, B, E, R ^N , R ² and R ³ are defined and formula (I) The definitions are consistent.

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

The compound (IV-B) can be purchased or prepared by a method such as WO2012075456, WO2011014973 or WO2012084794; the compound (IV-C) can be prepared by referring to the method of the present invention, WO0208241 or WO2013052094, etc.; the compound (IV) is passed through the compound (IV-C). It is obtained by transesterification with a thiol; R ^{4 is} selected as H or C _1-6 alkyl; and A, B, E, R ¹ , R ² and R ³ are as defined in the formula (IV).

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). The NMR was measured using a (Bruker Avance III 400 and Bruker Avance 300) nuclear magnetic apparatus, and the solvent was deuterated dimethyl hydrazine (DMSO-d ₆ ), deuterated chloroform (CDCl ₃ ), deuterated methanol (CD ₃ OD). ), the internal standard is tetramethyl decane (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-C18 100 x 4.6 mm).

The thin layer chromatography gelatin plate uses Yantai Huanghai HSGF254 or Qingdao GF254 gelatin plate. The specification of the 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 Silicone 200-300 mesh silicone 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, Anike Chemical, Shanghai Demo, 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 optimum reaction temperature at room temperature is 20 ° C to 30 ° C.

Example 1

(2S)-2-[[(1R)-2-(6-Amino嘌呤-9-yl)-1-methylethoxy]methyl]phenoxyphosphonium aminopropionic 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-(t-butoxycarbonyl) aminopropionic acid thioisopropyl ester (1B)

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

N-Tertioxycarbonyl-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. ), stirred 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), the organic layer was combined, dried over anhydrous sodium sulfate and evaporated. v/v)=1:0~9:1), the title compound (S)-2-(t-butoxycarbonyl)aminopropionic acid thioisopropyl ester (1B), light yellow liquid (4g, 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-(Tertiaryoxycarbonyl)aminopropionic acid thioisopropyl ester (1B) (4 g, 16.2 mmol) was dissolved in dichloromethane (10 mL). Stir at room temperature for 4 hours. Concentration under reduced pressure to dryness afforded crude product (S), EtOAc, EtOAc (EtOAc)

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, let stand overnight, filter, filter cake washed with water (10 mL), collect filter cake, and dry The title compound [[(1R)-2-(6-aminofluoren-9-yl)-1-methylethoxy]methyl]phenoxy hypophosphorous acid (1E), (3.5 g, yield 56%) .

^{1 H NMR (400MHz, DMSO)} δ 8.16 (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: 9-[(2R)-2-[[chloro(phenoxy)phosphonium]methoxy]propyl]6-aminopurine (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, hydrazine 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]phenoxyphosphonium aminopropionic 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 9-[(2R)-2-[[chloro(phenoxy)phosphonium]methoxy]propyl]6 was added dropwise. A suspension of aminoguanidine (1F) (2.1 g, 5.5 mmol) in methylene chloride (20 mL) was obtained. Water (20 mL) was added to the reaction mixture, and the mixture was evaporated, evaporated, evaporated, evaporated, evaporated /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 (50mL) was added, and the mixture was adjusted to pH 8 with a saturated aqueous solution of sodium hydrogencarbonate. The layers were separated and the aqueous layer was extracted with dichloromethane (20 mL) Washed with saturated sodium chloride (10 mL), dried over anhydrous sodium sulfate Methyl]phenoxyphosphonium aminopropionic acid thioisopropyl ester (Compound 1) (300 mg, yield 11%).

_{^{1 H NMR (400MHz, 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.

Separation 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, MG II 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; 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]phenoxyphosphonium aminopropionic acid thioate 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 (400MHz, 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]phenoxyphosphonium aminopropionic 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-(t-butoxycarbonylamino)propionic acid thioethyl ester (2B)

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

N-Tertioxycarbonyl-L-alanine (2A) (50,264 mmol) was dissolved in tetrahydrofuran (400 mL), and N,N'-carbonyldiimidazole (CDI) (47 g, 291 mmol) was added at room temperature. After stirring for 1 hour, ethanethiol (18 g, 291 mmol) was added and allowed to react at room temperature overnight. Water (100 mL) was added to the reaction mixture, and the mixture was evaporated. EtOAcjjjjjjjjjjjjjj The title compound (S)-2-(t-butoxycarbonylamino)propionic acid thioethyl ester was obtained (yield: petroleum ether: ethyl acetate = 1:0 to 9:1). 2B), white solid (46 g, yield 74.6%).

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 4.97 (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-(Tertiaryoxycarbonylamino)propionic acid thioethyl ester (1B) (23 g, 98.57 mmol) was dissolved in dichloromethane (20 mL). The reaction was stirred at room temperature for 2 hr and concentrated under reduced vacuo to dryness (2S) of ethyl thioacetate (2C) (25 g).

The third step (2S)-2-[[(1R)-2-(6-aminofluoren-9-yl)-1-methylethoxy]methyl]phenoxyphosphonium aminopropionic 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), cooled under nitrogen, cooled to -50 ° C with dry ice-ethanol, dropwise Triethylamine (36.6 g, 361.6 mmol), [[(1R)-2-(6-aminofluoren-9-yl)-1-methylethoxy]methyl]phenoxyphosphonium chloride (1F) (23 g, 60.24 mmol), and naturally heated to room temperature for 1 hour. After completion of the reaction, water (50 mL) was added, and the mixture was evaporated. mjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj The mixture was concentrated with EtOAc (EtOAc m.jjjjjjjjj -(6-Aminofluoren-9-yl)-1-methylethoxy]methyl]phenoxyphosphonium aminopropionic acid thioethyl ester (Compound 2) (8 g, yield 27.7%).

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 8.33 (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.

Segregation of Compound 2

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, preparative separation method: instrument: Thar 200 prepararive 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 is dissolved in ethanol to prepare sample concentration 60 mg/ml solution, 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 (400MHz, 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}} ) δ 8.35 (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, and then evaporated. The mixture was further 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)} δ 13.11 (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) Adding fumaric acid (0.471 g, 4.06 mmol) and anhydrous methanol (2 mL), heating to 50 ° C, stirring until the solution is completely clarified, then cooling to room temperature and stirring for 3 hours, filtering, and filtering the cake with ethyl acetate ( 20 mL) and dichloromethane (20 mL) were washed, and the filter cake was collected and dried under reduced pressure to give the compound 1-2 of the fumarate (1.5 g, yield 66.7%).

^{1 H NMR (400MHz, DMSO)} δ 13.11 (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-phosphonyl]amino]-2-methylpropane 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-phosphonium]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)phosphonium]methoxy]propyl]phosphonium-6-ammonia (16 g, 41.99 mmol), and react at room temperature for 2 hours. The reaction solution was washed with water (20 mL), and then washed with saturated sodium hydrogen sulfate (20 mL×2). =50:1) The title compound 2-[[[(1R)-2-(6-aminopurin-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphonium] Amino]-2-methyl-propionic acid ethyl ester (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-phosphonyl]amino]-2 - thioisopropyl methacrylate (Compound 5)

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

Dissolving trimethylaluminum (2 mol/L) (25.2 mL, 50 mmol, 2 mol/L) In methyl chloride (50 mL), diisopropyl mercaptan (3.8 g, 50 mmol) was added under ice-cooling with ice-cooling, and stirred for 30 minutes in ice-bath, and 2-[[[(1R)-2-(6-amino) was added.嘌呤-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphonium]amino]-2-methyl-propionic acid ethyl ester (5B) (3 g, 6.3 mmol), room After warming for 4 days, a saturated solution of ammonium chloride (50 mL) was added to the reaction mixture, and the mixture was filtered and evaporated. The aqueous phase was extracted with dichloromethane (50mL×3), and the organic phase was combined and the organic phase was washed with water (50mL×2) Drying over anhydrous sodium sulfate and concentrating to give the title compound 2-[[[(1(R))-(2-(6-aminopurin-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphine Thio[]amino]-2-methylpropionic acid thioisopropyl ester (Compound 5), red solid 3.1 g, yield 100%).

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

Separation 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, preparative separation method: instrument: Thar 200 prepararive SFC, column: ChiralCel OD-10u, 300 × 50 mm I. D, mobile phase: A is CO ₂ and B For 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 (400MHz, 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}} ) δ 8.34 (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-phosphonyl]amino]-2- Thioethyl methacrylate (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-phosphonium]amino]-2-methyl-propionic acid ethyl ester (5B) (8.0 g, 16.8 mmol), mp. After being added to a saturated aqueous solution of ammonium chloride, the mixture was evaporated. EtOAcjjjjjjjj 6-Amino-7H-indol-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphonium]amino]-2-methylpropionic acid thioethyl ester (Compound 6) , yellow oil (4.47 g, yield 54.4%).

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

Segregation 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 × 30mmI.D. mobile phase :A is CO2 and B is isopropanol, gradient: 30%, flow rate: 60mL/min, back pressure: 100bar, column temperature: 38°C, wavelength: 220nm, period: ~4min, sample preparation: compound 6 is dissolved in methanol Medium, injection: 2.1 ml per needle, 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-phosphonium]amino]-cyclopropyl formate 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-phosphonyl]amino]-cyclo Ethyl propyl formate (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 EtOAcjjjj:

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

Second step: 1-[[[(1R)-2-(6-aminofluoren-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphonyl]amino]-cyclo Thiopropyl propyl 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 was combined, EtOAcjjjjjjjjjjjj

Separation of compound 7

Preparation separation method: Instrument: waters SFC, Column: Chiralpak AS-3 (4.6 × 100mm), mobile phase: A and B is methanol 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 (400 MHz, 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 and the EC ₅₀ (half effective inhibitory concentration) was determined by qPCR using 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 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:

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 standard curve concentrations were 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.

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.

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 (11)

A compound of the formula (A), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: A is selected from a 6 to 10 membered aromatic ring, and the aromatic 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, Substituted by a substituent of a trifluoromethyl group, a C _1-4 alkoxy group or a -C(=O)OC _1-4 alkyl group; 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 H or C _1-6 alkyl; 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 or C _1-6 alkyl, said alkyl Further, it is optionally substituted with 0 to 5 substituents selected from H, F, Cl, Br, I, CN, amino, hydroxy, carboxy, C _1-4 alkyl or C _1-4 alkoxy. The compound, stereoisomer, or pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is selected from the group consisting of the compound 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} selected from H or C _1-6 alkyl; and R ³ is a C _1-6 alkyl group. The compound according to claim 2, a stereoisomer thereof, or a pharmaceutically acceptable salt 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 H or methyl; and R ³ is C _1-4 alkyl. The compound according to claim 3, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, 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. The compound according to claim 1, wherein the compound is selected from the group consisting of a compound represented by 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. The compound according to claim 5, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: A is selected from a substituted or unsubstituted phenyl or naphthyl group, and when substituted, optionally further Substituted by 1 to 3 substituents selected from H, F, Cl, Br, I, CN, methoxy, methyl, trifluoromethyl or ethoxycarbonyl; 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. The compound, stereoisomer or pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is: The compound, stereoisomer or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein the salt is a fumarate. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 8, a stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable Accepted carrier or excipient. A use of a compound according to any one of claims 1 to 8, a stereoisomer thereof or a pharmaceutically acceptable salt thereof for the preparation of a medicament for treating a viral infectious disease. 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.