TWI654198B - Aryl substituted phosphonium derivatives and their application in medicine - Google Patents

Abstract

This invention relates to an aryl substituted phosphonium amine derivative and its use in medicine. Specifically, the aryl-substituted phosphonamine derivative represented by the formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, a process for producing the same, and a pharmaceutical composition containing the same, and a preparation for treating a viral infection Use in medicines in sexual diseases.

Description

Aryl substituted phosphonium derivatives and their application in medicine

The present invention relates to an aryl-substituted phosphonamine derivative represented by the formula (I), a stereoisomer or a pharmaceutically acceptable salt thereof, a process for the preparation thereof, and a pharmaceutical composition containing the same, and a preparation treatment Use in medicines in viral infectious diseases.

B liver is one of the world's diseases, it is caused by hepatitis B virus. One-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. B liver virus can cause acute and chronic infections, acute infections are usually accompanied by liver inflammation, vomiting, jaundice, and very few can cause death, and chronic infection may induce cirrhosis and liver cancer. Although it is possible to prevent hepatitis B virus infection by vaccine, there is still no effective method for treating chronic hepatitis B disease.

Hepatitis B virus is a hepadnavirus 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 B hepatic virus encodes four known genes - C, X, P and S. Gene C encodes nuclear protein (HBcAg), gene S encodes surface antigen (HBsAg), and gene P encodes DNA polymerase. The function of protein encoded by gene X is unclear, but it is thought to be related to the occurrence of liver cancer because it starts A gene that induces cell proliferation and disables 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 chaperones. In the nucleus, hepatitis B virus converts part of the double-stranded DNA into intact double-stranded DNA by the host cell's DNA polymerase, and changes its 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 serve as templates, are transported into the cytoplasm, and are 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 nucleocapsid protein and a viral DNA polymerase. At the same time, this 3.5 kb long RNA will reverse transcribe the antisense strand of B hepatitis virus DNA, followed by 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 B-hepatic RNA and DNA is dependent on the B-hepatic DNA polymerase, which is essential for viral replication. The polymerase has four domains: a terminal protein, a spacer protein, a reverse transcriptase, and an RNaseH domain for degrading the pre-genomic RNA template that are important for the initiation of B-hepatic viral replication and assembly of the nucleocapsid. Despite this, the lack of proofreading results in a high mutation rate of the B-hepatic DNA polymerase.

The use of DNA polymerase inhibitors as anti-B hepatitis virus drugs has become an attractive option. A particular viral polymerase inhibitor belongs to the family of nucleoside analogs. The treatment of patients with chronic B liver has been improved by oral administration of anti-B hepatitis virus nucleoside analog drugs. In serum, nucleoside analogs can rapidly reduce B-hepatic 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, there are five nucleoside analogues B hepatic viral DNA polymerase inhibitors that are marketed in the United States and Europe for the treatment of chronic hepatitis B, including: lamivudine, adefovir dipivoxil, entecavir, telbivudine, tenofovir disoproxil fumarate, famciclovir and Clevudine, there are several other drugs in the research stage, such as tenofovir alafenamide (compound A), pradefovir, besifovir and so on. 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 creates a need for the development of new antiviral drugs.

Tenofovir, chemical name [(1R)-2-(6-aminofluoren-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 (Tenofovir disoproxil 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 prodrug of tenofovir has become a research hotspot.

European Patent EP206459 describes a 9-(phosphomethoxyalkyl) adenine derivative comprising a structure of tenofovir and its use in antiviral drugs, the structural formula of which is as follows:

Wherein R ^{1 is} selected from the group consisting of hydrogen, methyl, and hydroxymethyl, and R ^{2 is} selected from the group consisting of substituted or unsubstituted ethylene, methylene, propylene, and the like. The specific description of this patent should not be construed as limiting the invention.

EP 481214 describes a novel oral phosphate nucleoside analog-like prodrug comprising adefovir, and its antiviral medical use, particularly anti-RNA, DNA virus, and can also be used to treat tumors, etc. Its structure is as follows:

Wherein B is selected from the group consisting of purine, cytosine, uracil, thymine, and avidin, and R ^{3 is} selected from a substituted or unsubstituted C ₁ -C ₂₀ alkyl group, and R ¹ and 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 C Water H or CH ₂ OR ⁵ , R ^{5 is} selected from C ₄ -C ₂₀ alkyl, aryl or aryl-alkane unsubstituted or substituted by hydroxy, oxo, nitro, halogen Further, R ¹ and R ² may form a ring. The specific description of this patent should not be construed as limiting the invention.

WO0208241 describes an adenine derivative comprising tenofovir and tenofovir structure, which has the following structural formula:

Wherein R ^{1 is} selected from hydrogen and methyl. The specific description of this patent should not be construed as limiting the invention.

WO02057288 describes acyclic nucleoside phosphate compounds and their use in antiviral drugs, the structural formula of which is as follows:

Wherein Q is selected from the group consisting of purine or pyrimidine, R ⁴ and R ^{5 are} independently selected from hydrogen, alkyl, aryl, etc., and R ¹ , R ² , R ³ , R ⁷ and R ^{8 are} independently selected from the group consisting of hydroxyl, halogen, hydrogen, Amino group, alkyl group, alkoxy group, alkylamino group and the like. The specific description of this patent should not be construed as limiting the invention.

CN200410024276.X describes a 9-((phosphate) methoxyalkyl) adenine derivative and its use for antiviral drugs, the structural formula of which is as follows:

Wherein R ¹ and R ^{2 are} independently selected from hydrogen or substituted biphenylmethyl. The specific description of this patent should not be construed as limiting the invention.

CN200710041280.0 describes acyclic nucleoside phosphate compounds and their use for antiviral drugs, the structural formula of which is as follows;

Wherein R ^{1 is} selected from the group consisting of hydrogen, halogen, amino, cyclopropylamino, methoxy, ethoxy, etc., R ^{2 is} selected from hydrogen or amino, R ^{5 is} selected from methyl or hydrogen, and R ³ and R ^{4 are} independently selected. From (substituted aminocarbonyloxy)alkyl. The specific description of this patent should not be construed as limiting the invention.

CN200410088840.4 describes acyclic nucleoside phosphate compounds and their use for antiviral drugs, the structural formula is as follows

Wherein R is hydrogen or methyl, R ^{2 is} selected from hydrogen or camphorinyl, and R ^{1 is} selected from cycloalkyl having 3-8 carbons, unsaturated chain hydrocarbons of 3-8 carbons, and 3-8 carbons. An unsaturated cycloalkyl group or an aromatic hydrocarbon of 6-10 carbons. The specific description of this patent should not be construed as limiting the invention.

WO2011069322 describes acyclic nucleoside phosphate derivatives and their use for the treatment and prevention of diseases associated with viral infections, the structural formula of which is as follows:

Wherein R ^{1 is} selected from hydrogen or methyl, R ^{2 is} selected from -R ³ or -OR ³ , and R ^{3 is} selected from C _1-8 alkyl, C _3-8 cycloalkyl. The specific description of this patent should not be construed as limiting the invention.

The present invention is to design a compound represented by the general formula (I) on the basis of tenofovir disoproxil to provide a novel acyclic nucleoside phenyl phosphate-amine derivative, a stereoisomer thereof or a pharmaceutically acceptable substance. The accepted salt can be used to treat viral infectious diseases, including infectious diseases caused by hepatitis B virus and HIV virus.

The present invention relates to a compound of the formula (I) (an aryl-substituted phosphonamine derivative), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein:

Ar is selected from phenyl or naphthyl, and the phenyl and naphthyl optionally further selected from 0 to 5 are selected from the group consisting of H, F, Cl, Br, I, hydroxy, decyl, amino, cyano, carboxy, C ₁ Substituted by a substituent of _-4 alkyl or C _1-4 alkoxy; R ^{1 is} selected from H or methyl; R ^{2 is} selected from -(CH ₂ -CH ₂ -O) _n -R ^2a ; R ^{2a is} selected from H, C ₁ - ₆ alkyl, C _3-6 carbocyclic or 3 to 6 membered heterocyclic, said heterocyclic group containing at least 1 to 4 heteroatoms selected from N, O or S, said Further, the alkyl group, the carbocyclic group and the heterocyclic group are further selected from 0 to 5 selected from the group consisting of H, F, Cl, Br, I, hydroxy, decyl, amino, cyano, carboxy, C _1-4 alkyl or C ₃ Substituted by a substituent of _-6 cycloalkyl; n is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

Preferably, the present invention provides a compound of the formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein: Ar is selected from phenyl or naphthyl, preferably phenyl; And the naphthyl group is optionally further substituted with from 0 to 5 substituents selected from the group consisting of H, F, Cl, hydroxy, amino, carboxy, methyl, ethyl, methoxy; R ^{1 is} selected from H or methyl; R ^{2 is} selected from -(CH ₂ -CH ₂ -O) _n -R ^2a ; R ^{2a is} selected from H, C _{1 -6} alkyl, C ₃ carbocyclyl, C ₄ carbocyclyl, C ₅ carbocyclyl, a C ₆ carbocyclic group, a 3-membered heterocyclic group, a 4-membered heterocyclic group, a 5-membered heterocyclic group or a 6-membered heterocyclic group, preferably H, C ₁ - ₆ alkyl, C ₅ carbocyclic, C ₆ carbocyclic group, a 5 or 6-membered heterocyclic group a heterocyclic group, more preferably H or C ₁ - ₆ alkyl; said heterocyclic group containing at least 1 to 4 heteroatoms selected from N, O or S, said The alkyl group, the carbocyclic group and the heterocyclic group are optionally further substituted with 0 to 5 substituents selected from H, F, Cl, hydroxy, amino, methyl or ethyl; n is selected from 2, 3, 4 or 5.

Preferably, the present invention provides a compound of the formula (I), a stereoisomer or a pharmaceutically acceptable salt thereof, wherein: Ar is selected from phenyl or naphthyl, preferably phenyl; R ^{1 is} selected from H or methyl, preferably methyl; R ^{2 is} selected from -(CH ₂ -CH ₂ -O) _n -R ^2a ; R ^{2a is} selected from H or C _{1 -6} alkyl, said alkyl optionally further Substituted to 5 substituents selected from H, F, Cl, hydroxy, amino, methyl or ethyl; n is selected from 2, 3, 4 or 5.

Preferably, the present invention provides a compound of the formula (I), a stereoisomer or a pharmaceutically acceptable salt thereof, wherein: Ar is selected from phenyl or naphthyl, preferably phenyl; R ^{1 is} selected from H or methyl, preferably methyl; R ^{2 is} selected from -(CH ₂ -CH ₂ -O) _n -R ^2a ; R ^{2a is} selected from H or C _1-4 alkyl, preferably H, methyl, ethyl or Isopropyl, further preferably H or methyl; n is selected from 2, 3, 4 or 5, preferably 2 or 3.

Preferably, the present invention provides a compound of the formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein: Ar is a phenyl group; and R ^{1 is} selected from H or a methyl group, preferably a methyl group. ; R ^{2 is} selected from -(CH ₂ -CH ₂ -O) ₂ -H, -(CH ₂ -CH ₂ -O) ₂ -CH ₃ , -(CH ₂ -CH ₂ -O) ₃ -H or -( CH ₂ -CH ₂ -O) ₃ -CH ₃ .

Preferably, the present invention provides a compound of the formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein the compound is a compound of the formula (II): Wherein R ^{2 is} selected from -(CH ₂ -CH ₂ -O) ₂ -H, -(CH ₂ -CH ₂ -O) ₂ -CH ₃ , -(CH ₂ -CH ₂ -O) ₃ -H or - (CH ₂ -CH ₂ -O) ₃ -CH ₃ .

Preferably, the present invention provides a compound of the formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein the compound is selected from one of the following structures:

The present invention relates to a compound of the formula (I) or a pharmaceutically acceptable salt thereof, wherein the salt is selected from the group consisting of a hydrochloride, a sulfate, a phosphate, an acetate, a maleate, a succinate, Fumarate, malate, oxalate, tartrate, benzoate, cinnamate, p-toluenesulfonate, besylate, methanesulfonate, triflate or their Combinations, preferably maleate, fumarate or triflate.

The invention relates to a pharmaceutical composition comprising a therapeutically effective amount of at least one compound of the formula (I) according to the invention, or a stereoisomer or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable Carrier or excipient.

Further, the present invention relates to a compound of the formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and the use of the pharmaceutical composition for the preparation of a medicament for the treatment of a viral infectious disease.

Preferably, the viral infectious diseases of the present invention include infectious diseases caused by hepatitis B virus, hepatitis C virus or HIV.

Preferably, the viral infectious disease of the present invention is selected from the infectivity caused by hepatitis B virus. The disease is preferably chronic B liver disease.

Unless otherwise stated, the terms used in the present invention have the following meanings.

When substituted by a plurality of substituents, each substituent may be the same or different.

When a plurality of hetero atoms are contained, each hetero atom 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 1 to 5 of their corresponding isotopes, including ¹² C, ¹³ C and ¹⁴ C. The isotopes of hydrogen include helium (H) and 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, a fluorenyl group, and various 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, hydroxy, amino, cyano, carbonyl, carboxy, Non-limiting examples of C _1-10 alkyl, C _1-10 alkoxy, C _1-4 fluorenylamine, C _3-10 carbocyclyl or C _3-10 heterocyclyl, substituents, including H, F, Cl, Br, I, hydroxy, decyl, amino, cyano, carbonyl, carboxyl, C _1-10 alkyl, C _1-10 alkoxy, C _1-4 alkoxy-C _{1- 4} alkoxy, C _2-8 alkenyl, C _1-4 fluorenylamino, -C(=O)-OC _1-10 alkyl, -OC(=O)-C _1-10 alkyl, C _{3 -10} carbocyclyl or C _3-10 heterocyclyl, non-limiting examples of substituents including H, F, Cl, Br, I, hydroxy, thiol, amino, cyano, carbonyl, carboxy, methyl, B Base, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, methoxy, ethoxy, propoxy, isopropoxy, butoxy, first-butoxy, two Butoxy, tert-butoxy, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cycloheptyl, methoxyethoxy, methoxyethoxyethoxy, vinyl, A Amino, acetyl amino, acyl amino propyl, isopropyl acyl group, a butoxy group acyl, phenyl, substituted phenyl, naphthyl, substituted naphthyl.

"Alkoxy" means an -O-alkyl group wherein alkyl is as defined herein above. The alkoxy group may be substituted or unsubstituted, and non-limiting examples thereof include methoxy, ethoxy, propoxy, isopropoxy, butoxy, tert-butoxy, pentyloxy Or hexyloxy, preferably having from 1 to 12 members of alkoxy groups. When substituted, the substituent is preferably from 1 to 5, independently selected from the group consisting of H, F, Cl, Br, I, hydroxy, amino, cyano, carbonyl, carboxy, C _1-10 alkyl, C _1- a non-limiting example of a ₁₀ alkoxy group, a C _1-4 fluorenylamino group, a C _3-10 carbocyclic group or a C _3-10 heterocyclic group, including H, F, Cl, Br, I, hydroxy , mercapto, amino, cyano, carbonyl, carboxyl, C _1-10 alkyl, C _1-10 alkoxy, C _1-4 alkoxy-C _1-4 alkoxy, C _2-8 alkenyl , C _1-4醯 醯, -C(=O)-OC _1-10 alkyl, -OC(=O)-C _1-10 alkyl, C _3-10 carbocyclyl or C _3-10 Cycloalkyl, non-limiting examples of substituents include H, F, Cl, Br, I, hydroxy, thiol, amino, cyano, carbonyl, carboxy, methyl, ethyl, n-propyl, isopropyl, positive Butyl, isobutyl, tert-butyl, methoxy, ethoxy, propoxy, isopropoxy, butoxy, first-butoxy, di-butoxy, tert-butoxy, Cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cycloheptyl, methoxyethoxy, methoxyethoxyethoxy, vinyl, formamidine, ethyl hydrazine, propyl hydrazine Amine, Propan-acyl group, a butoxy group acyl, phenyl, substituted phenyl, naphthyl, substituted naphthyl.

"Alkenyl" means an alkyl group as defined in the invention comprising at least one carbon-carbon double bond, said alkenyl group having 2 to 20 carbon atoms, preferably 2 to 12 carbon atoms, further preferably 2 to 8 carbon atoms atom. Non-limiting examples of alkenyl groups include substituted or unsubstituted vinyl, 2-propenyl, 3-butenyl, 2-butenyl, 4-pentenyl, 3-pentenyl, 2-hexenyl 3-hexenyl, 2-heptene a group, a 3-heptenyl group, a 4-heptenyl group, a 3-enheptyl group, a 3-decenyl group or a 4-decenyl group, etc., when substituted, preferably has 1 to 5 substituents, independently selected From F, Cl, Br, I, =O, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, fluorenyl, hydroxy, nitro, cyano, amino, alkyl decylamino A cycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylindenyl, hydroxyalkyl, carboxylic acid, carboxylic acid ester or heterocycloalkyl fluorenyl group.

"Alkynyl" means an alkyl group as defined in the invention comprising at least one carbon-carbon triple bond, said alkynyl group having 2 to 20 carbon atoms, preferably 2 to 12 carbon atoms, further preferably 2 to 8 carbons atom. Non-limiting examples of alkynyl groups include substituted or unsubstituted ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 4-pentyne , 3-pentynyl, 2-hexynyl, 3-hexynyl, 3-butynyl, 2-heptynyl, 3-heptynyl, 4-heptynyl, 3-octynyl, 3-decynyl or 4-nonynyl, etc., when substituted, preferably 1 to 5, independently selected from F, Cl, Br, I, =O, alkyl, alkenyl, alkynyl , alkoxy, alkylthio, alkylamino, fluorenyl, hydroxy, nitro, cyano, amino, alkyl decylamino, cycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkane An oxy group, a cycloalkyl fluorenyl group, a hydroxyalkyl group, a carboxylic acid, a carboxylic acid ester or a heterocycloalkyl fluorenyl group.

"Cycloalkyl" means a substituted or unsubstituted 3 to 8 membered all-carbon monocyclic group which may be attached to a bridged or spiro ring, wherein 1 to 5 may contain 1 to 5 double bonds, but none of the rings have A fully conjugated π-electron system. Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo[3.2.1]octyl, bicyclo[5.2.0]decyl, tricyclic [5.3.1.1] Dodecyl, adamantyl, spiro[3.3]heptanyl, spiro[2.4]heptanyl, spiro[2.5]octyl or spiro[2.3]hexane and the like. When substituted, the substituent is preferably from 1 to 5, independently selected from the group consisting of H, F, Cl, Br, I, hydroxy, amino, cyano, carbonyl, carboxy, C _1-10 alkyl, C _1- a non-limiting example of a ₁₀ alkoxy group, a C _1-4 fluorenylamino group, a C _3-10 carbocyclic group or a C _3-10 heterocyclic group, including H, F, Cl, Br, I, hydroxy , mercapto, amino, cyano, carbonyl, carboxyl, C _1-10 alkyl, C _1-10 alkoxy, C _1-4 alkoxy-C _1-4 alkoxy, C _2-8 alkenyl , C _1-4醯 醯, -C(=O)-OC _1-10 alkyl, -OC(=O)-C _1-10 alkyl, C _3-10 carbocyclyl or C _3-10 Cycloalkyl, non-limiting examples of substituents include H, F, Cl, Br, I, hydroxy, thiol, amino, cyano, carbonyl, carboxy, methyl, ethyl, n-propyl, isopropyl, positive Butyl, isobutyl, tert-butyl, methoxy, ethoxy, propoxy, isopropoxy, butoxy, first-butoxy, di-butoxy, tert-butoxy, Cyclopropane, cyclobutyl, cyclohexane, cyclopentyl, cycloheptyl, methoxyethoxy, methoxyethoxyethoxy, vinyl, formamide, B Amidoxime, Acyl group, an isopropyl group acyl, acyl-butoxy group, a phenyl group, a substituted phenyl group, a naphthyl group, a substituted naphthyl group.

"Carbocyclyl" means a substituted or unsubstituted saturated or unsaturated aromatic ring group or a non-aromatic ring group, and the aromatic ring group or non-aromatic group may be a single ring of 3 to 8 members, a 4 to 12 membered double ring or A 10 to 15 membered tricyclic system, the carbocyclic ring may be attached to a bridged ring or a spiro ring, and non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclopentane. Alkenyl, cyclohexadienyl, cycloheptatrienyl, phenyl, naphthyl, benzocyclopentyl, bicyclo[3.2.1]octyl, bicyclo[5.2.0]decyl, three Ring [5.3.1.1] dodecyl, adamantyl or spiro[3.3]heptyl and the like. When substituted, the substituent is preferably from 1 to 5. When substituted, the substituent is preferably from 1 to 5, independently selected from the group consisting of H, F, Cl, Br, I, hydroxy, amino, cyano, carbonyl, carboxy, C _1-10 alkyl, C _1- a non-limiting example of a ₁₀ alkoxy group, a C _1-4 fluorenylamino group, a C _3-10 carbocyclic ring or a C _3-10 heterocyclic ring, including H, F, Cl, Br, I, hydroxy, fluorenyl , amino, cyano, carbonyl, carboxyl, C _1-10 alkyl, C _1-10 alkoxy, C _1-4 alkoxy-C _1-4 alkoxy, C _2-8 alkenyl, C _1-4醯 醯, -C(=O)-OC _1-10 alkyl, -OC(=O)-C _1-10 alkyl, C _3-10 carbocyclyl or C _3-10 heterocyclyl Non-limiting examples of substituents include H, F, Cl, Br, I, hydroxy, thiol, amino, cyano, carbonyl, carboxy, methyl, ethyl, n-propyl, isopropyl, n-butyl , isobutyl, tert-butyl, methoxy, ethoxy, propoxy, isopropoxy, butoxy, first-butoxy, di-butoxy, tert-butoxy, cyclopropane Base, cyclobutyl, cyclohexyl, cyclopentyl, cycloheptyl, methoxyethoxy, methoxyethoxyethoxy, vinyl, formamidine, ethylamino, propylamine Isopropyl Group, a butoxy group acyl, phenyl, substituted phenyl, naphthyl, substituted naphthyl.

"Heterocyclyl" means a substituted or unsubstituted saturated or unsaturated aromatic ring group, a non-aromatic ring group, and the aromatic ring, non-aromatic ring may be a single ring of 3 to 8 members, a 4 to 12 membered double ring or 10 Up to a 15-membered ring system, and consisting of at least one hetero atom selected from N, O or S, preferably a 3 to 10 membered heterocyclic ring, and optionally substituted N, S in the heterocyclic ring can be oxidized to various oxidation states . The heterocyclic ring may be attached to a hetero atom or a carbon atom, and the heterocyclic ring may be attached to a bridged ring or a spiro ring. Non-limiting examples include epoxyethyl, azacyclopropenyl, oxetanyl, azetidinyl, azepanyl, oxirane, oxetanyl, Azetidinyl, thietyl, oxacyclohexyl, thiacyclohexyl, 1,3-dioxolanyl, 1,4-dioxolanyl, 1,2-diaza Cyclopentyl, 1,2-oxazacyclopentyl, 1,2-oxathiolanyl, 1,3-oxazacyclopentyl, 1.3-oxathiolane, 1,2- Diazacyclohexyl, 1,2-oxazacyclohexyl, 1,2-oxothiohexyl, 1,3-oxazacyclohexyl, 1,3-thiazepine, 1.3-diaza Heterocyclohexyl, 1,3-dioxohexyl, 1,4-dioxohexyl, 1.3-oxathiacyclohexyl, 1.4-diazacyclohexyl, 1.4-oxathiamethylene, 1.4- Dithiacyclohexyl, spiro[3.3]heptanyl, spiro[2.3]hexane, azaspiro[3.3]heptanyl,oxaspiro[3.3]heptanyl, azaspiro[2.3]hexane , oxaspiro[2.3]hexane, spiro[2.4]heptanyl, azaspiro[2.4]heptanyl,oxaspiro[2.4]heptanyl,spiro[2.5]octyl, aza Spiro[2.5]octyl, oxaspiro[2.5]octyl, azabicyclo [3.2.1] Octyl, azabicyclo[5.2.0]decyl, oxatricyclo[5.3.1.1]dodecyl, azamantyl, pyridyl, furyl, thienyl , pyridyl, pyranyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, acridinyl, fluorenyl, morpholinyl, thiomorpholinyl, 1,3 -dithiane dihydrofuranyl, dihydropyranyl, dithiapentanyl, tetrahydrofuranyl, tetrahydropyrrolyl, tetrahydroimidazolyl, tetrahydrothiazolyl, hexahydropyranyl, benzimidazolyl, a benzopyridinyl group, a pyrrolopyridyl group, a benzodihydrofuranyl group or the like; when substituted, the substituent is preferably from 1 to 5, independently selected from the group consisting of H, F, Cl, Br, I, a hydroxyl group, an amino group, Cyano, carbonyl, carboxyl, C _1-10 alkyl, C _1-10 alkoxy, C _1-4 alkoxy-C _1-4 alkoxy, C _1-4 decylamino, C _{3- 10} carbocyclyl or C _3-10 heterocyclyl, non-limiting examples of substituents including H, F, Cl, Br, I, hydroxy, thiol, amino, cyano, carbonyl, carboxy, C _1-10 Alkyl, C _1-10 alkoxy, C _2-8 alkenyl, C _1-4 fluorenylamino, -C(=O)-OC _1-10 alkyl, -OC(=O)-C _{1 -10} alkyl, C _3-10 carbocyclyl or C _3-10 heterocyclyl, non-limiting examples of substituents including H, F, Cl, Br, I, hydroxy, thiol, amino, cyano, carbonyl ,carboxy, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, methoxy, ethoxy, propoxy, isopropoxy, butoxy , first-order butoxy, second-butoxy, tert-butoxy, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cycloheptyl, methoxyethoxy, methoxyethoxy Ethoxy, vinyl, formamidine, ethenylamino, propylamino, isopropylamino, butylamino, phenyl, substituted phenyl, naphthyl, substituted naphthyl.

"Aryl" means a substituted or unsubstituted 6 to 15 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 and 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 benzene And furan, benzocyclopentyl, benzothiazole and the like. When substituted, the substituent is preferably from 1 to 5, independently selected from the group consisting of H, F, Cl, Br, I, hydroxy, amino, cyano, carbonyl, carboxy, C _1-10 alkyl, C _{1- 10} alkoxy, C _1-4 alkoxy-C _1-4 alkoxy, C _1-4 fluorenylamino, C _3-10 carbocyclyl or C _3-10 heterocyclyl, unrestricted substituent Examples include H, F, Cl, Br, I, hydroxy, decyl, amino, cyano, carbonyl, carboxyl, C _1-10 alkyl, C _1-10 alkoxy, C _2-8 alkenyl , C _1-4醯 醯, -C(=O)-OC _1-10 alkyl, -OC(=O)-C _1-10 alkyl, C _3-10 carbocyclyl or C _3-10 Cycloalkyl, non-limiting examples of substituents include H, F, Cl, Br, I, hydroxy, thiol, amino, cyano, carbonyl, carboxy, methyl, ethyl, n-propyl, isopropyl, positive Butyl, isobutyl, tert-butyl, methoxy, ethoxy, propoxy, isopropoxy, butoxy, first-butoxy, di-butoxy, tert-butoxy, Cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cycloheptyl, methoxyethoxy, methoxyethoxyethoxy, vinyl, formamidine, ethyl hydrazine, propyl hydrazine Amine, Propan-acyl group, a butoxy group acyl, phenyl, substituted phenyl, naphthyl, substituted naphthyl.

"Heteroaryl" means a substituted or unsubstituted 5 to 15 membered aromatic ring group, and contains 1 to 3 heteroatoms selected from N, O or S, preferably 5 to 10 membered aromatic rings, and heteroaryl groups. Limitative examples include pyridyl, furyl, thienyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, benzofuran, benzimidazole, benzopyridine or pyrrolopyridine Wait. When substituted, the substituent is preferably from 1 to 5, independently selected from the group consisting of H, F, Cl, Br, I, hydroxy, amino, cyano, carbonyl, carboxy, C _1-10 alkyl, C _1- a non-limiting example of a ₁₀ alkoxy group, a C _1-4 fluorenylamino group, a C _3-10 carbocyclic group or a C _3-10 heterocyclic group, including H, F, Cl, Br, I, hydroxy , mercapto, amino, cyano, carbonyl, carboxyl, C _1-10 alkyl, C _1-10 alkoxy, C _1-4 alkoxy-C _1-4 alkoxy, C _2-8 alkenyl , C _1-4醯 醯, -C(=O)-OC _1-10 alkyl, -OC(=O)-C _1-10 alkyl, C _3-10 carbocyclyl or C _3-10 Cycloalkyl, non-limiting examples of substituents include H, F, Cl, Br, I, hydroxy, thiol, amino, cyano, carbonyl, carboxy, methyl, ethyl, n-propyl, isopropyl, positive Butyl, isobutyl, tert-butyl, methoxy, ethoxy, propoxy, isopropoxy, butoxy, first-butoxy, di-butoxy, tert-butoxy, Cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cycloheptyl, methoxyethoxy, methoxyethoxyethoxy, vinyl, formamidine, ethyl hydrazine, propyl hydrazine Amine, Propan-acyl group, a butoxy group acyl, phenyl, substituted phenyl, naphthyl, substituted naphthyl.

"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, alkane. Thio group, hydroxyl group, amino group, alkylamino group, alkyl nonylamino group, heterocycloalkyl group, cycloalkyl group, aryl group, heteroaryl group, cycloalkoxy group, heterocycloalkoxy group, hydroxyalkyl group, carboxylic acid Or a carboxylic acid ester, a non-limiting example of a substituent, including F, Cl, Br, I, hydroxy, amino, cyano, carboxy, methyl, ethyl, n-propyl, isopropyl, n-butyl, Isobutyl, tert-butyl, methoxy, ethoxy, propoxy, isopropoxy, butoxy, first-butoxy, di-butoxy, tert-butoxy, cyclopropyl , cyclobutyl, cyclohexyl, cyclopentyl, cycloheptyl, formamide, acetamino, propylamine, isopropylamine, butylamino, phenyl, substituted phenyl, naphthyl , substituted naphthyl.

The "=O" of the present invention is generally used in the art and refers to an oxygen atom bonded by a double bond, such as a double bond oxygen atom to a carbon atom in a carbonyl group.

"Stereoisomers" refers to isomers produced by the different arrangement of atoms in a molecule, including cis-trans isomers, para-isomers, and structural isomers.

"Pharmaceutical composition" means a mixture of one or more of the above compounds or a physiologically/pharmaceutically acceptable salt or prodrug thereof with other chemical ingredients, such as physiological/pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate the administration of a compound to an organism.

"Optional" or "optionally" means that the event or environment described subsequently may, but need not, occur, including where the event or environment occurs or does not occur. For example, "the aryl group is optionally substituted by an alkyl group" means that an alkyl group may be, but is not necessarily, 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 case where 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.

"Substitution" means a 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, -C water CH ₃ , -CH ₂ SH, -CH ₂ CH ₂ CN, -CH ₂ NH ₂ , -NHOH, -NHCH ₃ , -OCH ₂ Cl, -OC water CH ₂ CH ₃ , -OCH ₂ CH ₂ NH ₂ , -OCH ₂ CH ₂ SH, -OCH ₂ C water H, 1-hydroxycyclopropyl, 2-hydroxycyclopropyl, 2-aminocyclopropyl, 4-methyl Furanyl, 2-hydroxyphenyl, 4-aminophenyl, phenyl.

Method for synthesizing the compound of the present invention

[[2-(6-Aminofluoren-9-yl)-1-methylethoxy]methyl]phosphoric acid is reacted with the corresponding substrate under basic conditions to give compound (IB), compound (IB) and ester The alanine reaction gives the compound (I); if the esterified alanine contains a group such as an amino group or a hydroxyl group, it can be protected first and then reacted with the compound (IB), and then the protecting group is removed to obtain the compound (I); Wherein the protecting group includes, but is not limited to, tertiary butoxycarbonyl, benzyloxycarbonyl, fluorenylmethoxycarbonyl, allyloxycarbonyl, trichloroethoxycarbonyl, trimethyldecylethoxycarbonyl, methoxy Carbonyl, ethoxycarbonyl, 2-biphenyl-2-propoxycarbonyl, tert-butoxy, phthalic acid, p-toluenesulfonyl, o-nitrophenylsulfonyl, p-nitrobenzenesulfonate Mercapto, tert-amyl, carbenyl, trifluoroethenyl, benzhydryl, benzyl, trityl, p-methoxybenzyl or 2,4-dimethoxybenzyl, preferably H or a tertiary butoxycarbonyl group; wherein Ar, R ^{2 are} as defined in the definition of the compound of formula (I).

Hereinafter, the implementation process of the present invention and the beneficial effects produced will be described in detail by way of specific embodiments. To help the reader better understand the essence and characteristics of the present invention, it is not intended to limit the scope of the present invention.

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 III400 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.

Pipe column chromatography generally uses Yantai Huanghai Tanji 200~300 mesh silicone as 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.

If not specified in the examples, the solution means an aqueous solution.

Unless otherwise stated in the examples, the temperature of the reaction is room temperature.

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

Intermediate 1: [(1R)-2-(6-Aminofluoren-9-yl)-1-methyl-ethoxy]methyl-phenoxy-monophosphate

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

[[(1R)-2-(6-Aminofluoren-9-yl)-1-methylethoxy]methyl]phosphonic acid (PMPA) (10 g, 0.035 mol) was dissolved in acetonitrile (80 mL) and added 4-Dimethylaminopyridine (4.26 g, 0.035 mol) and triphenyl phosphite (16.21 g, 0.052 mol) were heated to reflux for 48 hours. The reaction solution was cooled to room temperature, and concentrated under reduced pressure. ethyl acetate (20 mL) and water (30 mL) was then evaporated. The mixture was partitioned. The aqueous phase was extracted with concentrated ethyl acetate (20 mL×2). 3, the aqueous phase was filtered, and the filter cake was washed with methanol and dried under reduced pressure to give the title compound [(1R)-2-(6-aminopurin-9-yl)-1-methyl-ethoxy]methyl-phenoxy Base-monophosphoric acid ( Intermediate 1 ), pale yellow solid (7.5 g, yield 60%).

^{1 H NMR (400MHz, DMSO)} δ 8.16 (s, 1H), 8.14 (s, 1H), 7.55 (s, 2H), 7.31-7.27 (m, 2H), 7.12-7.05 (m, 3H), 4.30 ( Dd, 1H), 4.19 (dd, 1H), 3.97 (m, 1H), 3.88-3.72 (m, 2H), 1.04 (d, 3H).

Example 1 : (2S)-2-[[[(1R)-2-(6-Aminofluoren-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphonium] Amino]propionic acid-2-(2-methoxyethoxy)ethyl ester ( Compound 1 )

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

First step: (2S)-2-(tertiary butoxyamino)propionic acid-2-(2-methoxyethoxy)ethyl ester (1B)

2-(2-methoxyethoxy)ethyl(2S)-2-(tert-butoxycarbonylamino)propanoate

Add (2S)-2-(tertiary butoxyamino)propionic acid (6.0 g, 0.05 mol), 2-(2-methoxyethoxy)ethanol (9.5 g, 0.05 mol) to toluene dichloride (150 mL), then dicyclohexylcarbodiimide (10.3 g, 0.05 mol) and 4-dimethylaminopyridine (6.1 g, 0.05 mol) were added, and the reaction was stirred at room temperature overnight under a nitrogen atmosphere. The temperature was raised overnight. The reaction mixture was filtered, and the filter cake was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, filtered, and evaporated. 1) The title product (2S)-2-(tert-butoxyamino)propanoic acid-2-(2-methoxyethoxy)ethyl ester ( 1B ), mp. 100%).

The second step: (2S)-2-aminopropionic acid-2-(2-methoxyethoxy)ethyl ester (1C)

2-(2-methoxyethoxy)ethyl(2S)-2-aminopropanoate

(2S)-2-(tertiary butoxyamino)propanoic acid-2-(2-methoxyethoxy)ethyl ester ( 1B ) was dissolved in dichloromethane (100 mL) and trifluoroacetic acid was added ( 58.7 g, 0.5 mol), reacted at room temperature for 4 hours. The reaction mixture was concentrated under reduced pressure. dichloromethane (dichloromethanol) (EtOAc) (2-Methoxyethoxy)ethyl ester ( 1C ), yellow oil (5.0 g, yield 52.3%).

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 4.29 (t, 2H), 3.72 (t, 2H), 3.65-3.61 (m, 3H), 3.56-3.54 (m, 2H), 3.38 (s, 3H), 2.72 (s, 2H), 1.37 (d, 3H).

The third step: (2S)-2-[[[(1R)-2-(6-aminofluoren-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphonium] Amino]propionic acid-2-(2-methoxyethoxy)ethyl ester (Compound 1)

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

Dissolving [(1R)-2-(6-aminofluoren-9-yl)-1-methyl-ethoxy]methyl-phenoxy-monophosphate ( Intermediate 1 ) (1.29 g, 3.55 mmol) Dichlorohydrazine (1.7 g, 14.2 mmol) was added to acetonitrile (30 mL), and the mixture was heated to 60 ° C for 5 hours. Compound (2S)-2-Aminopropionic acid-2-(2-methoxyethoxy)ethyl ester ( 1C ) (0.8 g, 4.26 mmol) and triethylamine (1.4 g, 14.2 mmol) were dissolved in dichloro In methane (20 mL), it was cooled to 0 ° C, and the intermediate phosphine was added dropwise to the reaction mixture, and the mixture was allowed to react to room temperature for 1 hour. Dichloromethane (50 mL) and water (50 mL) were added to the mixture, and the mixture was evaporated. Methyl chloride / methanol (v / v) = 1: 0 ~ 95: 5) to give the title product (2S)-2-[[[(1R)-2-(6-aminopurin-9-yl)-1-yl 2-Ethyloxy]methyl-phenoxy-phosphonyl]amino]propionic acid-2-(2-methoxyethoxy)ethyl ester ( Compound 1 ).

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 8.33 (d, 1H), 8.03 (d, 1H), 7.30 (d, 1H), 7.14 (m, 3H), 6.95 (d, 1H), 5.98 (d, 2H ), 4.47-4.29 (m, 1H), 4.27 (m, 1H), 4.25-4.03 (m, 3H), 4.03-3.85 (m, 2H), 3.74-3.57 (m, 6H), 3.56-3.44 (m , 2H), 3.38-3.28 (d, 3H), 1.33-1.21 (m, 6H).

MS M/Z (ESI): 537.2 (M + 1).

³¹ P NMR (162 MHz, CDCl ₃ ) δ 23.81, 22.23.

Example 2: Resolution/Resolution of Compound 1

(2S)-2-[[[(1R)-2-(6-Aminofluoren-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphonyl]amino]propyl Acid-2-(2-methoxyethoxy)ethyl ester ( Compound 1 ) (200 mg) was used for resolution.

Preparation conditions: instrument: Gilson GX-281; column: CHIRALPAK AD-H, 20 × 250 mm, 5 μm; mobile phase: A: n-hexane, B: isopropanol; equal ratio: A: B = 50: 50 (v/v); flow rate: 6 mL/min; wavelength: 260 nm; period: 50 min; sample preparation: Compound 1 was dissolved in n-hexane: isopropanol (15 mL, v/v = 1:1) to obtain 13.33 mg. /ml; Injection: 20 μL / needle.

After separation, the fraction was concentrated and dried by a rotary evaporator at 40 ° C to give two optical isomer compound 1-1 (50 mg), Compound 1-2 (45 mg).

Compound 1-1

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 8.32 (s, 1H), 7.97 (s, 1H), 7.34-7.28 (m, 2H), 7.17-7.14 (m, 3H), 5.80 (s, 2H), 4.43 (dd, 1H), 4.26-4.14 (m, 3H), 4.13-4.05 (m, 1H), 4.05-4.00 (m, 1H), 4.00-3.94 (m, 1H), 3.69-3.61 (m, 5H) , 3.57-3.52 (m, 2H), 3.44 (t, 1H), 3.37 (s, 3H), 1.25 (d, 3H), 1.21 (s, 3H).

MS M/Z (ESI): 537.3 (M + 1).

³¹ P NMR (162 MHz, CDCl ₃ ) δ 23.82.

Compound 1-2

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 8.34 (s, 1H), 8.03 (s, 1H), 7.20 (t, 2H), 7.09 (t, 1H), 6.95 (d, 2H), 5.93 (s, 2H ), 4.35-4.24 (m, 3H), 4.14 (dd, 1H), 4.05 (m, 1H), 3.91 (m, 2H), 3.73-3.67 (m, 3H), 3.65-3.59 (m, 3H), 3.52-3.44 (m, 2H), 3.30 (s, 3H), 1.31 (d, 3H), 1.20 (d, 3H)

MS M/Z (ESI): 537.3 (M + 1).

³¹ P NMR (162 MHz, CDCl ₃ ) δ 18.87.

Example 3: (2S)-2-[[[(1R)-2-(6-Aminofluoren-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphonium] Amino]propionic acid-2-(2-hydroxyethoxy)ethyl ester (Compound 2)

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

First step: 2-(2-benzyloxyethoxy)ethanol (2B)

2-(2-benzyloxyethoxy)ethanol

2-(2-Methoxyethoxy)ethanol (2A) (21.2 g, 0.2 mol) and cesium carbonate (8.55 g, 0.05 mol) were dissolved in N,N-dimethylformamide (50 mL) Benzyl bromide (27.7 g, 0.085 mol) was added and stirred at room temperature for 3 days. Water (300 mL) and ethyl acetate (100 mL×2) were added to the mixture, and the mixture was evaporated. Ether/ethyl acetate (v/v) = 1:0 to 5:1) to give the title product 2-(2-benzyloxyethoxy)ethanol ( 2B ) as a pale yellow oil (4.0 g, yield 40%).

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 7.38-7.32 (m, 4H), 7.30 (m, 1H), 4.57 (s, 2H), 3.74-3.69 (m, 4H), 3.67-3.60 (m, 4H) , 2.16(s, 1H).

Second step: (2S)-2-(tertiary butoxycarbonylamino)propionic acid-2-(2-benzyloxyethoxy)ethyl ester (2C)

2-(2-benzyloxyethoxy)ethyl(2S)-2-(tert-butoxycarbonylamino)propanoate

2-(2-Benzyloxyethoxy)ethanol ( 2B ) (4.0 g, 0.02 mol) and N-tertiary butoxycarbonylalanine (3.8 g, 0.02 mol) were dissolved in dichloromethane (100 mL) Among them, dicyclohexylcarbodiimide (4.12 g, 0.02 mol) and 4-dimethylaminopyridine (2.44 g, 0.02 mol) were added, and the reaction was stirred at room temperature overnight under a nitrogen atmosphere. The reaction mixture was filtered, and the filtrate was evaporated, evaporated, mjjjjjjjjjjjjjjjjjjj Butyloxycarbonylamino)propanoic acid-2-(2-benzyloxyethoxy)ethyl ester ( 2C ), mp (yield: 68.5%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.33 (m, 4H), 7.31 - 7.26 (m, 1H), 5.04 (s, 1H), 4.57 (s, 2H), 4.39-4.23 (m, 3H), 3.71 (t, 2H), 3.69 - 3.65 (m, 2H), 3.64 - 3.60 (m, 2H), 1.44 (s, 9H), 1.37 (d, 3H).

The third step: (2S)-2-aminopropionic acid-2-(2-benzyloxyethoxy)ethyl ester (2D)

2-(2-benzyloxyethoxy)ethyl(2S)-2-aminopropanoate

(2S)-2-(tertiary butoxycarbonylamino)propanoic acid-2-(2-benzyloxyethoxy)ethyl ester ( 2C ) was dissolved in dichloromethane (50 mL) and trifluoroacetic acid was added. (15.5 g, 0.136 mol), reacted at room temperature overnight. The reaction solution was concentrated under reduced pressure, dichloromethane (100 mL), EtOAc EtOAc EtOAc EtOAc EtOAc The title product (2S)-2-aminopropionic acid-2-(2-benzyloxyethoxy)ethyl ester ( 2D ), yellow oil (3.0 g, yield) The rate is 82.6%).

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 7.38-7.32 (m, 4H), 7.29 (m, 1H), 4.57 (s, 2H), 4.29 (t, 2H), 3.75-3.70 (m, 2H), 3.70 -3.66 (m, 2H), 3.65-3.61 (m, 2H), 3.56 (m, 1H), 2.02 (d, 2H), 1.35 (dd, 3H).

MS M/Z (ESI): 268.3 (M + 1).

Fourth step: (2S)-2-[[[(1R)-2-(6-aminofluoren-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphonium] Amino]propionic acid-2-(2-benzyloxyethoxy)ethyl ester (2E)

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

Dissolving [(1R)-2-(6-aminofluoren-9-yl)-1-methyl-ethoxy]methyl-phenoxy-monophosphate ( Intermediate 1 ) (1.29 g, 3.55 mmol) Dichlorohydrazine (1.7 g, 14.2 mmol) was added to acetonitrile (30 mL), and the mixture was heated to 60 ° C for 5 hours. The intermediate phosphine chloride was dissolved in dichloromethane (20 mL) and cooled to 0 ° C to give (2S)-2-aminopropionic acid-2-(2-benzyloxyethoxy)ethyl ester ( 2D ) (1.14 g, 4.26 mmol) was added to the reaction mixture, and triethylamine (1.4 g, 14.2 mmol) was added dropwise to the reaction mixture, and the mixture was stirred at room temperature for 1 hour. Dichloromethane (50 mL) and water (100 mL) were added to the mixture, and the mixture was evaporated. Methyl chloride / methanol (v / v) = 1: 0 ~ 95: 5) to give the title product (2S)-2-[[[(1R)-2-(6-aminopurin-9-yl)-1-yl 2-(2-Benzyloxyethoxy)ethyl 2-(2-benzyloxyethoxy)ethyl ester ( 2E ), light yellow oil (750 mg, yield 34.5%).

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 8.33 (d, 1H), 8.00 (d, 1H), 7.34-7.27 (m, 6H), 7.22-7.06 (m, 3H), 6.94 (m, 1H), 5.78 (d, 2H), 4.55 (s, 1H), 4.48 (s, 1H), 4.33-4.26 (m, 2H), 4.25-4.04 (m, 3H), 4.03-3.88 (m, 2H), 3.71-3.53 (m, 8H), 1.33-1.19 (m, 6H).

MS M/Z (ESI): 613.4 (M + 1).

The fifth step: (2S)-2-[[[(1R)-2-(6-aminofluoren-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphonium] Amino]propionic acid-2-(2-hydroxyethoxy)ethyl ester (Compound 2)

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

(2S)-2-[[[(1R)-2-(6-Aminofluoren-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphonyl]amino]propyl Acid-2-(2-benzyloxyethoxy)ethyl ester ( 2E ) (750 mg, 0.98 mmol) was dissolved in tetrahydrofuran (30 mL) and methanol (20 mL), and 4N hydrochloric acid (0.5 mL) Palladium on carbon (30 mg) was reacted overnight at room temperature under a hydrogen atmosphere. The reaction liquid was filtered, the filter cake was washed with dichloromethane, and the filtrate was concentrated under reduced pressure. The residue was purified by methylene chloride column (dichloromethane/methanol (v/v) = 1:0 to 95:5) to give the title product. (2S)-2-[[[(1R)-2-(6-aminofluoren-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphonyl]amino]propionic acid 2-(2-Hydroxyethoxy)ethyl ester ( Compound 2 ), pale yellow solid (250 mg, yield 50%).

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 8.33 (d, 1H), 8.11 (d, 1H), 7.33-7.28 (m, 1H), 7.14 (m, 3H), 6.92 (d, 1H), 6.31 (d , 2H), 4.43-4.31 (m, 2H), 4.27-4.10 (m, 2H), 4.08-4.02 (m, 1H), 3.99-3.91 (m, 3H), 3.78-3.59 (m, 8H), 1.32 -1.23 (m, 6H).

MS M/Z (ESI): 52.

³¹ P NMR (162 MHz, CDCl ₃ ) δ 23.70, 23.02.

Example 4: Resolution/Resolution of Compound 2

(2S)-2-[[[(1R)-2-(6-Aminofluoren-9-yl)-1-methyl-ethoxy]methyl-phenoxy-phosphonyl]amino]propyl Acid-2-(2-hydroxyethoxy)ethyl ester ( Compound 2 ) (200 mg) was used for resolution.

Preparation conditions: instrument: Gilson GX-281; column: CHIRALPAK AD-H, 20 × 250 mm, 5 μm; mobile phase: A: n-hexane, B: isopropanol; equal ratio: A: B = 50: 50 (v/v); flow rate: 10 mL/min; wavelength: 260 nm; period: 50 min; sample preparation: Compound 2 was dissolved in n-hexane: isopropanol (v/v = 7:3).

After separation, the fraction was concentrated and dried at 40 ° C, and evaporated to give two optical isomer compound 2-1 (40 mg), Compound 2-2 (20 mg).

Compound 2-1

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.33 (s, 1H), 8.06 (s, 1H), 7.33-7.28 (m, 2H), 7.16 (t, 3H), 6.53 (s, 2H), 4.42 (dd , 1H), 4.36-4.28 (m, 1H), 4.26-4.12 (m, 2H), 4.08-3.92 (m, 3H), 3.81-3.57 (m, 8H), 3.20 (t, 1H), 1.25 (d , 3H), 1.21 (d, 3H).

MS M/Z (ESI): 523.3 (M + 1).

³¹ P NMR (162 MHz, CDCl3) δ 23.62.

Compound 2-2

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 8.34 (s, 1H), 8.04 (s, 1H), 7.22 (t, 2H), 7.11 (t, 1H), 6.93 (d, 2H), 6.50 (s, 2H ), 4.41-4.33 (m, 2H), 4.25 (dt, 1H), 4.16-4.03 (m, 2H), 3.93 (dd, 2H), 3.72-3.58 (m, 8H), 1.33 (d, 3H), 1.27 (d, 3H).

MS M/Z (ESI): 523.3 (M + 1).

³¹ P NMR (162 MHz, CDCl ₃ ) δ 22.84.

Example 5: Fumarate of Compound 1-1

Compound 1-1 (1.29 g, 2.4 mmol) was dissolved in ethyl acetate (13 ml), and then evaporated, and then evaporated to 50 ° C for 1 hour, then cooled to room temperature and stirred for 3 hours. . Filtration, the filter cake was washed with ethyl acetate (20 ml), and the filtered cake was dried under reduced pressure to give compound 1-1 as a compound as a white solid (1.3 g, yield 82.8%).

1H NMR (400MHz, DMSO) δ 12.87 (s, 2H), 8.12 (d, 2H), 7.34 (t, 2H), 7.14 (m, 5H), 6.63 (s, 2H), 5.55 (dd, 1H), 4.37-4.19 (m, 2H), 4.20-3.75 (m, 6H), 3.62-3.46 (m, 4H), 3.40 (dd, 2H), 3.22 (s, 3H) 1.15 (d, 3H), 1.05 (d) , 3H).

31P NMR (162 MHz, DMSO) δ 24.62.

LCMS (m/z): 537.1.

Example 6: Fumarate of Compound 1-2

Compound 1-2 (1.82 g, 3.39 mmol) was dissolved in a mixed solvent of 18 ml of ethyl acetate/methanol (v/v=9/1), and fumaric acid (0.393 g, 3.39 mmol) was added and heated to 50 ° C. The reaction was carried out for 1 hour, and further cooled to room temperature and stirred for 3 hours. After filtration, the filter cake was washed with ethyl acetate (20 ml), and the filtered cake was evaporated to dryness to give the compound 1-2 as a white solid (1.6 g, yield 72.7%).

1H NMR (400MHz, DMSO) δ 13.10 (s, 2H), 8.14 (s, 1H), 8.09 (s, 1H), 7.28 (t, 2H), 7.19 (s, 2H), 7.13 (t, 1H), 7.01(d,2H), 6.63(s,2H), 5.69(t,1H), 4.26(dd,1H), 4.15-4.08(m,3H),4.01-3.82(m,3H),3.72(dd, 1H), 3.59 (t, 2H), 3.56-3.46 (m, 2H), 3.45-3.34 (m, 2H), 3.19 (s, 3H), 1.15 (d, 3H), 1.07 (d, 3H).

³¹ P NMR (162 MHz, DMSO) δ 23.47.

LCMS (m/z): 537.2 [M+1].

Test case

1, anti-B liver virus activity test

Each compound was dissolved to 20 mM using DMSO and stored at -20 °C. A 20 mM stock solution of each compound was diluted with DMSO to a 200x stock solution. The 200× stock solution was diluted 3 times with DMSO for a total of 9 concentrations. It was further diluted 200-fold with RPMI 1640 medium containing 2.0% FBS. QPCR experiments and in cytotoxicity experiments, EC ₅₀ for each compound tested the highest final concentration of 10 M (preferably a positive control pulse to pull the ingot (LAM, Lamivudine) of 0.1μM), CC ₅₀ was the highest tested at a final concentration 100μM.

Compound anti-B hepatic virus activity was determined by qPCR method and EC ₅₀ : HepG2.2.15 cells (4 x 104 cells/well) were plated into 96-well plates and cultured overnight at 37 ° C, 5% CO ₂ ; the next day, Dilute the compound, add different concentrations of the compound to the culture well, and the final concentration of DMSO in the culture solution is 0.5%; on the fifth day, replace the fresh culture solution containing the compound; on the eighth day, collect the culture solution in the culture well and extract the DNA. The experimental procedure is described in the QIAamp 96 DNA Blood Kit (QIAGEN 51161) instructions. The qPCR reaction mixture was prepared, 15 μl of the reaction mixture was added to each well, and then 10 μl of sample or standard was added to each well, PCR: 50 ° C, 2 min; 95 ° C, 10 min; 95 ° C, 15 sec, 60 ° C, 1 min, 40 cycles. Analytical data and calculated percent inhibition: The percentage of inhibition was calculated using the following formula: % Inh. = [(HBV quantity of DMSO control-HBV quantity of sample) / HBV quantity of DMSO control] × 100. EC ₅₀ values of the compounds of the last calculated using GraphPad Prism software.

The cytotoxicity of the compounds was determined by the Cell-titer blue method and the CC ₅₀ was calculated: the highest test final concentration of the compound was 100 μM, and cell culture and compound treatment were consistent with the anti-B hepatic virus activity assay method. On the eighth day, 20 μl of Cell-titer Blue reagent was added to each well, incubated at 37 ° C for 3 hours, and the fluorescence value (560 Ex/590 Em) was read. Analysis of data and calculation of relative cell viability: The percentage of cell activity was calculated using the following formula: % cell viability = (fluorescence of sample-fluorescence of medium control) / (fluorescence of DMSO control-fluorescence of medium control) x 100. Finally, the CC ₅₀ value of the compound was calculated using GraphPad Prism software. 50% of each compound the effective concentration of HBV (EC ₅₀₎ and 50% of the cell cytotoxic concentration (CC ₅₀₎ are shown in Table 1.

Conclusion: The compounds of the present invention show certain anti-B hepatic virus activity in the tested concentration range. Compared with compound A and Lamai superior ingot, compounds 1-1 , 1-2 and their fumarate show Higher anti-B hepatic virus activity, and the compounds of the invention did not exhibit cytotoxicity over the range of concentrations tested.

Claims (9)

A compound of the formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein: Ar is a phenyl group; R ^{1 is} selected from H or methyl; R ^{2 is} selected from -(CH ₂ -CH ₂ -O) ₂ -H, -(CH ₂ -CH ₂ -O) ₂ -CH ₃ , -(CH ₂ -CH ₂ -O) ₃ -H or -(CH ₂ -CH ₂ -O) ₃ -CH ₃ . The compound, stereoisomer or pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is represented by the formula (II): The compound, stereoisomer or pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is selected from one of the following structures: The compound according to any one of claims 1 to 3, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the salt is selected from the group consisting of a hydrochloride, a sulfate, a phosphate, and an acetic acid. Salt, maleate, succinate, fumarate, malate, oxalate, tartrate, benzoate, cinnamate, p-toluenesulfonate, besylate, methanesulfonic acid a salt, a triflate salt or a combination thereof. A use of a compound according to any one of claims 1 to 4, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, for use in the preparation of a medicament for the treatment of a viral infectious disease. The use according to claim 5, wherein the viral infectious disease comprises an infectious disease caused by hepatitis B virus, hepatitis C virus or HIV. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 4, a stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable Carrier or excipient. A use of the pharmaceutical composition according to claim 7 for the preparation of a medicament for the treatment of a viral infectious disease. The use according to claim 8, wherein the viral infectious disease comprises an infectious disease caused by hepatitis B virus, hepatitis C virus or HIV.