CN108558883A - A kind of nucleic acid base compound or its pharmaceutically acceptable salt and its preparation method and application - Google Patents

A kind of nucleic acid base compound or its pharmaceutically acceptable salt and its preparation method and application Download PDF

Info

Publication number
CN108558883A
CN108558883A CN201810493926.7A CN201810493926A CN108558883A CN 108558883 A CN108558883 A CN 108558883A CN 201810493926 A CN201810493926 A CN 201810493926A CN 108558883 A CN108558883 A CN 108558883A
Authority
CN
China
Prior art keywords
structure shown
formula
compound
bases
amine derivative
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201810493926.7A
Other languages
Chinese (zh)
Other versions
CN108558883B (en
Inventor
王玉成
朱梅
王菊仙
白晓光
张国宁
董飚
彭宗根
岑山
王宇佳
王明华
赵跃
杜潇楠
张煊笛
邵端阳
李云鸽
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Institute of Medicinal Biotechnology of CAMS
Original Assignee
Institute of Medicinal Biotechnology of CAMS
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Institute of Medicinal Biotechnology of CAMS filed Critical Institute of Medicinal Biotechnology of CAMS
Priority to CN201810493926.7A priority Critical patent/CN108558883B/en
Publication of CN108558883A publication Critical patent/CN108558883A/en
Application granted granted Critical
Publication of CN108558883B publication Critical patent/CN108558883B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D473/00—Heterocyclic compounds containing purine ring systems
    • C07D473/40—Heterocyclic compounds containing purine ring systems with halogen atoms or perhalogeno-alkyl radicals directly attached in position 2 or 6
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
    • C07D239/52—Two oxygen atoms
    • C07D239/54—Two oxygen atoms as doubly bound oxygen atoms or as unsubstituted hydroxy radicals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
    • C07D239/52—Two oxygen atoms
    • C07D239/54—Two oxygen atoms as doubly bound oxygen atoms or as unsubstituted hydroxy radicals
    • C07D239/545—Two oxygen atoms as doubly bound oxygen atoms or as unsubstituted hydroxy radicals with other hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/553—Two oxygen atoms as doubly bound oxygen atoms or as unsubstituted hydroxy radicals with other hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms with halogen atoms or nitro radicals directly attached to ring carbon atoms, e.g. fluorouracil
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/04—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D473/00—Heterocyclic compounds containing purine ring systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic System
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/645—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having two nitrogen atoms as the only ring hetero atoms
    • C07F9/6509—Six-membered rings
    • C07F9/6512—Six-membered rings having the nitrogen atoms in positions 1 and 3

Abstract

The present invention provides a kind of nucleic acid base compound or its pharmaceutically acceptable salt, the compound or its pharmaceutically acceptable salt have the apparent activity for inhibiting hiv protease and/or reverse transcriptase；Toxicity research shows it with good druggability, shows that such compound has a good application prospect as anti-AIDS drug.Experimental data according to the embodiment is it is found that the compound of the present invention has inhibitory activity to 1 protease of HIV and 1 reverse transcriptases of HIV, and all has lower cytotoxicity.The nucleic acid base compound or its pharmaceutically acceptable salt of the present invention is expected to become while inhibiting double target spot inhibitor of hiv protease and reverse transcriptase.

Description

A kind of nucleic acid base compound or its pharmaceutically acceptable salt and preparation method thereof and Using
Technical field
The present invention relates to nucleic acid base technical field more particularly to a kind of nucleic acid base compound or its is pharmaceutically acceptable Salt and its preparation method and application.
Background technology
Acquired immunodeficiency syndrome (Acquired Immune Deficiency Syndrome, AIDS), also known as ends Disease is grown, being the mankind leads to immune lack because of infection immunity defective virus (Human Immunodeficiency Virus, HIV) It falls into, and causes a series of syndrome of opportunistic infections and tumour.HIV is to be currently known differentiation, the highest virus of degree of variation, According to serological reaction, gene order difference and Characteristics of Geographical Distribution, two hypotypes can be divided into：HIV-1 and HIV-2.HIV-1 is The pathogen for causing AIDS Global prevalence accounts for 95% absolute predominance on number of the infected；HIV-2 primary limitations are in Africa Portion and some western areas, number of the infected are relatively fewer.
It goes through formally to be used for clinic AIDS medicines from Zidovudine (zidovudine, AZT) conduct in 1987 is first Since object, has more than 30 kind anti-AIDS drugs at present and be applied to clinic, including a kind of entry inhibitors, 15 kinds of reverse transcriptions Enzyme (RT) inhibitor, 10 kinds of protease (PR) inhibitor, a kind of integrase inhibitor, a kind of fusion inhibitor and 5 kinds of complexing agents Deng.But prolonged application list target drug easily causes crossing drug resistant and serious toxic side effect.Chinese descendant in America's science in 1996 What big one proposes " cocktail therapy " i.e. highly effective antiretroviral therapy (HAART) for family, reduces what single drug was also easy to produce Drug resistance inhibits the duplication of virus, greatly improves the quality of life of patient to the maximum extent.However this therapy dosage Greatly, toxic side effect is strong, drug interaction is complicated, patient compliance is poor, and the severe situation faced above forces people constantly to visit Rope finds novel inverase.
Multiple target point drug design (multitarget-directed ligands, MT-DLs) is due to its uniform pharmacokinetics Property reduces drug interaction and improves the advantages such as therapeutic effect, it has also become the Disciplinary Frontiers of current medical design, for many The treatment of difficult diseases brings new hope.Reverse transcriptase inhibitor and protease inhibitors are the head in " cocktail therapy " Drug is selected, therefore the research of the bis- target spot inhibitor of HIV PR/RT has great importance.
HIV-1 reverse transcriptase (reverse transcriptase, RT) plays very in the reproduction process of inhibition of HIV Important role is catalyzed RNA reverse transcription synthetic dsdnas, and double-stranded DNA enters under the action of integrase in nucleus, and turns Viral RNA is recorded, a part for host cell is become.Therefore, RT is the important target spot of inverase, while being also HIV earliest Study target spot.
HIV-1 protease is the species specificity aspartyl protease by HIV gene codes, and active form is two The homodimer of identical peptide chain composition, every peptide chain are made of 99 amino acid residues, active site be located at two peptide chains it Between.HIV-1 protease inhibitors is made infected thin by preventing shearing of the protease to viral gag gag-pol genes Born of the same parents can only generate it is immature, do not have infective virus.Therefore, HIV-1PR is the important target spot for researching and developing inverase.
So far, there is not yet HIV-1 proteinase activities and reverse transcriptase activity can be inhibited and applied to treatment Chinese mugwort Grow the relevant report of the derivative of disease.
Invention content
The purpose of the present invention is to provide a kind of active nucleic acid bases that can inhibit HIV-1 protease and reverse transcriptase Derivative or its pharmaceutically acceptable salt.
In order to achieve the above-mentioned object of the invention, the present invention provides following technical scheme：
The present invention provides a kind of nucleic acid base compound or its pharmaceutically acceptable salts, have structure shown in Formulas I：
In Formulas I, R1For amino, methoxyl group, nitro, aminomethyl or methylol；
X is-CH2,-O- ,-S- or-NH-；
Y is
When X is-CH2When, R Ra, Rb, Rc or Rd；
When X is-O- ,-S- or-NH-, R is Rc or Rd；
The Ra isThe Rb isThe Rc isThe Rd For
Wherein, R2For hydrogen, hydroxyl, amino or halogen；R3For hydrogen, hydroxyl, amino, halogen or methyl；R4For hydrogen or methyl； R5For hydrogen, hydroxyl, amino or phosphate group；
When Z is nitrogen, R6For hydrogen, hydroxyl, amino or phosphate group；When Z is oxygen or sulphur, R6It is not present.
Preferably, the nucleic acid base compound includes 2- (the chloro- 9H- purine -9- bases of 6-)-N- ((2S, 3R) -3- hydroxyls - 4- (N- isobutyl group -4- methoxyphenyl sulphonyls amido) -1- phenyl butane -2- bases)-acetamide, 2- (8- methyl -9H- purine - 9- yls)-N- [(2S, 3R) -3- hydroxyls -4- (N- isobutyl group -4- methoxyphenyl sulphoxide imines base) -1- phenyl butyl -2- bases] - Acetamide, 2- (2,4- dioxo -2H-3,4- dihydro-pyrimidin -1- bases)-N- ((2S, 3R) -3- hydroxyls -4- (N- isobutyl group -4- first Phenyl sulfoamido) -1- phenyl butane -2- bases)-acetamide, 2- (the fluoro- 2,4- dioxos -2H-3,4- dihydro-pyrimidins-of 5- 1- yls)-N- ((2S, 3R) -3- hydroxyls -4- (N- isobutyl group -4- methoxyphenyl sulphonyls amido) -1- phenyl butane -2- bases)-second Amide, ((2S, 5R) -5- (- 1 base of 1H-2- oxo -4- aminopyrimidines)-tetrahydrofuran -2- bases) methyl N-((2S, 3R) -3- hydroxyls Base -4- (N- isobutyl group -4- aminocarbonyl phenyls sulfoamido) -1- phenyl butane -2- bases))-carbamate or N- ((2R, 3S) - 3- (2- (the fluoro- 2,4- dioxos -2H-3,4- dihydro-pyrimidins -2H-1- bases of 5-) acetamido) -2- hydroxy-4-phenyl butyl-N- Isobutyl group-P- (4- methoxyphenyls) phosphamic acid.
The present invention provides the preparation methods of the nucleic acid base compound described in above-mentioned technical proposal, include the following steps：
When X is-CH in the compound of structure shown in Formulas I2, when R is Ra or Rb, by the change with structure shown in Formula II -1 It closes object and carries out condensation reaction under the action of catalyst with amine derivative, obtain the compound with structure shown in Formulas I；
In Formula II -1, R is Ra or Rb, and the Ra isThe Rb isWherein, R2 For hydrogen, hydroxyl, amino or halogen；R3For hydrogen, hydroxyl, amino, halogen or methyl；R4For hydrogen or methyl；
When X is-CH in the compound of structure shown in Formulas I2,-O- ,-S- or-NH-, R be when being Rc or Rd, will have Formula II- Compound, carbonic acid trichloromethyl ester and the amine derivative of structure shown in 2 carry out condensation reaction under the action of catalyst, are had The compound of structure shown in Formulas I；
Formula II -2 R-XH,
In Formula II -2, X is-CH2,-O- ,-S- or-NH-；R is Rc or Rd, and the Rc isThe Rd isWherein, R2For hydrogen, hydroxyl, amino or halogen, R3For hydrogen, hydroxyl, amino, halogen or methyl；R4For hydrogen Or methyl；R5For hydrogen, hydroxyl, amino or phosphate group；When Z is nitrogen, R6For hydrogen, hydroxyl, amino or phosphate group；When Z is oxygen Or when sulphur, R6It is not any atom or group；
The amine derivative has structure shown in formula III：
In formula III, R1For amino, methoxyl group, nitro, aminomethyl or methylol；Y is
Preferably, the compound and amine derivative with structure shown in Formula II -1 are in carbodiimide hydrochloride, 1- Condensation reaction is carried out under the catalytic action of hydroxybenzotriazole and 4-dimethylaminopyridine；
The compound with structure shown in Formula II -2 is with carbonic acid trichloromethyl ester and amine derivative in N, N- diisopropyls Condensation reaction is carried out under the catalytic action of base ethamine.
Preferably, the preparation method of the compound with structure shown in Formula II -1, includes the following steps:
Substituted pyrimidines base or substituted purin base are subjected to substitution reaction with bromoacetic acid or bromoacetate, replaced Product；
The substitution product is hydrolyzed, the compound with structure shown in Formula II -1 is obtained；
The substituted pyrimidines base has structure shown in formula a：
The substituted purin base has structure shown in formula b：
Preferably, when in the amine derivative Y beWhen, the amine derivative has structure shown in formula III -1：
The preparation method of the amine derivative with structure shown in formula III -1, includes the following steps：
[(S) -1- ((S)-ethylene oxide -2- bases) -2- benzene ethylaminos t-butyl formate] is carried out nucleophilic with isobutyl amine to take Generation reaction, obtains (1S, 2R) -1- benzyl -2- hydroxyls -3- (isobutyl amine) t-butyl carbamate；
The R of (1S, 2R) -1- benzyls -2- hydroxyls -3- (isobutyl amine) t-butyl carbamates and structure shown in formula d by described in1 Substituted phenylsulfonyl chloride carries out nucleophilic substitution, obtains the intermediate of structure shown in formula c；
By the intermediate Deprotection of structure shown in the formula c, the amine derivative with structure shown in formula III -1 is obtained；
Preferably, when in the amine derivative Y beWhen, the amine derivative has structure shown in formula III -2：
The preparation method of the amine derivative with structure shown in formula III -2, includes the following steps：
[(S) -1- ((S)-ethylene oxide -2- bases) -2- benzene ethylaminos t-butyl formate] is carried out nucleophilic with isobutyl amine to take Generation reaction, obtains (1S, 2R) -1- benzyl -2- hydroxyls -3- (isobutyl amine) t-butyl carbamate；
The R of (1S, 2R) -1- benzyls -2- hydroxyls -3- (isobutyl amine) t-butyl carbamates and structure shown in formula d by described in1 Substituted phenylsulfonyl chloride carries out nucleophilic substitution, obtains the intermediate of structure shown in formula c；
The intermediate Yu Sodium azide of structure shown in the formula c are subjected to imidization, obtain the centre of structure shown in formula e Body；
By the intermediate Deprotection of structure shown in the formula e, the amine derivative with structure shown in formula III -2 is obtained.
Preferably, when in the amine derivative Y beWhen, the amine derivative has structure shown in formula III -3：
The preparation method of the amine derivative with structure shown in formula III -3, includes the following steps：
[(S) -1- ((S)-ethylene oxide -2- bases) -2- benzene ethylaminos t-butyl formate] is carried out nucleophilic with isobutyl amine to take Generation reaction, obtains (1S, 2R) -1- benzyl -2- hydroxyls -3- (isobutyl amine) t-butyl carbamate；
By 4- substituted-phenyls phosphonyl dichloride, benzyl alcohol and (1S, the 2R) -1- benzyl -2- hydroxyls with structure shown in formula f Base -3- (isobutyl amine) t-butyl carbamate carries out the intermediate that structure shown in formula g is obtained by the reaction in Arbuzov；
By the intermediate and H of structure shown in the formula g2Reduction reaction is carried out, the intermediate of structure shown in formula h is obtained；
By the intermediate Deprotection of structure shown in the formula h, the amine derivative with structure shown in formula III -3 is obtained.
The present invention provides nucleic acid base compounds described in above-mentioned technical proposal or its pharmaceutically acceptable salt to prepare Application in hiv inhibitor, the hiv inhibitor is using hiv protease and reverse transcriptase as target spot.
Preferably, the dosage of the nucleic acid base compound or its pharmaceutically acceptable salt in hiv inhibitor is 0.01~100nM.
The present invention provides a kind of nucleic acid base compound or its pharmaceutically acceptable salt, the compound or its pharmaceutically Acceptable salt has the apparent activity for inhibiting hiv protease and/or reverse transcriptase；Toxicity research shows it with good Druggability shows that such compound has a good application prospect as anti-AIDS drug.Experimental data according to the embodiment It is found that the compound of the present invention has inhibitory activity (table 1) to HIV-1 protease and HIV-1 reverse transcriptase, and all have Lower cytotoxicity (table 2).The nucleic acid base compound or its pharmaceutically acceptable salt of the present invention is expected to become while pressing down Double target spot inhibitor of hiv protease and reverse transcriptase processed.
Description of the drawings
The compound of Fig. 1 Formulas I structures；
The synthetic route chart of Fig. 2 compounds 1；
The synthetic route chart of Fig. 3 compounds 2；
The synthetic route chart of Fig. 4 compounds 3；
The synthetic route chart of Fig. 5 compounds 4；
The synthetic route chart of Fig. 6 compounds 5；
The synthetic route chart of Fig. 7 compounds 6.
Specific implementation mode
The present invention provides a kind of nucleic acid base compound or its pharmaceutically acceptable salts, have structure shown in Formulas I：
In Formulas I, R1For amino, methoxyl group, nitro, aminomethyl or methylol；
X is-CH2,-O- ,-S- or-NH-；
Y is
When X is-CH2When, R Ra, Rb, Rc or Rd；
When X is-O- ,-S- or-NH-, R is Rc or Rd；
The Ra isThe Rb isThe Rc isThe Rd For
Wherein, R2For hydrogen, hydroxyl, amino or halogen；R3For hydrogen, hydroxyl, amino, halogen or methyl；R4For hydrogen or methyl； R5For hydrogen, hydroxyl, amino or phosphate group；
When Z is nitrogen, R6For hydrogen, hydroxyl, amino or phosphate group；When Z is oxygen or sulphur, R6It is not present.
In the present invention, the nucleic acid base compound preferably include 2- (the chloro- 9H- purine -9- bases of 6-)-N- ((2S, 3R) -3- hydroxyls -4- (N- isobutyl group -4- methoxyphenyl sulphonyls amido) -1- phenyl butane -2- bases)-acetamide, 2- (8- first Base -9H- purine -9- bases)-N- [(2S, 3R) -3- hydroxyls -4- (N- isobutyl group -4- methoxyphenyl sulphoxide imines base) -1- phenyl Butyl -2- bases]-acetamide, 2- (2,4- dioxo -2H-3,4- dihydro-pyrimidin -1- bases)-N- ((2S, 3R) -3- hydroxyl -4- (N- Isobutyl group -4- methoxyphenyl sulphonyls amido) -1- phenyl butane -2- bases)-acetamide, 2- (the fluoro- 2,4- dioxos -2H-3 of 5-, 4- dihydro-pyrimidin -1- bases)-N- ((2S, 3R) -3- hydroxyls -4- (N- isobutyl group -4- methoxyphenyl sulphonyls amido) -1- phenyl fourths Alkane -2- bases)-acetamide, ((2S, 5R) -5- (- 1 base of 1H-2- oxo -4- aminopyrimidines)-tetrahydrofuran -2- bases) methyl N - ((2S, 3R) -3- hydroxyls -4- (N- isobutyl group -4- aminocarbonyl phenyls sulfoamido) -1- phenyl butane -2- bases))-carbamate Or N- ((2R, 3S) -3- (2- (the fluoro- 2,4- dioxos -2H-3,4- dihydro-pyrimidins -2H-1- bases of 5-) acetamido) -2- hydroxyls - 4- phenyl butyl-N- isobutyl groups-P- (4- methoxyphenyls) phosphamic acid.
The present invention provides the preparation methods of the nucleic acid base compound described in above-mentioned technical proposal, include the following steps：
When X is-CH in the compound of structure shown in Formulas I2, when R is Ra or Rb, by the change with structure shown in Formula II -1 It closes object and carries out condensation reaction under the action of catalyst with amine derivative, obtain the compound with structure shown in Formulas I；
In Formula II -1, R is Ra or Rb, and the Ra isThe Rb isWherein, R2 For hydrogen, hydroxyl, amino or halogen；R3For hydrogen, hydroxyl, amino, halogen or methyl；R4For hydrogen or methyl；
When X is-CH in the compound of structure shown in Formulas I2,-O- ,-S- or-NH-, R be when being Rc or Rd, will have Formula II- Compound, carbonic acid trichloromethyl ester and the amine derivative of structure shown in 2 carry out condensation reaction under the action of catalyst, are had The compound of structure shown in Formulas I；
Formula II -2 R-XH,
In Formula II -2, X is-CH2,-O- ,-S- or-NH-；R is Rc or Rd, and the Rc isThe Rd isWherein, R2For hydrogen, hydroxyl, amino or halogen, R3For hydrogen, hydroxyl, amino, halogen or methyl；R4For hydrogen Or methyl；R5For hydrogen, hydroxyl, amino or phosphate group；When Z is nitrogen, R6For hydrogen, hydroxyl, amino or phosphate group；When Z is oxygen Or when sulphur, R6It is not any atom or group；
The amine derivative has structure shown in formula III：
In formula III, R1For amino, methoxyl group, nitro, aminomethyl or methylol；Y is
In the present invention, the compound and amine derivative with structure shown in Formula II -1 are preferably in carbodiimides salt Condensation reaction is carried out under the catalytic action of hydrochlorate, I-hydroxybenzotriazole and 4-dimethylaminopyridine, is obtained with knot shown in Formulas I The compound of structure, reaction process are as follows：
In the present invention, the compound with structure shown in Formula II -1, amine derivative, carbodiimide hydrochloride, The molar ratio of I-hydroxybenzotriazole and 4-dimethylaminopyridine is preferably 1：(1.0~1.1)：(1.4~1.6)：(1.0~ 1.2)：(0.19~0.21), more preferably 1:1.05:1.5:1.1:0.2.In the present invention, the carbodiimide hydrochloride Preferably 1- ethyls-(3- dimethylaminopropyls) carbodiimide hydrochloride.In the present invention, the condensation reaction preferably exists It is carried out under the conditions of organic solvent is existing；The present invention does not have the type of the organic solvent special restriction, using ability The organic solvent that known to field technique personnel condensation reaction can be smoothed out, it is specific such as dimethylformamide (DMF). In the present invention, the condensation reaction preferably includes following steps：
Compound, amine derivative and DMF with structure shown in Formula II -1 are mixed, mixed to gained under condition of ice bath Carbodiimide hydrochloride and I-hydroxybenzotriazole are added in solution, carries out first 0.5~1.5h of condensation reaction at room temperature； Then 4-dimethylaminopyridine is added in resulting material, carries out second 1.5~2.5h of condensation reaction at room temperature.
After completing the condensation reaction, the present invention preferably post-processes gained condensation material, obtains with shown in Formulas I The compound of structure.In the present invention, the post-processing preferably includes following steps：
It carries out gained condensation material that removal DMF is concentrated under reduced pressure, residue be mixed with water, using ethyl acetate to gained Mixture is extracted, and gained organic phase is dried with anhydrous sodium sulfate, the organic phase after drying is concentrated to give crude product, The crude product is prepared thin-layer chromatography through silica gel to isolate and purify, obtains the compound with structure shown in Formulas I.
In the present invention, the preparation method of the compound with structure shown in Formula II -1, preferably includes following steps:
Substituted pyrimidines base or substituted purin base are subjected to substitution reaction with bromoacetic acid or bromoacetate, replaced Product；
The substitution product is hydrolyzed, the compound with structure shown in Formula II -1 is obtained；
The substituted pyrimidines base has structure shown in formula a：
The substituted purin base has structure shown in formula b：
Substituted pyrimidines base or substituted purin base preferably with bromoacetic acid or bromoacetate replace anti-by the present invention It answers, obtains substitution product.In the present invention, the substituted pyrimidines base with structure shown in formula a and with structure shown in formula b Substituted purin base be preferably commercial product.In the present invention, the substituted pyrimidines base or substituted purin base and bromine second The molar ratio of acid or bromoacetate preferably stands alone as 10：(10~12), more preferably 10：11.In the present invention, the substitution It is carried out under the conditions of reaction preferably existing for catalyst and organic solvent；Kind of the present invention for the catalyst and organic solvent Class does not have special restriction, using the catalyst well known to those skilled in the art that substitution reaction can be made to be smoothed out and organic Solvent, it is specific if catalyst is sodium hydride, organic solvent DMF.
In the present invention, the substitution reaction preferably includes following steps：
Substituted pyrimidines base or substituted purin base are mixed with DMF, under condition of ice bath, sodium hydride is added drop-wise to gained In mixture, 0.5~1.5h is reacted into line replacement at room temperature, bromoacetic acid or bromoacetate, which are added drop-wise to gained, replaces reactant In material, 0.5~1.5h of substitution reaction is carried out at room temperature.
After completing the substitution reaction, gained is preferably replaced material to post-process by the present invention, obtains substitution product. In the present invention, the post-processing preferably includes following steps：
Replace material to be mixed with water gained, extracted with ethyl acetate, gained organic phase is done with anhydrous sodium sulfate It is dry, the organic phase after drying is concentrated to give crude product, the crude product is prepared thin-layer chromatography through silica gel to be isolated and purified, Obtain substitution product.
In the present invention, the structural formula of the substitution product is preferably
After obtaining substitution product, the present invention preferably hydrolyzes the substitution product, obtains having structure shown in Formula II -1 Compound.In the present invention, it is described hydrolysis preferably existing for sodium hydroxide solution under the conditions of carry out；The sodium hydroxide solution Concentration be preferably 3mol/L；The molar ratio of the substitution product and sodium hydroxide is preferably 1:3.In the present invention, described The temperature of hydrolysis is preferably 25~35 DEG C, can be specifically room temperature；The time of the hydrolysis is preferably 0.5~ 1.5h, more preferably 1h.
After completing the hydrolysis, the present invention preferably post-processes gained hydrolysis material, obtains with Formula II -1 The compound of shown structure.In the present invention, the post-processing preferably includes following steps：
Using hydrochloric acid solution by gained hydrolyze material pH value be adjusted to 3.5~4.5, under condition of ice bath stir 25~ Solid is precipitated in 35min, filters, and filter cake is washed with water, dries, and obtains the compound with structure shown in Formula II -1.
In the present invention, when in the amine derivative Y beWhen, the amine derivative has knot shown in formula III -1 Structure：
The preparation method of the amine derivative with structure shown in formula III -1, preferably includes following steps：
[(S) -1- ((S)-ethylene oxide -2- bases) -2- benzene ethylaminos t-butyl formate] is carried out nucleophilic with isobutyl amine to take Generation reaction, obtains (1S, 2R) -1- benzyl -2- hydroxyls -3- (isobutyl amine) t-butyl carbamate；
The R of (1S, 2R) -1- benzyls -2- hydroxyls -3- (isobutyl amine) t-butyl carbamates and structure shown in formula d by described in1 Substituted phenylsulfonyl chloride carries out nucleophilic substitution, obtains the intermediate of structure shown in formula c；
By the intermediate Deprotection of structure shown in the formula c, the amine derivative with structure shown in formula III -1 is obtained；
The present invention is preferably by [(S) -1- ((S)-ethylene oxide -2- bases) -2- benzene ethylaminos t-butyl formate] and isobutyl amine Nucleophilic substitution is carried out, (1S, 2R) -1- benzyl -2- hydroxyls -3- (isobutyl amine) t-butyl carbamate is obtained.In the present invention In, 3 [(S) -1- ((S)-ethylene oxide -2- bases) -2- benzene ethylaminos t-butyl formates] are preferred with the molar ratio of isobutyl amine For (73~78)：(187~192), more preferably (75~76)：(189~190), most preferably 76：189.In the present invention, 3 [(S) -1- ((S)-ethylene oxide -2- bases) -2- benzene ethylaminos t-butyl formates] are preferably commercial product, specifically may be used To derive from lark prestige Science and Technology Ltd..In the present invention, the nucleophilic displacement of fluorine preferably existing for organic solvent under the conditions of It carries out, the organic solvent is preferably acetonitrile.In the present invention, the temperature of the nucleophilic substitution is preferably 75~85 DEG C, More preferably 80 DEG C；The time of reaction is preferably 4.5~5.5h, more preferably 5h.After completing the nucleophilic substitution, this hair It is bright that preferably gained nucleophilic displacement of fluorine material is cooled down, concentrated and recrystallized successively, obtain (1S, 2R) -1- benzyl -2- hydroxyls - 3- (isobutyl amine) t-butyl carbamate.In the present invention, recrystallization reagent used by the recrystallization is preferably acetic acid second Ester and n-hexane；The volume ratio of ethyl acetate and n-hexane is preferably 1 in the recrystallization reagent：(8~10), more preferably 1: 9。
After obtaining (1S, 2R) -1- benzyl -2- hydroxyls -3- (isobutyl amine) t-butyl carbamate, the present invention preferably will be described The R of (1S, 2R) -1- benzyls -2- hydroxyls -3- (isobutyl amine) t-butyl carbamates and structure shown in formula d1Substituted phenylsulfonyl chloride into Row nucleophilic substitution obtains the intermediate of structure shown in formula c.In the present invention, described (1S, the 2R) -1- benzyl -2- hydroxyls - 3- (isobutyl amine) t-butyl carbamates and N, N- diisopropylethylamine, 4-dimethylaminopyridine and R1Substituted phenylsulfonyl chloride rubs You are than preferably 14~15:16~17:1~2:16~17, more preferably 14.86mmol：16.34mmol：1.49mmol： 16.34mmol.In the present invention, the nucleophilic substitution preferably existing for organic solvent and catalyst under the conditions of carry out, The organic solvent is preferably tetrahydrofuran and THF, and the catalyst is preferably n,N-diisopropylethylamine and 4- dimethylaminos Pyridine.The present invention is preferably first mixed by (1S, 2R) -1- benzyls -2- hydroxyls -3- (isobutyl amine) t-butyl carbamates and tetrahydrofuran It closes, n,N-diisopropylethylamine, 4-dimethylaminopyridine, R is then added under condition of ice bath1Substituted phenylsulfonyl chloride and THF, into Row nucleophilic substitution.In the present invention, the temperature of the nucleophilic substitution is preferably 0 DEG C of ice bath, and the nucleophilic displacement of fluorine is anti- The time answered preferably 0.2~1h, more preferably 0.5h.The present invention preferably detects the extent of reaction by TLC.The nucleophilic to be done After substitution reaction, gained nucleophilic displacement of fluorine material is preferably concentrated under reduced pressure and removes THF by the present invention, then adopts and is extracted with ethyl acetate, Extraction gained organic phase is concentrated, by concentration gained crude product through flash column purifications, obtains the intermediate of structure shown in formula c. In the present invention, eluant, eluent used by the flash column purifications is preferably ethyl acetate and n-hexane, second in the eluant, eluent The volume ratio of acetoacetic ester and n-hexane is preferably 1:5.
After obtaining the intermediate of structure shown in formula c, the intermediate of structure shown in the formula c is preferably deprotected by the present invention Base obtains the amine derivative with structure shown in formula III -1.In the present invention, the Deprotection preferably in organic solvent and It is carried out under the conditions of catalyst is existing, the organic solvent is preferably CH2Cl2, the catalyst is preferably trifluoroacetic acid.At this In invention, the amount and CH of the substance of the intermediate of structure shown in the formula c2Cl2Volume ratio with trifluoroacetic acid is preferably 8~ 10mmol：9~11mL：9~11mL, more preferably 9.87mmol：10mL：10mL.In the present invention, the deprotection reaction Temperature be preferably 25~35 DEG C, be specifically as follows room temperature；The time of the deprotection reaction is preferably 2~4h, more preferably For 3h.After completing the deprotection reaction, then it is super that saturated sodium bicarbonate solution is added in the preferably first concentration of reaction solution of the present invention Solid is precipitated in sound, stirring, filters, and will filter products therefrom and carries out column chromatography for separation, obtains having structure shown in formula III -1 Amine derivative.In the present invention, eluant, eluent used by the column chromatography for separation is preferably CH2Cl2And MeOH, the eluant, eluent Middle CH2Cl2Volume ratio with MeOH is preferably 10:1.
In the present invention, when in the amine derivative Y beWhen, the amine derivative has knot shown in formula III -2 Structure：
The preparation method of the amine derivative with structure shown in formula III -2, preferably includes following steps：
3 [(S) -1- ((S)-ethylene oxide -2- bases) -2- benzene ethylaminos t-butyl formates] are carried out nucleophilic with isobutyl amine to take Generation reaction, obtains (1S, 2R) -1- benzyl -2- hydroxyls -3- (isobutyl amine) t-butyl carbamate；
The R of (1S, 2R) -1- benzyls -2- hydroxyls -3- (isobutyl amine) t-butyl carbamates and structure shown in formula d by described in1 Substituted phenylsulfonyl chloride carries out nucleophilic substitution, obtains the intermediate of structure shown in formula c；
The intermediate Yu Sodium azide of structure shown in the formula c are subjected to imidization, obtain the centre of structure shown in formula e Body；
By the intermediate Deprotection of structure shown in the formula e, the amine derivative with structure shown in formula III -2 is obtained；
In the present invention, the specific preparation method of the intermediate of structure shown in the formula c is same as mentioned above, herein not It repeats again.
After obtaining the intermediate of structure shown in formula c, the present invention is preferably by the intermediate and Sodium azide of structure shown in the formula c Imidization is carried out, the intermediate of structure shown in formula e is obtained.In the present invention, the intermediate of structure shown in the formula c with it is folded The molar ratio of nitrogen sodium is preferably 6~7:6~7, more preferably 6.30：6.90.In the present invention, the imidization preferably exists It is carried out under the conditions of organic solvent and catalyst are existing, the organic solvent is preferably anhydrous chloroform, and the catalyst is excellent It is selected as the concentrated sulfuric acid.In the imidization, the present invention is preferably first by the intermediate of structure shown in formula c and anhydrous chloroform Mixing, is then added Sodium azide under protection of argon gas, and the concentrated sulfuric acid then is added dropwise under the conditions of 0 DEG C of ice bath.The present invention preferably exists The process that the concentrated sulfuric acid is added dropwise is completed in 5min.In the present invention, the temperature of the imidization is preferably 40~50 DEG C, more excellent It is selected as 45 DEG C；The time of the imidization is preferably 10~12h.Present invention preferably employs TIC to monitor reaction process.It has waited for After the imidization, present invention preferably uses chloroforms to extract gained reactant material, then uses saturated common salt water washing Organic phase concentrates organic phase after drying, is purified through silica gel column chromatography to enriched product, obtains the intermediate of structure shown in formula e. In the present invention, eluant, eluent used by silica gel column chromatography purifying is preferably ethyl acetate and methanol, in the eluant, eluent The volume ratio of ethyl acetate and methanol is preferably 30:1.
After obtaining the intermediate of structure shown in formula e, the present invention obtains the intermediate Deprotection of structure shown in the formula e To the amine derivative with structure shown in formula III -2.The intermediate of structure shown in formula e is carried out the specific of Deprotection by the present invention Method with it is above-mentioned that the method for intermediate Deprotection of structure shown in formula c is identical, details are not described herein.
In the present invention, when in the amine derivative Y beWhen, the amine derivative has knot shown in formula III -3 Structure：
The preparation method of the amine derivative with structure shown in formula III -3, preferably includes following steps：
[(S) -1- ((S)-ethylene oxide -2- bases) -2- benzene ethylaminos t-butyl formate] is carried out nucleophilic with isobutyl amine to take Generation reaction, obtains (1S, 2R) -1- benzyl -2- hydroxyls -3- (isobutyl amine) t-butyl carbamate；
By 4- substituted-phenyls phosphonyl dichloride, benzyl alcohol and (1S, the 2R) -1- benzyl -2- hydroxyls with structure shown in formula f Base -3- (isobutyl amine) t-butyl carbamate carries out Arbuzov reactions, obtains the intermediate of structure shown in formula g；
By the intermediate and H of structure shown in the formula g2Reduction reaction is carried out, the intermediate of structure shown in formula h is obtained；
By the intermediate Deprotection of structure shown in the formula h, the amine derivative with structure shown in formula III -3 is obtained；
The present invention carries out [(S) -1- ((S)-ethylene oxide -2- bases) -2- benzene ethylaminos t-butyl formate] and isobutyl amine Nucleophilic substitution, obtain the specific method of (1S, 2R) -1- benzyl -2- hydroxyls -3- (isobutyl amine) t-butyl carbamate with it is upper It is identical to state content, details are not described herein.
After obtaining (1S, 2R) -1- benzyl -2- hydroxyls -3- (isobutyl amine) t-butyl carbamate, the present invention will preferably have 4- substituted-phenyls phosphonyl dichloride, benzyl alcohol and (1S, the 2R) -1- benzyl -2- hydroxyls -3- (isobutyl amine) of structure shown in formula f T-butyl carbamate carries out Arbuzov reactions, obtains the intermediate of structure shown in formula g.In the present invention, described that there is formula f The 4- substituted-phenyls phosphonyl dichloride and (1S, 2R) -1- benzyl -2- hydroxyls -3- (isobutyl amine) t-butyl carbamate of shown structure The amount of substance and the volume ratio of benzyl alcohol be preferably 4~5mol：3~4mL：3~4mol, more preferably 4.16:3.78: 3.78.In the present invention, it is carried out under the conditions of the Arbuzov reacts preferably existing for organic solvent and catalyst；It is described to have Solvent is preferably benzene, and the catalyst is selected as n,N-diisopropylethylamine (DIEA) and 1-H- tetrazoles.In the present invention, institute It states Arbuzov reactions and preferably includes following steps：
4- substituted-phenyls phosphonyl dichloride, 1-H- tetrazoles with structure shown in formula f are dissolved in benzene, under condition of ice bath to Be added dropwise benzyl alcohol, n,N-diisopropylethylamine (DIEA) in gained mixed solution successively, carry out the first ice bath reaction 25~ 35min；After completing the reaction of the first ice bath, first 2~3h of room temperature reaction is carried out at room temperature；After completing the first room temperature reaction, (1S, 2R) -1- benzyl -2- hydroxyls -3- (isobutyl amine) carbamic acid uncle is sequentially added under condition of ice bath into gained reactant material Butyl ester and DIEA carry out the second ice bath and react 25~35min；After completing the reaction of the second ice bath, the second room temperature is carried out at room temperature React 3~4h.
After completing Arbuzov reaction, present invention preferably employs reversed-phase column chromatography to gained Arbuzov reaction masses into Row isolates and purifies, and obtains the intermediate of structure shown in formula g.In the present invention, the mobile phase of the reversed-phase column chromatography is preferably second The mixed liquor of alcohol and water, the volume ratio of ethyl alcohol and water is preferably 5 in the mobile phase:1.
After obtaining the intermediate of structure shown in formula g, the present invention is preferably by the intermediate and H of structure shown in the formula g2It carries out Reduction reaction obtains the intermediate of structure shown in formula h.In the present invention, the reduction reaction is preferably in organic solvent and catalysis It is carried out under the conditions of agent is existing, the organic solvent is preferably tetrahydrofuran, and the catalyst is preferably Pd/C.In the reduction In reaction, the present invention is preferably under protection of argon gas by intermediate Yu tetrahydrofuran, Pd/C, K of structure shown in formula g2CO3It is mixed with water It closes, then replaces the argon gas using hydrogen, carry out the reduction reaction at room temperature.In the present invention, the reduction reaction Time be preferably 3~5h, the temperature of more preferably 4h, the reduction reaction are preferably 25~35 DEG C, can be specifically room Temperature.After completing the reduction reaction, present invention preferably uses miillpore filters to be filtered reaction solution obtained by reduction reaction, in ice Under the conditions of bath, the pH value of gained filtrate is adjusted to 5.0 using hydrochloric acid, is then extracted using ethyl acetate, with anhydrous sulphur Sour sodium dries organic phase, concentrates organic phase, obtains the intermediate of structure shown in formula h.
After obtaining the intermediate of structure shown in formula h, the intermediate of structure shown in the formula h is preferably deprotected by the present invention Base obtains the amine derivative with structure shown in formula III -3.The intermediate of structure shown in formula h is carried out Deprotection by the present invention Specific method with it is above-mentioned that the method for intermediate Deprotection of structure shown in formula c is identical, details are not described herein.
In the present invention, the compound and carbonic acid trichloromethyl ester and amine derivative with structure shown in Formula II -2 are excellent It is selected under the catalytic action of n,N-diisopropylethylamine and carries out condensation reaction, obtain the compound with structure shown in Formulas I, react Process is as follows：
In the present invention, compound, carbonic acid trichloromethyl ester, amine derivative and the N with structure shown in Formula II -2, The molar ratio of N- diisopropylethylamine is preferably 0.5：(0.45~0.55)：(0.5~0.6)：(0.5~0.6), more preferably 0.5:0.5:0.55:0.55.In the present invention, the condensation reaction preferably existing for organic solvent under the conditions of carry out；It is described Organic solvent is preferably anhydrous organic solvent.The present invention does not have the type of the organic solvent special restriction, using this The organic solvent that known to field technology personnel condensation reaction can be smoothed out, it is specific such as dichloromethane and/or four Hydrogen furans.In the present invention, the condensation reaction preferably includes following steps：
Carbonic acid trichloromethyl ester is mixed with dichloromethane under the conditions of ice bath, nitrogen protection, obtains the first mixed solution； Compound, n,N-diisopropylethylamine with structure shown in Formula II -2 are mixed with tetrahydrofuran, obtain the second mixed solution； Second mixed solution is added drop-wise in first mixed solution under condition of ice bath, it is anti-to carry out the first condensation at room temperature 25~35min is answered, the first condensation material is obtained；
Amine derivative is mixed with dichloromethane, n,N-diisopropylethylamine is added drop-wise in gained mixed solution, in room Temperature is lower to carry out second 4~6min of condensation reaction, obtains the second condensation material；
First condensation material is mixed with the second condensation material, carries out 0.5~1.5h of condensation reaction at room temperature.
After completing the condensation reaction, the present invention preferably post-processes gained condensation material, obtains with shown in Formulas I The compound of structure.In the present invention, the post-processing preferably includes following steps：
Gained condensation material is mixed with dichloromethane, is washed with saturated ammonium chloride solution, gained organic phase is used Anhydrous sodium sulfate is dried, and the organic phase after drying is concentrated to give crude product, the crude product is prepared thin-layer chromatography through silica gel It is isolated and purified, obtains the compound with structure shown in Formulas I.In the present invention, the method for the thin-layer chromatography with it is above-mentioned Scheme is identical, and details are not described herein.
In the present invention, the compound with structure shown in Formula II -2 is preferably commercially available substituted nucleosides or deoxidation core Glycosides.
In the present invention, the preparation method of the amine derivative is identical as said program, and details are not described herein.
The present invention provides nucleic acid base compounds described in above-mentioned technical proposal or its pharmaceutically acceptable salt to prepare Application in hiv inhibitor, the hiv inhibitor is using hiv protease and reverse transcriptase as target spot.
Preferably, the dosage of the nucleic acid base compound or its pharmaceutically acceptable salt in hiv inhibitor is 0.01-100nM。
Nucleic acid base compound provided by the invention or its pharmaceutically acceptable salt are carried out with reference to embodiment detailed Thin explanation, but they cannot be interpreted as limiting the scope of the present invention.
Embodiment 1
6-chloropurine (0.62g, 4mmol) and Anhydrous potassium carbonate (1.66g, 12mmol) are added in 10mL eggplant-shape bottles, Anhydrous solvent n,N-Dimethylformamide (DMF) 2mL is added, room temperature is vigorously stirred reaction 1h under nitrogen protection；By bromoacetic acid (0.64g, 4.6mmol) is dissolved in anhydrous solvent n,N-Dimethylformamide (DMF) 1mL, is slowly dropped in reaction solution, is added It finishes, continues to be stirred at room temperature reaction overnight.5mL distilled water is added into reaction solution, with diatomite filtering solution, washing obtains Yellow clarified solution body adjusts pH to 3.0 with 4M HCl, stirs 10min under ice bath, stand, filter under ice bath stirring state, Filter cake is washed with a small amount, dry, obtains target product yellow greenish powder solid i.e. intermediate 1 (0.37g, 43.4%).Intermediate 1 LC-MS (ESI, M+H+)m/z 213.4。
By (S) -1- ((S)-ethylene oxide -2- bases) -2- benzene ethylaminos t-butyl formates (20.0g, 75.94mmol), second Nitrile 80mL and isobutyl amine (19.02mL, 189.46mmol) are added in 200mL eggplant-shape bottles, and 80 DEG C of mixture stirs 5 hours.Reaction After reaction solution be cooled to room temperature, be concentrated under reduced pressure remove solvent.Crude product ethyl acetate/n-hexane (1:After 9) recrystallizing White object product (21.2g, 83%), the white object product are intermediate b1, LC-MS (ESI, M+H+)m/z 337.2。
Intermediate b1 (5.0g, 14.86mmol), tetrahydrofuran (THF) (40mL) are added in 250mL eggplant-shape bottles, ice bath Under be slowly added to n,N-diisopropylethylamine (DIEA) (3.68mL, 16.34mmol) and 4-dimethylaminopyridine (DMAP) (0.18g, 1.49mmol), is then added 4- Methoxybenzenesulfonyl chlorides (3.38g, 16.34mmol) and the mixing of THF (10mL) is molten Liquid.It is stirred 0.5 hour under ice bath and moves to room temperature.TLC detections are concentrated under reduced pressure after completion of the reaction removes THF, ethyl acetate extraction (30 × 3mL), organic phase concentration.Crude product is through flash column purifications ethyl acetate-hexane (1:5) in obtaining target product white solid i.e. LC-MS (ESI, M+H+) m/z 507.0 of mesosome b2 (6.14g, 82%), intermediate b2.
Intermediate b2 (5.0g, 9.87mmol) is added in 100mL eggplant-shape bottles, CH is added at room temperature2Cl2(10mL) and three Fluoroacetic acid (10mL).It finishes, reacts at room temperature 3h.Concentration of reaction solution after completion of the reaction, and 200mL saturated sodium bicarbonate solutions are added Ultrasound, stirring, have solid precipitation, filter to obtain crude product, column chromatography CH2Cl2-MeOH(10:1) in obtaining target product white solid i.e. Mesosome B1a (2.71g, 68%).LC-MS (ESI, M+H+) m/z 407.3 of intermediate B 1a.
Intermediate 1 (0.21g, 1mmol), intermediate B 1a (0.43g, 1.05mmol) are added in 10mL eggplant-shape bottles, are added 2mL anhydrous DMFs, are placed in ice bath, are slowly added to 1- ethyls-(3- dimethylaminopropyls) carbodiimide hydrochloride hydrochloride (EDCHCl) (0.29g, 1.5mmol), I-hydroxybenzotriazole (HOBt) (0.15g, 1.1mmol), finishes and is transferred to room temperature It is stirred to react 1h, DMAP (0.024g, 0.2mmol) is added, continues to be stirred to react 2h.DMF is concentrated under reduced pressure after completion of the reaction, remains 6mL water is added in object, is extracted with ethyl acetate (6 × 3mL), and organic phase is dried with anhydrous sodium sulfate, is concentrated to give crude product.Crude product is through silicon Glue prepare thin-layer chromatography isolated and purified (solvent be ethyl acetate and methanol, by volume, ethyl acetate：Methanol=8： 1) white powder solid (0.54g, 89.3%) to get to compound 1：2- (the chloro- 9H- purine -9- bases of 6-)-N- ((2S, 3R) -3- hydroxyls -4- (N- isobutyl group -4- methoxyphenyl sulphonyls amido) -1- phenyl butane -2- bases)-acetamide.
The synthetic route of compound 1 is shown in that Fig. 2, the analysis result of compound 1 are as follows：LC-MS(ESI,M+H+)m/z 601.7。1H NMR(400MHz,CDCl3)δ7.81(s,1H),7.62(s,1H),7.18-7.12(m,7H),7.01(brs, 2H),4.50(m,2H),4.25-4.23(m,1H),4.06(m.1H),3.90(s,3H),3.37(m,2H),3.04-2.88(m, 4H),1.92(m,1H),0.82(s,6H).13CNMR(101MHz,CDCl3)δ174.2,165.3,162.8,153.6,147.4, 138.1,137.0,130.3,129.5,128.3,127.0,114.3,71.6,57.5,55.7,54.6,47.6,34.5,26.8, 20.0。
Embodiment 2
8- methyl purines (1.34g, 10mmol) are added in 50mL eggplant-shape bottles, 10mL anhydrous DMFs are added, are placed in ice bath In, sodium hydride (0.44g, 11mmol) is slowly added portionwise, finishes and is transferred to room temperature and carries out reaction 1h, bromoacetic acid second is slowly added dropwise Ester (1.84g, 11mmol), the reaction was continued 1h.Reaction is terminated, 10mL water is added, is extracted with ethyl acetate (15 × 3mL), it is organic It is mutually dried with anhydrous sodium sulfate, is concentrated to give yellow oily crude product.Through flash column purifications, (eluant, eluent used is ethyl acetate to crude product And hexane, by volume, ethyl acetate：Hexane=1：2) colorless needle crystals, that is, intermediate 2 (1.53g, 69.6%) is obtained.It is intermediate LC-MS (ESI, the M+H of body 2+)m/z 221.4。
NaOH (0.65g, 16.2mmol) is dissolved in 5mL water, is added to containing intermediate 2 (1.19g, 5.4mmol) In 100mL eggplant-shape bottle, reaction 1h is stirred at room temperature, eggplant-shape bottle is placed in ice bath, under stirring with 4M HCl adjust pH to 4.0,30min is stirred under ice bath, a large amount of solids are precipitated, is stood, is filtered, and filter cake is washed with water, dry, obtains target product white powder Last solid, that is, intermediate 3 (0.58g, 56.3%).LC-MS (ESI, the M-H of intermediate 3-)m/z 191.4。
Intermediate b2 (3.19g, 6.30mmol) is added in the there-necked flask of 50mL dryings, the anhydrous chloroforms of 8mL is added, It stirs evenly, is slowly added to Sodium azide (0.45g, 6.90mmol) under protection of argon gas, reaction bulb is placed in ice bath, at 0 DEG C It is lower to be slowly added dropwise into the 1.53mL concentrated sulfuric acids with dropping funel, it is added dropwise in 5min.Then reaction bulb is transferred to 45 DEG C of conditions Under be stirred to react overnight.TIC monitors reaction process, and reaction is quenched with water, and chloroform extracts (25 × 3mL), merges organic phase, uses 20mL saturated common salt water washings, dry, organic phase concentration.Crude product is through silica gel column chromatography purifying ethyl acetate-methanol (30:1) Target product white solid, that is, intermediate b3 (2.16g, 68%), LC-MS (ESI, M+H+) m/z 506.4 of intermediate b3.
The synthesis of intermediate B 2a differs only in reference to the synthesis of the intermediate B 1a of embodiment 1 and replaces intermediate b2 For equimolar intermediate b3, intermediate B 2a, yield 54.3%, LC-MS (ESI, M+H+) m/z of intermediate B 2a are obtained 406.3。
Compound 2 synthesis with reference to 1 compound 1 of embodiment synthesis, with the synthesis of compound 1 differ only in will in Mesosome 1 replaces with equimolar intermediate 3, and B1a is replaced with equimolar B2a, obtains compound 2：(8- methyl -9H- is fast by 2- Purine -9- bases)-N- [(2S, 3R) -3- hydroxyls -4- (N- isobutyl group -4- methoxyphenyl sulphoxide imines base) -1- phenyl butyls -2- Base]-acetamide, yield 68.7%.
The synthetic route of compound 2 is shown in that Fig. 3, the analysis result of compound 2 are as follows：LC-MS(ESI,M+H+)m/z 583.7。1H NMR (600MHz, Acetone) δ 7.84 (s, 1H), 7.73 (d, J=8.8Hz, 2H), 7.63 (s, 1H), 7.26- 7.24 (m, 2H), 7.21-7.18 (m, 3H), 7.03 (d, J=8.8Hz, 2H), 4.04 (td, J=9.2,4.8Hz, 1H), 3.84 (s, 3H), 3.79 (m, 1H), 3.24 (dd, J=15.0,3.3Hz, 1H), 3.01 (ddd, J=23.4,14.4,6.0Hz, 2H), 2.91-2.86 (m, 2H), 2.67 (dd, J=13.8,10.6Hz, 1H), 1.92-1.88 (m, 1H), 0.86 (d, J=6.6Hz, 6H), 0.84 (d, J=6.6Hz, 6H) .13C NMR (151MHz, Acetone) δ 166.8,163.7,157.3,149.5, 145.9,141.3,139.4,132.0,130.2,130.0129.5,129.0,126.9,115.0,72.7,58.5,56.2, 55.1,53.5,46.6,35.4,27.5,20.1,20.0。
Embodiment 3
The synthesis of synthesis reference 2 intermediate 2 of embodiment of intermediate 4, with differing only in 8- for the synthesis of intermediate 2 Methyl purine replaces with equimolar uracil, obtains 4 white powder solid of intermediate, yield 49.1%, structural formula such as Fig. 3 It is shown.LC-MS (ESI, the M+H of intermediate 4+)m/z 199.3。
Intermediate 5 synthesis with reference to 2 intermediate 3 of embodiment synthesis, with the synthesis of intermediate 3 differ only in will in Mesosome 2 replaces with equimolar intermediate 4, obtains 5 white powder solid of intermediate, yield 61.2%.The LC-MS of intermediate 5 (ESI,M-H)m/z 169.4。
Compound 3 synthesis with reference to 1 compound 1 of embodiment synthesis, with the synthesis of compound 1 differ only in will in Mesosome 1 replaces with equimolar intermediate 5, obtains compound 3：2- (2,4- dioxo -2H-3,4- dihydro-pyrimidin -1- bases)-N- ((2S, 3R) -3- hydroxyls -4- (N- isobutyl group -4- methoxyphenyl sulphonyls amido) -1- phenyl butane -2- bases)-acetamide is received Rate 97.5%.
The synthetic route of compound 3 is shown in that Fig. 4, the analysis result of compound 3 are as follows：LC-MS(ESI,M+H+)m/z 559.71H NMR(500MHz,CDCl3) δ 8.89 (s, 1H), 7.75 (d, J=8.6Hz, 2H), 7.29-7.27 (m, 3H), 7.23-7.22 (m, 2H), 7.01 (d, J=8.6Hz, 2H), 6.70 (d, J=7.8Hz, 1H), 5.68 (d, J=7.8Hz, 1H), 4.26 (d, J=15.4Hz, 2H), 4.17 (d, J=15.7Hz, 1H), 3.99-3.98 (m, 1H), 3.90 (s, 3H), 3.13 (d, J =5.7Hz, 2H), 2.94-2.86 (m, 4H), 1.93-1.84 (m, 1H), 0.90 (d, J=6.0Hz, 6H) .13C NMR (101MHz,CDCl3)δ166.6,163.9,163.1,151.3,144.9,137.9,130.0,129.5,129.4,128.5, 126.5,114.4,102.6,72.3,58.3,55.7,54.1,53.0,50.4,34.7,27.0,20.1,20.0。
Embodiment 4
Anhydrous solvent n,N-Dimethylformamide (DMF) 5mL is added in 25mL eggplant-shape bottles, under stirring slowly 5 FU 5 fluorouracil (1.05g, 8.0mmol, Beijing are coupled Science and Technology Ltd.) is added, stirs evenly and is placed in ice bath, slowly Sodium hydride (0.23g, 9.6mmol) is added portionwise, finishes to be transferred to and reacts 1h at room temperature, be slowly added dropwise into bromoacetate (1.60g, 9.6mmol), the reaction was continued 2h.Filtering, is washed three times, merging filtrate with water (5 × 3mL), and 0.96g hydroxides are added Sodium (24.0mmol) is stirred at room temperature reaction 1h, is placed in ice bath, adjusts pH to 3.0 with 4M HCl, 30min is stirred under ice bath, quiet It sets, filters, filter cake is washed with water, dry, obtains target product white powder solid i.e. intermediate 6 (0.58g, 38.7%).It is intermediate LC-MS (ESI, the M-H of body 6-)m/z 187.4。
Compound 4 synthesis with reference to 1 compound 1 of embodiment synthesis, with the synthesis of compound 1 differ only in will in Mesosome 1 replaces with equimolar intermediate 6, obtains compound 4：2- (the fluoro- 2,4- dioxos -2H-3,4- dihydro-pyrimidins -1- of 5- Base)-N- ((2S, 3R) -3- hydroxyls -4- (N- isobutyl group -4- methoxyphenyl sulphonyls amido) -1- phenyl butane -2- bases)-acetyl Amine, yield 96.0%.
The synthetic route of compound 4 is shown in that Fig. 5, the analysis result of compound 4 are as follows：LC-MS(ESI,M+H+)m/z 577.7。1H NMR(500MHz,CDCl3) δ 10.44 (s, 1H), 8.07 (s, 1H), 7.79 (d, J=7.4Hz, 2H), 7.44- 7.38 (m, 1H), 7.33-7.24 (m, 1H), 7.15-7.14 (m, 2H), 7.06 (d, J=7.4Hz, 1H), 6.98 (d, J= 7.4Hz, 2H), 4.33 (td, J=9.4,4.8Hz, 1H), 4.29-4.20 (m, 2H), 4.16-4.13 (m, 1H), 4.01 (m, 1H),3.85(s,3H),3.29-3.26(m,1H),3.20-3.10(m,2H),2.97-2.96(m,1H),2.93-2.90(m, 2H),1.99-1.92(m,1H),0.90(brs,6H).13C NMR(101MHz,CDCl3)δ166.5,162.9,157.9, 150.0,139.1,138.0,130.0,129.5,129.4,128.4,126.4,114.4,72.3,58.1,55.7,54.1, 52.9,50.1,36.6,26.9,20.1,20.0。
Embodiment 5
Triphosgene (0.15g, 0.50mmol) is weighed in the eggplant-shape bottle that 50mL is dried, is placed in ice bath, in nitrogen protection Under be slowly added to 4mL anhydrous methylene chloride solvents, stirred under ice bath.By Zalcitabine (0.106g, 0.50mmol) It is dissolved in 4mL anhydrous tetrahydro furan solvents with n,N-diisopropylethylamine (0.065g, 0.50mmol).Under condition of ice bath slowly It is added dropwise in the dichloromethane solution for filling triphosgene, finishes, be transferred to room temperature reaction 30min.
The synthesis of intermediate b4 is similar with the synthesis of intermediate b2, differs only in and replaces with 4- Methoxybenzenesulfonyl chlorides Equimolar 4- nitrobenzene sulfonyl chlorides；
The synthesis of intermediate B 1b is similar with the synthesis of B1a, differs only in intermediate b2 replacing with equimolar centre Body b4；
Intermediate B 1b (0.23g, 0.55mmol) is dissolved in 4mL anhydrous methylene chloride solvents, N, N- diisopropyls are added dropwise to Base ethamine (0.071g, 0.55mmol) is stirred at room temperature reaction 5min, is then slowly added dropwise under condition of ice bath into three chloromethane of carbonic acid Base ester, Zalcitabine reaction solution in, finish, be transferred to room temperature reaction 1h.Reaction is terminated, is added into reaction solution 10mL dichloromethane washs (20 × 3mL) with saturated ammonium chloride solution, and organic phase is dried with anhydrous sodium sulfate, is concentrated to give crude product. Crude product through silica gel prepare thin-layer chromatography isolated and purified (solvent be ethyl acetate, hexane and ammonium hydroxide, by volume, acetic acid Ethyl ester：Hexane：NH3·H2O=2：3：0.1%) white powder solid, that is, intermediate 7 (0.078g, 23.7%) is obtained.Intermediate 9 LC-MS(ESI,M+H+)m/z 659.6。
Intermediate 7 (0.040g, 0.06mmol) is dissolved in the in the mixed solvent of 1mL methanol and 1mL ethyl acetate, first is added Sour ammonium (0.031g, 0.49mmol), 10%Pd/C (0.040g, humidity 36.68%), heating reflux reaction one hour, while hot mistake Filter reaction solution, methanol washing, used after concentration prepare thin-layer chromatography isolated and purified (solvent for ethyl acetate and methanol, By volume, ethyl acetate：Methanol=4：1) white powder solid (0.035g, 91.4%), i.e. compound 5, are obtained：((2S, 5R) -5- (- 1 base of 1H-2- oxo -4- aminopyrimidines)-tetrahydrofuran -2- bases) ((N- is different by (2S, 3R) -3- hydroxyls -4- for methyl N - Butyl -4- aminocarbonyl phenyls sulfoamido) -1- phenyl butane -2- bases))-carbamate.
The synthetic route of compound 5 is shown in that Fig. 6, the analysis result of compound 5 are as follows：LC-MS(ESI,M+H+)m/z 629.7。1HNMR(500MHz,CDCl3) δ 9.02 (s, 1H), 7.65 (d, J=8.6Hz, 2H), 7.29-7.27 (m, 3H), 7.23- 7.22 (m, 2H), 6.61 (d, J=8.6Hz, 2H), 5.48 (d, J=7.8Hz, 1H), 5.23-5.19 (m, 1H), 4.80-4.75 (m, 1H), 4.52-4.47 (m, 2H), 4.17 (d, J=15.7Hz, 1H), 3.99-3.98 (m, 1H), 3.89 (s, 3H), 3.13 (d, J=5.7Hz, 2H), 2.99-2.84 (m, 6H), 1.93-1.84 (m, 2H), 1.71-1.68 (m, 1H), 0.93 (d, J= 6.6Hz, 3H), 0.91 (d, J=6.6Hz, 3H) .13C NMR (101MHz, MeOD) δ 169.6,168.2,164.5,158.9, 147.8,140.0,132.1,130.7,130.5,129.4,129.1,111.6,97.3,95.7,81.2,73.5,70.5, 56.2,54.0,52.4,36.3,30.2,28.1,20.6,20.5.
Embodiment 6
Compound 6 synthesis with reference to 1 compound 1 of embodiment synthesis, with the synthesis of compound 1 differ only in will in Mesosome 1 replaces with equimolar intermediate 6, and intermediate B 1a is replaced with equimolar intermediate B 3a, obtains compound 6：N- ((2R, 3S) -3- (2- (the fluoro- 2,4- dioxos -2H-3,4- dihydro-pyrimidins -2H-1- bases of 5-) acetamido) -2- hydroxyl -4- benzene Base butyl-N- isobutyl groups-P- (4- methoxyphenyls) phosphamic acid, yield 70.2%.
The building-up process of intermediate B 3a is：
4- methoxyphenyls phosphonyl dichloride (0.85g, 4.16mmol), 1-H- tetrazoles (0.027g, 0.038mmol) is molten In 8mL drying benzene in, reaction bulb is placed in ice bath, be slowly added dropwise successively under protection of argon gas into benzyl alcohol (0.39mL, 3.78mmol), DIEA (0.72mL, 4.16mmol).Finish, 30min be stirred to react in ice bath, be then transferred at room temperature after Continuous reaction 2.5 hours.Reaction bulb is placed in ice bath, be slowly added to successively 4 (1.27g, 3.78mmol), DIEA (0.72mL, 4.16mmol), be stirred to react 30min in ice bath, after be transferred to that the reaction was continued at room temperature 3.5 hours.Filtering is adopted after filtrate concentration (MeOH/H is isolated and purified with reversed-phase column chromatography2O:5/1) white powder i.e. intermediate b5 (0.95g, 42.3%), is obtained, in LC-MS (ESI, M+H+) m/z 597.7 of mesosome b5；
By intermediate b5 (0.10g, 0.17mmol), 1mL tetrahydrofurans, 0.010g 10%Pd/C, K2CO3(0.038g, 0.27mmol) and 1mL water is added in dry hydrogenation bottle, is vigorously stirred under protection of argon gas uniformly, is then replaced 3 with hydrogen It is secondary, room temperature catalytic hydrogenation 4 hours under 40psi pressure.Reaction solution filtering with microporous membrane, filtrate are placed in ice bath, use 2M HCl adjust pH to 5.0, stir 30min, are extracted with ethyl acetate (5 × 3mL), organic phase is dried with anhydrous sodium sulfate, dense Contracting, obtains white powder solid i.e. intermediate b6 (0.072g, 83.9%), LC-MS (ESI, the M+H of intermediate b6+)m/z 507.7；
The synthesis of intermediate B 3a differs only in reference to the synthesis of the intermediate B 1a of embodiment 1 and replaces intermediate b2 For equimolar intermediate b6, intermediate B 3a, yield 54.3%, LC-MS (ESI, M+H+) m/ of intermediate B 3a are obtained z406.7。
The synthetic route of compound 6 is shown in that Fig. 7, the analysis result of compound 6 are as follows：LC-MS(ESI,M+H+)m/z 577.7。1H NMR(500MHz,CDCl3) δ 10.21 (s, 1H), 8.04 (s, 1H), 7.75 (d, J=7.4Hz, 2H), 7.44- 7.38 (m, 1H), 7.33-7.24 (m, 2H), 7.03-6.99 (m, 2H), 6.98 (d, J=7.4Hz, 2H), 4.28 (td, J= 9.4,5.0Hz,1H),4.28-4.21(m,2H),4.19-4.15(m,1H),4.03(m,1H),3.85(s,3H),3.29-3.26 (m,1H),2.97-2.96(m,1H),2.93-2.90(m,2H),2.63-2.52(m,2H),1.95-1.87(m,1H),0.84 (d, J=6.6Hz, 3H), 0.79 (d, J=6.6Hz, 3H) .13C NMR (101MHz, CDCl3)δ168.2,163.3,157.7, 150.0,139.0,138.2,133.5,133.2,131.1,129.5,128.4,126.4,72.3,58.1,56.1,55.7, 52.9,48.5,36.2,27.1,20.1,20.0。
Embodiment 7
Compound 1~6 prepared by Examples 1 to 6 is dissolved with DMSO, is used in combination distilled water to carry out gradient dilution and obtains not Solution with concentration measures above compound to HIV-1 protease inhibiting activities and cell toxicant by the following method as sample Property.
According to document (Dong Biao, Zhang Tian, foundation [J] China of pottery pendant treasure's high throughput fluorescence substrate HIV-1 Protease models AIDS venereal disease, 2006 (05):Method 402-405.) carries out compound 1~6 test of HIV-1 protease inhibiting activities：
Substrate is (Arg-Glu (EDANS)-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys (DABCYL)- Arg) (AnaSpec), substrate point of contact both sides mark Edans and Dabcyl chromophories respectively.The fluorescence hair color spectrum of Edans with The absorption spectrum of Dabcyl is overlapped, and fluorescent quenching is generated by fluorescence resonance energy transfer in distance close enough, is made complete Substrate almost without fluorescence.When fluorogenic substrate is after hiv protease is cut, Edans chromophories are far from Dabcyl groups, fluorescence Quenched conditions disappear, and at this moment Edans just generates fluorescence under the exciting light of 340nm at 490nm, after untested compound is added, Then substrate product is reduced when compound is strong to enzyme inhibition activity, and fluorescence intensity reduces, otherwise fluorescence intensity increases.
According to document (Dong Biao, Zhang Tian, foundation [J] China of pottery pendant treasure's high throughput fluorescence substrate HIV-1 Protease models AIDS venereal disease, 2006 (05):Method 402-405) carries out sample with 96 orifice plates the measurement of HIV-1PR inhibitory activity, often Substrate (5 μM) and 185 μ L of buffer solution is added in hole, and 5 μ L sample solution are added, and measures blank and absorbs, 10 μ L HIV-1PR are added, incubate The absorbance for measuring 490nm wavelength after 5min is educated, the inhibiting rate of sample under each concentration is calculated, is calculated with Graphpad softwares Obtain IC50Value is the positive with DRV (Darunavir) (being purchased from U.S. ARP (American Research Product) company) Control.
Wherein, HIV-1PR is according to (expression, purifying and its inhibitor in-vitro screening side of the .HIV-1 protease such as Wang Yunhua Method establishes CHINA virus March phase .2006 of volume 21 2) in method in expression in escherichia coli and purify, HIV-1PR Use PD-10 column desalinations.
With HIV-1 protease inhibitors DRV (Darunavir) be positive control, according to the method described above measure embodiment 1~ 1~6 pair of protease (PR) inhibitory activity of compound prepared by 6 and cytotoxicity.The result shows that compound 2, compound 5 and change It closes object 6 and positive control medicine HIV-1 protease inhibitors DRV is better than to the inhibitory activity of HIV-1 protease；Compound 1, chemical combination Object 3 and compound 4 live to the inhibitory activity of HIV-1 protease and the inhibition of positive control medicine HIV-1 protease inhibitors DRV Property it is close or quite (table 1).
The inhibitory activity of 1 1~6 pair of HIV-1 protease of compound of table
Compound PRIC50(nM)
Compound 1 2.11±0.37
Compound 2 0.94±0.32
Compound 3 2.53±0.42
Compound 4 1.95±0.32
Compound 5 0.46±0.22
Compound 6 0.032±0.019
DRV 1.12±0.59
The test method of cytotoxicity：
Compound Cytotoxicity is measured using kit Cell Counting Kit-8 (CCK-8 kits).With 96 orifice plates Cytotoxicity test is carried out to sample, 20,000,293T cells are added per hole, incubation is added 1 μ L samples, continues to be incubated afterwards for 24 hours For 24 hours, be added 10 μ L CCK-8,2h after with measure absorbance under 450nm, calculate the percentage of each concentration survivaling cell, use CC is calculated in Graphpad softwares50Value, using DMSO as blank control, with DRV (Darunavir) for positive control.
The cytotoxicity of 2 compound 1~6 of table
The result shows that above compound 1~6 all has lower cytotoxicity (table 2).
Embodiment 8
Compound 1~6 is dissolved with DMSO, is used in combination distilled water to carry out gradient dilution and obtains the solution conduct of various concentration Sample measures above compound to HIV-1 albumen enzyme inhibition rates by the following method.
Using 293T cells as virus host, determination sample inhibits pseudotype virus reporter gene uciferase activity.It adopts With envelope glycoprotein pHCMV-G (VSV-G) the cotransfections 293T of pNL-Luc-E- Strain and expression vesicular stomatitis virus Generate VSV-G-HIV.
According to document, [Wang Ping, Chen Huan, Luo Ronghua wait .VSVG/HIV-1NL4-3Luc pseudovirus screens anti-HIV-1 medicines Condition optimizing and application [J] Chinese Pharmacological Bulletins, 2016,32 (3)：433-438.] method carry out HIV-1 protease suppressions The test of rate processed：
Envelope glycoprotein pHCMV-G (VSV-G) plasmid of plasmid pNL-Luc-E- and expression vesicular stomatitis virus is total It transfects 293T cells and prepares VSV-G-HIV pseudovirus, a certain concentration protease inhibitor sample is added after transfecting 5h, sets 5% CO2, 37C cultures 48 hours.10 μ l inoculation 293T cells of supernatant, 96 well culture plate is taken per hole later, after cultivating 48 hours, Uciferase activity in infection cell is measured, the inhibiting rate of each sample is calculated.
With HIV-1 protease inhibitors DRV (Darunavir) for positive control, according to the method described above under 10nM concentration Determine the inhibiting rate of 1~6 pair of hiv protease (PR) of above compound.The result shows that above-mentioned 6 compounds all have it is higher Inhibiting rate, wherein compound 5 and 6 inhibiting rate of compound (table 3) be higher than positive control.As shown in table 3.
The inhibiting rate of 3 1~6 pair of HIV-1 protease of compound of table
Embodiment 9
Compound 1~6 is dissolved with DMSO, is used in combination distilled water to carry out gradient dilution and obtains the solution conduct of various concentration Sample measures above compound to HIV-1 reverse transcriptase inhibiting rate by the following method.
The test method of HIV-1 reverse transcriptase inhibitory activity is as follows：
Primer is 5 ＇-CAG CAG TACAAATGG CAG TATTC-3 ＇, is marked with Cyanine 5 (Cy5) in the positions T19 Note, with 3 ＇-TGT CGT CAT GTT TAC CGT CATAAG TAGGTG TTACTAGTC CGATTT CCC CTAGTC CGACCCATG-5 ＇ are template, are marked with carboxytetramethylrhodamine (TMR) in the positions T2, TMR excitation wavelengths For 540nm, launch wavelength 580nm, using FRET as donor.The measurement of HIV-1RT inhibitory activity is carried out to sample with 96 orifice plates, 100nM HIV-1RT are mixed with the bis- index object/template composites of 100nM, and kinetics of polymerization reaction is opened by 100 μM of dNTPs are added It is dynamic.HIV-1RT and double index object/templates is added, the absorbance of 580nm wavelength is measured after incubation 5min, calculates each concentration IC is calculated with Graphpad softwares in the inhibiting rate of lower sample50Value, with EFV (Efavirenz) for positive control.If without spy Different explanation, all tests are carried out in buffer solution at 20 DEG C.Buffer solution 4M hydrochloric acid (pH7.5), 10mM KCl and 6mM MgCl is configured.
With HIV-1 reverse transcriptase inhibitor EFV (Efavirenz) for positive control, determined under 10nM concentration above-mentioned The inhibiting rate of 1~6 pair of hiv reverse transcriptase (RT) of compound.The result shows that above-mentioned 6 compounds all have higher inhibiting rate, Wherein compound 5 and 6 inhibiting rate of compound (table 4) is suitable with positive control inhibiting rate.As shown in table 4.
The inhibiting rate of 4 1~6 pair of HIV-1 reverse transcriptase of compound of table
As seen from the above embodiment, the present invention provides a kind of nucleic acid base compound or its pharmaceutically acceptable salt, The compound or its pharmaceutically acceptable salt have the apparent activity for inhibiting hiv protease and/or reverse transcriptase；Toxicity is ground Study carefully and shows that it, with good druggability, shows that such compound has a good application prospect as anti-AIDS drug.Root According to the experimental data of embodiment it is found that the compound of the present invention has inhibition to live in HIV-1 protease and HIV-1 reverse transcriptase Property (table 1), and all have lower cytotoxicity (table 2).The nucleic acid base compound of the present invention or its is pharmaceutically acceptable Salt be expected to become while inhibiting double target spot inhibitor of hiv protease and reverse transcriptase.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered It is considered as protection scope of the present invention.

Claims (10)

1. a kind of nucleic acid base compound or its pharmaceutically acceptable salt have structure shown in Formulas I：
In Formulas I, R1For amino, methoxyl group, nitro, aminomethyl or methylol；
X is-CH2,-O- ,-S- or-NH-；
Y is
When X is-CH2When, R Ra, Rb, Rc or Rd；
When X is-O- ,-S- or-NH-, R is Rc or Rd；
The Ra isThe Rb isThe Rc isThe Rd is
Wherein, R2For hydrogen, hydroxyl, amino or halogen；R3For hydrogen, hydroxyl, amino, halogen or methyl；R4For hydrogen or methyl；R5For Hydrogen, hydroxyl, amino or phosphate group；
When Z is nitrogen, R6For hydrogen, hydroxyl, amino or phosphate group；When Z is oxygen or sulphur, R6It is not present.
2. nucleic acid base compound according to claim 1 or its pharmaceutically acceptable salt, which is characterized in that the core Soda acid based compound includes 2- (the chloro- 9H- purine -9- bases of 6-)-N- ((2S, 3R) -3- hydroxyls -4- (N- isobutyl group -4- methoxyl groups PhenylSulphon amido) -1- phenyl butane -2- bases)-acetamide, 2- (8- methyl -9H- purine -9- bases)-N- [(2S, 3R) -3- hydroxyls Base -4- (N- isobutyl group -4- methoxyphenyl sulphoxide imines base) -1- phenyl butyl -2- bases]-acetamide, 2- (2,4- dioxos - 2H-3,4- dihydro-pyrimidin -1- bases)-N- ((2S, 3R) -3- hydroxyls -4- (N- isobutyl group -4- methoxyphenyl sulphonyls amido) -1- Phenyl butane -2- bases)-acetamide, 2- (the fluoro- 2,4- dioxos -2H-3,4- dihydro-pyrimidins -1- bases of 5-)-N- ((2S, 3R) -3- Hydroxyl -4- (N- isobutyl group -4- methoxyphenyl sulphonyls amido) -1- phenyl butane -2- bases)-acetamide, ((2S, 5R) -5- (- 1 base of 1H-2- oxo -4- aminopyrimidines)-tetrahydrofuran -2- bases) methyl N-((2S, 3R) -3- hydroxyls -4- (N- isobutyl groups -4- Aminocarbonyl phenyl sulfoamido) -1- phenyl butane -2- bases))-carbamate or N- ((2R, 3S) -3- (2- (fluoro- 2,4- of 5- bis- Oxo -2H-3,4- dihydro-pyrimidin -2H-1- bases) acetamido) -2- hydroxy-4-phenyl butyl-N- isobutyl groups-P- (4- methoxyl groups Phenyl) phosphamic acid.
3. the preparation method of nucleic acid base compound as claimed in claim 1 or 2, includes the following steps：
When X is-CH in the compound of structure shown in Formulas I2, R be Ra or Rb when, by with structure shown in Formula II -1 compound with Amine derivative carries out condensation reaction under the action of catalyst, obtains the compound with structure shown in Formulas I；
In Formula II -1, R is Ra or Rb, and the Ra isThe Rb isWherein, R2For Hydrogen, hydroxyl, amino or halogen；R3For hydrogen, hydroxyl, amino, halogen or methyl；R4For hydrogen or methyl；
When X is-CH in the compound of structure shown in Formulas I2,-O-,-S- or-NH-, R be when being Rc or Rd, will have shown in Formula II -2 Compound, carbonic acid trichloromethyl ester and the amine derivative of structure carry out condensation reaction under the action of catalyst, obtain with Formulas I institute Show the compound of structure；
Formula II -2 R-XH,
In Formula II -2, X is-CH2,-O- ,-S- or-NH-；R is Rc or Rd, and the Rc isThe Rd isWherein, R2For hydrogen, hydroxyl, amino or halogen, R3For hydrogen, hydroxyl, amino, halogen or methyl；R4For hydrogen Or methyl；R5For hydrogen, hydroxyl, amino or phosphate group；When Z is nitrogen, R6For hydrogen, hydroxyl, amino or phosphate group；When Z is oxygen Or when sulphur, R6It is not any atom or group；
The amine derivative has structure shown in formula III：
In formula III, R1For amino, methoxyl group, nitro, aminomethyl or methylol；Y isOr
4. preparation method according to claim 3, which is characterized in that the compound with structure shown in Formula II -1 with Amine derivative is condensed under the catalytic action of carbodiimide hydrochloride, I-hydroxybenzotriazole and 4-dimethylaminopyridine Reaction；
The compound with structure shown in Formula II -2 is with carbonic acid trichloromethyl ester and amine derivative in N, N- diisopropyl second Condensation reaction is carried out under the catalytic action of amine.
5. preparation method according to claim 3, which is characterized in that the compound with structure shown in Formula II -1 Preparation method includes the following steps:
Substituted pyrimidines base or substituted purin base are subjected to substitution reaction with bromoacetic acid or bromoacetate, obtain substitution production Object；
The substitution product is hydrolyzed, the compound with structure shown in Formula II -1 is obtained；
The substituted pyrimidines base has structure shown in formula a：
The substituted purin base has structure shown in formula b：
6. preparation method according to claim 3, which is characterized in that when Y is in the amine derivativeWhen, it is described Amine derivative has structure shown in formula III -1：
The preparation method of the amine derivative with structure shown in formula III -1, includes the following steps：
[(S) -1- ((S)-ethylene oxide -2- bases) -2- benzene ethylaminos t-butyl formate] and isobutyl amine progress nucleophilic displacement of fluorine is anti- It answers, obtains (1S, 2R) -1- benzyl -2- hydroxyls -3- (isobutyl amine) t-butyl carbamate；
The R of (1S, 2R) -1- benzyls -2- hydroxyls -3- (isobutyl amine) t-butyl carbamates and structure shown in formula d by described in1Substitution Benzene sulfonyl chloride carries out nucleophilic substitution, obtains the intermediate of structure shown in formula c；
By the intermediate Deprotection of structure shown in the formula c, the amine derivative with structure shown in formula III -1 is obtained；
7. preparation method according to claim 3, which is characterized in that when Y is in the amine derivativeWhen, institute Amine derivative is stated with structure shown in formula III -2：
The preparation method of the amine derivative with structure shown in formula III -2, includes the following steps：
[(S) -1- ((S)-ethylene oxide -2- bases) -2- benzene ethylaminos t-butyl formate] and isobutyl amine progress nucleophilic displacement of fluorine is anti- It answers, obtains (1S, 2R) -1- benzyl -2- hydroxyls -3- (isobutyl amine) t-butyl carbamate；
The R of (1S, 2R) -1- benzyls -2- hydroxyls -3- (isobutyl amine) t-butyl carbamates and structure shown in formula d by described in1Substitution Benzene sulfonyl chloride carries out nucleophilic substitution, obtains the intermediate of structure shown in formula c；
The intermediate Yu Sodium azide of structure shown in the formula c are subjected to imidization, obtain the intermediate of structure shown in formula e；
By the intermediate Deprotection of structure shown in the formula e, the amine derivative with structure shown in formula III -2 is obtained.
8. preparation method according to claim 3, which is characterized in that when Y is in the amine derivativeWhen, institute Amine derivative is stated with structure shown in formula III -3：
The preparation method of the amine derivative with structure shown in formula III -3, includes the following steps：
[(S) -1- ((S)-ethylene oxide -2- bases) -2- benzene ethylaminos t-butyl formate] and isobutyl amine progress nucleophilic displacement of fluorine is anti- It answers, obtains (1S, 2R) -1- benzyl -2- hydroxyls -3- (isobutyl amine) t-butyl carbamate；
By 4- substituted-phenyls phosphonyl dichloride, benzyl alcohol and (1S, the 2R) -1- benzyl -2- hydroxyls-with structure shown in formula f 3- (isobutyl amine) t-butyl carbamate carries out the intermediate that structure shown in formula g is obtained by the reaction in Arbuzov；
By the intermediate and H of structure shown in the formula g2Reduction reaction is carried out, the intermediate of structure shown in formula h is obtained；
By the intermediate Deprotection of structure shown in the formula h, the amine derivative with structure shown in formula III -3 is obtained.
9. nucleic acid base compound described in claims 1 or 2 or its pharmaceutically acceptable salt answering in preparing hiv inhibitor With the hiv inhibitor is using hiv protease and reverse transcriptase as target spot.
10. application according to claim 9, which is characterized in that the nucleic acid base compound or its is pharmaceutically acceptable Dosage of the salt in hiv inhibitor be 0.01~100nM.
CN201810493926.7A 2018-05-22 2018-05-22 A kind of nucleic acid base compound or its pharmaceutically acceptable salt and its preparation method and application Active CN108558883B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201810493926.7A CN108558883B (en) 2018-05-22 2018-05-22 A kind of nucleic acid base compound or its pharmaceutically acceptable salt and its preparation method and application

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201810493926.7A CN108558883B (en) 2018-05-22 2018-05-22 A kind of nucleic acid base compound or its pharmaceutically acceptable salt and its preparation method and application

Publications (2)

Publication Number Publication Date
CN108558883A true CN108558883A (en) 2018-09-21
CN108558883B CN108558883B (en) 2019-10-15

Family

ID=63539595

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201810493926.7A Active CN108558883B (en) 2018-05-22 2018-05-22 A kind of nucleic acid base compound or its pharmaceutically acceptable salt and its preparation method and application

Country Status (1)

Country Link
CN (1) CN108558883B (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113277991A (en) * 2021-02-03 2021-08-20 中国医学科学院医药生物技术研究所 Nitrogen heterocyclic ring amino derivative, preparation method thereof and anti-HIV-1 drug

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101062909A (en) * 2006-04-28 2007-10-31 北京大学 HIV-1prolease peptide mimic new inhibitor having biological activity, preparation method and usage
US20110003827A1 (en) * 2003-12-11 2011-01-06 Flentge Charles A HIV protease inhibiting compounds
WO2018064071A1 (en) * 2016-09-27 2018-04-05 Gilead Sciences, Inc. Therapeutic compositions for treatment of human immunodeficiency virus

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20110003827A1 (en) * 2003-12-11 2011-01-06 Flentge Charles A HIV protease inhibiting compounds
CN101062909A (en) * 2006-04-28 2007-10-31 北京大学 HIV-1prolease peptide mimic new inhibitor having biological activity, preparation method and usage
WO2018064071A1 (en) * 2016-09-27 2018-04-05 Gilead Sciences, Inc. Therapeutic compositions for treatment of human immunodeficiency virus

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113277991A (en) * 2021-02-03 2021-08-20 中国医学科学院医药生物技术研究所 Nitrogen heterocyclic ring amino derivative, preparation method thereof and anti-HIV-1 drug
CN113277991B (en) * 2021-02-03 2022-07-12 中国医学科学院医药生物技术研究所 Nitrogen heterocyclic ring amino derivative, preparation method thereof and anti-HIV-1 drug

Also Published As

Publication number Publication date
CN108558883B (en) 2019-10-15

Similar Documents

Publication Publication Date Title
US5296604A (en) Proline derivatives and compositions for their use as inhibitors of HIV protease
CN104530078B (en) Thieno [3, 2-d] pyrimidine derivative and preparation method and application thereof
CZ291054B6 (en) Pharmaceutical compositions comprising protease inhibitors, i.e. sulfonamides containing tetrahydrofuran ring
NZ500592A (en) 4,4-disubstituted-3,4-dihydro-2(1H)-quinazolinones useful as HIV reverse transcriptase inhibitors
CN101367749B (en) A set of mesidino mesitoyl derivant, preparation and application thereof
CN112920208B (en) Boric acid-containing indole aryl sulfone derivative and preparation method and application thereof
EP0484071A2 (en) Synergism of HIV reverse transcriptase inhibitors
CN112028836B (en) Diarylpyrimidine derivative containing six-membered nitrogen heterocycle and preparation method and application thereof
CN108218890A (en) A kind of five yuan of non-aromatic ring miazines HIV-1 reverse transcriptase inhibitor and its preparation method and application
CN108558883B (en) A kind of nucleic acid base compound or its pharmaceutically acceptable salt and its preparation method and application
CN108558808B (en) Amide derivative or pharmaceutically acceptable salt thereof, and preparation method and application thereof
CN108676009B (en) Pyrimidine derivatives as HER2 tyrosine kinase inhibitors and uses thereof
CN109369623B (en) Substituted 1,2,3 triazole diaryl pyrimidine derivative and preparation method and application thereof
CN112898193B (en) Indole aryl sulfone derivative and preparation method and application thereof
US10266489B2 (en) Pyrrolic amide compound and preparation method and application thereof
CN111205206B (en) Carbonyl compound containing amino acid connecting chain or pharmaceutically acceptable salt thereof, and preparation method and application thereof
CN108440500B (en) Quinazoline HIV-1 inhibitor and preparation method and application thereof
CN108409698A (en) Bis- target spot hiv inhibitors of RT/PR and its preparation method and application
CN108586482A (en) A kind of Diarylmiazines HIV-1 inhibitor of the ring containing triazole and its preparation method and application
WO2005087759A1 (en) Qiunolin-4-ones as inhibitors of retroviral integrase for the treatment of hiv, aids and aids related complex (arc)
CN114230630B (en) Triptolide derivative and application thereof
CN113620903A (en) 4- (benzothiazole-2-yl) -N-substituted aniline compound and preparation method and application thereof
CN112961081B (en) Bibenzamide urea compound and preparation method and application thereof
CN111647034B (en) 2-mercaptobenzamide thioester compound and preparation method and application thereof
US6313110B1 (en) Substituted 2H-1,3-diazapin-2-one useful as an HIV protease inhibitor

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant