An HIV inhibitor

Abstract

The present invention discloses a HIV inhibitor, which is a compound prepared by condensing BHA-Lys-Lys2-Lys4-Lys8-Lys16-32TFA and a capping group compound protected by tert-butyloxycarbonyl, and then removing the tert-butyloxycarbonyl protecting group. The compound shows in an in vitro anti-HIV-1 activity test that the compound has good solubility and anti-HIV-1 activity, and has no obvious cytotoxicity. In addition, the compound is based on simulating the CD4 receptor binding to the gp120 key domain Phe43 cavity, achieving antiviral effects on the key protein specific binding of the virus invading the cell. The compound can not only greatly avoid the disadvantage that the polyanionic polymer will promote the formation of amyloid fibers in semen, but also because it is a macromolecule, the transdermal effect is poor and it will not be absorbed by the mucous membrane into the blood circulation.

Description

Technical Field

[0001] The invention belongs to the field of medicine, and in particular relates to an HIV inhibitor. Background Art

[0002] The HIV-1 entry inhibitor targeting the gp120 target currently on the market is Rukobia (fostemsavir) from ViiV Healthcare, which was approved by the FDA in July 2020. Another series of compounds in the research and optimization stage are NBD-556 and NBD-557 and their derivatives. Currently, this type of compound has not yet entered clinical research.

[0003]

[0004] Patent document CN108676067A discloses compound IV, and its structural formula is shown below:

[0005]

[0006] It is an anion-terminated dendrimer compound, which also showed excellent anti-HIV-1 activity in in vitro activity tests. However, in clinical trials of HIV-1 infection caused by sexual transmission, this type of compound (anionic polymer) was found to be ineffective in preventing HIV-1 and may even increase the HIV-1 infection rate. The main reason is that anionic compounds can promote the formation of semen amyloid fibers, and semen amyloid fibers have a great role in promoting HIV-1 infection of target cells such as CD4+T cells. Summary of the invention

[0007] In order to overcome the problems existing in the prior art, one of the objectives of the present invention is to provide a compound represented by formula (I).

[0008] A second object of the present invention is to provide a method for preparing the compound represented by formula (I).

[0009] A third object of the present invention is to provide an HIV inhibitor.

[0010] A fourth object of the present invention is to provide a use of a compound represented by formula (I) in the preparation of a drug for preventing or treating sexually transmitted diseases.

[0011] In order to achieve the above object, the technical solution adopted by the present invention is:

[0012] The first aspect of the present invention is to provide a compound represented by formula (I) or a pharmaceutically acceptable salt, isomer, or solvate thereof:

[0013]

[0014] R is:

[0015]

[0016] Wherein, R1, R2 or R3 are independently selected from -H, C 1-6 Hydrocarbon, C 2-6 Unsaturated hydrocarbon, -CN, C 3-6 heterocyclic group or halogen.

[0017] Preferably, the C 1-6 Hydrocarbon groups include C 1-6 Straight chain hydrocarbon, C 3-6 Branched chain hydrocarbon, C 3-6 Cyclic hydrocarbon group.

[0018] Preferably, the C 2-6 Unsaturated hydrocarbon groups include C 2-6 Alkenyl, C 2-6 Alkynyl, phenyl.

[0019] Preferably, the C 3-6 Heterocyclic groups include C 3-6 Heterocyclic aromatic or C 3-6 Non-heterocyclic aromatic group; further preferably, the C 3-6 Heterocyclic groups include C 3-6 Heterocyclic aromatic or C 3-6 Non-heterocyclic aromatic group; said C 3-6 The heterocyclic ring of the heterocyclic group contains N, S or O atoms.

[0020] Preferably, the C 1-6 The hydrocarbon groups are -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)2, -C(CH3)3, -CH2CH2CH2CH3, -CH2CH(CH3)2, -CH(CH3)CH2CH3,

[0021] Preferably, the C 2-6 Unsaturated hydrocarbon groups are: -CH=CH2, -CH2=CHCH3, -CH2=C(CH3)2, -C≡CCH3 or

[0022] Preferably, the C 3-6 Heterocyclic groups include

[0023] Preferably, the compound is:

[0024]

[0025] Among them, R is

[0026] Preferably, the pharmaceutically acceptable salt of the compound comprises hydrochloride or hydrobromide.

[0027] The second aspect of the present invention is to provide a method for preparing the compound of formula (I) provided in the first aspect of the present invention, comprising the following steps:

[0028] The compound represented by formula (II) is subjected to a condensation reaction with the compound represented by formula (III), and then the tert-butyloxycarbonyl protecting group is removed by reduction to obtain the compound represented by formula (I);

[0029] The formula (II) is:

[0030] The formula (III) is:

[0031] Preferably, the condensation reaction is specifically: reacting the compound represented by formula (II) with the compound represented by formula (III) in the presence of an organic base and a condensation agent.

[0032] Preferably, the organic base comprises triethylamine or diisopropylethylamine.

[0033] Preferably, the condensing agent includes 1H-benzotriazol-1-yloxytripyrrolidino hexafluorophosphate.

[0034] Preferably, the condensation reaction further comprises the step of adding a solvent.

[0035] Preferably, the solvent comprises N,N-dimethylformamide or dimethyl sulfoxide.

[0036] Preferably, the preparation method further comprises a refining step; further preferably, the refining step comprises washing and recrystallization.

[0037] Preferably, the step of reducing and removing the tert-butyloxycarbonyl protecting group is specifically: removing the tert-butyloxycarbonyl protecting group on the amino group by hydrogen chloride or hydrogen bromide gas.

[0038] The third aspect of the present invention is to provide an HIV inhibitor, comprising the compound provided in the first aspect of the present invention or a pharmaceutically acceptable salt, isomer, or solvate thereof.

[0039] The compounds of the present invention exist in geometric isomers and stereoisomers. According to the specific examples of the "isomers" described in the present invention, for example, cis-trans isomers, enantiomers, diastereomers, tautomers, and racemic mixtures and other mixtures thereof, all of which belong to the scope of the present invention. The term "enantiomer" refers to stereoisomers that are mirror images of each other. The term "tautomer" refers to a type of functional group isomer that has different connection points through one or more double bond displacements, for example, a ketone and its enol form are keto-enol tautomers. The term "diastereomer" refers to a stereoisomer in which a molecule has two or more chiral centers and the molecules are non-mirror-image stereoisomers. The term "cis-trans isomer" refers to different configurations formed by different spatial arrangements in which double bonds or single bonds of ring carbon atoms in a molecule cannot rotate freely.

[0040] Preferably, the HIV inhibitor further comprises a pharmaceutically acceptable adjuvant; further preferably, the pharmaceutically acceptable adjuvant is a substance that is non-toxic, compatible with the active ingredient and otherwise biologically suitable for use in an organism. The choice of a specific adjuvant will depend on the mode of administration or the type and state of the disease used to treat a specific patient. Examples of pharmaceutically acceptable adjuvants include, but are not limited to, conventional solvents, diluents, dispersants, suspending agents, surfactants, isotonic agents, thickeners, emulsifiers, adhesives, lubricants, stabilizers, hydrating agents, emulsification accelerators, buffers, absorbents, colorants, ion exchangers, release agents, coating agents, flavoring agents, and antioxidants, etc., which are conventional in the pharmaceutical field. If necessary, flavoring agents, preservatives, sweeteners, etc., may also be added to the pharmaceutical composition.

[0041] Preferably, the HIV includes HIV-1 and HIV-2.

[0042] The fourth aspect of the present invention is to provide the use of the compound of formula (I) in the preparation of a drug for preventing or treating sexually transmitted diseases.

[0043] Preferably, the sexually transmitted disease is a vaginal or rectal sexually transmitted disease; further preferably, the sexually transmitted disease includes HIV virus infection; and even further preferably, the HIV virus includes HIV-1 and HIV-2.

[0044] The beneficial effects of the present invention are as follows: the compounds represented by formula (I) of the present invention have good solubility and anti-HIV-1 activity (EC 50 =10-500nmol / L) and no obvious cytotoxicity (CC 50>300 μmol / L). In addition, the compound in formula (I) simulates the CD4 receptor binding to the gp120 key domain Phe43 cavity as a basis, and achieves specific binding to the key protein of virus invasion into cells to exert antiviral effect. This type of compound can not only greatly avoid the disadvantage of polyanionic polymers that promotes the formation of amyloid fibers in semen, but also because it is a large molecule, the transdermal effect is poor, and it will not be absorbed by the mucous membrane into the blood circulation. In the clinical trial of local medication, it has shown the potential to effectively reduce the risk of HIV-1 infection. DETAILED DESCRIPTION

[0045] The specific implementation of the present invention is further described in detail below in conjunction with examples, but the implementation and protection of the present invention are not limited thereto. It should be noted that if there are processes that are not particularly described in detail below, they can be implemented or understood by those skilled in the art with reference to the prior art. If the manufacturer of the reagents or instruments used is not indicated, they are deemed to be conventional products that can be purchased commercially.

[0046] The general formula of the compound of formula (I) of the present invention is:

[0047]

[0048] Among them, R is

[0049]

[0050] wherein each R1, R2 or R3 is independently selected from -H, C 1-6 Hydrocarbon, C 2-6 Unsaturated hydrocarbon, -CN, C 3-6 Heterocyclyl, halogen.

[0051] The preparation method of the polylysine trifluoroacetate with diphenylamine as the core (Formula (II), the mother core) used to synthesize the compound of formula (I) is as follows: BHA-Lys-Lys2-Lys4-Lys8-Lys16-32TFA (BHA: diphenylamine, Lys: lysine, TFA: trifluoroacetic acid, the number in the lower right corner of Lys represents the number of lysines connected). The structural formula (II) is as follows:

[0052]

[0053] The precursor compounds of the end-capping group R used to synthesize the compound of formula (I) are listed below, which are respectively the formula (III) with amino group protected by tert-butyloxycarbonyl:

[0054]

[0055] The preparation method of the compound of formula (I) of the present invention is as follows: adding the compound of formula (II) and the compound of formula (III) into a solvent N,N-dimethylformamide (DMF) or dimethyl sulfoxide (DMSO), then adding an organic base triethylamine (Et3N) or diisopropylethylamine (DIPEA), and causing a condensation reaction under the action of a condensation agent 1H-benzotriazole-1-yloxytripyrrolidino hexafluorophosphate (PyBOP) to generate an amide bond, and separating the amide crude product through post-treatment, and then dispersing the amide crude product in methanol or ethanol, and introducing hydrogen chloride or hydrogen bromide gas to remove the tert-butyloxycarbonyl protection on the amino group to obtain the corresponding hydrochloride or hydrobromide, and refining the amide crude product to obtain the target compound of formula (I). The specific reaction formula is as follows:

[0056]

[0057] The end-capping group R can be prepared by the following preparation method, and the specific reaction formula is as follows:

[0058]

[0059] At room temperature, in N,N-dimethylformamide (DMF) solvent, phenylalanine derivatives with Boc-protected amino groups and leucine methyl ester hydrochloride or serine methyl ester hydrochloride are added in sequence, and then diisopropylethylamine (DIPEA) and 1H-benzotriazol-1-yloxytripyrrolidino hexafluorophosphate (PyBOP) are added, and the reaction is carried out at room temperature for 3-10 hours. After monitoring that the phenylalanine derivatives with Boc-protected amino groups are completely converted, water and dichloromethane are added to the system, and the liquids are separated to separate the aqueous phase and the organic phase, and the aqueous phase is extracted twice with dichloromethane, and the organic phases are combined and washed twice with saturated brine, and the solvent is removed by rotary evaporation under reduced pressure to obtain a crude condensation product of the phenylalanine derivatives with Boc-protected amino groups and leucine methyl ester or serine methyl ester. The crude product was dissolved in methanol, and sodium hydroxide aqueous solution (1 mol / L) was added, and hydrolysis was carried out at room temperature for 1-3 hours. After the hydrolysis of the crude leucine methyl ester or serine methyl ester was completed, dilute hydrochloric acid (0.5 mol / L) was used to adjust the pH of the reaction system to 4-5 (pH test paper detection), and dichloromethane was added. The liquid was separated to separate the aqueous phase and the organic phase. The aqueous phase was extracted twice with dichloromethane, and the organic phases were combined and washed twice with saturated brine. The solvent was removed by rotary evaporation under reduced pressure to obtain the target dipeptide crude product, which was then purified by silica gel column chromatography to obtain The total yield of the dipeptide end-capping group obtained by the above two-step synthesis method is 34%-77%, and the purity is 90%-98%.

[0060] When the end-capping group R is: When the compound in formula (I) is a compound, a solvate, a pharmaceutically acceptable salt or an isomer thereof in Table 1, wherein the pharmaceutically acceptable salt includes a hydrochloride or a hydrobromide, as shown in Table 1:

[0061] Table 1 Compound Series 1

[0062]

[0063]

[0064] When the end-capping group R is: When the compound in formula (I) is a compound, a solvate, a pharmaceutically acceptable salt or an isomer thereof in Table 2, wherein the pharmaceutically acceptable salt includes a hydrochloride or a hydrobromide, as shown in Table 2:

[0065] Table 2 Compound Series 2

[0066]

[0067]

[0068]

[0069] When the end-capping group R is: When the compound in formula (I) is a compound, a solvate, a pharmaceutically acceptable salt or an isomer thereof in Table 3, wherein the pharmaceutically acceptable salt includes a hydrochloride or a hydrobromide, as shown in Table 3:

[0070] Table 3 Compound Series 3

[0071]

[0072]

[0073] The present invention is further described in detail below with reference to embodiments:

[0074] Example 1. Preparation of Compound 1

[0075] The end-capping group R in compound 1 is: That is, formula (III) is Compound 1 can be prepared by the following reaction formula:

[0076]

[0077] Take compound formula (II) BHA-Lys-Lys2-Lys4-Lys8-Lys16-32TFA (1 mmol), dissolve it in N,N-dimethylformamide (DMF, 100 mL), add triethylamine (Et3N, 80 mmol), and stir to obtain solution A; take compound formula (III) (40 mmol) and 1H-benzotriazole-1-yloxytripyrrolidino hexafluorophosphate (PyBOP) (40 mmol), add N,N-dimethylformamide (DMF, 150 mL) to dissolve, and stir to obtain solution B. Add solution A to solution B at room temperature (20-35°C), and stir at room temperature (20-35°C) for 1 hour. Water (800 mL) was added to the reaction mixture to precipitate a white solid, which was filtered off with suction. The filter cake was washed with water (200 mL×3) several times. After drying, the wet product was dried in a vacuum oven at 60°C to constant weight to obtain a white solid crude product. The crude product was added to methanol (200 mL) and dispersed. The mixture was stirred at room temperature and HCl gas was continuously introduced for 1 hour. After the aeration was stopped, the reaction was allowed to react at room temperature overnight under nitrogen protection.

[0078] Methyl tert-butyl ether (800 mL) was dropped into the stirred reaction solution to precipitate a large amount of white solid, which was filtered off with suction. The filter cake was washed with methyl tert-butyl ether (100 mL×3). After the solid was dried, it was moved into an oven at 60° C. and dried under vacuum to constant weight to obtain a crude off-white solid of compound 1.

[0079] The off-white solid crude product of compound 1 was added to methanol (150 mL) and dissolved; tetrahydrofuran (150 mL) was added dropwise to the methanol solution of compound 1 with stirring at room temperature to precipitate a white solid, which was stirred for 15 minutes and then filtered off with suction. The filter cake was dried in an oven at 60° C. and dried under vacuum to constant weight to obtain 7.80 g of a refined product of compound 1 (MW: 10032.75 g / mol (hydrochloride salt), 8866.03 g / mol (free base)) with a purity of 99% and a yield of 78%.

[0080] Compound 1 was tested by H NMR spectrum, the instrument was Bruker AVANCE III 500 MHz, the solvent was heavy water, the obtained H spectrum data were: 1 H-NMR (D2O, 500MHz): δ6.92-7.30(m,170H), 5.96(s,1H), 4.05-4.51(m,63H), 2.73-3.32(m,126H), 1.45-1.83(m,186H).

[0081] Compound 1 was detected by mass spectrometry using DIONEX UltiMate 3000HPLC and Thermo QExactive. The data obtained were: LC-Ms(ESI): the molecule was multiply charged, [M+18H] 18+, [M+17H] 17+ , [M+16H] 16+ , [M+15H] 15+ ......[M+3H] 3+ , the molecular weight after deconvolution is: 8866.0519.

[0082] Compounds 2 to 93 can all be prepared by referring to the preparation method of compound 1, and the purity of the obtained products is 95-99%, and the yield is 60-92%.

[0083] Example 2. Preparation of Compound 6

[0084] The end-capping group R of compound 6 is: That is, formula (II) is Compound 6 is prepared by the following reaction formula:

[0085]

[0086] Compound (II) BHA-Lys-Lys2-Lys4-Lys8-Lys16-32TFA (0.5 mmol) was dissolved in dimethyl sulfoxide (DMSO, 50 mL), and diisopropylethylamine (DIPEA, 42 mmol) was added, and solution A was obtained under stirring; compound (III) (22 mmol) and 1H-benzotriazole-1-yloxytripyrrolidino hexafluorophosphate (PyBOP) (22 mmol) were added into dimethyl sulfoxide (DMSO, 80 mL) to dissolve, and solution B was obtained under stirring. Solution A was added into solution B at room temperature (20-35° C.), and the mixture was stirred at room temperature (20-35° C.) for 2 hours. Water (400 mL) was added to the reaction mixture to precipitate a white solid, which was filtered off with suction. The filter cake was washed with water (100 mL×3) several times. After drying, the wet product was dried in a vacuum oven at 60°C to constant weight to obtain a white solid crude product. The crude product was dispersed in ethanol (120 mL), stirred at room temperature and HCl gas was continuously introduced for 1 hour. After stopping the aeration, the reaction was allowed to react at room temperature overnight under nitrogen protection.

[0087] Methyl tert-butyl ether (450 mL) was dropped into the stirred reaction solution to precipitate a large amount of white solid, which was filtered off with suction. The filter cake was washed with methyl tert-butyl ether (60 mL×3). After the solid was dried, it was moved into an oven at 60°C and dried in vacuum to constant weight to obtain a crude off-white solid of compound 6.

[0088] The off-white solid crude product of compound 6 was added to methanol (100 mL) and dissolved; tetrahydrofuran (100 mL) was added dropwise to the methanol solution of compound 6 with stirring at room temperature to precipitate a white solid, which was stirred for 30 minutes and then filtered off with suction. The filter cake was dried in an oven at 60°C and dried under vacuum to constant weight to obtain 5.25 g of a refined product of compound 6 (MW: 11828.27 g / mol (hydrochloride salt), 10661.55 g / mol (free base)) with a purity of 97% and a yield of 89%.

[0089] Compound 6 was tested by H NMR spectrum, the instrument was Bruker AVANCE III 500 MHz, the solvent was heavy water, the obtained H spectrum data were: 1 H-NMR (D2O, 500MHz): δ6.98-7.30(m,138H), 5.99(s,1H), 4.01-4.55(m,63H) ), 2.70-3.30(m,126H), 1.46-1.80(m,186H), 1.35(s,144H),1.28(s,144H).

[0090] Compound 6 was detected by mass spectrometry using DIONEX UltiMate 3000HPLC and Thermo QExactive. The data obtained were as follows: LC-Ms (ESI): the molecule was multiply charged, [M+19H]19+, [M+18H]18+, [M+17H]17+, [M+16H]16+, [M+15H]15+......[M+4H]4+, and the molecular weight after deconvolution was 10661.5329.

[0091] Example 3. Preparation of Compound 25

[0092] The end-capping group R of compound 25 is: That is, formula (II) is Compound 25 was prepared by the following reaction formula:

[0093]

[0094] Take BHA-Lys-Lys2-Lys4-Lys8-Lys16-32TFA (0.5 mmol) of formula (II), dissolve it in dimethyl sulfoxide (DMSO, 50 mL), add diisopropylethylamine (DIPEA, 40 mmol), and stir to obtain solution A; take compound (III) (21 mmol) and 1H-benzotriazole-1-yloxytripyrrolidino hexafluorophosphate (PyBOP) (21 mmol), add dimethyl sulfoxide (DMSO, 70 mL) to dissolve, and stir to obtain solution B. Add solution A to solution B at room temperature (20-35°C), and stir at room temperature (20-35°C) for 1.2 hours. Water (400 mL) was added to the reaction mixture to precipitate a white solid, which was filtered off with suction. The filter cake was washed with water (90 mL×3) several times. After drying, the wet product was dried in a vacuum oven at 60°C to constant weight to obtain a white solid crude product. The crude product was added to ethanol (100 mL) and dispersed. The mixture was stirred at room temperature and HCl gas was continuously introduced for 1.5 hours. After the aeration was stopped, the reaction was allowed to react overnight at room temperature under nitrogen protection.

[0095] Methyl tert-butyl ether (400 mL) was dropped into the stirred reaction solution to precipitate a large amount of white solid, which was filtered off with suction and the filter cake was washed with methyl tert-butyl ether (50 mL×3). After the solid was dried by suction, it was moved into an oven at 60°C and dried in vacuum to constant weight to obtain a crude off-white solid of compound 25.

[0096] The off-white solid crude product of compound 25 was added to methanol (100 mL) and dissolved; tetrahydrofuran (120 mL) was added dropwise to the methanol solution of compound 25 with stirring at room temperature to precipitate a white solid, which was stirred for 45 minutes and then filtered off with suction. The filter cake was dried in an oven at 60° C. and dried under vacuum to constant weight to obtain 5.14 g of compound 25 (MW: 12403.95 g / mol (hydrochloride salt), 11237.23 g / mol (free base)) with a purity of 96% and a yield of 83%.

[0097] Compound 25 was detected by H NMR spectrum using Bruker AVANCE III 500 MHz and heavy water as solvent. The obtained H NMR spectrum data are as follows: 1 H-NMR (D2O, 500MHz): δ7.03-7.42(m,106H), 5.95(s,1H), 4.01-4.55(m,63H) ), 2.73-3.32(m,126H), 1.45-1.83(m,186H), 1.36(s,144H), 1.30(s,144H).

[0098] Compound 25 was detected by mass spectrometry using DIONEX UltiMate 3000HPLC and Thermo QExactive. The data obtained were: LC-Ms(ESI): the molecule was multiply charged [M+17H] 17+ , [M+16H] 16+ , [M+15H] 15+ ......[M+4H] 4+ , [M+3H] 3+ ,[M+2H]2 + , the molecular weight after deconvolution is: 11237.2264.

[0099] Example 4. Preparation of Compound 27

[0100] The end-capping group R of compound 27 is: That is, formula (II) is Compound 27 was prepared by the following reaction formula:

[0101]

[0102] Compound (II) BHA-Lys-Lys2-Lys4-Lys8-Lys16-32TFA (0.5 mmol) was dissolved in dimethyl sulfoxide (DMSO, 50 mL), and diisopropylethylamine (DIPEA, 43 mmol) was added, and solution A was obtained under stirring; compound (III) (22 mmol) and 1H-benzotriazole-1-yloxytripyrrolidino hexafluorophosphate (PyBOP) (22 mmol) were added into dimethyl sulfoxide (DMSO, 80 mL) to dissolve, and solution B was obtained under stirring. Solution A was added into solution B at room temperature (20-35° C.), and the mixture was stirred at room temperature (20-35° C.) for 1 hour. Water (400 mL) was added to the reaction mixture to precipitate a white solid, which was filtered off with suction. The filter cake was washed with water (100 mL×3) several times. After drying, the wet product was dried in a vacuum oven at 60°C to constant weight to obtain a white solid crude product. The crude product was added to ethanol (100 mL) and dispersed. The mixture was stirred at room temperature and HCl gas was continuously introduced for 1 hour. After the aeration was stopped, the reaction was allowed to react at room temperature overnight under nitrogen protection.

[0103] Methyl tert-butyl ether (400 mL) was dropped into the stirred reaction solution to precipitate a large amount of white solid, which was filtered off with suction and the filter cake was washed with methyl tert-butyl ether (50 mL×3). After the solid was dried by suction, it was moved into an oven at 60°C and dried in vacuum to constant weight to obtain a crude off-white solid of compound 27.

[0104] The off-white solid crude product of compound 27 was added to methanol (100 mL) and dissolved; tetrahydrofuran (110 ml) was added dropwise to the methanol solution of compound 27 with stirring at room temperature to precipitate a white solid, which was stirred for 20 minutes and then filtered off with suction. The filter cake was dried in an oven at 60° C. and dried under vacuum to constant weight to obtain 5.48 g of compound 27 (MW: 12628.59 g / mol (hydrochloride salt), 11461.87 g / mol (free base)) with a purity of 98% and a yield of 87%.

[0105] Compound 27 was detected by H NMR spectrum, the instrument was: Bruker AVANCE III 500MHz, the solvent was deuterated water, the H NMR spectrum data was: 1 H-NMR (D2O, 500MHz): δ7.00-7.51(m,106H), 5.97(s,1H), 3.98-4.53(m,63H) ), 2.71-3.33(m,126H), 1.42-1.80(m,186H), 1.37(s,144H), 1.29(s,144H).

[0106] Compound 27 was detected by mass spectrometry using DIONEX UltiMate 3000HPLC and Thermo QExactive. The data obtained were: LC-Ms(ESI): the molecule was multiply charged [M+18H] 18+ , [M+17H] 17+ , [M+16H] 16+ , [M+15H] 15+ ......[M+4H] 4+ , [M+3H] 3+ , the molecular weight after deconvolution is: 11461.8876.

[0107] Example 5. Preparation of Compound 29

[0108] The end-capping group R of compound 29 is: That is, formula (II) is Compound 29 was prepared by the following reaction formula:

[0109]

[0110] Compound (II) BHA-Lys-Lys2-Lys4-Lys8-Lys16-32TFA (0.4 mmol) was dissolved in dimethyl sulfoxide (DMSO, 50 mL), and diisopropylethylamine (DIPEA, 40 mmol) was added, and solution A was obtained under stirring; compound (III) (19 mmol) and 1H-benzotriazole-1-yloxytripyrrolidino hexafluorophosphate (PyBOP) (19 mmol) were added to dimethyl sulfoxide (DMSO, 75 mL) to dissolve, and solution B was obtained under stirring. Solution A was added to solution B at room temperature (20-35° C.), and the mixture was stirred at room temperature (20-35° C.) for 5 hours. Water (380 mL) was added to the reaction mixture to precipitate a white solid, which was filtered off with suction. The filter cake was washed with water (100 mL×3) several times. After drying, the wet product was dried in a vacuum oven at 60°C to constant weight to obtain a white solid crude product. The crude product was dispersed in ethanol (90 mL), stirred at room temperature and HCl gas was continuously introduced for 2 hours. After stopping the aeration, the reaction was allowed to react at room temperature overnight under nitrogen protection.

[0111] Methyl tert-butyl ether (380 mL) was dropped into the stirred reaction solution to precipitate a large amount of white solid, which was filtered off with suction. The filter cake was washed with methyl tert-butyl ether (50 mL×3). After the solid was dried by suction, it was moved into an oven at 60°C and dried in vacuo to constant weight to obtain a crude off-white solid of compound 29.

[0112] The off-white solid crude product of compound 29 was added to methanol (80 mL) and dissolved; tetrahydrofuran (120 mL) was added dropwise to the methanol solution of compound 29 with stirring at room temperature to precipitate a white solid, which was stirred for 20 minutes and then filtered off with suction. The filter cake was dried in an oven at 60° C. and dried under vacuum to constant weight to obtain 4.35 g of compound 29 (MW: 13895.88 g / mol (hydrochloride salt), 12729.16 g / mol (free base)) with a purity of 99% and a yield of 78%.

[0113] Compound 29 was detected by H NMR spectrum using Bruker AVANCE III 500 MHz and heavy water as solvent. The obtained H NMR spectrum data are as follows: 1 H-NMR (D2O, 500MHz): δ6.92-7.38(m,106H), 5.95(s,1H), 3.95-4.48(m,63H) ), 2.68-3.30(m,126H), 1.40-1.81(m,186H), 1.33(s,288H),1.28(s,288H).

[0114] Compound 29 was detected by mass spectrometry using DIONEX UltiMate 3000HPLC and Thermo QExactive. The data obtained were: LC-Ms(ESI): the molecule was multiply charged [M+17H] 17+ , [M+16H] 16+ , [M+15H] 15+ ......[M+4H] 4+ , [M+3H] 3+ , the molecular weight after deconvolution is: 12729.1585.

[0115] Example 6. Preparation of Compound 37

[0116] The end-capping group R of compound 37 is: That is, formula (II) is Compound 37 was prepared by the following reaction formula, as follows:

[0117]

[0118] Compound (II) BHA-Lys-Lys2-Lys4-Lys8-Lys16-32TFA (0.5 mmol) was dissolved in N,N-dimethylformamide (DMF, 50 mL), and triethylamine (Et3N, 40 mmol) was added, and solution A was obtained under stirring; compound (III) (20 mmol) and 1H-benzotriazole-1-yloxytripyrrolidino hexafluorophosphate (PyBOP) (20 mmol) were added to N,N-dimethylformamide (DMF, 90 mL) to dissolve, and solution B was obtained under stirring. Solution A was added to solution B at room temperature (20-35°C), and the mixture was stirred at room temperature (20-35°C) for 1 hour. Water (400 mL) was added to the reaction mixture to precipitate a white solid, which was filtered off with suction. The filter cake was washed with water (100 mL×3) several times. After drying, the wet product was dried in a vacuum oven at 60°C to constant weight to obtain a white solid crude product. The crude product was dispersed in methanol (120 mL), stirred at room temperature and HBr gas was continuously introduced for 1 hour. After stopping the aeration, the reaction was allowed to react at room temperature overnight under nitrogen protection.

[0119] Methyl tert-butyl ether (400 mL) was dropped into the stirred reaction solution to precipitate a large amount of white solid, which was filtered off with suction and the filter cake was washed with methyl tert-butyl ether (60 mL×3). After the solid was dried by suction, it was moved into an oven at 60°C and dried under vacuum to constant weight to obtain a crude off-white solid of compound 37.

[0120] The off-white solid crude product of compound 37 was added to methanol (80 mL) and dissolved; tetrahydrofuran (85 mL) was added dropwise to the methanol solution of compound 37 with stirring at room temperature to precipitate a white solid, which was stirred for 25 minutes and then filtered off with suction. The filter cake was dried and moved into an oven at 60°C under vacuum to constant weight to obtain 5.31 g of compound 37 (MW: 16871.79 g / mol (hydrobromide salt), 14282.67 g / mol (free base)) with a purity of 95% and a yield of 63%.

[0121] Compound 37 was detected by H NMR spectrum, the instrument was Bruker AVANCE III 500 MHz, the solvent was deuterated water, and the H NMR spectrum data were: 1 H-NMR (D2O, 500MHz): δ6.95-7.40(m,138H), 5.98(s,1H), 3.97-4.63(m,95H), 2.68-3. 30(m,126H), 1.40-1.81(m,282H), 1.34(s,144H), 1.30(s,144H), 0.59-0.72(m,192H).

[0122] Compound 37 was detected by mass spectrometry using DIONEX UltiMate 3000HPLC and Thermo QExactive. The data obtained were: LC-Ms(ESI): the molecule was multiply charged [M+16H] 16+ , [M+15H] 15+ ......[M+4H] 4+ , [M+3H] 3+ , the molecular weight after deconvolution is: 14282.6573.

[0123] Example 7. Preparation of Compound 56

[0124] The end-capping group R of compound 56 is: That is, formula (II) is Compound 56 is prepared by the following reaction formula, specifically:

[0125]

[0126] Compound (II) BHA-Lys-Lys2-Lys4-Lys8-Lys16-32TFA (0.3 mmol) was dissolved in N,N-dimethylformamide (DMF, 40 mL), and triethylamine (Et3N, 30 mmol) was added, and solution A was obtained under stirring; compound (III) (13 mmol) and 1H-benzotriazole-1-yloxytripyrrolidino hexafluorophosphate (PyBOP) (13 mmol) were added to N,N-dimethylformamide (DMF, 60 mL) to dissolve, and solution B was obtained under stirring. Solution A was added to solution B at room temperature (20-35°C), and the mixture was stirred at room temperature (20-35°C) for 1 hour. Water (300 mL) was added to the reaction mixture to precipitate a white solid, which was filtered off with suction. The filter cake was washed with water (70 mL×3) several times. After drying, the wet product was dried in a vacuum oven at 60°C to constant weight to obtain a white solid crude product. The crude product was added to methanol (80 mL) and dispersed. The mixture was stirred at room temperature and HBr gas was continuously introduced for 1 hour. After the aeration was stopped, the reaction was allowed to react overnight at room temperature under nitrogen protection.

[0127] Methyl tert-butyl ether (300 mL) was dropped into the stirred reaction solution to precipitate a large amount of white solid, which was filtered off and the filter cake was washed with methyl tert-butyl ether (50 mL×3). After the solid was dried by suction, it was moved into an oven at 60°C and dried in vacuo to constant weight to obtain a crude off-white solid of compound 56.

[0128] The off-white solid crude product of compound 56 was added to methanol (50 mL) and dissolved; tetrahydrofuran (60 mL) was added dropwise to the methanol solution of compound 56 with stirring at room temperature to precipitate a white solid, which was stirred for 30 minutes and then filtered off. The filter cake was dried in an oven at 60°C and dried under vacuum to constant weight to obtain 3.45 g of compound 56 (MW: 17447.47 g / mol (hydrobromide salt), 14858.35 g / mol (free base)) with a purity of 97% and a yield of 66%.

[0129] Compound 56 was detected by mass spectrometry using DIONEX UltiMate 3000HPLC and Thermo QExactive. The data obtained were: LC-Ms(ESI): the molecule was multiply charged [M+18H] 18+ , [M+17H] 17+ , [M+16H] 16+ , [M+15H] 15+ ......[M+4H] 4+ , [M+3H] 3+ , the molecular weight after deconvolution is: 14858.3649.

[0130] Example 8. Preparation of Compound 60

[0131] The end-capping group R of compound 60 is: That is, formula (II) is Compound 60 is prepared by the following reaction formula, specifically:

[0132]

[0133] Compound (II) BHA-Lys-Lys2-Lys4-Lys8-Lys16-32TFA (0.3 mmol) was dissolved in N,N-dimethylformamide (DMF, 40 mL), and triethylamine (Et3N, 30 mmol) was added, and solution A was obtained under stirring; compound (III) (12 mmol) and 1H-benzotriazole-1-yloxytripyrrolidino hexafluorophosphate (PyBOP) (12 mmol) were added to N,N-dimethylformamide (DMF, 50 mL) to dissolve, and solution B was obtained under stirring. Solution A was added to solution B at room temperature (20-35°C), and the mixture was stirred at room temperature (20-35°C) for 1 hour. Water (300 mL) was added to the reaction mixture to precipitate a white solid, which was filtered off with suction. The filter cake was washed with water (70 mL×3) several times. After drying, the wet product was dried in a vacuum oven at 60°C to constant weight to obtain a white solid crude product. The crude product was added to methanol (80 mL) and dispersed. The mixture was stirred at room temperature and HBr gas was continuously introduced for 1 hour. After the aeration was stopped, the reaction was allowed to react overnight at room temperature under nitrogen protection.

[0134] Methyl tert-butyl ether (300 mL) was dropped into the stirred reaction solution to precipitate a large amount of white solid, which was filtered off and the filter cake was washed with methyl tert-butyl ether (50 mL×3). After the solid was dried by suction, it was moved into an oven at 60°C and dried in vacuo to constant weight to obtain a crude off-white solid of compound 60.

[0135] The off-white solid crude product of compound 60 was added to methanol (55 mL) and dissolved; tetrahydrofuran (70 mL) was added dropwise to the methanol solution of compound 60 with stirring at room temperature to precipitate a white solid. The mixture was stirred for 20 minutes and then filtered off. The filter cake was dried in an oven at 60°C and dried under vacuum to constant weight to obtain 3.81 g of compound 60 (MW: 18666.99 g / mol (hydrobromide salt), 16077.87 g / mol (free base)) with a purity of 96% and a yield of 67%.

[0136] Compound 60 was detected by mass spectrometry using DIONEX UltiMate 3000HPLC and Thermo QExactive. The data obtained were: LC-Ms(ESI): the molecule was multiply charged [M+19H] 19+ , [M+18H] 18+ , [M+17H] 17+ , [M+16H] 16+, [M+15H] 15+ ......[M+4H] 4+ , the molecular weight after deconvolution is: 16077.8546.

[0137] Example 9. Preparation of Compound 89

[0138] The end-capping group R of compound 89 is: That is, formula (II) is Compound 89 is prepared by the following reaction formula, specifically:

[0139]

[0140] Compound (II) BHA-Lys-Lys2-Lys4-Lys8-Lys16-32TFA (0.4 mmol) was dissolved in dimethyl sulfoxide (DMSO, 50 mL), and diisopropylethylamine (DIPEA, 45 mmol) was added, and solution A was obtained under stirring; compound (III) (20 mmol) and 1H-benzotriazole-1-yloxytripyrrolidino hexafluorophosphate (PyBOP) (20 mmol) were added to dimethyl sulfoxide (DMSO, 80 mL) to dissolve, and solution B was obtained under stirring. Solution A was added to solution B at room temperature (20-35° C.), and the mixture was stirred at room temperature (20-35° C.) for 1.8 hours. Water (400 mL) was added to the reaction mixture to precipitate a white solid, which was filtered off with suction. The filter cake was washed with water (100 mL×3) several times. After drying, the wet product was dried in a vacuum oven at 60°C to constant weight to obtain a white solid crude product. The crude product was dispersed in ethanol (95 mL), stirred at room temperature and HCl gas was continuously introduced for 1.6 hours. After stopping the aeration, the reaction was allowed to react at room temperature overnight under nitrogen protection.

[0141] Methyl tert-butyl ether (400 mL) was dropped into the stirred reaction solution to precipitate a large amount of white solid, which was filtered off with suction and the filter cake was washed with methyl tert-butyl ether (50 mL×3). After the solid was dried by suction, it was moved into an oven at 60°C and dried in vacuum to constant weight to obtain a crude off-white solid of compound 89.

[0142] The off-white solid crude product of compound 89 was added to methanol (70 mL) and dissolved; tetrahydrofuran (115 mL) was added dropwise to the methanol solution of compound 89 with stirring at room temperature to precipitate a white solid. The mixture was stirred for 12 hours and then filtered off with suction. The filter cake was dried in an oven at 60°C under vacuum to constant weight to obtain 4.43 g of compound 89 (MW: 16581.87 g / mol (hydrochloride salt), 15415.15 g / mol (free base)) with a purity of 96% and a yield of 67%.

[0143] Compound 89 was detected by H NMR spectrum using Bruker AVANCE III 500 MHz and heavy water as solvent. The obtained H NMR spectrum data are as follows: 1 H-NMR (D2O, 500MHz): δ7.08-7.55(m,106H), 5.99(s,1H), 4.02-4.71(m,159 H), 2.93-3.28(m,64H), 1.48-1.77(m,186H), 1.35(s,144H), 1.31(s,144H).

[0144] Compound 89 was detected by mass spectrometry using DIONEX UltiMate 3000HPLC and Thermo QExactive. The data obtained were: LC-Ms(ESI): the molecule was multiply charged [M+19H] 19+ , [M+18H] 18+ , [M+17H] 17+ , [M+16H] 16+ , [M+15H] 15+ ......[M+4H] 4+ , [M+3H] 3+ , the molecular weight after deconvolution is: 15415.1396.

[0145] Example 10. Preparation of Compound 90

[0146] The end-capping group R of compound 90 is: That is, formula (II) is Compound 90 is prepared by the following reaction formula, specifically:

[0147]

[0148] Compound (II) BHA-Lys-Lys2-Lys4-Lys8-Lys16-32TFA (0.4 mmol) was dissolved in dimethyl sulfoxide (DMSO, 50 mL), and diisopropylethylamine (DIPEA, 42 mmol) was added, and solution A was obtained under stirring; compound (III) (22 mmol) and 1H-benzotriazole-1-yloxytripyrrolidino hexafluorophosphate (PyBOP) (22 mmol) were added to dimethyl sulfoxide (DMSO, 80 mL) to dissolve, and solution B was obtained under stirring. Solution A was added to solution B at room temperature (20-35° C.), and the mixture was stirred at room temperature (20-35° C.) for 2.5 hours. Water (400 mL) was added to the reaction mixture to precipitate a white solid, which was filtered off with suction. The filter cake was washed with water (100 mL×3) several times. After drying, the wet product was dried in a vacuum oven at 60°C to constant weight to obtain a white solid crude product. The crude product was dispersed in ethanol (90 mL), stirred at room temperature and HCl gas was continuously introduced for 2 hours. After stopping the aeration, the reaction was allowed to react at room temperature under nitrogen protection overnight.

[0149] Methyl tert-butyl ether (400 mL) was dropped into the stirred reaction solution to precipitate a large amount of white solid, which was filtered off with suction and the filter cake was washed with methyl tert-butyl ether (50 mL×3). After the solid was dried by suction, it was moved into an oven at 60°C and dried in vacuum to constant weight to obtain a crude off-white solid of compound 90.

[0150] The off-white solid crude product of compound 90 was added to methanol (80 mL) and dissolved; tetrahydrofuran (120 mL) was added dropwise to the methanol solution of compound 90 with stirring at room temperature to precipitate a white solid. The mixture was stirred for 16 hours and then filtered off with suction. The filter cake was dried in an oven at 60°C under vacuum to constant weight to obtain 4.91 g of compound 90 (MW: 17128.11 g / mol (hydrochloride salt), 15961.39 g / mol (free base)) with a purity of 98% and a yield of 72%.

[0151] Compound 89 was detected by mass spectrometry using DIONEX UltiMate 3000 HPLC and Thermo QExactive. The data obtained were: LC-Ms(ESI): the molecule was multiply charged [M+18H] 18+ , [M+17H] 17+ , [M+16H] 16+ , [M+15H] 15+ ......[M+4H] 4+ , [M+3H] 3+ , the molecular weight after deconvolution is: 15961.3982.

[0152] Example 11. Preparation of Compound 91

[0153] The end-capping group R of compound 91 is: That is, formula (II) is Compound 91 is prepared by the following reaction formula, specifically:

[0154]

[0155] Compound (II) BHA-Lys-Lys2-Lys4-Lys8-Lys16-32TFA (0.2 mmol) was dissolved in dimethyl sulfoxide (DMSO, 30 mL), and then diisopropylethylamine (DIPEA, 20 mmol) was added, and solution A was obtained under stirring; compound (III) (12 mmol) and 1H-benzotriazole-1-yloxytripyrrolidino hexafluorophosphate (PyBOP) (12 mmol) were added to dimethyl sulfoxide (DMSO, 40 mL) to dissolve, and solution B was obtained under stirring. Solution A was added to solution B at room temperature (20-35° C.), and the mixture was stirred at room temperature (20-35° C.) for 2 hours. Water (200 mL) was added to the reaction mixture to precipitate a white solid, which was filtered off with suction. The filter cake was washed with water (50 mL×3) several times. After drying, the wet product was dried in a vacuum oven at 60°C to constant weight to obtain a white solid crude product. The crude product was dispersed in ethanol (60 mL), stirred at room temperature and HCl gas was continuously introduced for 1.6 hours. After stopping the aeration, the reaction was allowed to react at room temperature overnight under nitrogen protection.

[0156] Methyl tert-butyl ether (200 mL) was dropped into the stirred reaction solution to precipitate a large amount of white solid, which was filtered off and the filter cake was washed with methyl tert-butyl ether (30 mL×3). After the solid was dried by suction, it was moved into an oven at 60°C and dried in vacuo to constant weight to obtain a crude off-white solid of compound 91.

[0157] The off-white solid crude product of compound 91 was added to methanol (50 mL) and dissolved; tetrahydrofuran (100 mL) was added dropwise to the methanol solution of compound 91 with stirring at room temperature to precipitate a white solid. The mixture was stirred for 14 hours and filtered off with suction. The filter cake was dried by evaporation and moved into an oven at 60°C under vacuum to constant weight to obtain 2.72 g of compound 91 (MW: 17576.75 g / mol (hydrochloride salt), 16410.03 g / mol (free base)) with a purity of 99% and a yield of 77%.

[0158] Compound 89 was detected by mass spectrometry using DIONEX UltiMate 3000 HPLC and Thermo QExactive. The data obtained were: LC-Ms(ESI): The molecule was multiply charged [M+19H] 19+ , [M+18H] 18+ , [M+17H] 17+ , [M+16H] 16+, [M+15H] 15+ ......[M+4H] 4+ , [M+3H] 3+ , the molecular weight after deconvolution is: 16410.0166.

[0159] Test Example 1: Anti-HIV-1 Activity and Cytotoxicity Test

[0160] 1. Materials

[0161] 1. Compound pretreatment

[0162] The test compounds 1-93 were dissolved in serum-free DMEM to prepare solutions with a concentration of 2.5 mg / mL, filtered through a 0.22 μm filter membrane for sterilization, and stored at 4° C. after being aliquoted.

[0163] 2. Reagents and Solutions

[0164] (1) Reagents

[0165] MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), penicillin, sodium chloride, potassium chloride, potassium dihydrogen phosphate, and disodium hydrogen phosphate are products of Sigma-Aldrich; SDS (sodium dodecyl sulfate), streptomycin sulfate, and trypsin are products of Amresco; DMF (N,N-dimethylformamide) is a product of Xilong Chemical Co., Ltd.; DMEM and fetal bovine serum are products of Invitrogen; and Bright-Glo luciferase analysis system is a product of Promega.

[0166] (2) Culture medium

[0167] DMEM complete medium contains 10% fetal bovine serum, 2mmol / L L-glutamine, 10mmol / L HEPES (4-hydroxyethylpiperazineethanesulfonic acid), 50μmol / L 2-mercaptoethanol, 100IU penicillin / ml, and 100μg / ml streptomycin.

[0168] 3. Cells and viruses

[0169] TZM-bl cells and HIV-1NL4.3 were obtained from the National Institutes of Health (NIH) of the United States; the cells were cultured in DMEM complete medium containing 10% fetal bovine serum. HIV-1 NL4-3 , titrate and calculate the TCID50 of the virus. The virus stock solution was packaged and stored at -70℃. Cells and viruses were frozen and revived according to conventional methods.

[0170] 2. Toxicity of compounds to TZM-bl cells

[0171] 2x10 4 TZM-b1 cells / well were inoculated into a 96-well cell culture plate and cultured at 37°C and 5% CO2 for 16 hours. The culture medium was discarded and the drug prepared in DMEM complete medium was added. Six dilutions were set with a dilution ratio of 5 times and three replicate wells for each dilution. Control wells without drugs and blank control wells were also set. The cells were cultured at 37°C and 5% CO2 for 3 days, and the cytotoxicity was detected by MTT colorimetry. The OD value was determined by ELx800 microplate reader, and the measurement wavelength was 570 / 630nm. CC was calculated. 50 value.

[0172] III. Effects of Compounds on HIV-1 NL4.3 Inhibition experiments of replication in TZM-bl cells

[0173] 2x10 4 TZM-b1 cells / well were inoculated into 96-well cell culture plates and cultured at 37°C, 5% CO2 for 16 hours. Drugs were prepared in DMEM complete medium, with 6 dilutions and a dilution ratio of 5 times. 2 hours before infection, TZM-b1 cell culture supernatant was discarded, and 100 μL of DMEM-10 medium containing drugs diluted to the appropriate concentration was added. Three replicate wells were set for each concentration, and negative control wells and positive control wells were set. The wells were incubated at 37°C, 5% CO2 for 2 hours.

[0174] In the P3 laboratory, add 100 μL of HIV-1 NL4.3 Virus diluted supernatant (3000TCID50), 37 ℃, 5% CO2 continued to culture for 48h, discarded culture supernatant, 200μL / well PBS washed cells twice, then added cell lysis buffer, 100μL / well, 4 ℃ lysis for 30min; transferred cell lysate to a white 96-well plate, 80μL / well, added pre-prepared luciferase substrate, 80μL / well, used FLEX Station 3 microplate reader to detect the relative fluorescence unit (RLU) of each well, and calculated EC 50 .

[0175] 4. Calculation formula

[0176] Cell growth survival rate (%) = (OD value of experimental well - OD value of blank well) / (OD value of control well - OD value of blank well) × 100%;

[0177] HIV-1 inhibition rate (%) = (1-(RLU value of experimental well-RLU value of negative control well) / (RLU value of positive control well-RLU value of negative control well))×100%.

[0178] According to the cell growth survival rate and HIV-1 inhibition rate, the Origin2016 software was used to draw the fitting curve, and the 50% effective concentration (EC 50 ), 50% inhibitory concentration of cell growth (CC 50 ).

[0179] 5. Test results:

[0180] Compounds 1-93 were tested against HIV-1 virus strains according to the above method. NL4.3 The effective concentration (EC 50 ) and the 50% inhibitory concentration (CC 50 ), the specific test results are shown in Table 4 below:

[0181] Table 4 Anti-HIV-1 activity and cytotoxicity of compounds

[0182]

[0183]

[0184]

[0185] Test Example 2: Solubility Test

[0186] Referring to the test method recorded in the 2020 edition of the Chinese Pharmacopoeia, the solubility of compounds 1-93 obtained in the present invention was tested respectively, and the selected test solvents were water and ethanol. The specific test method was:

[0187] Weigh 1.00-1.05g of ground compound 1 to compound 93, add to a 50mL stoppered volumetric flask at 25±2°C, add solvent water, or add solvent ethanol, the amount of solvent added each time is 2mL, shake vigorously for 30 seconds after the solvent is added, and continue to increase the amount of solvent by 2mL if it is not dissolved after standing for 30 minutes, repeat the operation until there are no solute particles visually, which is considered to be completely dissolved. According to statistics, the compounds 1-93 prepared in the present invention can be completely dissolved in 2-10mL water, and all compounds can be completely dissolved in 14-28mL ethanol. It can be seen that the compound of formula (I) in the present invention is easily soluble in water and soluble in ethanol.

[0188] Test Example 3: Transdermal Test

[0189] In vitro permeation experiments were performed on the abdominal and dorsal skin of isolated mice for 12 hours and 24 hours, respectively, and no permeation of compounds 1 to 93 was observed.

[0190] Therefore, based on the EC in Table 4 of the present invention50 Test data, screened for HIV-1 NL4.3 The compounds of formula (I) with good inhibitory activity are shown in Table 5.

[0191] Table 5 Preferred compounds of formula (I) of the present invention

[0192]

[0193]

[0194] In summary, the compound of formula (I) of the present invention has excellent solubility and activity against HIV-1 virus infection, and has little toxicity and side effects on normal cells and poor transdermal effect, and can be used to prepare HIV inhibitors for topical administration.

[0195] The above is a detailed description of the embodiments of the present invention, but the present invention is not limited to the above embodiments. Various changes can be made within the knowledge of ordinary technicians in the relevant technical field without departing from the purpose of the present invention. In addition, the embodiments of the present invention and the features in the embodiments can be combined with each other without conflict.

Claims

1. A compound represented by formula (I) or a pharmaceutically acceptable salt thereof, R is: in, R1 is selected from H, F, -CN, -CH(CH3)2 or -C(CH3)3; R2 is selected from H, -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)2, -C(CH3)3, -CH2CH2CH2CH3, -CH2CH(CH3)2, -CH(CH3)CH2CH3, -CH=CH2, -CH=CHCH3, -CH=C(CH3)2, -C≡CCH3, -CN, F, Cl or Br; R3 is selected from H, F or -CN.

2. The compound according to claim 1, characterized in that: The compound represented by the formula (I) is any one of the compounds numbered 1 to 31 in Table 1 of the specification.

3. The compound according to claim 1, characterized in that: The compound represented by the formula (I) is any one of the compounds numbered 32 to 62 in Table 1 of the specification.

4. The compound according to claim 1, characterized in that: The compound represented by the formula (I) is any one of the compounds numbered 63 to 93 in Table 1 of the specification.

5. The compound according to claim 1, characterized in that: The compound is: Among them, R is 6. The compound according to claim 1, characterized in that: Pharmaceutically acceptable salts of the compounds include hydrochloride or hydrobromide.

7. The method for preparing the compound according to any one of claims 1 to 6, characterized in that: The following steps are involved: The compound represented by formula (II) is subjected to a condensation reaction with the compound represented by formula (III), and then the tert-butyloxycarbonyl protecting group is removed by reduction to obtain the compound represented by formula (I); The formula (II) is: The formula (III) is:

8. The method for preparing the compound according to claim 7, characterized in that: The condensation reaction is specifically: reacting the compound represented by formula (II) with the compound represented by formula (III) in the presence of an organic base and a condensing agent.

9. An HIV inhibitor, characterized in that: The invention comprises the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable auxiliary material.

10. Use of the compound according to any one of claims 1 to 6 in the preparation of a medicament for preventing or treating sexually transmitted diseases caused by HIV-1.

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