A triptolide derivative and its preparation method and application

Abstract

The present invention provides a triptolide derivative and a preparation method and application thereof, which belong to the field of pharmaceutical chemistry technology. The triptolide derivative provided by the present invention has a structure shown in formula (I). Pharmacological activity experiments show that the triptolide derivative provided by the present invention and its pharmaceutically acceptable salt or precursor have good inhibitory activity against HIV-1 protease and can be used as an active ingredient of anti-AIDS drugs. The present invention provides a method for preparing the above-mentioned triptolide derivative, which is simple to operate and easy to achieve industrial batch production. Furthermore, the present invention provides an anti-AIDS pharmaceutical composition, comprising an active ingredient and a drug carrier, wherein the active ingredient is the above-mentioned triptolide derivative and its pharmaceutically acceptable salt or precursor. This pharmaceutical composition can be used as an HIV-1 protease inhibitor.

Description

Technical Field

[0001] The present invention relates to the technical field of medicinal chemistry, and in particular to a triptolide derivative and a preparation method and application thereof. Background Art

[0002] Acquired Immune Deficiency Syndrome (AIDS) is a syndrome in which human immunodeficiency virus (HIV) infection causes immune deficiency, leading to a series of opportunistic infections and tumors. Although various approaches to HIV prevention and treatment are currently available, the development of new anti-HIV drugs remains the most effective way to treat AIDS.

[0003] In the early stages of HIV treatment, only single drugs, such as nucleoside reverse transcriptase inhibitors (NTIs) such as lamivudine and zidovudine, were used clinically. The 1995 launch of saquinavir, the first HIV protease inhibitor, marked the beginning of highly active antiretroviral therapy (HAART). HAART can maximally suppress viral replication, slow disease progression, and significantly prolong patients' lives and improve their quality of life.

[0004] The increasing number of HIV / AIDS patients has placed a tremendous burden on society, jeopardizing socioeconomic development and stability. Currently, HIV / AIDS is still treated with medication. However, with the frequent use of various anti-HIV drugs, drug resistance has increasingly developed, limiting their clinical application. HIV-1PR is a specific aspartyl protease encoded by the HIV gene. Inhibiting the protease's activity can limit infected cells to producing only immature, non-infectious viruses. Therefore, HIV-1PR is an important target for the development of anti-HIV drugs. Summary of the Invention

[0005] In view of this, the present invention aims to provide a triptolide derivative and its preparation method and application. The triptolide derivative and its pharmaceutically acceptable salt or precursor provided by the present invention have good HIV-1 protein kinase inhibitory activity.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The present invention provides a triptolide derivative and a pharmaceutically acceptable salt or precursor thereof, characterized in that the triptolide derivative has a structure represented by formula (I):

[0008]

[0009] In formula (I), R1 is H, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted 6-10 membered aryl, or substituted or unsubstituted 5-10 membered heteroaryl;

[0010] R2 is H, halogen, -CN, -CF3, -OCF3, -OR 21 、-CH2OR 21 、-SR 21 、-CH2SR 21 、-NR 21 R 22 , C1~C4 alkyl or C3~C6 cycloalkyl; wherein R 21 、R 22 are independently H or C1-C4 alkyl;

[0011] R3 is R or S type; R3 is selected from H, C1~C4 alkyl, C3~C6 cycloalkyl, -CH2OR 31 、-CH2SR 31 or -CH2CONR 31 R 32 ; where R 31 、R 32 are independently H or C1-C4 alkyl.

[0012] Preferably, the number of substituents in the substituted C1-C4 alkyl, substituted C3-C6 cycloalkyl, substituted 6-10 membered aryl or substituted 5-10 membered heteroaryl is 1 to 3; and the substituents are independently hydrogen, halogen, hydroxyl, amino, trifluoromethyl, trifluoromethoxy, mercapto, C1-C4 alkyl, C1-C4 alkylthio or C1-C4 alkoxy.

[0013] Preferably, R1 is H,

[0014] R2 is H, halogen, -CN, -CF3, -OCF3, -OR 21 、-SR 21 or -NR 21 R 22 ; where R 21 、R 22 are independently H or C1-C4 alkyl.

[0015] Preferably, R3 is H, -CH3 or -CH2CONH2.

[0016] Preferably, it has a structure shown in any one of Formulas 1 to 31:

[0017]

[0018]

[0019]

[0020] The present invention provides a method for preparing a triptolide derivative and a pharmaceutically acceptable salt or precursor thereof. The method for preparing the triptolide derivative comprises the following steps:

[0021] Under the action of pyridine, triptolide having a structure represented by Formula A undergoes a substitution reaction with p-nitrophenyl chloroformate having a structure represented by Formula B to obtain a compound having a structure represented by Formula C;

[0022]

[0023] A compound having a structure represented by Formula D is subjected to a condensation reaction with a compound having a structure represented by Formula E, and after de-Boc protection, a compound having a structure represented by Formula F is obtained;

[0024]

[0025] Under the action of triethylamine, the compound having the structure represented by formula C and the compound having the structure represented by formula F undergo an acylation reaction to obtain a triptolide derivative.

[0026] Preferably, the temperature of the substitution reaction is 0-25°C and the time is 2-4 hours;

[0027] The condensation reaction temperature is 0-25°C and the time is 2-5h;

[0028] The temperature for removing Boc protection is 25° C. and the time is 2 to 5 hours.

[0029] Preferably, the molar ratio of the compound having the structure represented by Formula C to triethylamine is 1:2-3;

[0030] The acylation reaction is carried out at a temperature of 25° C. and for a period of 2 to 5 hours.

[0031] The present invention provides the use of the above-mentioned triptolide derivatives and pharmaceutically acceptable salts or precursors thereof in the preparation of anti-AIDS drugs.

[0032] The present invention provides an anti-AIDS pharmaceutical composition comprising an active ingredient and a pharmaceutical carrier, wherein the active ingredient comprises the above-mentioned triptolide derivative and a pharmaceutically acceptable salt or precursor thereof.

[0033] The present invention provides triptolide derivatives and pharmaceutically acceptable salts or precursors thereof. The triptolide derivatives have the structure represented by formula (I). Pharmacological activity experiments have shown that the triptolide derivatives and pharmaceutically acceptable salts or precursors provided by the present invention have good inhibitory activity against HIV-1 protease and can be used as active ingredients in anti-AIDS drugs.

[0034] The present invention provides a method for preparing the triptolide derivative, which is simple to operate and easy to realize industrial batch production.

[0035] Furthermore, the present invention provides an anti-AIDS pharmaceutical composition comprising an active ingredient and a pharmaceutical carrier, wherein the active ingredient is the aforementioned triptolide derivative and a pharmaceutically acceptable salt or precursor thereof. This pharmaceutical composition can be used as an HIV-1 protease inhibitor. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 The synthetic route of triptolide derivatives. DETAILED DESCRIPTION

[0037] The present invention provides triptolide derivatives and pharmaceutically acceptable salts or precursors thereof. The triptolide derivatives have a structure shown in formula (I):

[0038]

[0039] In formula (I), R1 is H, a substituted or unsubstituted C1-C4 alkyl group, a substituted or unsubstituted C3-C6 cycloalkyl group, a substituted or unsubstituted 6- to 10-membered aryl group, or a substituted or unsubstituted 5- to 10-membered heteroaryl group. In the present invention, the heteroatom in the substituted or unsubstituted 6- to 10-membered aryl group or the substituted or unsubstituted 5- to 10-membered heteroaryl group is independently O, N, or S; and the number of the heteroatoms is preferably 1 to 3.

[0040] In the present invention, the substituents in the substituted C1-C4 alkyl, substituted C3-C6 cycloalkyl, substituted 6-10 membered aryl or substituted 5-10 membered heteroaryl are denoted by R 11 , the number of the substituents is preferably 1 to 3; the substituents are independently preferably hydrogen, halogen, hydroxy, amino, trifluoromethyl, trifluoromethoxy, mercapto, C1~C4 alkyl, C1~C4 alkylthio or C1~C4 alkoxy.

[0041] In the present invention, R1 is preferably H,

[0042] In the present invention, R2 is H, halogen, -CN, -CF3, -OCF3, -OR 21 、-CH2OR 21、-SR 21 、-CH2SR 21 、-NR 21 R 22 , C1~C4 alkyl or C3~C6 cycloalkyl; wherein R 21 、R 22 are independently H or C1-C4 alkyl; in the present invention, R 21 、R 22 Same or different, R 21 、R 22 R2 is independently H or C1-C4 alkyl. Further, R2 is preferably H, halogen, -CN, -CF3, -OCF3, -OR 21 、-SR 21 、-NR 21 R 22 .

[0043] In the present invention, R3 is R or S type; R3 is selected from H, C1~C4 alkyl, C3~C6 cycloalkyl, -CH2OR 31 、-CH2SR 31 or -CH2CONR 31 R 32 In the present invention, R 31 、R 32 Same or different, R 31 、R 32 are independently H or C1-C4 alkyl.

[0044] In the present invention, R3 is preferably H, -CH3 or -CH2CONH2.

[0045] In the present invention, the C1-C4 alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl; the C3-C6 cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

[0046] In the present invention, the halogen is fluorine, chlorine, bromine or iodine; the aryl is a monocyclic or polycyclic aromatic system; and the heteroaryl is a monocyclic or polycyclic aromatic system containing one or more heteroatoms selected from N, O and S.

[0047] In the present invention, the triptolide derivative preferably has the structure shown in Table 1.

[0048] Table 1 Structures and names of triptolide derivatives

[0049]

[0050]

[0051]

[0052]

[0053]

[0054]

[0055]

[0056] In the present invention, in the triptolide derivative and its pharmaceutically acceptable salt or precursor, the pharmaceutically acceptable salt is a salt obtained by adding an acid to the triptolide derivative. In the present invention, the acid is an inorganic acid and / or an organic acid, and is preferably one or more of hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalene disulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid and benzoic acid.

[0057] The present invention also provides precursors of triptolide derivatives, i.e., prodrugs. The prodrugs of triptolide derivatives of the present invention are derivatives of formula (I), which may have weak or even no activity themselves, but after administration, are converted into the corresponding biologically active form under physiological conditions (e.g., by metabolism, solvolysis, or other means).

[0058] The present invention provides a method for preparing the above-mentioned triptolide derivative and a pharmaceutically acceptable salt or precursor thereof. The method for preparing the triptolide derivative comprises the following steps:

[0059] Under the action of pyridine, triptolide having a structure represented by Formula A undergoes a substitution reaction with p-nitrophenyl chloroformate having a structure represented by Formula B to obtain a compound having a structure represented by Formula C;

[0060]

[0061] A compound having a structure represented by Formula D is subjected to a condensation reaction with a compound having a structure represented by Formula E, and after de-Boc protection, a compound having a structure represented by Formula F is obtained;

[0062]

[0063] Under the action of triethylamine, the compound having the structure represented by formula C and the compound having the structure represented by formula F undergo an acylation reaction to obtain a triptolide derivative.

[0064] In the present invention, under the action of pyridine, triptolide having the structure represented by Formula A undergoes a substitution reaction with p-nitrophenyl chloroformate having the structure represented by Formula B to obtain a compound having the structure represented by Formula C. In the present invention, the molar ratio of triptolide having the structure represented by Formula A to pyridine is preferably 1:2. In the present invention, the substitution reaction is preferably carried out in an organic solvent, and the organic solvent is preferably dichloromethane.

[0065] In the present invention, the temperature of the substitution reaction is preferably 0-25° C., and the time is preferably 2-4 h, more preferably 3 h.

[0066] After the substitution reaction, the present invention preferably performs post-treatment on the obtained substitution reaction liquid. In the present invention, the post-treatment preferably includes the following steps:

[0067] The substitution reaction liquid is mixed with water, the organic phases are combined, the organic solvent is removed, and column chromatography is performed to obtain a pure compound having the structure shown in Formula C.

[0068] In the present invention, the method of removing the organic solvent is preferably concentration under reduced pressure; the mobile phase of the column chromatography separation is cyclohexane and ethyl acetate, and the volume ratio of the cyclohexane and ethyl acetate is 1:1.

[0069] In the present invention, a compound having a structure represented by Formula D is subjected to a condensation reaction with a compound having a structure represented by Formula E, and after deprotection with the Boc group, a compound having a structure represented by Formula F is obtained. In the present invention, the temperature of the condensation reaction is preferably 0 to 25° C., and the time is preferably 2 to 5 hours, more preferably 3 to 4 hours; the temperature of the deprotection with the Boc group is preferably 25° C., and the time is preferably 2 to 5 hours, more preferably 3 to 4 hours.

[0070] In the present invention, the preparation method of the compound having the structure represented by Formula F refers to Bioorganic Medicinal Chemistry Letters 25 (2015) 1880–1883; European Journal of Medicinal Chemistry 137 (2017) 30-44; Bioorganic & Medicinal Chemistry 28 (2020) 115623.

[0071] In the present invention, a compound having a structure represented by Formula C and a compound having a structure represented by Formula F undergo an acylation reaction in the presence of triethylamine to obtain a triptolide derivative. In the present invention, the molar ratio of the compound having a structure represented by Formula C to triethylamine is preferably 1:2-3, more preferably 1:2.29. In the present invention, the acylation reaction is preferably carried out in an organic solvent, preferably acetonitrile.

[0072] In the present invention, the temperature of the acylation reaction is preferably 25° C., and the time is preferably 2 to 5 hours, more preferably 3 to 4 hours.

[0073] After the acylation reaction, the present invention preferably performs post-treatment on the obtained acylation reaction solution. In the present invention, the post-treatment preferably includes the following steps:

[0074] The organic solvent of the acylation reaction solution is removed, and the organic phase is extracted, dried and separated by column chromatography to obtain a pure product of the triptolide derivative.

[0075] In the present invention, the method for removing the organic solvent is preferably concentration under reduced pressure. In the present invention, the extractant used in the extraction is preferably ethyl acetate, and the desiccant used for drying is preferably anhydrous sodium sulfate. In the present invention, the column chromatography separation is preferably performed using a silica gel column, and the mobile phase of the column chromatography is preferably dichloromethane and methanol, and the volume ratio of the dichloromethane and methanol is preferably 50:1.

[0076] In the present invention, the synthetic route of the triptolide derivative is as follows: Figure 1 shown.

[0077] The present invention provides the use of the above-mentioned triptolide derivatives and pharmaceutically acceptable salts or precursors thereof in the preparation of anti-AIDS drugs.

[0078] The present invention provides an anti-AIDS pharmaceutical composition comprising an active ingredient and a pharmaceutical carrier, wherein the active ingredient comprises the above-mentioned triptolide derivative and a pharmaceutically acceptable salt or precursor thereof.

[0079] In the present invention, the drug carrier is preferably one or more of microcapsules, microspheres, nanoparticles and liposomes.

[0080] The triptolide derivatives provided by the present invention, their preparation methods and applications are described in detail below with reference to the examples, but they should not be construed as limiting the scope of protection of the present invention.

[0081] The following examples are representative of the synthesis methods. The triptolide derivatives of the structure represented by formula (I) of the present invention (especially compounds 1 to 31 in Table 1) can be synthesized by referring to the synthesis routes and preparation methods of the examples.

[0082] Example 1 Preparation of Compound 1: Triptolide-(2-(((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methoxyphenyl)sulfonamido)-1-phenylbutan-2-yl)amino)-2-oxoethyl)amino acid methyl ester

[0083]

[0084] Step A: Preparation of Triptolide-(4-nitrophenyl)formate (C)

[0085]

[0086] Triptolide (0.30 g, 0.83 mmol) was added to a 50 mL eggplant-shaped flask containing 15 mL of dichloromethane. p-Nitrophenyl chloroformate (0.25 g, 1.25 mmol) was slowly added dropwise under an ice bath, followed by the slow addition of pyridine (0.13 mL, 1.66 mmol). After the addition, the mixture was transferred to room temperature and reacted for 2 hours. After TLC detection, the reaction was complete. 45 mL of water was added three times, the organic phases were combined, and the dichloromethane was removed by concentration under reduced pressure. The crude product was purified by column chromatography (mobile phase: n-hexane: ethyl acetate = 1:1) to obtain the target product as a white solid, 0.32 g (73.4%).

[0087] 1 H NMR(400MHz,Chloroform-d)δ8.29(d,J=9.0Hz,2H),7.44(d,J=9.0Hz,2H),4.91(s,1H ),4.75–4.66(m,2H),3.88(d,J=3.0Hz,1H),3.61(d,J=3.0Hz,1H),3.56(d,J=5.4Hz,1H ),2.73(d,J=12.8Hz,1H),2.37–2.32(m,1H),2.26–2.16(m,2H),2.07–1.93(m,2H),1.6 8(s,1H),1.63–1.59(m,1H),1.11(s,3H),1.03(d,J=7.0Hz,3H),0.91(d,J=7.0Hz,3H);

[0088] 13 C NMR(101MHz,Chloroform-d)δ173.17,159.70,155.45,152.50,145.41,125.71,125.27,121.72,69.98,6 3.39,62.78,61.53,59.61,55.33,55.18,40.30,35.66,29.83,28.25,23.36,17.52,17.05,16.67,13.82;

[0089] LC-MS (ESI) [M+H] + m / z 525.2.

[0090] Step B: Preparation of Compound 1

[0091] Intermediate C (0.20 g, 0.38 mmol), Et3N (0.12 ml, 0.87 mmol), and 2-amino-N-((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methoxyphenyl)sulfonamide)-1-phenylbutan-2-yl)acetamide (0.16 g, 0.34 mmol) were added to a 50 mL eggplant flask containing 10 mL of acetonitrile. The reaction was allowed to react at room temperature for 2.5 hours. The reaction was detected to be complete by TLC. The reaction system was concentrated under reduced pressure to remove excess acetonitrile and extracted with 30 mL (3×10 mL) of ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and subjected to silica gel column chromatography using a mobile phase of CH2Cl2:MeOH=50:1 to obtain 10.17 g (53.1%) of the white solid compound.

[0092] 1 HNMR(400MHz,MeOD)δ7.77(d,J=8.8Hz,2H),7.24–7.19(m,4H),7.15–7.11(m,1H), 7.05(d,J=8.8Hz,2H),4.90(s,1H),4.79–4.72(m,2H),4.01(ddd,J=10.6,6.8,3.6H z,1H),3.96(d,J=3.2Hz,1H),3.84(s,3H),3.82–3.80(m,1H),3.60–3.58(m,2H),3 .51(d,J=5.6Hz,1H),3.39(dd,J=15.0,3.2Hz,1H),3.18(dd,J=14.0,3.6Hz,1H),3. 00(dd,J=13.6,8.4Hz,1H),2.85(dd,J=15.0,8.8Hz,1H),2.77(dd,J=13.6,6.6Hz, 2H),2.61(dd,J=14.0,10.8Hz,1H),2.28–2.19(m,2H),2.06(d,J=8.8Hz,1H),2.00– 1.88(m,3H),1.48(dd,J=12.4,4.8Hz,1H),1.38–1.27(m,2H),1.05(s,3H),0.92(d ,J=7.0Hz,3H),0.88(d,J=7.0Hz,3H),0.82(d,J=4.8Hz,3H),0.80(d,J=4.8Hz,3H).

[0093] 13C NMR(101MHz,MeOD)δ174.6,170.2,163.1,162.3,157.1,138.4,130.6,129.3,128.9,127.9,125.9,124.1,113.9,72.7,72.6,70.5,63 .5,62.9,61.9,60.5,57.9,55.7,54.8,54.3,52.7,43.6,40.0,35.6,35.4,29.4,28.2,26.5,22.8,19.1,19.0,16.6,16.5,15.7,13.0.

[0094] HRMS(ESI)m / z calcd.for C 44 H 55 N3O 12 S([M+Na] + ):872.3404,found872.3391.

[0095] Example 2 Preparation of Compound 2: Triptolide-(2-(((2S,3R)-3-hydroxy-4-((N-isobutyl-4-aminophenyl)sulfonamido)-1-phenylbutan-2-yl)amino)-2-oxoethyl)amino acid methyl ester

[0096]

[0097] The preparation method refers to Example 1, wherein in step B, 2-amino-N-((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methoxyphenyl)sulfonamide)-1-phenylbutan-2-yl)acetamide is replaced by 2-amino-N-((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methoxyphenyl)sulfonamide)-1-phenylbutan-2-yl)acetamide.

[0098] 1H NMR(400MHz,MeOD)δ7.50(d,J=8.8Hz,2H),7.24–7.19(m,4H),7.15–7.11(m,1H),6.67(d,J=8.8Hz,2H),4.90(s,1H),4.80–4.72(m,2H),4.03(ddd,J=10.4,6.4,3.6Hz,1H),3.95(d,J=3.2Hz,1H),3.83–3.79(m,1H),3.62(d,J=17.0Hz,1H),3.58(d,J=2.8Hz,1H),3.52(d,J=17.0Hz,1H),3.49(d,J=5.6Hz,1H),3.37–3.32(m,1H),3.16(dd,J=14.0,3.6Hz,1H),2.93(dd,J=13.6,8.0Hz,1H),2.84(dd,J=15.0,8.4Hz,1H),2.73(dd,J=13.6,6.8Hz,2H),2.62(dd,J=14.0,10.8Hz,1H),2.27–2.22(m,1H),2.20(d,J=5.4Hz,1H),2.06(d,J=8.8Hz,1H),1.98–1.87(m,3H),1.47(dd,J=12.4,4.8Hz,1H),1.30(td,J=11.8,5.6Hz,1H),1.04(s,3H),0.92(d,J=7.0Hz,3H),0.88(d,J=6.8Hz,3H),0.82(t,J=6.8Hz,6H).

[0099] 13 C NMR(101MHz,MeOD)δ174.6,170.2,162.4,157.0,152.8,138.5,129.2,128.9,127.9,125.8,124.5,124.1,113.1,72.7,70.6,63.5,63.0,61.9,60.4,58.1,55.7,54.7,54.3,52.8,43.6,40.0,35.4,29.4,28.2,26.6,22.8,19.2,19.1,16.6,16.5,15.7,13.0.

[0100] HRMS(ESI)m / z calcd.for C 43 H 54 N4O 11 S([M+Na] + ):857.3407,857.3440.

[0101] Example 3 Preparation of Compound 3: Triptolide-(2-(((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methylthiophenyl)sulfonamido)-1-phenylbutan-2-yl)amino)-2-oxoethyl)amino acid methyl ester

[0102]

[0103] The preparation method refers to Example 1, wherein in step B, 2-amino-N-((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methoxyphenyl)sulfonamide)-1-phenylbutan-2-yl)acetamide is replaced by 2-amino-N-((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methylthiophenyl)sulfonamide)-1-phenylbutan-2-yl)acetamide.

[0104] 1 H NMR(400MHz,MeOD)δ7.73(d,J=8.6Hz,2H),7.36(d,J=8.6Hz,2H),7.24–7.18(m,4H ),7.15–7.11(m,1H),4.90(s,1H),4.79–4.72(m,2H),4.03–4.00(m,1H),3.95(d,J =3.2Hz,1H),3.84–3.79(m,1H),3.59–3.58(m,2H),3.55(s,1H),3.51–3.50(m,1H) ,3.41(dd,J=15.0,3.2Hz,1H),3.18(dd,J=14.0,3.6Hz,1H),3.02(dd,J=13.6,8.4 Hz,1H),2.87(dd,J=15.0,9.0Hz,1H),2.80–2.73(m,2H),2.60(dd,J=14.0,10.8Hz ,1H),2.50(s,3H),2.28–2.19(m,2H),2.06(dd,J=14.8,5.6Hz,1H),1.99(dd,J=14 .0,7.2Hz,1H),1.94–1.87(m,2H),1.48(dd,J=12.4,4.8Hz,1H),1.35–1.27(m,1H) ,1.04(s,3H),0.92(d,J=7.0Hz,3H),0.89(d,J=7.0Hz,3H),0.81(t,J=7.0Hz,6H).

[0105] 13C NMR(101MHz,MeOD)δ174.6,170.2,162.3,157.1,145.8,138.4,134.7,128.9,127.9,127.6,125.9,125.1,124.1,72.7,72.6,70.5,63.5, 62.9,61.9,60.5,57.8,55.7,54.7,54.4,52.7,43.6,40.0,35.7,35.4,29.4,28.2,26.5,22.8,19.1,19.0,16.6,16.5,15.7,13.2,13.1.

[0106] HRMS(ESI)m / z calcd.for C 44 H 55 N3O 11 S2([M+Na] + ):888.3176,found888.3210.

[0107] Example 4 Preparation of Compound 6: Triptolide-(2-(((2S,3R)-3-hydroxy-4-((N-cyclopropyl-4-methoxyphenyl)sulfonamido)-1-phenylbutan-2-yl)amino)-2-oxoethyl)amino acid methyl ester

[0108]

[0109] The preparation method refers to Example 1, wherein in step B, 2-amino-N-((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methoxyphenyl)sulfonamide)-1-phenylbutan-2-yl)acetamide is replaced by 2-amino-N-((2S,3R)-3-hydroxy-4-((N-cyclopropyl-4-methoxyphenyl)sulfonamide)-1-phenylbutan-2-yl)acetamide.

[0110] 1H NMR(400MHz,CDCl3)δ7.85(d,J=8.8Hz,2H),7.35–7.32(m,2H),7.29–7.23(m,3H),7.04(d,J=8.8Hz,2H),5.00(s,1H),4.79–4.69(m,2H),4.18(tt,J=9.6,4.8Hz,1H),4.03(dd,J=8.4,4.4Hz,1H),3.99–3.96(m,1H),3.91(d,J=5.6Hz,4H),3.61–3.55(m,2H),3.52(d,J=5.6Hz,1H),3.47(dd,J=14.4,4.4Hz,1H),3.24(dd,J=14.0,4.0Hz,1H),3.04(dd,J=14.4,8.4Hz,1H),2.82(dd,J=14.0,10.4Hz,1H),2.71(d,J=12.8Hz,1H),2.37(d,J=16.0Hz,1H),2.21(dt,J=14.8,5.6Hz,2H),2.07–1.99(m,3H),1.61(dd,J=12.0,4.8Hz,1H),1.30–1.26(m,1H),1.14(s,3H),1.04(d,J=7.0Hz,3H),0.99(d,J=6.4Hz,1H),0.91(d,J=7.0Hz,3H),0.88–0.81(m,1H),0.79–0.74(m,1H),0.71–0.64(m,1H).

[0111] 13 C NMR(101MHz,CDCl3)δ173.3,169.3,163.3,160.1,156.3,138.2,130.1,129.4,128.5,128.4,126.5,125.5,114.3,72.9,72.6,70.2,63.5,63.3,62.6,60.9,56.0,55.7,55.2,54.9,54.3,44.6,40.3,35.7,35.5,32.9,30.0,28.5,23.4,17.6,17.1,16.7,14.1,8.12,7.15.

[0112] HRMS(ESI)m / z calcd.for C 43 H 51 N3O 12 S([M+Na] + ):856.3091,found856.3115.

[0113] Example 5 Preparation of Compound 11: Triptolide ((S)-1-(((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methoxyphenyl)sulfonamido)-1-phenylbutan-2-yl)amino)-1-oxopropan-2-yl)amino acid methyl ester

[0114]

[0115] The preparation method refers to Example 1, wherein in step B, 2-amino-N-((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methoxyphenyl)sulfonamide)-1-phenylbutan-2-yl)acetamide is replaced by (S)-2-amino-N-((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methoxyphenyl)sulfonamide)-1-phenylbutan-2-yl)propionamide.

[0116] 1 H NMR(400MHz,MeOD)δ7.81(d,J=8.8Hz,2H),7.25–7.19(m,4H),7.13(t,J=7.0Hz,1H) ,7.03(d,J=8.8Hz,2H),4.93(s,1H),4.81–4.73(m,2H),3.99(d,J=3.2Hz,1H),3.95( dd,J=7.2,3.6Hz,1H),3.90(d,J=7.2Hz,1H),3.84(d,J=11.2Hz,4H),3.61–3.60(m, 2H),3.41(dd,J=15.0,3.0Hz,1H),3.23(d,J=3.6Hz,1H),3.00(dd,J=13.6,8.8Hz,1H ),2.84(dd,J=15.0,9.2Hz,1H),2.75(dd,J=13.6,6.4Hz,2H),2.60(dd,J=13.8,11. 2Hz,1H),2.30–2.20(m,2H),1.99–1.87(m,3H),1.50(dd,J=12.4,4.8Hz,1H),1.35(d d,J=12.0,6.0Hz,1H),1.31–1.27(m,1H),1.10(s,3H),0.99(d,J=7.2Hz,3H),0.96( d,J=7.2Hz,3H),0.89(d,J=6.8Hz,3H),0.85(d,J=6.8Hz,3H),0.80(d,J=6.8Hz,3H).

[0117] 13C NMR(101MHz,MeOD)δ174.6,173.5,163.0,162.3,156.3,138.4,130.7,129.4,129.0,127.9,125.9,124.1,113.9,72.8,72.7,70.5,63.5, 62.9,62.5,60.9,58.1,56.0,54.8,54.7,54.5,52.9,51.1,39.9,36.1,35.4,29.5,28.6,26.5,22.8,19.1,16.6,16.5,16.2,15.7,13.2.

[0118] HRMS(ESI)m / z calcd.for C 45 H57N3O 12 S([M+Na] + ):886.3561,found886.3564.

[0119] Example 6 Preparation of Compound 12: Triptolide ((S)-1-(((2S,3R)-3-hydroxy-4-((N-isobutyl-4-aminophenyl)sulfonamido)-1-phenylbutan-2-yl)amino)-1-oxopropan-2-yl)amino acid methyl ester

[0120]

[0121] The preparation method refers to Example 1, wherein in step B, 2-amino-N-((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methoxyphenyl)sulfonamide)-1-phenylbutan-2-yl)acetamide is replaced by (S)-2-amino-N-((2S,3R)-3-hydroxy-4-((N-isobutyl-4-aminophenyl)sulfonamide)-1-phenylbutan-2-yl)propionamide.

[0122] 1H NMR(400MHz,MeOD)δ7.53(d,J=8.8Hz,2H),7.25–7.19(m,4H),7.14–7.11(m,1H),6.66(d,J=8.8Hz,2H),4.92(s,1H),4.80–4.73(m,2H),3.98(d,J=3.2Hz,1H),3.97–3.93(m,1H),3.91(d,J=7.2Hz,1H),3.85–3.80(m,1H),3.60(d,J=2.8Hz,1H),3.58(d,J=5.6Hz,1H),3.36(dd,J=15.0,3.2Hz,1H),3.23(dd,J=14.0,3.2Hz,1H),2.94(dd,J=13.6,8.4Hz,1H),2.82(dd,J=15.0,9.2Hz,1H),2.71(dd,J=13.6,6.4Hz,2H),2.61(dd,J=13.8,11.2Hz,1H),2.28–2.22(m,1H),2.20(s,1H),2.07(d,J=8.8Hz,1H),1.98–1.86(m,3H),1.49(dd,J=12.4,4.8Hz,1H),1.33(dd,J=12.0,5.6Hz,1H),1.08(s,3H),1.01(d,J=7.2Hz,3H),0.95(d,J=7.0Hz,3H),0.88(d,J=6.8Hz,3H),0.85(d,J=6.8Hz,3H),0.82(d,J=6.8Hz,3H).

[0123] 13 C NMR(101MHz,MeOD)δ174.6,173.4,162.3,156.2,152.8,138.4,129.2,129.0,127.9,125.9,124.7,124.1,113.1,72.7,70.6,63.5,62.9,62.4,60.9,58.2,56.0,54.8,54.4,53.0,51.0,39.9,35.9,35.4,29.5,28.6,26.6,22.8,19.2,19.1,16.6,16.5,16.3,15.7,13.2.

[0124] HRMS(ESI)m / z calcd.for C 44 H 56 N4O 11 S([M+Na] + ):871.3564,found871.3589.

[0125] Example 7 Preparation of Compound 13: Triptolide ((S)-1-(((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methylthiophenyl)sulfonamido)-1-phenylbutan-2-yl)amino)-1-oxopropan-2-yl)amino acid methyl ester

[0126]

[0127] The preparation method refers to Example 1, wherein in step B, 2-amino-N-((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methoxyphenyl)sulfonamide)-1-phenylbutan-2-yl)acetamide is replaced by (S)-2-amino-N-((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methylthiophenyl)sulfonamide)-1-phenylbutan-2-yl)propionamide.

[0128] 1 H NMR(400MHz,MeOD)δ7.77(d,J=8.6Hz,2H),7.35(d,J=8.6Hz,2H),7.25–7.22( m,2H),7.20–7.18(m,2H),7.15–7.11(m,1H),4.93(s,1H),4.81–4.73(m,2H), 3.99(d,J=3.2Hz,1H),3.93–3.89(m,2H),3.87–3.81(m,1H),3.61–3.59(m,2H ),3.42(dd,J=15.0,3.0Hz,1H),3.23(d,J=3.6Hz,1H),3.02(dd,J=13.6,8.8Hz ,1H),2.86(dd,J=15.0,9.6Hz,1H),2.79–2.75(m,2H),2.60(dd,J=14.0,11.2 Hz,1H),2.49(s,3H),2.28–2.21(m,2H),2.00–1.87(m,3H),1.50(dd,J=12.4,4 .8Hz,1H),1.37–1.26(m,2H),1.09(s,3H),0.99(d,J=7.2Hz,3H),0.96(d,J=7 .2Hz,3H),0.89(d,J=6.8Hz,3H),0.85(d,J=6.8Hz,3H),0.81(d,J=6.8Hz,3H).

[0129] 13C NMR(101MHz,MeOD)δ174.6,173.6,162.3,156.3,145.7,138.4,134.8,128.9,128.0,127.6,125.9,125.0,124.1,72.8,72.7,70.5,63.5, 62.9,62.5,61.0,58.0,56.0,54.8,54.5,52.9,51.1,39.9,36.1,35.4,29.5,28.6,26.5,22.8,19.1,16.6,16.5,16.2,15.7,13.3,13.2.

[0130] HRMS(ESI)m / z calcd.for C 45 H 57 N3O 11 S2([M+Na] + ):902.3332,found902.3340.

[0131] Example 8 Preparation of Compound 21: Triptolide ((R)-1-(((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methoxyphenyl)sulfonamido)-1-phenylbutan-2-yl)amino)-1-oxopropan-2-yl)amino acid methyl ester

[0132]

[0133] The preparation method refers to Example 1, wherein in step B, 2-amino-N-((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methoxyphenyl)sulfonamide)-1-phenylbutan-2-yl)acetamide is replaced by (R)-2-amino-N-((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methoxyphenyl)sulfonamide)-1-phenylbutan-2-yl)propionamide.

[0134] 1H NMR(400MHz,MeOD)δ7.74(d,J=8.8Hz,2H),7.22–7.18(m,4H),7.14–7.10(m,1H),7.05(d,J=8.8Hz,2H),4.78–4.71(m,3H),3.98(ddd,J=11.2,7.6,3.6Hz,1H),3.91(d,J=3.2Hz,1H),3.88(d,J=7.2Hz,1H),3.85(s,3H),3.72(td,J=8.8,2.8Hz,1H),3.51(d,J=2.8Hz,1H),3.42–3.35(m,2H),3.23(dd,J=14.0,3.6Hz,1H),2.98–2.92(m,2H),2.83(dd,J=13.2,7.2Hz,1H),2.74–2.71(m,1H),2.54(dd,J=14.0,11.2Hz,1H),2.25–2.19(m,2H),2.06–1.93(m,2H),1.84(dt,J=13.6,10.0Hz,2H),1.45(dd,J=12.4,4.4Hz,1H),1.32–1.27(m,1H),0.99(s,3H),0.88(t,J=6.8Hz,6H),0.84(d,J=6.8Hz,3H),0.74(d,J=6.8Hz,3H),0.64(d,J=6.8Hz,3H).

[0135] 13 C NMR(101MHz,MeOD)δ174.6,173.7,163.0,162.4,155.8,138.6,130.7,129.3,128.9,127.8,125.8,124.1,114.0,73.3,72.0,70.5,63.6,63.1,61.0,59.7,57.8,55.5,54.8,54.6,53.6,52.8,50.7,40.0,35.8,35.4,29.3,27.6,26.5,22.7,19.1,19.0,16.7,16.6,16.4,15.6,12.7.

[0136] HRMS(ESI)m / z calcd.for C 45 H 57 N3O 12 S([M+Na] + ):886.3561,found886.3572.

[0137] Example 9 Preparation of Compound 26: Triptolide ((R)-1-(((2S,3R)-3-hydroxy-4-((N-((S)-2-hydroxypropyl)-4-methoxyphenyl)sulfonamido)-1-phenylbutan-2-yl)amino)-1-oxopropan-2-yl)amino acid methyl ester

[0138]

[0139] The preparation method refers to Example 1, wherein in step B, 2-amino-N-((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methoxyphenyl)sulfonamido)-1-phenylbutan-2-yl)acetamide is replaced by (R)-2-amino-N-((2S,3R)-3-hydroxy-4-((N-((S)-2-hydroxypropyl)-4-methoxyphenyl)sulfonamido)-1-phenylbutan-2-yl)propionamide.

[0140] 1 H NMR(600MHz, CDCl3)δ7.73(d,J=8.8Hz,2H),7.27–7.25(m,2H),7.22–7.18(m ,3H),6.99(d,J=8.8Hz,2H),4.83(s,1H),4.69–4.63(m,2H),4.26–4.21(m,1H ),4.11–4.09(m,1H),3.95(t,J=7.0Hz,2H),3.87(s,3H),3.82(d,J=2.8Hz,1 H),3.49(d,J=2.4Hz,1H),3.45(d,J=5.6Hz,1H),3.21(dd,J=15.0,7.2Hz,1H) ,3.14–3.08(m,3H),2.90(dd,J=14.0,9.0Hz,1H),2.86–2.81(m,1H),2.67(d ,J=12.6Hz,1H),2.31(d,J=16.4Hz,1H),2.17–2.13(m,2H),1.93–1.89(m,2H) ,1.55(dd,J=12.0,5.0Hz,1H),1.23–1.19(m,1H),1.17(d,J=7.2Hz,3H),1.1 3(d,J=7.2Hz,3H),1.05(s,3H),0.86(d,J=6.8Hz,3H),0.77(d,J=6.8Hz,3H).

[0141] 13C NMR (101MHz, CDCl3) δ173.3,172.2,163.2,160.1,155.8,137.8,129.6,129.3,129.2,128.5,126.5,125.6,114.5,73.0,72.3,70.1,67.0 ,63.7,63.5,61.2,59.9,58.5,55.7,55.6,54.9,54.2,53.6,51.1,40 .4,36.0,35.7,29.9,28.0,23.4,20.8,18.0,17.5,17.1,16.6,13.8.

[0142] HRMS(ESI)m / z calcd.for C 44 H 55 N3O 13 S([M+Na] + ):888.3353,found888.3380.

[0143] Example 10 Preparation of Compound 28: Triptolide ((R)-1-(((2S,3R)-3-hydroxy-4-((N-((R)-2-hydroxypropyl)-4-methoxyphenyl)sulfonamido)-1-phenylbutan-2-yl)amino)-1-oxopropan-2-yl)amino acid methyl ester

[0144]

[0145] The preparation method refers to Example 1, wherein in step B, 2-amino-N-((2S,3R)-3-hydroxy-4-((N-isobutyl-4-methoxyphenyl)sulfonamido)-1-phenylbutan-2-yl)acetamide is replaced by (R)-2-amino-N-((2S,3R)-3-hydroxy-4-((N-((R)-2-hydroxypropyl)-4-methoxyphenyl)sulfonamido)-1-phenylbutan-2-yl)propionamide.

[0146] 1H NMR(600MHz, CDCl3)δ7.72(d,J=8.8Hz,2H),7.25(d,J=7.2Hz,2H),7.21–7.1 7(m,3H),6.99(d,J=8.8Hz,2H),4.84(s,1H),4.70–4.64(m,2H),4.22–4.14( m,2H),4.04(t,J=6.0Hz,1H),3.94–3.92(m,1H),3.87(s,3H),3.82(d,J=2.4 Hz,1H),3.46(dd,J=17.4,12.6Hz,3H),3.24(d,J=16.2Hz,1H),3.12(dd,J=1 4.4,4.8Hz,1H),2.93–2.85(m,2H),2.76(dd,J=12.6,11.4Hz,1H),2.68–2.6 6(m,1H),2.31(d,J=18.0Hz,1H),2.17–2.10(m,2H),1.90(t,J=15.6Hz,2H), 1.56(dd,J=12.6,5.4Hz,1H),1.23–1.19(m,1H),1.16(d,J=6.6Hz,3H),1.13 (d,J=7.0Hz,3H),1.06(s,3H),0.88(d,J=6.6Hz,3H),0.76(d,J=6.6Hz,3H).

[0147] 13 C NMR (101MHz, CDCl3) δ173.3,172.2,163.2,160.1,155.6,137.8,129.6,129.5,129.3,128.5,126.5,125.6,114.5,74.1,72.3,70.0,68 .1,63.7,63.5,61.2,59.9,59.5,55.8,55.7,54.9,52.9,51.0,40.4,35.7,35.4,29.9,28.0,23.4,20.5,18.0,17.5,17.1,16.6,13.9.

[0148] HRMS(ESI)m / z calcd.for C 44 H 55 N3O 13 S([M+Na] + ):888.3353,found888.3376.

[0149] Test case pharmacological activity test

[0150] The substrate used in the compound described in the present invention is (Arg-Glu(EDANS)-Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gln-Lys(DABCYL)-Arg)(AnaSpec), and the substrate cleavage point is labeled with Edans and Dabcyl chromophores on both sides. The fluorescence color spectrum of Edans overlaps with the absorption spectrum of Dabcyl, and fluorescence quenching occurs through fluorescence resonance energy transfer within a sufficiently close distance, so that the intact substrate has almost no fluorescence. When the fluorescent substrate is cleaved by HIV protease, the Edans chromophore moves away from the Dabcyl group, and the fluorescence quenching condition is reduced. At this time, Edans generates fluorescence at 490nm under 340nm excitation light. After the test compound is added, if the compound has strong enzyme inhibitory activity, the substrate product decreases and the fluorescence intensity decreases, and vice versa, the fluorescence intensity increases.

[0151] HIV-1 protease was expressed in Escherichia coli and purified. The protease was desalted using a PD-10 column. Activity was assayed in a pH 4.7 buffer solution composed of 0.1 M sodium acetate, 1 M sodium chloride, 1 mM EDTA, 1 mM DTT, 2% DMSO, and 1 mg / mL bovine serum albumin.

[0152] HIV-1 protease inhibitory activity was determined using a 96-well plate. Substrate (5 μM) and 185 μL of buffer were added to each well. 5 μL of sample solution was added, blank absorption was measured, 10 μL of HIV-1 protease was added, and absorbance at a wavelength of 490 nm was measured after incubation for 5 minutes. The inhibition rate of the sample at each concentration was calculated, and the IC was obtained using Graphpad software. 50 The results are shown in Table 2.

[0153] Table 2 HIV-1 protease inhibitory pharmacological activity test results of compounds 1 to 31

[0154]

[0155]

[0156]

[0157]

[0158]

[0159] The preliminary pharmacological activity results in Table 2 show that the triptolide derivatives provided by the present invention have good HIV-1 protease inhibitory activity, among which the IC 50The value is 0.16nM, which is better than the positive control Darunavir 0.82nM. Since there are no reports on the introduction of triptolide into the design of HIV-1 protease, the triptolide derivative with the structure shown in formula (I) has good prospects for further research and development.

[0160] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A triptolide derivative and a pharmaceutically acceptable salt thereof, characterized in that: The triptolide derivative has a structure shown in Formula 1, Formula 2, Formula 4, Formula 6, Formula 8, Formula 9, Formula 10, Formula 11, Formula 12, Formula 13, Formula 14, Formula 15, Formula 16, Formula 18, Formula 19, Formula 20, Formula 21, Formula 22, Formula 23, Formula 24, Formula 25, Formula 28, Formula 29, Formula 30 or Formula 31:

2. The method for preparing the triptolide derivative and the pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The preparation method of the triptolide derivative comprises the following steps: Under the action of pyridine, triptolide having a structure represented by Formula A undergoes a substitution reaction with p-nitrophenyl chloroformate having a structure represented by Formula B to obtain a compound having a structure represented by Formula C; A compound having a structure represented by Formula D is subjected to a condensation reaction with a compound having a structure represented by Formula E, and after de-Boc protection, a compound having a structure represented by Formula F is obtained; Under the action of triethylamine, the compound having the structure represented by formula C and the compound having the structure represented by formula F undergo an acylation reaction to obtain a triptolide derivative; The triptolide derivative is the triptolide derivative according to claim 1 having a structure shown in Formula 1, Formula 2, Formula 4, Formula 6, Formula 8, Formula 9, Formula 10, Formula 11, Formula 12, Formula 13, Formula 14, Formula 15, Formula 16, Formula 18, Formula 19, Formula 20, Formula 21, Formula 22, Formula 23, Formula 24, Formula 25, Formula 28, Formula 29, Formula 30 or Formula 31.

3. The preparation method according to claim 2, characterized in that The temperature of the substitution reaction is 0-25°C and the time is 2-4 hours; The condensation reaction temperature is 0-25°C and the time is 2-5h; The temperature for removing Boc protection is 25° C. and the time is 2 to 5 hours.

4. The preparation method according to claim 2 or 3, characterized in that The molar ratio of the compound having the structure represented by formula C to triethylamine is 1:2-3; The acylation reaction is carried out at a temperature of 25° C. and for a period of 2 to 5 hours.

5. Use of the triptolide derivative and pharmaceutically acceptable salt thereof according to claim 1 in the preparation of anti-AIDS drugs.

6. An anti-AIDS pharmaceutical composition comprising an active ingredient and a drug carrier, characterized in that: The active ingredient includes the triptolide derivative according to claim 1 and a pharmaceutically acceptable salt thereof.

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