A steroidal compound, its preparation method and application

Abstract

The present application discloses a kind of steroid compound and its preparation method and application, the steroid compound has the structure shown in formula I or formula II: Wherein, X is selected from O or N, when selected from O, R3 is not present;R1 is selected from C 1～10 Alkyl or C 1～10 Alkyl substituted with one of hydroxyl, acetoxy, C 2～10 Alkenyl;R2 is selected from H, halogen, C 1～6 Alkyl, C 1～6 Alkoxy;Or R2 is selected from phenyl or phenyl substituted with hydroxyl, C 1～6 Alkyl, and with adjacent benzoquinone form fused ring;R3 is selected from H, C 1～6 Alkyl, phenyl, benzyl.The present application provides a series of novel structure steroid compound has anti-inflammatory effect.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to a steroid compound, its preparation method, and its application. Background Technology

[0002] Steroids are widely found in plants, animals, and microorganisms and are an important type of small molecule drug. Currently, over 300 steroidal compounds have been developed for clinical disease treatment, with a market size exceeding $10 billion. However, reports on steroids based on ring skeleton transformations (ring expansion / contraction) are scarce.

[0003] 6 / 6 / 6 / 6 / 5 type pentacyclic steroids are a rare class of steroidal compounds. Only about 20 such natural products have been reported to date, distributed in marine fungi and plant endophytes. The preparation of these steroidal compounds mainly involves isolation and purification from fungi; no synthetic methods have been reported. However, the low abundance of 6 / 6 / 6 / 6 / 5 type pentacyclic steroids in natural resources greatly hinders in-depth research into their biological functions and mechanisms of action. There is an urgent need to develop efficient and concise synthetic methods and to explore their biological functions. Summary of the Invention

[0004] The present invention aims to at least solve one of the technical problems existing in the prior art. To this end, the first aspect of the present invention provides a steroidal compound with anti-inflammatory effects.

[0005] A second aspect of the present invention also provides a method for preparing a steroidal compound.

[0006] A third aspect of the present invention also provides an application of a steroid compound.

[0007] The steroidal compound or a pharmaceutically acceptable salt thereof provided according to a first aspect of the present invention has the structure shown in Formula I or Formula II:

[0008]

[0009] Where X is selected from O or N, and when it is selected from O, R3 does not exist;

[0010] R1 is selected from C 1～10 The C-aryl group or the C-aryl group substituted with one of the hydroxyl or acetoxy groups. 1～10 alkyl, C 2～10 alkenyl groups;

[0011] R2 is selected from H, halogens, and C. 1～6 alkyl, C 1～6 The alkoxy group; or R2 is selected from phenyl or hydroxyl, C 1～6 The alkyl-substituted phenyl group forms a fused ring with the adjacent benzoquinone;

[0012] R3 is selected from H and C. 1～6 Alkyl, phenyl, benzyl.

[0013] The steroidal compounds according to embodiments of the present invention have at least the following beneficial effects:

[0014] The novel carrier compounds provided by this invention can reduce the expression level of related inflammatory factor mRNA at the cellular level induced by LPS, and have anti-inflammatory effects.

[0015] According to some embodiments of the present invention, R1 is selected from one of the following structural formulas:

[0016]

[0017] According to some embodiments of the present invention, the steroid compound or a pharmaceutically acceptable salt thereof is selected from one of the following structural formulas:

[0018]

[0019] The method for preparing the steroid compound according to a second aspect embodiment of the present invention includes the following steps:

[0020] Compound i, compound ii, an oxidant, and a solvent are mixed and reacted to obtain;

[0021] The structural formula of compound i is as follows:

[0022] R4 is selected from hydroxyl, C 2～5 Acyloxy group, C 1～6 One of the following: alkoxy, azide, alkylamine, amide, or sulfonamide groups;

[0023] Compound ii is selected from one of the following structural formulas:

[0024]

[0025] The method for preparing steroidal compounds according to embodiments of the present invention has at least the following beneficial effects:

[0026] This invention uses compound i as a raw material to prepare steroidal compounds in a one-step reaction under oxidant conditions. This method has the characteristics of high selectivity, scalability, and no need for protection in key steps.

[0027] According to some embodiments of the present invention, the oxidant is selected from at least one of p-benzoquinone, o-benzoquinone, dichlorocyanobenzoquinone, tetrachlorobenzoquinone, 1,4-naphthoquinone, 1,2-naphthoquinone, 2,6-naphthoquinone, o-phenanthrenequinone, p-phenanthrenequinone, anthraquinone, alizarin, silver oxide, copper acetate, copper chloride, copper trifluoromethanesulfonate, copper bromide, and oxygen.

[0028] According to some embodiments of the present invention, the solvent is selected from at least one of formic acid, acetic acid, propionic acid, trifluoroacetic acid, hydrochloric acid, sulfuric acid, acetic anhydride, chloroform, dichloromethane, ethyl acetate, acetonitrile, tetrahydrofuran, 1,4-dioxane, diethyl ether, dimethyl sulfoxide, N,N-dimethylformamide, methanol, ethanol, and n-heptane.

[0029] According to some embodiments of the present invention, the solvent is selected from a mixture of n-heptane and formic acid, acetic acid, propionic acid, trifluoroacetic acid, hydrochloric acid, sulfuric acid, acetic anhydride, chloroform, dichloromethane, ethyl acetate, acetonitrile, tetrahydrofuran, 1,4-dioxane, diethyl ether, dimethyl sulfoxide, N,N-dimethylformamide, methanol, or ethanol.

[0030] According to some embodiments of the present invention, the reaction temperature is 40°C to 130°C.

[0031] According to some embodiments of the present invention, the reaction time is 1 to 6 hours.

[0032] According to some embodiments of the present invention, the molar ratio of compound i, compound ii and oxidant is 1:(0.001-1):(0.5-5).

[0033] According to some embodiments of the present invention, the molar ratio of compound i, compound ii and oxidant is 1:(0.01-1):(0.5-5).

[0034] According to some embodiments of the present invention, the present invention further includes a purification step after the reaction.

[0035] According to some embodiments of the present invention, the purification includes dissolution, filtration, column chromatography, extraction, and recrystallization.

[0036] A third aspect of the present invention provides the use of the steroidal compound described herein in the preparation of a medicament for treating and / or preventing inflammatory diseases.

[0037] According to some embodiments of the present invention, the steroidal compounds of the present invention are used in the preparation of medicaments for treating and / or preventing inflammatory diseases caused by inflammatory factors.

[0038] According to some embodiments of the present invention, the inflammatory factors include TNF-α, IL-1β, and IL-6.

[0039] According to some embodiments of the present invention, the inflammatory diseases include acute lung injury, sepsis, rheumatoid arthritis, and chronic nephritis.

[0040] Definitions and general terms

[0041] “C1-6 "alkyl" indicates an alkyl group with a total number of 1-6 carbon atoms, including C64. 1-6 straight-chain alkyl, C 1-6 Branched alkyl groups and C 3-6 The cycloalkyl group can be, for example, a straight-chain alkyl group with a total of 1, 2, 3, 4, 5, or 6 carbon atoms; a branched-chain alkyl group with a total of 1, 2, 3, 4, 5, or 6 carbon atoms; or a cycloalkyl group with a total of 3, 4, 5, or 6 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, n-hexyl, cyclopropyl, methylcyclopropyl, ethylcyclopropyl, cyclopentyl, methylcyclopentyl, cyclohexyl, etc. Regarding "C 1-10 "alkyl" has a similar interpretation, except that the number of carbon atoms is different.

[0042] “C 1-6 "alkoxy group" refers to an alkoxy group with a total number of 1-6 carbon atoms, including C64 and C64. 1-6 straight-chain alkoxy, C 1-6 Branched alkoxy groups and C 2-6 The cycloalkoxy group can be, for example, a straight-chain alkoxy group with a total number of carbon atoms of 1, 2, 3, 4, 5 or 6, a branched-chain alkoxy group with a total number of carbon atoms of 1, 2, 3, 4, 5 or 6, or a cycloalkoxy group with a total number of carbon atoms of 2, 3, 4, 5 or 6, such as methoxy, ethoxy, n-propoxy, isopropoxy, etc.

[0043] “C 2-10 "Alkenyl" refers to a straight-chain or branched hydrocarbon group having one or more double bonds, and the total number of carbon atoms in the alkenyl group is 2-10, and the double bonds in the group can be in any position.

[0044] "Halogen" includes any one or more of fluorine, chlorine, bromine, and iodine.

[0045] Other features and advantages of the invention will be set forth in the description which follows, and will be apparent in part from the description, or may be learned by practicing the invention. Attached Figure Description

[0046] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the description of the embodiments taken in conjunction with the following drawings, in which:

[0047] Figure 1 This is a cytotoxicity diagram of the steroidal compounds prepared in the embodiments of the present invention;

[0048] Figure 2 This is a graph showing the effect of the steroidal compounds prepared in the embodiments of the present invention on inflammatory factors. Detailed Implementation

[0049] The following are specific embodiments of the present invention, and the technical solutions of the present invention will be further described in conjunction with the embodiments, but the present invention is not limited to these embodiments.

[0050] Unless otherwise specified, the reagents, methods and equipment used in this invention are all conventional reagents, methods and equipment in this technical field.

[0051] Example 1

[0052] This example provides a steroidal compound, the reaction equation of which and its preparation method are as follows:

[0053]

[0054] At 60 °C, compound i (ergosterol, CAS No. 57-87-4; 0.5 mmol, 1.0 eq.) and 1,4-benzoquinone (3 mmol, 6 eq.), followed by acetic acid (5.0 mL), were added sequentially to a reaction flask equipped with a magnetic stir bar. The mixture was stirred at this temperature for 3 h after the addition was complete. After the reaction was complete as monitored by TLC, it was cooled to room temperature. The reaction solution was diluted with ethyl acetate and washed sequentially with saturated sodium bicarbonate solution, water, and saturated sodium chloride aqueous solution, followed by concentration under reduced pressure. The crude product was purified by column chromatography (petroleum ether:EtOAc = 50:1) to give 130 mg of the red product 2a, with a yield of 54%.

[0055] Its proton NMR data are as follows:

[0056] 1 H NMR (500MHz, CDCl3)δ H7.63(s,1H),6.76(d,J=10.2Hz,1H),6.73(d,J=10.2Hz,1H),6.27(d,J=9.7Hz,1H),6.02–5.98(m,1H),5.40(dd,J=15.3,7.9Hz,1 H),5.29(dd,J=15.3,8.4Hz,1H),3.37(dd,J=18.7,7.2Hz,1H),2.87(dd,J=18.7,11.2Hz,1H),2.59–2.56(m,1H),2.41(dq,J=15.1 ,6.6Hz,1H),2.29(m,2H),2.24(dt,J=12.4,3.0Hz,1H),1.94–1.86(m,4H),1.72(td,J=10.7,7.3Hz,1H),1.52(ddt,J=19.4,12.8 ,5.6Hz,3H),1.11(d,J=6.6Hz,3H),1.04(s,3H),1.02(d,J=6.8Hz,3H),0.89(d,J=6.8Hz,3H),0.87(d,J=6.8Hz,3H),0.83(s,3H).

[0057] Example 2

[0058] This example provides a steroidal compound with the following structural formula, prepared by the following method:

[0059]

[0060] The preparation method is basically the same as that in Example 1, except that compound ii is different. Compound ii in Example 2 is... Preparation of 162 mg of product 2b 1 and 2b 2 The mixture, in a ratio of 1:1.5, yielded a total yield of 63%.

[0061] Its NMR data are as follows:

[0062] 1 H NMR (400MHz, CDCl3)δ H7.65&7.63(s,2H),7.04&7.03(s,2H),6.28(d,J=9.4Hz,2H),6.08–6.01(m,2H) ,5.36(m,4H),3.39(m,2H),2.90(m,2H),2.61(t,J=9.0Hz,2H),2.47–2.36(m,2H ),2.27(m,6H),1.96–1.86(m,8H),1.80–1.69(m,2H),1.53(m,6H),1.12(d,J=6 .6Hz, 6H), 1.06 (s, 6H), 1.01 (d, J=6.8Hz, 6H), 0.90–0.85 (m, 12H), 0.85 (s, 6H).

[0063] Example 3

[0064] This example provides a steroidal compound with the following structural formula:

[0065]

[0066] The preparation method is basically the same as that in Example 1, except that compound ii is different. Compound ii in Example 3 is... The prepared 118.5 mg product 2c 1 and 2c 2 The mixture, in a ratio of 1:1.7, yielded a total output of 48%.

[0067] Its NMR data are as follows:

[0068] 1 H NMR (400MHz, CDCl3)δ H 7.66&7.62(s,2H),6.65&6.63(s,2H),6.27(d,J=8.6Hz,2H),6.03–5.95(m,2H),5.44–5.2 5(m,4H),3.39(m,2H),2.89(m,2H),2.59(t,J=9.0Hz,2H),2.47–2.34(m,2H),2.31–2.21( m,6H),2.10&2.07(d,J=1.5Hz,6H),1.96–1.84(m,8H),1.73(m,2H),1.52(m,6H),1.12&1. 11(d,J=6.8,6H),1.05(s,6H),1.02&1.01(d,J=6.8,6H),0.90–0.85(m,12H),0.84(s,6H).

[0069] Example 4

[0070] This example provides a steroidal compound with the following structural formula:

[0071]

[0072] The preparation method is basically the same as that in Example 1, except that compound ii is different. Compound ii in Example 4 is... 118.0 mg of product was obtained 2 days ago. 1 and 2D 2 , 2d 1 :2d 2 =1:1.3, 48% total yield.

[0073] Compound 2d 1 NMR data (21%):

[0074] 1 H NMR (600MHz, CDCl3)δ H 7.36(s,1H),6.63(s,1H),6.29(d,J=10.3Hz,1H),5.98(m,1H),5.39(dd,J=15.3,7.9Hz,1H),5.30(dd,J=15.6,8.6Hz ,1H),3.33(dd,J=18.5,7.1Hz,1H),2.88(dd,J=18.5,11.2Hz,1H),2.59–2.56(m,1H),2.44–2.38(m,1H),2.32–2.27( m,2H),2.25(dt,J=12.8,3.2Hz,1H),1.94–1.87(m,4H),1.72(td,J=10.8,7.2Hz,1H),1.56–1.49(m,3H),1.32(s,9H) ,1.11(d,J=6.6Hz,3H),1.05(s,3H),1.01(d,J=6.8Hz,3H),0.89(d,J=6.8Hz,3H),0.87(d,J=6.9Hz,3H),0.86(s,3H).

[0075] Compound 2d 2 NMR data (27%):

[0076] 1 H NMR (600MHz, CDCl3)δ H7.60(s,1H),6.64(s,1H),6.28(d,J=9.8Hz,1H),5.98(m,1H),5.40(dd,J=15.3,7.6Hz,1H),5.34(dd,J=15.4,7.9Hz,1H),3 .32(dd,J=18.4,7.1Hz,1H),2.96(dd,J=18.4,11.2Hz,1H),2.58(dd,J=10.5,7.6Hz,1H),2.42(ddt,J=14.3,9.9,6.7Hz,1H) ,2.32–2.26(m,2H),2.24(dt,J=12.7,3.2Hz,1H),1.97–1.86(m,4H),1.71(td,J=10.7,7.1Hz,1H),1.57–1.48(m,3H),1.30( s,9H),1.12(d,J=6.6Hz,3H),1.05(s,3H),1.01(d,J=6.8Hz,3H),0.89(d,J=6.8Hz,3H),0.87(d,J=6.8Hz,3H),0.83(s,3H).

[0077] Example 5

[0078] This example provides a steroidal compound with the following structural formula:

[0079]

[0080] The preparation method is basically the same as that in Example 1, except that compound ii is different. Compound ii in Example 5 is... 106.8 mg of product 2e was obtained 1 and 2e 2 ,2e 1 :2e 2 =1:1.3, 42% total yield.

[0081] Compound 2e 1 NMR data (18%):

[0082] 1 H NMR (500MHz, CDCl3)δ H7.63(s,1H),6.26(d,J=9.7Hz,1H),6.04–5.99(m,1H),5.98(s,1H),5.40(dd,J=15.3,7.7Hz,1H),5. 31(dd,J=15.0,8.0Hz,1H),3.81(s,3H),3.42(dd,J=18.6,7.2Hz,1H),2.89(dd,J=18.6,11.2Hz,1H) ,2.61(t,J=9.0Hz,1H),2.47–2.37(m,1H),2.31–2.22(m,3H),1.96–1.86(m,4H),1.74(td,J=10.7,7 .3Hz,1H),1.59(m,3H),1.12(d,J=6.6Hz,3H),1.06(s,3H),1.02(d,J=6.8Hz,3H),0.91–0.84(m,9H).

[0083] Compound 2e 2 NMR data (24%):

[0084] 1 H NMR (500MHz, CDCl3)δ H 7.70(s,1H),6.29(d,J=9.9Hz,1H),5.98–6.02(s,2H),5.39(dd,J=15.2,7.9Hz,1H),5.29(dd,J =16.5,7.0Hz,1H),3.82(s,3H),3.39(dd,J=18.8,7.1Hz,1H),2.90(dd,J=18.7,11.1Hz,1H),2.6 1(t,J=9.1Hz,1H),2.44–2.39(m,1H),2.32–2.23(m,3H),1.93–1.88(m,4H),1.79–1.70(m,1H), 1.62–1.57(m,3H),1.12(d,J=6.5Hz,3H),1.06(s,3H),1.01(d,J=6.8Hz,3H),0.91–0.83(m,9H).

[0085] Example 6

[0086] This example provides a steroidal compound with the following structural formula:

[0087]

[0088] The preparation method is basically the same as that in Example 1, except that compound i is different. Compound i (7-dehydrocholesterol, CAS number: 434-16-2) in Example 6 yielded 147.2 mg of product, with a yield of 63%.

[0089] Its NMR data are as follows:

[0090] 1 H NMR (400MHz, CDCl3)δ H 7.63(s,1H),6.78(d,J=10.2Hz,1H),6.74(d,J=10.2Hz,1H),6.27(d,J=9.8Hz,1H),6.00(dt,J=9.0,4. 0Hz,1H),3.49(dd,J=18.3,6.9Hz,1H),2.90(dd,J=18.3,10.7Hz,1H),2.60–2.55(m,1H),2.29–2.25(m ,3H),1.93–1.85(m,3H),1.79–1.76(m,1H),1.70(td,J=10.6,7.0Hz,1H),1.62–1.49(m,4H),1.40–1.3 6(m,1H),1.23–1.16(m,4H),1.03(s,3H),1.01(d,J=6.4Hz,3H),0.88(s,3H),0.86(s,3H),0.83(s,3H).

[0091] Example 7

[0092] This example provides a steroidal compound with the following structural formula:

[0093]

[0094] The preparation method was basically the same as that in Example 1, except that compound ii was different; in Example 7, compound ii was 1,4-naphthoquinone. 159.1 mg of product 2f was obtained from 200 mg of starting material compound 1, in a 30% yield.

[0095] Its NMR data are as follows:

[0096] 1 H NMR (400MHz, CDCl3)δ H8.19–8.13(m,2H),7.72(s,1H),7.71–7.64(m,2H),6.32(d,J=9.8Hz,1H),6.05–5.96(m,1H), 5.46–5.31(m,2H),3.52(dd,J=18.6,7.2Hz,1H),3.02(dd,J=18.6,11.2Hz,1H),2.66–2.57(m, 1H),2.50–2.39(m,1H),2.33–2.24(m,3H),1.96–1.89(m,4H),1.77(td,J=10.8,7.2Hz,1H),1. 56–1.48(m,3H),1.13(d,J=6.6Hz,3H),1.08(s,3H),1.05(d,J=6.8Hz,3H),0.93–0.79(m,9H).

[0097] Example 8

[0098] This example provides a steroidal compound with the following structural formula:

[0099]

[0100] The preparation method is basically the same as that in Example 1, except that compound ii is different. Compound ii in Example 8 is... 65 mg of product was obtained in 2 hours. 1 and 2h 2 The mixture, in a ratio of 1:1.6, yielded a total yield of 24%.

[0101] Its NMR data are as follows:

[0102] 1 H NMR (600MHz, CDCl3)δ H13.02(s,2H),7.77(s,2H),7.67(dd,J=7.5,1.1Hz,2H),7.57–7.53(m,2H),7.19(dd,J=8.3,1.1Hz,2H),6.32(dd ,J=9.9,1.9Hz,2H),6.04(ddd,J=8.9,5.5,2.6Hz,2H),5.40–5.35(m,4H),3.52(dd,J=18.6,7.2Hz,2H),2.99(dd ,J=18.6,11.2Hz,2H),2.63–2.57(m,2H),2.49–2.39(m,2H),2.34–2.24(m,6H),1.96–1.89(m,8H),1.77(td,J=1 0.8,7.2Hz,2H),1.57–1.49(m,6H),1.14(d,J=6.6Hz,6H),1.07(s,6H),1.04(d,J=6.8Hz,6H),0.90–0.87(m,9H).

[0103] Example 9

[0104] This example provides a steroidal compound with the following structural formula:

[0105]

[0106] At -40°C, compound 1 (CAS No. 2418-45-3; 0.5 mmol, 1.0 eq.), DDQ (0.6 mmol, 1.2 eq.), and DCM (1 mL) were added sequentially to a reaction flask equipped with a magnetic stir bar. After the addition was complete, the mixture was stirred at this temperature for 3-5 min until the solution turned pink. Maleimide (1.1 mmol, 2.2 eq.) and HOAc (5 mL) were added, and the mixture was stirred at 60°C for 1.5 h. DDQ (0.6 mmol, 1.2 eq.) was added again, and the mixture was stirred at this temperature for another 1.5 h. The mixture was then cooled to room temperature, diluted with ethyl acetate, and washed sequentially with saturated sodium bicarbonate solution, water, and saturated sodium chloride aqueous solution. After concentration under reduced pressure, the mixture was subjected to column chromatography (petroleum ether: EtOAc = 10:1) to give 110.2 mg of product, 47% yield.

[0107] The structural formula of compound 1 is as follows:

[0108]

[0109] The NMR data for product compound 3a are as follows:

[0110] 1 H NMR (400MHz, CDCl3)δ H7.71(s,1H),7.10(s,1H),6.24(d,J=9.8Hz,1H),6.01(dt,J=9.0,4.0Hz,1H),5.36(dd,J=15.3,7.6Hz,1H ),5.27(dd,J=15.3,7.9Hz,1H),3.16(dd,J=17.5,7.0Hz,1H),2.77(dd,J=17.5,11.2Hz,1H),2.64(dd,J=1 0.6,7.3Hz,1H),2.46–2.36(m,1H),2.33–2.22(m,3H),1.91(ddd,J=13.1,8.1,4.8Hz,4H),1.80(td,J=10 .9,7.1Hz,1H),1.52(m,3H),1.11(d,J=6.6Hz,3H),1.07(s,3H),0.97(d,J=6.8Hz,3H),0.87–0.82(m,9H).

[0111] Example 10

[0112] This example provides a steroidal compound with the following structural formula:

[0113]

[0114] The preparation method is basically the same as in Example 9, except that compound ii is different; compound ii in Example 10 is... 158.6 mg of product was obtained, with a yield of 66%.

[0115] The NMR data of the product are as follows:

[0116] 1 H NMR (500MHz, CDCl3)δ H7.11(s,1H),6.24(d,J=9.8Hz,1H),5.99(dt,J=8.8,3.8Hz,1H),5.36(dd,J=15.3,7.8Hz,1H),5.2 7(s,1H),3.15(dd,J=17.5,6.9Hz,1H),3.06(s,3H),2.75(dd,J=17.5,11.3Hz,1H),2.63(dd,J=11. 0,6.9Hz,1H),2.45–2.37(m,1H),2.33–2.21(m,3H),1.95–1.83(m,4H),1.79(td,J=10.8,7.1Hz,1H ),1.59–1.46(m,3H),1.11(d,J=6.6Hz,3H),1.06(s,3H),0.97(d,J=6.8Hz,3H),0.89–0.82(m,9H).

[0117] Example 11

[0118] This example provides a steroidal compound with the following structural formula:

[0119]

[0120] The preparation method is basically the same as in Example 9, except that compound ii is different. Compound ii in Example 11 is... 130 mg of product was obtained, with a yield of 48%.

[0121] Its NMR data are as follows:

[0122] 1 H NMR (400MHz, CDCl3)δ H7.48–7.42(m,2H),7.40–7.31(m,3H),7.20(s,1H),6.24(d,J=9.8Hz,1H),6.02(dt,J=9.0,4.0Hz,1H),5.37( dd,J=15.3,7.8Hz,1H),5.26(dd,J=15.3,8.3Hz,1H),3.22(dd,J=17.6,7.0Hz,1H),2.81(dd,J=17.6,11.2Hz ,1H),2.69(dd,J=10.7,7.2Hz,1H),2.46–2.38(m,1H),2.28(dd,J=13.2,3.2Hz,3H),1.95–1.83(m,5H),1.59 –1.47(m,3H),1.12(d,J=6.6Hz,3H),1.10(s,3H),0.96(d,J=6.8Hz,3H),0.88(s,3H),0.84(t,J=6.5Hz,6H).

[0123] Example 12

[0124] This example provides a steroidal compound with the following structural formula:

[0125]

[0126] The preparation method is basically the same as in Example 9, except that compound ii is different; compound ii in Example 12 is... 117.2 mg of product was obtained, with a yield of 42%.

[0127] Its NMR data are as follows:

[0128] 1 H NMR (400MHz, CDCl3)δ H7.37(d,J=7.2Hz,2H),7.22(m,3H),7.11(s,1H),6.20(d,J=9.8Hz,1H),5.97(dt,J=8.9,3.9Hz,1H),5.32(dd, J=15.3,7.7Hz,1H),5.24(dd,J=15.3,8.0Hz,1H),4.73(s,2H),3.15(dd,J=17.5,6.9Hz,1H),2.73(dd,J=17.5, 11.2Hz,1H),2.60(dd,J=10.7,7.2Hz,1H),2.43–2.31(m,1H),2.29–2.18(m,3H),1.86(m,4H),1.76(td,J=10. 8,7.1Hz,1H),1.54–1.42(m,3H),1.08(d,J=6.6Hz,3H),1.03(s,3H),0.95(d,J=6.8Hz,3H),0.87–0.79(m,9H).

[0129] Example 12

[0130] This example provides a steroidal compound with the following structural formula:

[0131]

[0132] The preparation method is basically the same as in Example 9, except that the compound ii is different. In Example 12, compound ii is maleic anhydride, and 85 mg of product was obtained, with a yield of 36%.

[0133] 1 H NMR (400MHz, CDCl3)δ H 6.99(s,1H),6.25(d,J=9.8Hz,1H),6.08(dt,J=9.1,4.1Hz,1H),5.37(dd,J=15.3,7.8H z,1H),5.26(dd,J=15.3,8.3Hz,1H),3.15(dd,J=17.7,7.0Hz,1H),2.80(dd,J=17.7,11. 3Hz,1H),2.72–2.63(m,1H),2.46–2.37(m,1H),2.29(m,3H),1.99–1.80(m,5H),1.57–1. 44(m,3H),1.12(d,J=6.6Hz,3H),1.09(s,3H),0.97(d,J=6.8Hz,3H),0.88–0.82(m,9H).

[0134] Example 13

[0135] This example provides a series of steroidal compounds, whose reaction equations and preparation methods are as follows:

[0136]

[0137] At -40°C, compound 1 (0.3 mmol, 1 eq.), DDQ (0.36 mmol, 1.2 eq.), and DCM (0.6 mL) were added sequentially to a reaction flask equipped with a magnetic stir bar. After the addition was complete, the mixture was stirred at this temperature for 3-5 min until the solution turned pink. Then, a dienophile (0.66 mmol, 2.2 eq.) and HOAc (3 mL) were added, and the mixture was stirred at 60°C for 1.5 h. DDQ (0.36 mmol, 1.2 eq.) was added again, and the mixture was stirred at this temperature for another 1.5 h. The mixture was then cooled to room temperature, diluted with ethyl acetate, and washed sequentially with saturated sodium bicarbonate solution, water, and saturated sodium chloride aqueous solution. After concentration under reduced pressure, the mixture was subjected to column chromatography to obtain a series of compounds 18.

[0138] In this embodiment, the yields of the products obtained when different substituents were selected for the maleimide compounds are shown in Table 1; the yields in Table 1 are calculated based on the yield of general formula compound 18.

[0139] In the preparation of compounds 18a to 18f in this embodiment, the amount of other materials used is adjusted proportionally according to the amount of raw material (compound i).

[0140] Table 1. Yields of products obtained by using different substituents for general formula compound 18 in this embodiment.

[0141]

[0142] Preparation of compound 18a: 65.8 mg of product was obtained from 100 mg of starting material compound 1 (CAS No.: 118746-72-8), with a yield of 53%.

[0143] 1 H NMR (400MHz, CDCl3)δ H7.62(s,1H),6.75(q,J=10.2Hz,2H),6.27(d,J=9.7Hz,1H),6.01(dt,J=9.0,4.2H z,1H),3.82(dd,J=10.5,3.2Hz,1H),3.62(dd,J=10.5,6.2Hz,1H),3.48(dd,J=18 .1,7.0Hz,1H),2.95(dd,J=18.1,10.8Hz,1H),2.58(t,J=9.0Hz,1H),2.32–2.20( m,3H),1.89(m,5H),1.53(m,3H),1.14(d,J=6.5Hz,3H),1.04(s,3H),0.83(s,3H).

[0144]

[0145] Preparation of compound 18b: 68 mg of product was obtained from 124 mg of starting material compound 1 (CAS No.: 911661-60-4), with a yield of 51%.

[0146] 1 H NMR (600MHz, CDCl3)δ H 7.10(s,1H),6.24(d,J=9.9Hz,1H),6.05–5.99(m,1H),4.24(dd,J=10.9,3.6Hz,1H),3.94(dd ,J=10.9,6.5Hz,1H),3.28(dd,J=16.9,7.0Hz,1H),2.87(dd,J=16.9,11.1Hz,1H),2.66(dd,J =10.8,7.2Hz,1H),2.38–2.20(m,3H),2.12(m,J=10.4,6.6,3.8Hz,1H),2.06(s,3H),1.96–1. 88(m,3H),1.58(m,J=35.3,12.5,5.5Hz,3H),1.11(d,J=6.7Hz,3H),1.08(s,3H),0.86(s,3H).

[0147]

[0148] Preparation of compound 18c: 71.2 mg of product was obtained from 124 mg of starting material compound 1 (CAS No.: 911661-60-4), with a yield of 52%.

[0149] 1 H NMR (400MHz, CDCl3)δ H7.13(s,1H),6.25(d,J=9.8Hz,1H),6.01(dt,J=8.9,4.0Hz,1H),4.23(dd,J=10.9,3.7H z,1H),3.95(dd,J=10.9,6.5Hz,1H),3.28(dd,J=16.9,7.0Hz,1H),3.08(s,3H),2.86(d d,J=16.9,11.1Hz,1H),2.69–2.62(m,1H),2.33–2.21(m,3H),2.12(m,1H),2.06(s,3H) ,1.98–1.84(m,3H),1.67–1.49(m,3H),1.11(d,J=6.6Hz,3H),1.07(s,3H),0.85(s,3H).

[0150]

[0151] Preparation of compound 18d: 68.5 mg of product was obtained from 124 mg of starting material compound 1 (CAS No.: 911661-60-4), with a yield of 44%.

[0152] 1 H NMR (400MHz, CDCl3)δ H 7.45(t,J=7.7Hz,2H),7.41–7.31(m,3H),7.21(s,1H),6.25(d,J=9.8Hz,1H),6.06–5 .99(m,1H),4.23(dd,J=10.9,3.8Hz,1H),3.36(dd,J=17.0,6.9Hz,1H),2.93(dd,J=17 .0,11.1Hz,1H),2.75–2.66(m,1H),2.35–2.25(m,3H),2.15(tt,J=12.6,6.4Hz,1H),2 .06(s,3H),1.99–1.92(m,3H),1.68–1.57(m,3H),1.12(d,J=6.4Hz,6H),0.89(s,3H).

[0153]

[0154] Preparation of compound 18e: 67 mg of product was obtained from 124 mg of starting material compound 1 (CAS No.: 911661-60-4), with a yield of 42%.

[0155] 1 H NMR (400MHz, CDCl3)δ H7.37(d,J=7.3Hz,2H),7.24–7.14(m,3H),7.11(s,1H),6.20(d,J=9.9Hz,1H),5.97(dt,J=8.9,3.9H z,1H),4.73(s,2H),4.19(dd,J=10.9,3.7Hz,1H),3.90(dd,J=10.9,6.4Hz,1H),3.26(dd,J=16.9,6 .9Hz,1H),2.83(dd,J=16.9,11.1Hz,1H),2.66–2.57(m,1H),2.29–2.19(m,3H),2.12–2.05(m,1H), 2.02(s,3H),1.92–1.83(m,3H),1.58–1.51(m,3H),1.07(d,J=6.6Hz,3H),1.04(s,3H),0.81(s,3H).

[0156]

[0157] Preparation of compound 18f: 54.5 mg of product was obtained from 124 mg of starting material compound 1 (CAS No.: 911661-60-4), with a yield of 41%.

[0158] 1 H NMR (500MHz, CDCl3)δ H 6.99(s,1H),6.25(d,J=9.8Hz,1H),6.09(dt,J=9.1,4.0Hz,1H),4.22(dd,J=10.9,3.7 Hz,1H),3.94(dd,J=10.9,6.3Hz,1H),3.28(dd,J=17.2,7.0Hz,1H),2.91(dd,J=17.2, 11.0Hz,1H),2.69(t,J=9.0Hz,1H),2.36–2.24(m,3H),2.18–2.09(m,1H),2.06(s,3H) ,1.94(td,J=10.2,9.4,3.1Hz,3H),1.70–1.51(m,3H),1.16–1.06(m,6H),0.86(s,3H).

[0159] Anti-inflammatory activity test

[0160] (1) The cytotoxicity of the steroidal compounds prepared in the embodiments of the present invention was determined by the MTT assay, including the following steps:

[0161] RAW264.7 cells in logarithmic growth phase were diluted with culture medium to a density of 3000 cells per well and seeded into 96-well plates. 100 μL of cell suspension was added to each well, and the cells were cultured at 37°C in a 5% CO2 incubator for 12 h. After cell attachment, drugs were administered according to concentration gradients and in three parallel wells for each concentration, and the cells were cultured in the drug-treated groups for another 24 h. The drug-treated culture medium in each well was discarded, and 100 μL of 10% MTT (thiazolyl blue) diluted in DMEM medium was added to each well. The cells were cultured at 37°C in a 5% CO2 incubator for approximately 4 h. After removing the supernatant, 100 μL of DMSO was added, and the absorbance of each well was measured at 490 nm using a microplate reader.

[0162] Based on the absorbance value of each well, calculate the cell viability using the formula below, plot the results, and the graph is shown below. Figure 1 As shown, the results indicate that this series of compounds has no significant toxicity to RAW264.7 cells.

[0163] Calculation formula: Cell viability (%) = (OD value of drug administration well - OD value of blank group) / (OD value of normal well - OD value of blank group) * 100%.

[0164] (2) The effects of the steroidal compounds provided in this invention on the expression of LPS-induced inflammatory factors TNF-α, IL-1β and IL-6 were detected in RAW264.7 cells, including the following specific steps:

[0165] Log-grown RAW264.7 cells were seeded evenly in 12-well plates at a density of 20,000 cells per well and cultured at 37°C in a 5% CO2 incubator for 12 hours. The following day, after cell attachment, cells were divided into groups and treated with LPS (Liquid Propionate) in triplicate, with each group cultured in 3 replicates for another 12 hours at 37°C in a 5% CO2 incubator. Cells were collected, total RNA was extracted, and the expression of TNF-α, IL-1β, and IL-6 mRNA was detected using qRT-PCR.

[0166] Table 2 qRT-PCR primer sequences

[0167]

[0168] The result is as follows Figure 2 As shown, where, Figure 2 A shows the effect of the inflammatory factor IL-6. Figure 2 B shows the effect of the inflammatory factor TNF-α. Figure 2 The figure shows the effect of the inflammatory factor IL-1β. It can be seen from the figure that the steroidal compound provided in the embodiments of the present invention can reduce the expression level of related inflammatory factor mRNA at the cellular level induced by LPS at 20 μM, indicating that the steroidal compound of the present invention has an anti-inflammatory effect.

[0169] The present invention has been described in detail above with reference to the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of the present invention.

Claims

1. A steroidal compound or a pharmaceutically acceptable salt thereof, characterized in that, It has the structure shown in Formula II: Where X is selected from O or N, and when X is selected from O, R3 does not exist; R1 is selected from one of the following structural formulas: ； R3 is selected from H.

2. The steroidal compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, Selected from the following structural formulas: 。 3. The method for preparing the steroidal compound according to claim 1 or 2, characterized in that, Includes the following steps: Compound i, compound ii, an oxidant, and a solvent are mixed and reacted to obtain; The structural formula of compound i is as follows: R4 is selected from hydroxyl, C 2~5 Acyloxy group, C 1~6 One of the following: alkoxy, azide, alkylamine, amide, or sulfonamide groups; Compound ii is selected from the following structural formulas: 。 4. The preparation method according to claim 3, characterized in that, The oxidant is selected from at least one of p-benzoquinone, o-benzoquinone, dichlorodicyanobenzoquinone, tetrachlorobenzoquinone, 1,4-naphthoquinone, 1,2-naphthoquinone, 2,6-naphthoquinone, o-phenanthrenequinone, p-phenanthrenequinone, anthraquinone, alizarin, silver oxide, copper acetate, copper chloride, copper trifluoromethanesulfonate, copper bromide, and oxygen.

5. The preparation method according to claim 3, characterized in that, The solvent is selected from at least one of formic acid, acetic acid, propionic acid, trifluoroacetic acid, hydrochloric acid, sulfuric acid, acetic anhydride, chloroform, dichloromethane, ethyl acetate, acetonitrile, tetrahydrofuran, 1,4-dioxane, diethyl ether, dimethyl sulfoxide, N,N-dimethylformamide, methanol, ethanol, and n-heptane.

6. The preparation method according to claim 3, characterized in that, The reaction temperature is -40 ℃ to 130 ℃.

7. The preparation method according to claim 3, characterized in that, The reaction time is 1 to 6 hours.

8. The preparation method according to claim 3, characterized in that, The molar ratio of compound i, compound ii and oxidant is 1:(0.001~1):(0.5~5).

9. The use of the steroidal compound according to claim 1 or 2 in the preparation of medicaments for the prevention and / or treatment of inflammatory diseases.

Images