A class of hard asafetida extracted compounds, and preparation method and application thereof

Abstract

This invention relates to a class of compounds extracted from the whole herb of Ferula assa-foetida, their preparation methods, and applications, belonging to the field of pharmaceutical technology. It also includes a class of compounds having the following chemical structural formula and their pharmaceutically acceptable salts. This invention provides for the first time a method for preparing and identifying six new compounds using whole Ferula as a raw material, and systematically evaluates their anti-inflammatory activity, clarifying their application in the development of anti-inflammatory drugs.

Description

Compounds extracted from hard Ferula assa-foetida, their preparation methods and applications Technical Field

[0001] This invention relates to a class of compounds extracted from Ferula assa-foetida, their preparation methods, and applications, belonging to the field of pharmaceutical technology. Background Technology

[0002] Ferula bungeana Kitagawa is a plant belonging to the genus Ferula L. of the family Umbelliferae. There are over 150 species of Ferula worldwide. In my country, there are 26 species and 1 variety of Ferula.

[0003] In traditional medicine, *Ferula assa-foetida* (hard asafoetida) is used medicinally for its resin and rhizome, possessing properties that dispel cold, relieve pain, reduce swelling, and detoxify. It is commonly used to treat rheumatic joint pain, flatulence, and colds. The main active components of *Ferula assa-foetida* include sesquiterpenes, coumarins, phenylpropanoids, flavonoids, and polysaccharides. These components exhibit various biological activities, such as anti-inflammatory, antibacterial, antioxidant, anticancer, and immunomodulatory effects. Modern pharmacological studies have shown that *Ferula assa-foetida* extract has significant effects in inhibiting the release of inflammatory mediators, regulating immune responses, and combating oxidative stress. It can be used as an adjunct treatment for chronic inflammatory diseases, infectious diseases, and immune-related disorders, and has the potential to be developed into a novel natural drug. Summary of the Invention

[0004] This invention provides six new compounds extracted from the whole herb of Ferula assa-foetida.

[0005] A class of compounds having the following chemical structural formula and their pharmaceutically acceptable salts, characterized in that,

[0006]

[0007] Another object of the present invention is to provide a method for preparing the above-mentioned compound extracted from the whole herb of Ferula assa-foetida.

[0008] A method for preparing compounds extracted from hard Ferula assa-foetida includes the following steps:

[0009] (1) The whole herb of Ferula bungeana K. was extracted by heating and reflux with ethanol or methanol solution, or by ultrasonic extraction, and the crude extract was obtained by recovering the extract.

[0010] (2) After the crude extract obtained in step (1) is dissolved in water, it is extracted with organic solvent at a volume ratio of 1:1 between the aqueous phase and the organic phase. The organic solvents used are petroleum ether or cyclohexane, dichloromethane or chloroform, ethyl acetate, and n-butanol, respectively. The extraction is performed 3 to 5 times to obtain extracts with different polarities.

[0011] (3) The extract obtained in step (2) was dissolved in an organic solvent, then mixed with silica gel and dried. It was separated by silica gel column chromatography using gradient elution with a mixture of petroleum ether and ethyl acetate, or a mixture of petroleum ether and acetone, or a mixture of dichloromethane and acetone, or a mixture of chloroform and acetone, or a mixture of chloroform and methanol, or a mixture of dichloromethane and methanol.

[0012] (4) The fraction obtained in step (3) above is separated by ODS column chromatography, with a gradient elution using a mixed solvent of methanol-water or acetonitrile as the mobile phase;

[0013] (5) The methanol and water, acetonitrile and water eluents obtained in step (4) above are further separated by HPLC and gradient elution is performed using a methanol and water mixed solvent or an acetonitrile and water mixed solvent as the mobile phase to obtain compounds 1, 2, 3, 4, 5 and 6.

[0014] Preferably, in step (1), *Ferula assa-foetida* is extracted by heating and reflux with ethanol or methanol solution 3-5 times, or by ultrasonic extraction 4-8 times. The volume concentration of the ethanol solution is 70%-95%, the volume concentration of the methanol solution is 80%-90%, and the solid-liquid ratio is 1:5-1:20 g / mL.

[0015] Furthermore, the volume concentration of the ethanol solution is 80%–95%, the volume concentration of the methanol solution is 70%–90%, and the material-to-liquid ratio is 1:10–1:15 g / mL.

[0016] Preferably, in step (2), the organic solvent extraction method involves sequentially extracting with petroleum ether or cyclohexane, dichloromethane or chloroform, ethyl acetate, and n-butanol 3 to 5 times according to a volume ratio of aqueous phase to organic phase of 1:1 to 1:5, and then recovering the organic solvents under reduced pressure.

[0017] Furthermore, the volume ratio of the aqueous phase to the organic phase is 1:1 to 1:3.

[0018] Further, the extraction was performed four times.

[0019] Preferably, in step (3), the volume ratio of petroleum ether to ethyl acetate is 100:5 to 1:1, the volume ratio of petroleum ether to acetone is 100:5 to 1:1, the volume ratio of dichloromethane to acetone is 100:1 to 1:1, the volume ratio of chloroform to acetone is 100:1 to 1:1, the volume ratio of dichloromethane to methanol is 100:1 to 1:1, and the volume ratio of chloroform to methanol is 100:1 to 1:1.

[0020] Furthermore, the volume ratio of petroleum ether to ethyl acetate is 100:10 to 1:1, and the volume ratio of petroleum ether to acetone is 100:10 to 1:1.

[0021] Furthermore, the volume ratio of the mixed solvent of dichloromethane and acetone, or chloroform and acetone, or dichloromethane and methanol, or chloroform and methanol is 100:1 to 3:1, preferably 100:3 to 2:1.

[0022] Preferably, in step (4), the volume ratio of methanol to water is 4:6 to 9:1, and the volume ratio of acetonitrile to water is 3:7 to 8:2.

[0023] Furthermore, the volume ratio of methanol to water is 1:9 to 9:1, and the volume ratio of acetonitrile to water is 1:9 to 9:1.

[0024] Preferably, in step (5), the volume ratio of methanol to water is 4:6 to 9:1, and the volume ratio of acetonitrile to water is 4:6 to 8:2.

[0025] Furthermore, the volume ratio of methanol to water is 5:5 to 9:1, and the volume ratio of acetonitrile to water is 4:6 to 9:1.

[0026] Another object of the present invention is to provide a pharmaceutical composition comprising the compounds extracted from the above-mentioned Ferula assa-foetida.

[0027] A pharmaceutical composition comprising a compound with the following chemical structure, a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0028]

[0029] Another object of the present invention is to provide the use of the above-mentioned compounds extracted from *Ferula assa-foetida* and their pharmaceutically acceptable salts or the above-mentioned pharmaceutical compositions containing the compounds extracted from *Ferula assa-foetida* in the preparation of anti-inflammatory drugs for prevention or treatment.

[0030] Furthermore, the present invention provides the use of the above-mentioned compounds extracted from *Ferula assa-foetida* and their pharmaceutically acceptable salts or the above-mentioned pharmaceutical compositions containing the compounds extracted from *Ferula assa-foetida* in the preparation of anti-inflammatory drugs.

[0031] The beneficial effects of this invention are as follows: This invention provides, for the first time, a method for preparing and identifying six new compounds using the whole herb of Ferula assa-foetida as raw material, and systematically evaluates their anti-inflammatory activity, clarifying their application in the development of anti-inflammatory drugs. This invention evaluated the anti-inflammatory activity of the preparation methods of the new compounds 1-6 using an LPS-activated RAW264.7 cell model. The results showed that new compounds 1-6 could inhibit the release of NO from LPS-activated RAW264.7 cells, exhibiting significant dose-dependent anti-inflammatory activity. Therefore, the new compounds prepared in this invention can be applied in the development of anti-inflammatory drugs. Detailed Implementation

[0032] The following non-limiting embodiments are intended to enable those skilled in the art to more fully understand the invention, but do not limit the invention in any way.

[0033] Unless otherwise specified, the experimental methods described in the following examples are conventional methods; the reagents and materials described are commercially available unless otherwise specified.

[0034] Example 1

[0035] (1) 1 kg of whole Ferula assa-foetida herb was extracted three times by heating and refluxing with 95% ethanol (dosage: 9 L), and the crude extract was obtained by vacuum recovery.

[0036] (2) After the crude extract obtained in step (1) above is dissolved in water, it is extracted three times in sequence with petroleum ether, ethyl acetate and n-butanol at a volume ratio of 1:1 between the aqueous phase and the organic phase to obtain extracts of different polarities.

[0037] (3) The ethyl acetate extract obtained in step (2) above was separated by silica gel column chromatography and eluted sequentially with a mixture of petroleum ether and ethyl acetate in the following ratios: 100:1, 100:3, 100:5, 10:1, 8:1, 5:1, 3:1, and 1:1.

[0038] (4) The petroleum ether: ethyl acetate fraction obtained in step (3) above was subjected to ODS chromatography and eluted with a gradient of mixed solvents of methanol-water 3:7, 5:5, 7:3, and 9:1.

[0039] (5) The methanol-water (7:3) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of acetonitrile:water = 67%:33%, to obtain compound 6 (t R =10.1min)(yield 0.00002‰), compound 1 (t R =32.1min)(yield 0.00026‰) and compound 2(t R =41.1 min)(yield rate 0.000012‰).

[0040] (6) The methanol-water (3:7) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of acetonitrile:water = 40%:60%, to obtain compound 3 (t R =21.1 min)(yield rate 0.000012‰).

[0041] (7) The methanol-water (9:1) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of methanol:water = 77%:24%, to obtain compound 4 (t R =9.1 min)(yield of 0.00003‰).

[0042] (8) The methanol-water (7:3) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of methanol:water = 66%:34%, to obtain compound 5 (t). R =22.0 min)(yield rate 0.00003‰).

[0043] The structures of compounds 1–6 were identified based on their physicochemical properties and spectral data.

[0044] The structural identification data of compound 1 are as follows:

[0045] Colorless oil (methanol). HR-ESI-MS yielded a quasi-molecular ion peak at m / z 495.2356 [M+Na]. + :(calcd.495.2359for C 27 H 36 O7Na), its molecular formula is presumed to be C. 27 H 36 O7 has an unsaturation degree of 10. 1 In H-NMR (600MHz, CDCl3), the low-field region exhibits characteristic hydrogen signals from the parent nucleus of scopolamine-type coumarin: δ H 7.61(1H,d,J=9.4Hz,H-4),6.89(1H,s,H-8),6.84(1H,s,H-5),6.29(1H,d,J=9.4Hz,H-3),3.87(3H,s,6-OC H 3); 1 group of hydroxymethylene hydrogen signals: δ H 4.43(1H,dd,J=9.4,4.5Hz,H a -11′), 4.25(1H,t,J=9.4Hz,H b -11′); 1 hydroxymethyl hydrogen signal: 4.66 (1H,t,J=2.8Hz,H-3′); 5 methyl hydrogen signals are given in the high field region: δ H 2.05(3H,s,C H 3-2″), 1.29(3H,s,C H 3-12′), 0.96(3H,s,C H 3-15′), 0.91(3H,s,CH 3-14′), 0.89(3H,s,C H 3-13′). 13 27 carbon signals are provided in C-NMR (150MHz, CDCl3): δ C 161.3(C-2),151.5(C-7),149.8(C-9),146.5(C-6),143.3(C-4),113.8(C-3),111.8(C-10),107.8(C-5),100.7(C-8),56.3(6-O C H3); 15 are sesquiterpene unit carbon signals, including 3 oxygen-linked carbon signals: δ C 77.5 (C-3′), 72.5 (C-8′), 68.1 (C-11′); 1 group of acetoxy carbon signals: δ C 170.6 (C-1″), 21.3 (C-2″). In the HMBC spectrum, δ H 1.29(C H 3-12′) and δ C 43.0(C-7′), 72.5(C-8′), and 57.7(C-9′) are related, indicating that CH3-12′ is connected at position C-8′; δ H 0.89(C H 3-13′), 0.91(C H 3-14′) and δ C The values ​​36.6 (C-4′), 77.5 (C-3′), and 49.5 (C-5′) are relevant, suggesting that CH3-13′ and CH3-14′ are connected at the C-4′ position; δ H 0.96(C H 3-15′) and δ C The values ​​49.5 (C-5′), 57.7 (C-9′), 37.2 (C-10′), and 33.8 (C-1′) are relevant, indicating that CH3-15′ is connected at position C-10′; δ H 4.66(H-3′) and δ C 33.8(C-1′), 49.5(C-5′), 170.6(C-1″) correlation, δ H 2.05(H-2″) and δ C 170.6(C-1″) is relevant, indicating that -OAc is connected in bit C-3′; δ H 4.43(H a -11′), 4.25(H b -11′) and δ CThe correlations of 151.5 (C-7), 37.2 (C-10′), 57.7 (C-9′), and 72.5 (C-8′) suggest that the sesquiterpene fragment is linked to the C-7 position of the coumarin core via an ether bond; δ H 3.87 (6-OC) H 3) with δ C 146.5 (C-6) correlation, indicating that 6-OCH3 is linked at the C-6 position. In the NOESY spectrum, δ H 4.25(H b -11′) and δ H 0.96(C H 3-15′), 1.29(C H (3-12′) related, δ H 1.53(H-5′) and δ H 2.08(H-9′) correlation, δ H 4.66(H-3′) and δ H 0.96(C H The correlation with 3-15′ suggests that H-3′, H2-11′, CH3-12′, and CH3-15′ are α-configurations, and H-5′ and H-9′ are β-configurations. The relative configurations of the chiral centers are inferred to be 3′S*, 5′R*, 8′S*, 9′R*, and 10′R*. By comparing measured and calculated ECD data, the absolute configuration was determined and named (3′S, 5′R, 8′S, 9′R, 10′R)-ferubungeanolI.

[0046] The structural identification data of compound 2 are as follows:

[0047] White powder (methanol). HR-ESI-MS yielded a quasi-molecular ion peak at m / z 413.2338 [M+H]. + :(calcd.413.2328for C 25 H 33 O5), whose molecular formula is presumed to be C. 25 H 32 O5 has an unsaturation degree of 10. 1 H-NMR (600MHz, CDCl3) in the low-field region revealed characteristic hydrogen signals from the parent nucleus of scopolamine-type coumarin: δ H 7.60(1H,d,J=9.4Hz,H-4),6.86(1H,s,H-8),6.84(1H,s,H-5),6.27(1H,d,J=9.4Hz,H-3),3.85(3H,s,6-OC H 3); 1 set of terminal double bond hydrogen signals: δ H 4.91(1H,br.s,H a-12′), 4.57(1H,br.s,H b -12′); 1 group of hydroxymethylene hydrogen signals: δ H 4.27(1H,dd,J=9.9,4.2Hz,H a -11′),4.23(1H,dd,J=9.9,7.4Hz,H b -11′); 1 hydroxymethyl hydrogen signal: 3.46 (1H,t,J=2.9Hz,H-3′); 3 methyl hydrogen signals are given in the high field region: δ H 0.98(3H,s,C H 3-14′), 0.87(3H,s,C H 3-13′), 0.86(3H,s,C H 3-15′). 13 The C-NMR (150MHz, CDCl3) spectrum provides 25 carbon signals, 10 of which are characteristic carbon signals of the parent nucleus of scopolamine-type coumarin: δ C 161.7(C-2),152.9(C-7),150.2(C-9),147.0(C-6),143.5(C-4),113.4(C-3),111.5(C-10),109.1(C-5),101.2(C-8),56.9(6-O C H3); 15 carbon signals are sesquiterpene units, including one set of terminal double bond carbon signals: δ C 146.8 (C-8′), 107.8 (C-12′); 2 oxygen-carbon signals: δ C 75.9 (C-3′), 66.7 (C-11′).

[0048] In the HMBC spectrum, δ H 0.87(C H 3-13′), 0.98(C H 3-14′) and δ C 37.9(C-4′), 75.9(C-3′), and 48.2(C-5′) are related, indicating that CH3-13′ and CH3-14′ are connected at the C-4′ position; δ H 0.86(C H 3-15′) and δ C The values ​​31.8 (C-1′), 39.0 (C-10′), 48.2 (C-5′), and 54.5 (C-9′) are relevant, indicating that CH3-15′ is connected at position C-10′; δ H 3.46(H-3′) and δ C The correlation at 31.8 (C-1′) and 48.2 (C-5′) suggests that the 3′-OH is attached at the C-3′ position; δH 4.91(H a -12′), 4.57(H b -12′) and δ C The values ​​37.7 (C-7′), 54.5 (C-9′), and 146.8 (C-8′) are relevant, indicating that the terminal double bond is connected at the C-8′ position; δ H 4.27(H a -11′), 4.23(H b -11′) and δ C The correlations of 39.0 (C-10′), 54.5 (C-9′), 146.8 (C-8′), and 152.9 (C-7′) suggest that the sesquiterpene fragment is linked to the C-7 position of the coumarin core via an ether bond; δ H 3.85 (6-OC) H 3) with δ C 147.0 (C-6) correlation indicates that 6-OCH3 is linked at the C-6 position. In the NOESY spectrum, δ H 4.27(H a -11′), 4.23(H b -11′) and δ H 0.86(C H (3-15′) related, δ H 1.65(H-5′) and δ H 2.41(H-9′), 0.98(C H (3-14′) related, δ H 3.46(H-3′) and δ H 0.98(C H The correlation between 3-14′ and 1.65(H-5′) suggests that H-3′, H-5′, and H-9′ are α-configurations, and CH3-15′ is a β-configuration. The relative configurations of the chiral centers are inferred to be 3′S*, 5′R*, 9′S*, and 10′S*. By comparing measured and calculated ECD data, the absolute configuration was determined and named (3′S, 5′R, 9′S, 10′S)-ferubungeanol J.

[0049] The structural identification data of compound 3 are as follows:

[0050] White powder (methanol). HR-ESI-MS yielded a quasi-molecular ion peak at m / z 421.1993 [M+Na]. + :(calcd.421.1991for C 24 H 30 O5Na), its molecular formula is presumed to be C. 24 H 30 O5 has an unsaturation degree of 10.1 H-NMR (600MHz, CDCl3) in the low field region gives the characteristic hydrogen signal of the 7-O-substituted coumarin nucleus: δ H 7.63 (1H, d, J = 9.5 Hz, H-4), 7.35 (1H, d, J = 9.2 Hz, H-5), 6.82 (2H, overlap, H-6, 8), 6.25 (1H, d, J = 9.5 Hz, H-3); 1 set of terminal double bond hydrogen signals: δ H 5.27(1H,br.s,H a -12′), 4.73(1H,br.s,H b -12′); 1 group of hydroxymethylene hydrogen signals: δ H 4.24 (2H, m, H2-11′); two hydroxymethyl hydrogen signals: 4.12 (1H, dd, J = 11.0, 5.5 Hz, H-7′), 3.49 (1H, t, J = 3.5 Hz, H-3′); three methyl hydrogen signals are given in the high-field region: δ H 1.00(3H,s,C H 3-13′), 0.88(3H,s,C H 3-14′), 0.85(3H,s,C H 3-15′). 13 The C-NMR (150 MHz, CDCl3) provides 24 carbon signals, 9 of which are characteristic carbon signals of the 7-O-substituted coumarin nucleus: δ C 162.2(C-7), 161.2(C-2), 155.9(C-9), 143.4(C-4), 128.8(C-5), 113.1(C-3,6), 112.5(C-10), 101.3(C-8); 15 carbon signals representing sesquiterpene units, including one set of terminal double bond carbon signals: δ C 148.8 (C-8′), 104.3 (C-12′); 3 oxygen-carbon signals: δ C 75.5 (C-3′), 73.3 (C-7′), 65.3 (C-11′). In the HMBC spectrum, δ H 1.00(C H 3-13′), 0.88(C H 3-14′) and δ C The values ​​37.7 (C-4′), 75.5 (C-3′), and 45.7 (C-5′) are relevant, suggesting that CH3-13′ and CH3-14′ are connected at the C-4′ position; δ H 0.85(C H 3-15′) and δ CCorrelation at 31.6(C-1′), 38.6(C-10′), 45.7(C-5′), and 53.1(C-9′) indicates that CH3-15′ is connected at position C-10′; δ H 3.49(H-3′) and δ C 31.6 (C-1′) and 45.7 (C-5′) are related, indicating that the 3′-OH is attached at the C-3′ position; δ H 5.27(H a -12′), 4.73(H b -12′) and δ C 53.1(C-9′), 73.3(C-7′), 148.8(C-8′) correlation, δ H 4.12(H-7′) and δ C 32.9 (C-6′) is relevant, indicating that the terminal double bond is attached at the C-8′ position and the 7′-OH is attached at the C-7′ position; δ H 4.24(H2-11′) and δ C The correlations at 38.6 (C-10′), 53.1 (C-9′), 148.8 (C-8′), and 162.2 (C-7′) suggest that the sesquiterpene fragment is linked to the C-7 position of the coumarin core via an ether bond. In the NOESY spectrum, δ... H 4.24(H2-11′) and δ H 0.85(C H (3-15′) related, δ H 2.28(H-9′) and δ H 1.75(H-5′), 4.12(H-7′) correlation, δ H 1.75(H-5′) and δ H 1.00(C H (3-13′) related, δ H 3.49(H-3′) and δ H 0.88(C H The correlation with 3-14′ suggests that CH3-15′ and H-3′ are α-configurations, while H-5′, H-7′, and H-9′ are β-configurations. The relative configurations of the chiral centers are inferred to be 3′S*, 5′S*, 7′R*, 9′S*, and 10′R*. The absolute configuration was determined by comparing measured and calculated ECD data and named (3′S, 5′S, 7′R, 9′S, 10′R)-ferubungeanol K.

[0051] Table 1. Attribution of 1H NMR and 1C NMR data for compounds 1–3

[0052]

[0053]

[0054] The structural identification data of compound 4 are as follows:

[0055] Colorless oil (methanol). HR-ESI-MS yielded a quasi-molecular ion peak at m / z 293.1397 [MH]. - :(calcd.293.1394for C 16 H 21 O5), whose molecular formula is presumed to be C. 16 H 22 O5 has an unsaturation degree of 6. 1 The characteristic ABX-coupled aromatic hydrogen signal of the benzene ring is given in H-NMR (600MHz, CDCl3): δ H 6.87 (1H, d, J = 8.0 Hz, H-5′), 6.85 (1H, d, J = 1.8 Hz, H-2′), 6.78 (1H, dd, J = 8.0, 1.8 Hz, H-6′); 1 group of angelica acyl hydrogen signals: δ H 6.03 (1H, qq, J = 7.3, 1.6 Hz, C) H 3-3″),1.97(3H,dq,J=7.3,1.6Hz,C H 3-4″), 1.90 (3H, p, J = 1.6Hz, C H 3-5″); 1 hydroxyl hydrogen signal: δ H 5.60(1H,s,3′-O H ); Two hydroxymethyl hydrogen signals: 5.20 (1H, dq, J = 7.0, 6.5 Hz, H-2), 4.09 (1H, d, J = 7.0 Hz, H-1), based on the coupling constant, the configurations of H-1 and H-2 are inferred to be 1R*, 2R*; Two methoxy hydrogen signals: δ H 3.89(3H,s,4′-OC H 3), 3.23(3H,s,1-OC) H 3). 13 The C-NMR (150MHz, CDCl3) spectrum provides 14 carbon signals, including 6 aromatic carbon signals: δ C 146.9(C-4′), 145.7(C-3′), 130.2(C-1′), 121.5(C-6′), 114.1(C-5′), 109.3(C-2′); Carbon signals of the five angelic acyl groups: δ C 167.9 (C-1″), 137.1 (C-3″), 128.5 (C-2″), 20.8 (C-5″), 15.8 (C-4″); 2 methoxy carbon signals: δC 57.1(1-O C H3), 56.1(4′-O C H3).

[0056] In the HMBC spectrum, δ H 3.89(4′-OC H 3) with δ C 146.9(C-4′) correlation, δ H 5.60(3′-OH) and δ C 145.7(C-3′) correlation, δ H 5.20(H-2) and δ C The correlation at 167.9 (C-1″) suggests that the methoxy, hydroxyl, and angelic acyl fragments are attached at the C-4′, C-3′, and C-2 positions, respectively; δ H 4.09(H-1) and δ C The correlations at 130.2 (C-1′), 121.5 (C-6′), and 109.3 (C-2′) suggest that C-1 is connected at the C-1′ position. In the NOESY spectrum, δ... H 1.90(C H 3-5″) and δ H The correlation of 6.03(H-3″) indicates that the C-2″ and C-3″ positions are double bonds of the Z type. Based on the coupling constant, the configuration of H-1 and H-2 is inferred to be 1R*, 2R*. By comparing the measured ECD and calculated ECD data, the absolute configuration was determined and named (1R, 2R)-bungeanolA.

[0057] The structural identification data of compound 5 are as follows:

[0058] Colorless oil (methanol). HR-ESI-MS yielded a quasi-molecular ion peak at m / z 399.1791 [M+Na]. + :(calcd.399.1784for C 21 H 28 O6Na), its molecular formula is presumed to be C. 21 H 28 O6 has an unsaturation degree of 8. 1 One set of ABX-coupled aromatic hydrogen signals was given in H-NMR (600MHz, CDCl3): δ H 6.96 (1H, dd, J = 8.2, 1.9 Hz, H-6′), 6.89 (1H, d, J = 1.9 Hz, H-2′), 6.84 (1H, d, J = 8.2 Hz, H-5′); 2 groups of angelica acyl hydrogen signals: δ H 6.08 (1H, qq, J = 7.2, 1.4 Hz, C) H3-3″), 1.96(3H,m,C H 3-4″), 1.88(3H,m,C H 3-5″) and δ H 6.05 (1H, qq, J = 7.2, 1.5 Hz, C) H 3-3″′), 1.95(3H,m,C H 3-4″′), 1.85(3H,m,C H 3-5″′); 2 hydroxymethyl hydrogen signals: δ H 5.84 (1H, d, J = 7.5 Hz, H⁻¹), 5.39 (1H, dq, J = 7.5, 6.4 Hz, H⁻²), based on the coupling constants, the configurations of H⁻¹ and H⁻² are deduced to be 1R*, 2R*.

[14] ; 2 groups of methoxy hydrogen signals: δ H 3.87(6H,s,3′-OC H 3,4′-OC H 3). 13 The C-NMR (150MHz, CDCl3) provides 21 carbon signals, including 6 aromatic carbon signals: δ C 149.3(C-3′), 149.1(C-4′), 129.9(C-1′), 120.3(C-6′), 111.1(C-5′), 110.4(C-2′); 10 angelica acyl carbon signals, including 2 carbonyl carbon signals: δ C 167.4 (C-1″′), 166.9 (C-1″), two sets of double bond carbon signals: δ C 138.8 (C-3″), 127.7 (C-2″), 138.3 (C-3″′), 128.0 (C-2″′); 2 methoxy carbon signals: δ C 56.0(3′-O C H3,4′-O C H3).

[0059] In the HMBC spectrum, δ H 3.87(3′-OC H 3) with δ C 149.3(C-3′) correlation, δ H 3.87(4′-OC H 3) with δ C 149.1 (C-4′) is relevant, suggesting that the two methoxy groups are attached at the C-3′ and C-4′ positions, respectively; δ H 5.84(H-1) and δ C 166.9(C-1″) correlation, δ H 5.39(H-2) and δ C167.4 (C-1″′) is relevant, suggesting that two angelic acyl fragments are substituted at the C-1 and C-2 positions, respectively; δ H 5.84(H-1) and δ C Correlation at 129.9 (C-1′), 120.3 (C-6′), and 110.4 (C-2′) indicates that C-1 is attached at the C-1′ position. This confirms the planar structure of the compound. In the NOESY spectrum, δ H 1.88(C H 3-5″) and δ H 6.08(H-3″) correlation, δ H 1.85(C H 3-5″′) and δ H The correlation of 6.05 (H-3″′) indicates that the double bonds at positions C-2″, C-3″ and C-2″′, C-3″′ are all of the Z-type. Based on the coupling constant, the configuration of H-1 and H-2 is inferred to be 1R*, 2R*. By comparing the measured ECD and calculated ECD data, the absolute configuration was determined and named (1R, 2R)-bungeanolA.

[0060] Table 2. Attribution of 1H NMR and 1C NMR data for compounds 4–5

[0061]

[0062]

[0063] *Overlapping signals

[0064] The structural identification data for compound 6 are as follows:

[0065] White powder (methanol). HR-ESI-MS yielded a quasi-molecular ion peak at m / z 433.2580 [M+H]. + :(calcd.433.2585for C 25 H 37 O6), whose molecular formula is presumed to be C. 25 H 36 O6 has an unsaturation degree of 8.

[0066] 1 In H-NMR (600MHz, CDCl3), two sets of angelic acyl hydrogen signals were given: δ H 6.05(1H,q,J=7.3Hz,H-3′),1.97(3H,d,J=7.3Hz,C H 3-4′), 1.83(3H,m,C H 3-5′) and δ H6.09(1H,q,J=7.3Hz,H-3″),1.98(3H,d,J=7.3Hz,C H 3-4″), 1.85(3H,m,C H 3-5″); 1 double-bonded hydrogen signal: δ H 5.69 (1H, d, J = 7.7 Hz, H⁻³); 1 hydroxymethyl hydrogen signal: δ H 5.39 (1H, d, J = 7.7 Hz, H⁻²); 1 group of hydroxymethylene hydrogen signals: δ H 4.10(1H,d,J=11.4Hz,H a -14), 4.28 (1H,d,J=11.4Hz,H) b -14); 1 hydroxyl hydrogen signal: δ H 2.26(1H,br.s,5-O H )

[24] In addition, there are 3 methyl hydrogen signals: δ H 1.74(3H,s,C H 3-15), 1.05(3H,d,J=6.8Hz,C H 3-13), 0.99(3H,d,J=6.8Hz,C H 3-12). 13 The C-NMR (150 MHz, CDCl3) provides 25 carbon signals, 15 of which are sesquiterpene unit carbon signals, including one carbonyl carbon signal: δ C 215.8 (C-1); Group 1 double-bonded carbon signal: δ C 148.1 (C-4), 119.2 (C-3); 3 oxygen-carbon signals: δ C 82.8 (C-5), 73.0 (C-2), 62.9 (C-10); 3 methyl carbon signals: δ C 26.3 ( C H3-15), 24.7 ( C H3-13), 20.6 ( C H3-12); 10 are angelica acyl carbon signals, including 2 carbonyl carbon signals: δ C 166.7 (C-1″), 165.9 (C-1′), two sets of double bond carbon signals: δ C 140.2 (C-3′), 126.7 (C-2′) and δ C 140.3 (C-3″), 126.8 (C-2″); 4 methyl carbon signals: δ C 20.9 ( C H3-5′,5″),15.9( C H3-4′,4″).

[0067] In the HMBC spectrum, δ H 0.99(C H 3-12) and δ C 26.4 (C-11), 50.9 (C-7) correlation, δ H 1.05(C H 3-13) and δ C The correlation values ​​of 20.6 (C-12), 26.4 (C-11), and 50.9 (C-7) suggest that the isopropyl group is attached at the C-7 position; δ H 1.74(C H 3-15) and δ C 119.2(C-3) and 148.1(C-4) are related, indicating that CH3-15 is connected at position C-4; δ H 2.26(5-O H ) and δ C The values ​​36.3 (C-6), 62.9 (C-10), and 82.8 (C-5) are relevant, indicating that -OH is connected at the C-5 position; δ H 4.10(H a -14), 4.28(H b -14) and δ C 62.9(C-10), 73.0(C-2), 166.7(C-1″), 215.8(C-1) are correlated, δ H 5.39(H-2) and δ C The values ​​82.8 (C-5), 119.2 (C-3), 148.1 (C-4), 165.9 (C-1′), and 215.8 (C-1) are related, with the hydroxymethylene group (C-14) attached at the C-10 position, and the two angelic acyl groups attached at the C-14 and C-2 positions, respectively. In the NOESY spectrum, δ... H 6.05(H-3′) and δ H 1.83(C H 3-5′) related, δ H 6.09(H-3″) and δ H 1.85(C H (3-5″) Related, indicating that the double bonds of C-2′,C-3′ and C-2″,C-3″ are all of the Z type. δ H 4.10(H a -14), 4.28(H b -14) and δ H 5.39(H-2), 2.26(5-O) H Related to δ H 2.26(5-O H ) and δ H 0.99(CH (3-12) Related to this, it is suggested that H-7 is α-configuration and H-2,5-OH is β-configuration, and the relative configuration of the chiral center is inferred to be 2S*,5S*,7R*,10S*. By comparing the measured ECD and calculated ECD data, the absolute configuration was determined and named (2S,5S,7R,10S)-bungeanol C.

[0068] Table 3. Attribution of 1H NMR and 1C NMR data for compound 6

[0069]

[0070] *Overlapping signals

[0071] Implement column 2

[0072] (1) 500g of whole Ferula assa-foetida herb was extracted three times by heating and reflux with 95% ethanol (volume: 6L), and the crude extract was obtained by vacuum recovery.

[0073] (2) The ethanol extract obtained in step (1) above is extracted with an organic solvent, and petroleum ether, ethyl acetate and n-butanol are extracted in sequence with a volume ratio of water phase and organic phase of 1:2. The extraction is carried out 3 times to obtain extracts of different polar fractions.

[0074] (3) The ethyl acetate extract obtained in step (2) above was separated by silica gel column chromatography and eluted sequentially with a mixed solvent of dichloromethane and acetone: 100:1, 100:3, 100:5, 10:1, 8:1, 5:1, 3:1.

[0075] (4) The dichloromethane:acetone fraction obtained in step (3) above was subjected to ODS chromatography and eluted with a gradient of mixed solvents of methanol-water 3:7, 5:5, 7:3, and 9:1.

[0076] (5) The methanol-water (7:3) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of methanol:water = 75%:25%, to obtain compound 6 (t). R =13.1min)(yield 0.00002‰), compound 1 (t R =19.7min)(yield 0.00022‰) and compound 2(t R =44.3 min)(yield rate 0.000012‰).

[0077] (6) The methanol-water (3:7) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of acetonitrile:water = 37%:63%, to obtain compound 3(t). R =29.7 min)(yield rate 0.000012‰).

[0078] (7) The methanol-water (7:3) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of methanol:water = 75%:24%, to obtain compound 4 (t R =11.1 min)(yield 0.00003‰) and compound 5 (t R =16.2min)(yield rate 0.00003‰).

[0079] The structural identification methods for new compounds 1–6 are described in Example 1.

[0080] Example 3

[0081] (1) 1000g of whole Ferula assa-foetida herb was extracted three times by heating and reflux with 80% ethanol (dosage: 15L), and the crude extract was obtained by vacuum recovery.

[0082] (2) The ethanol extract obtained in step (1) above was extracted with organic solvents, namely cyclohexane, ethyl acetate and n-butanol at a volume ratio of 1:3 for aqueous phase and organic phase, and extracted 5 times to obtain extracts of different polar fractions.

[0083] (3) The ethyl acetate extract obtained in step (2) above was separated by silica gel column chromatography and eluted sequentially with a mixture of chloroform and methanol at ratios of 100:1, 100:3, 100:5, 10:1, 8:1, 5:1, and 3:1.

[0084] (4) The chloroform:methanol 100:3 to 8:1 fraction obtained in step (3) above was subjected to ODS chromatography and eluted with a gradient of mixed solvents of acetonitrile-water 3:7, 5:5, 7:3, and 9:1.

[0085] (5) The methanol-water (7:3) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of methanol:water = 77%:23%, to obtain compound 6 (t R =10.0 min)(yield 0.00002‰), compound 1 (t R =13.1min)(yield 0.00022‰) and compound 2(t R =35.8 min)(yield rate 0.000012‰).

[0086] (6) The methanol-water (3:7) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of acetonitrile:water = 35%:65%, to obtain compound 3(t). R =37.6 min)(yield rate 0.000012‰).

[0087] (7) The methanol-water (7:3) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of acetonitrile:water = 70%:30%, to obtain compound 4 (t R =13.6 min)(yield 0.00003‰) and compound 5 (t R =12.4min)(yield rate 0.00003‰).

[0088] The structural identification methods for new compounds 1–6 are shown in Example 1.

[0089] Example 4

[0090] (1) 1500g of whole Ferula assa-foetida herb was extracted 4 times by heating and reflux with 80% methanol (volume: 20L), and the crude extract was obtained by vacuum recovery of the extract.

[0091] (2) The methanol extract obtained in step (1) above was extracted with an organic solvent, and then extracted with petroleum ether, dichloromethane, ethyl acetate and n-butanol in a volume ratio of 1:3 between the aqueous phase and the organic phase. The extraction was carried out 4 times to obtain extracts of different polar fractions.

[0092] (3) The ethyl acetate extract obtained in step (2) above was separated by silica gel column chromatography and eluted sequentially with a mixture of chloroform and acetone at ratios of 100:1, 100:3, 100:5, 10:1, 8:1, 5:1, and 3:1.

[0093] (4) The chloroform:acetone 100:3 to 8:1 fraction obtained in step (3) above was subjected to ODS chromatography and eluted with a gradient of mixed solvents of acetonitrile-water at ratios of 2:8, 4:6, 6:4, and 8:2:

[0094] (5) The acetonitrile-water (6:4) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of methanol:water = 70%:30%, to obtain compound 6 (t R =18.5min)(yield 0.00002‰)

[0095] (6) The acetonitrile-water (4:6) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of methanol:water = 50%:50%, to obtain compound 3 (t R =16.1min)(yield rate 0.000010‰).

[0096] (7) The acetonitrile-water (8:2) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of acetonitrile:water = 72%:28%, to obtain compound 1 (t R =15.0min)(yield 0.00025‰), compound 2 (t R =21.3min)(yield 0.000012‰) and compound 4(t R =11.2min)(yield 0.00003‰) and compound 5(t R =9.4 min)(yield rate 0.00003‰).

[0097] The structural identification methods for new compounds 1–6 are shown in Example 1.

[0098] Example 5

[0099] (1) 2000g of whole Ferula assa-foetida herb was extracted three times by heating and reflux with 80% methanol (volume: 20L), and the crude extract was obtained by vacuum recovery.

[0100] (2) The methanol extract obtained in step (1) above was extracted with organic solvents, and cyclohexane, chloroform and n-butanol were extracted in sequence with a volume ratio of 1:1 between the aqueous phase and the organic phase. The extraction was carried out 5 times to obtain extracts of different polar fractions.

[0101] (3) The chloroform extract obtained in step (2) above was separated by silica gel column chromatography and eluted sequentially with a mixture of petroleum ether and acetone in the following ratios: 100:5, 10:1, 8:1, 5:1, 3:1, and 1:1.

[0102] (4) The petroleum ether: acetone fraction obtained in step (3) above was subjected to ODS chromatography and eluted with a gradient of mixed solvents of methanol-water 3:7, 5:5, 7:3, 8:2, and 9:1.

[0103] (5) The acetonitrile-water (8:2) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of methanol:water = 68%:32%, to obtain compound 6 (t R =24.2min)(yield 0.00002‰) and compound 4(t R=26.7 min)(yield rate 0.00003‰).

[0104] (6) The acetonitrile-water (5:5) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of methanol:water = 45%:55%, to obtain compound 3 (t R =28.1 min)(yield rate 0.000011‰).

[0105] (7) The acetonitrile-water (7:3) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of acetonitrile:water = 74%:26%, to obtain compound 1 (t R =12.3min)(yield 0.00025‰), compound 2 (t R =14.8min)(yield 0.000015‰) and compound 5(t R =6.8 min)(yield rate 0.00001‰).

[0106] The structural identification methods for new compounds 1–6 are shown in Example 1.

[0107] Example 6

[0108] (1) 2500g of whole Ferula assa-foetida herb was extracted three times by heating and reflux with 90% methanol (volume: 25L), and the crude extract was obtained by vacuum recovery of the extract.

[0109] (2) The methanol extract obtained in step (1) above was extracted with an organic solvent, and then extracted with petroleum ether, ethyl acetate and n-butanol in a volume ratio of 1:2 between the aqueous phase and the organic phase, and extracted 4 times to obtain extracts of different polar fractions.

[0110] (3) The ethyl acetate extract obtained in step (2) above was separated by silica gel column chromatography and eluted sequentially with a mixture of chloroform and methanol at ratios of 100:1, 100:3, 100:5, 10:1, 8:1, 5:1, and 3:1.

[0111] (4) The chloroform:methanol 100:3 to 8:1 fraction obtained in step (3) above was subjected to ODS chromatography and eluted with a gradient of mixed solvents of acetonitrile-water 2:8, 4:6, 6:4, 8:2, and 9:1.

[0112] (5) The acetonitrile-water (8:2) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of methanol:water = 70%:30%, to obtain compound 4 (t R =7.3min)(yield rate 0.00003‰).

[0113] (6) The acetonitrile-water (5:5) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of methanol:water = 48%:52%, to obtain compound 3 (t R =20.6 min)(yield rate 0.000011‰).

[0114] (7) The acetonitrile-water (7:3) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of acetonitrile:water = 70%:30%, to obtain compound 1 (t R =25.9min)(yield 0.00025‰), compound 2 (t R =31.8 min)(yield rate 0.000015‰).

[0115] (8) The acetonitrile-water (7:3) fraction obtained in step (4) above was separated by HPLC-UV chromatography, with detection at 210 nm, a flow rate of 3 mL / min, and a mobile phase of acetonitrile:water = 72%:28%, to obtain compound 6 (t R =17.2min)(yield 0.00002‰) and compound 5(t R =18.6 min)(yield rate 0.00001‰).

[0116] The structural identification methods for new compounds 1–6 are shown in Example 1.

[0117] Example 7: Anti-inflammatory activity test of the new compounds 1-6 prepared in Examples 1-6.

[0118] (1) Experimental Principle: Abnormal activation of macrophages is an important part of the inflammatory response and is closely related to the occurrence and development of various inflammatory diseases. Therefore, inhibiting the excessive activation of macrophages may become an important target for drug development. LPS can activate macrophages to release inflammatory factors such as NO, TNF-α, IL-6, and IL-1β. In this experiment, an in vitro LPS-induced abnormally activated RAW264.7 mouse macrophage model was established, and the anti-inflammatory activity of new compounds 1-6 was evaluated using NO release as an indicator.

[0119] (2) Experimental methods:

[0120] ① Culture of RAW264.7 macrophages

[0121] All glassware and metal instruments used in the experiment (culture flasks, pipettes, solution bottles, etc.) were autoclaved at 121°C for 30 minutes to thoroughly remove LPS contamination. A cell culture medium containing 10% fetal bovine serum (FBS) was prepared using DMEM medium as a base. RAW264.7 cells were cultured and passaged at a concentration of approximately 2.0 × 10⁵ cells / mL at 37°C and 5% CO₂. When the cells covered 70%-80% of the bottom of the culture flask, the adherent cells were digested with trypsin and then passaged. RAW264.7 cells revived from cryopreservation at -80°C were used as the starting cells for the experiments, with cells from passages 3-8 selected for use.

[0122] ② Drug preparation method

[0123] All test compounds were dissolved in DMSO to prepare a stock solution (100 mM) and stored at -20°C. Before use, the solution was diluted with DMEM culture medium to 100 μM, 30 μM, 10 μM, and 1 μM, respectively. The final DMSO concentration was <1‰.

[0124] ③ Griess method for detecting the inhibitory effect of compounds on LPS-activated RAW264.7 cells

[0125] RAW264.7 cells in logarithmic growth phase were harvested and their density adjusted to 2.0 × 10⁶ cells / year using fresh DMEM medium containing 10% fetal bovine serum. 5 Cells were seeded at 100 μL / well in 96-well plates and cultured at 37°C and 5% CO2 for 24 hours until cell adhesion. The culture medium was then replaced with serum-free fresh medium, and compounds (concentrations of 100 μM, 30 μM, 10 μM, and 1 μM) were added in combination with LPS. The final LPS concentration was 100 ng / mL. A blank control group (without LPS or the compound) was included. The final LPS concentration in each treatment group was 100 ng / mL. After culturing for another 24 hours following drug administration, the supernatant was collected, and NO levels in the supernatant were determined using the Griess colorimetric method. 2- content.

[0126] ④ The effect of the compound on the survival rate of microglia was detected by MTT assay.

[0127] RAW264.7 cells cultured in the logarithmic growth phase were used to adjust the cell density to 2.0 × 10⁶ cells / year using fresh DMEM medium containing 10% fetal bovine serum. 5Cells were seeded at 100 μL / well in 96-well plates and cultured at 37°C in a 5% CO2 incubator. After 24 h of adherent culture, the medium was replaced with fresh medium, and the drug was added simultaneously. Compound doses of 100 μM, 30 μM, 10 μM, and 1 μM were used in combination with LPS. A blank control was also included. The final LPS concentration in each treatment group was 100 ng / mL. After drug addition, cells were cultured for another 24 h, then MTT solution (10 μL / well) was added to the cell culture medium. Cells were incubated with 0.25 mg / mL MTT at 37°C for 3 h. The culture medium was then removed, and 150 μL of DMSO solution was added. The optical density (OD) value was measured. Data processing was performed using microplate reader software. The average OD value of three wells for each sample was calculated, and the cell viability (CV%) was calculated using the average value according to the following formula.

[0128] Cell viability % = [Average OD value of sample group / Average OD value of blank control group] × 100%

[0129] ⑤ Statistical methods

[0130] All data were analyzed using SPSS (19.0) statistical software. Results are expressed as mean ± standard error. To assess overall differences, one-way ANOVA was used to analyze homogeneity of variance between groups, combined with Dunnett's test for inter-group comparisons. The Levene test was used to test homogeneity of variance in multiple samples. When p > 0.05, the variances were homogeneous, and Dunnett's two-tailed t-test was used to assess differences in means among groups. When p < 0.05, the variances were unequal, and Dunnett's T3 test was used to assess differences in means among groups.

[0131] ⑥IC 50 Calculation method

[0132] IC was calculated by fitting parameters such as dose and inhibition rate using nonlinear regression. 50 .

[0133] (3) Experimental results:

[0134] The experimental results are shown in Table 4.

[0135] Table 4. Effects of compounds 1-6 on LPS-activated NO release from RAW264.7 cells.

[0136]

[0137]

[0138] Note: *P<0.05, **P<0.01, ***P<0.001 compared with the LPS-induced group; ### P<0.001 compared with the control group.

[0139] The results showed that the new compounds 1 (10 μM, 30 μM, 100 μM), 2 (1 μM, 10 μM, 30 μM, 100 μM), 3 (10 μM, 30 μM, 100 μM), 4 (1 μM, 10 μM, 30 μM, 100 μM), 5 (10 μM, 30 μM, 100 μM), and 6 (1 μM, 10 μM, 30 μM, 100 μM) prepared in Implementation Regulations 1-6 could significantly inhibit LPS-induced NO release from RAW264.7 cells.

Claims

1. A class of compounds having the following chemical structural formula and their pharmaceutically acceptable salts, characterized in that, The chemical structural formula is: 。 2. The method for preparing the compound according to claim 1, characterized in that... The method includes the following steps: (1) The whole herb of Ferula bungeana K. is extracted by heating and reflux with ethanol or methanol solution, and the crude extract is recovered; (2) The crude extract obtained in step (1) is dissolved in water and then extracted with organic solvent at a volume ratio of 1:1 between the aqueous phase and the organic phase. The organic solvents used are petroleum ether, dichloromethane, ethyl acetate, and n-butanol, which are extracted 3 to 5 times to obtain extracts of different polarities; (3) The ethyl acetate extract obtained in step (2) is dissolved in organic solvent, dried with silica gel, and separated by silica gel column chromatography with gradient elution using a mixed solvent of chloroform and acetone; (4) The fraction obtained in step (3) above is separated by ODS column chromatography with gradient elution using a mixed solvent of acetonitrile and water as the mobile phase; (5) The acetonitrile-water eluent obtained in step (4) above is further separated by HPLC with gradient elution using a mixed solvent of methanol and water as the mobile phase to obtain compound 6.

3. The preparation method according to claim 2, characterized in that, In step (1), hard asafoetida is extracted by heating and refluxing with ethanol solution or methanol solution 3 to 5 times, wherein the volume concentration of ethanol solution is 70% to 95%, the volume concentration of methanol solution is 80% to 90%, and the material-to-liquid ratio is 1:5 to 1:20 g / mL.

4. The preparation method according to claim 2, characterized in that, In step (3), the volume ratio of chloroform to acetone is 100:1 to 1:

1.

5. The preparation method according to claim 2, characterized in that, In step (4), the volume ratio of acetonitrile to water is 1:9 to 9:

1.

6. The preparation method according to claim 2, characterized in that, In step (5), the volume ratio of methanol to water is 4:6 to 9:

1.

7. A pharmaceutical composition, characterized in that, It comprises the compound of claim 1, its pharmaceutically acceptable salt, and its pharmaceutically acceptable carrier.

8. The use of the compound of claim 1 and its pharmaceutically acceptable salt or the pharmaceutical composition of claim 7 in the preparation of a medicament for the prevention or treatment of inflammation.

Images