Preparation and Application of Two Iridoid Compounds with Anti-Inflammatory Activity Extracted from the Rhizomes of *Semen Pharbitidis*

Abstract

The present invention relates to two iridoid compounds Xylanchinoid A (1) and Xylanchinoid B (2) extracted from the rhizome of *Gelsemium elegans* and their preparation methods and applications, belonging to the field of pharmaceutical technology. Compounds 1 and 2 have inhibitory effects on the cytokines TNF- α , IL-6 and IL-1 β secreted by LPS-induced macrophages RAW264.7, can form a pharmaceutical composition with a pharmaceutically acceptable carrier, and can be used for the development of anti-inflammatory drugs.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceuticals, and particularly relates to the preparation and application of two iridoid compounds with anti-inflammatory activity extracted from the rhizomes of Xylanche himalaica Hook.f.et Thoms.. Background Art

[0002] Xylanche himalaica Hook.f.et Thoms., also known as Xylanche himalaica, Loquat Tuber (Shaanxi), Pinellia (Tibet), is a plant of the genus Xylanche in the family Orobanchaceae. It often parasitizes on the roots of plants of the genus Rhododendron and is mainly distributed in Yunnan, Qinghai, Tibet, Sichuan, Hubei and other regions. Its spherical tubers are used as medicine, with the effects of relieving cough and reducing phlegm, relieving distension and strengthening the stomach, and regulating qi and relieving pain. Some ethnic minorities use its tubers to treat schistosomiasis, irregular menstruation, rheumatic arthralgia, mumps and aconite poisoning and other diseases. This traditional Chinese medicine has good clinical value in traditional medicine and modern drug discovery. However, the research on its material basis is very weak. The main secondary metabolites isolated from this plant by domestic and foreign scholars are triterpenoids, phenyl ethanol glycosides and lignans. Iridoid components have not been reported from this plant, and the pharmacological activity research is relatively scarce. In order to explore the medicinal value of Xylanche himalaica, the present invention takes the active part of Xylanche himalaica as the research object, and uses various chromatographic means such as silica gel, RP 18 , MCI, etc. to separate and purify compounds, and uses various spectroscopic techniques such as NMR, HR-ESI-MS, IR, etc. to identify two novel iridoid compounds. Pharmacological activity screening shows that two compounds have inhibitory effects on the cytokines TNF-α, IL-6 and IL-1β secreted by LPS-induced macrophages RAW264.7, and can be used for the preparation of anti-inflammatory drugs. So far, there is no report on compound Xylanchinoid A (1) and Xylanchinoid B (2), nor on their medicinal activities, and even less on the anti-inflammatory effects of compounds 1-2 and their pharmaceutical compositions. Summary of the Invention

[0003] The purpose of the present invention is to provide a new class of two novel iridoid compounds with medicinal value, their preparation methods and applications. These compounds have good anti-inflammatory activity and can be used in the preparation of anti-inflammatory drugs. In the in-depth research of the applicant on the rhizomes of Xylanche himalaica, two previously unreported iridoid compounds were discovered. Therefore, the present invention provides work on their separation methods, structure identification, anti-inflammatory activity, medicinal prospects, etc. In order to achieve the above object of the present invention, the present invention provides the following technical solutions:

[0004] The present invention provides two novel iridoid compounds, and their structural formulas are shown as follows:

[0005]

[0006] Both of the two iridoid compounds are isolated from the rhizome of Flemingia philippinensis Merr. ex Rolfe, and the Flemingia philippinensis Merr. ex Rolfe used is Xylanche himalaica Hook.f.et Thoms of the family Orobanchaceae.

[0007] The preparation method of the two new iridoid compounds of the present invention is as follows:

[0008] Step 1: Extraction

[0009] After crushing the dried Flemingia philippinensis Merr. ex Rolfe medicinal materials, reflux extraction is carried out with water. The dosage of the extraction solvent is 8 - 15 times the mass of the raw materials, the extraction times are 2 - 4 times, 1 - 3 h each time. The extraction solution is filtered through a 200 - mesh filter, the water extraction solutions are combined, concentrated to a clear paste with a relative density of 0.8 - 1.5 (60 °C), cooled, 95% ethanol is added to make the alcohol content reach 65% - 75%, left standing for 24 hours, filtered, and the filtrate is concentrated to obtain a fluid extract.

[0010] Step 2: Segmenting

[0011] Take the fluid extract obtained in Step 1, carry out silica gel column chromatography separation, and use the chloroform:methanol solvent systems 15:1, 9:1, 8:2, 7:3, 6:4, 1:1 as eluents for gradient elution in turn. Collect the elution solutions, and monitor by TLC and combine them into 11 segments (Fr1 - Fr11).

[0012] Step 3: Isolation and purification

[0013] The extract of Fr7 segment is successively subjected to MCI column chromatography (water - 70% methanol), silica gel column chromatography (dichloromethane∶methanol∶water 7∶2.5∶1, 7∶3∶1, 6∶4∶1, v / v / v), RP 18 column chromatography (15% - 70% methanol) gradient elution, silica gel column chromatography dichloromethane∶methanol∶water 7∶2.5∶1 elution, silica gel column chromatography (ethyl acetate∶ethanol 10∶1) and Sephadex LH - 20 (methanol) purification to obtain Compound 1 and Compound 2.

[0014] The present invention provides a pharmaceutical composition, which comprises at least one of the iridoid compounds with anti - inflammatory activity in the rhizome of Flemingia philippinensis Merr. ex Rolfe and a pharmaceutically acceptable carrier; the pharmaceutical composition is divided into an oral - administration, external - use or injection - administration pharmaceutical composition according to the administration route. The present invention has no special limitation on the dosage form, and conventional dosage forms well - known in the art can be selected, including but not limited to tablets, capsules, granules, powders, oral liquids, sprays, plasters, powder injections or injections.

[0015] The present invention also provides the use of the iridoid compound or the pharmaceutically acceptable salt thereof having anti-inflammatory activity in the above-mentioned Qianjinzhui, or the use of the above-mentioned pharmaceutical composition in the preparation of anti-inflammatory drugs.

[0016] The embodiments of the present invention provide the in vitro anti-inflammatory activities of the two new iridoid compounds described above.

[0017] Advantages of the present invention:

[0018] The present invention reports for the first time two new iridoid compounds, Xylanchinoid A (1) and Xylanchinoid B (2), from the rhizomes of Qianjinzhui; Compounds 1-2 have inhibitory effects on the cytokines TNF-α, IL-6 and IL-1β secreted by LPS-induced macrophages RAW264.7, and can form a pharmaceutical composition with a pharmaceutical carrier for the preparation of anti-inflammatory drugs. The present invention further enriches the chemical substance basis of the active part of Qianjinzhui, provides active lead compounds for new drug development, and also provides a theoretical basis for the research and development of the drug property of Qianjinzhui. Description of the drawings

[0019] Figure 1 It is the result of the cytotoxicity experiment of Compound 1 and Compound 2.

[0020] Figure 2 It is the detection result of inflammation-related indexes of Compound 1 and Compound 2.

[0021] Figure 3 For Compound 1 1 H- 1 H COSY and HMBC correlations.

[0022] Figure 4 It is the comparison chart of the experimental and theoretical ECD of Compound 1.

[0023] Figure 5 For Compound 2 1 H- 1 H COSY and HMBC correlations. Detailed implementation manners

[0024] In order to better understand the essence of the present invention, the following combines the drawings, and uses the test examples and embodiments of the present invention to further illustrate the iridoid compounds, Xylanchinoid A-B (1-2) of the present invention, and their preparation methods, structure identification, and pharmacological effects, but the present invention is not limited by these test examples and embodiments.

[0025] The technical solutions of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0026] Example 1

[0027] Separation and Preparation of Compound 1 and Compound 2

[0028] Step 1: Extraction

[0029] 34 kg of the rhizome medicinal materials of Herba Lysimachiae christinae are crushed into coarse powder by a CSJ-60 type coarse crusher, divided into 2 batches, 17 kg per batch. Each batch is decocted twice with water, with 10 (170 L) times the amount of water added each time, decocted for 1.5 h. The extract is filtered through a 200-mesh filter screen, and the water extracts are combined and concentrated to a clear paste with a relative density of 1.1 (60 °C), cooled, and 95% ethanol is added to make the alcohol content reach 70%. It is left standing for 24 hours, filtered, and the filtrate is concentrated to obtain 13 kg of a fluid extract.

[0030] Step 2: Segmenting

[0031] Take 2 kg of the above-mentioned fluid extract of Herba Lysimachiae christinae, mix the sample with silica gel (100-200 mesh) according to a mass ratio of 1:1.2, and separate it by silica gel (200-300 mesh) column chromatography. Use chloroform-methanol with a gradient of 15:1 → 1:1 as the eluent for gradient elution. Each fraction is collected separately, and the same fractions are combined by TLC monitoring. A total of 11 component segments are obtained, named Fr1-Fr11 respectively.

[0032] Step 3: Separation and Purification

[0033] The extract of Fr7 segment (324.51 g) is subjected to MCI column chromatography (water-70% methanol), and the same fractions are combined by TLC detection and divided into 10 segments, Fr7.1-Fr7.10. Fr7.3 (200 g) is eluted by silica gel column chromatography (dichloromethane:methanol:water 7:2.5:1, 7:3:1, 6:4:1, v / v / v), and 8 segments are obtained by TLC monitoring and combining the same fractions, Fr7.3.1-Fr7.3.8. Fr7.3.5 (40 g) is subjected to RP 18Column chromatography (15%-70% methanol) with gradient elution was performed, and the same fractions were combined by TLC detection to obtain 9 sub-fractions: Fr7.3.5.1 - Fr7.3.5.9. Fr7.3.5.3 (4.6 g) was eluted with dichloromethane∶methanol∶water 7∶2.5∶1 by silica gel column chromatography, and the same fractions were combined by TLC monitoring to obtain Fr7.3.5.3.1 - Fr7.3.5.3.6; Fr7.3.5.3.1 (450 mg) was purified by sephadex LH-20 (methanol), and the same fractions were combined by TLC monitoring to obtain 2 segments, Fr7.3.5.3.1.1 - Fr7.3.5.3.1.2; Fr7.3.5.3.1.1 (210 mg) was purified by sephadex LH-20 (methanol) to obtain compound 2 (39.8 mg), and Fr7.3.5.3.1.2 (120 mg) was purified by silica gel column chromatography with ethyl acetate∶ethanol 10∶1 elution to obtain compound 1 (25 mg).

[0034] Example 2

[0035] Separation and preparation of compound 1 and compound 2

[0036] Step 1: Extraction

[0037] The rhizome medicinal materials of Fimbristylis peduncularis were crushed into coarse powder, placed in a multi-functional extraction tank, added with 8 times the amount of water, refluxed and extracted for 3 h, extracted 2 times in total. The extract was filtered through a 200-mesh filter screen, and the water extracts were combined, concentrated to a clear paste with a relative density of 1.5 (60 °C), cooled, added with 95% ethanol to make the alcohol content reach 65%, allowed to stand for 24 hours, filtered, and the filtrate was concentrated to obtain a fluid extract for standby.

[0038] Step 2: Segmenting and Step 3: Separation and purification were the same as in Example 1.

[0039] Example 3

[0040] Separation and preparation of compound 1 and compound 2

[0041] Step 1: Extraction

[0042] The medicinal materials of Fimbristylis peduncularis were crushed into coarse powder, placed in a multi-functional extraction tank, added with 15 times the amount of water each time, decocted for 1 h, extracted 2 times. The extract was filtered through a 200-mesh filter screen, and the water extracts were combined, concentrated to a clear paste with a relative density of 0.8 (60 °C), cooled, added with 95% ethanol to make the alcohol content reach 75%, allowed to stand for 24 hours, filtered, and the filtrate was concentrated to obtain a fluid extract for standby.

[0043] Step 2: Segmenting and Step 3: Separation and purification were the same as in Example 1.

[0044] Example 4

[0045] Structural Identification of Compound 1 and Compound 2

[0046] (1) Structural data of Compound 1.

[0047]

[0048] Xylanchinoid A (1)

[0049] Molecular formula: C 15 H 20 O9

[0050] Appearance: White solid

[0051] Optical rotation: 109.41 (c 0.170, methanol)

[0052] HRESIMS (+) m / z: Experimental value 389.1099 [M + COOH] + , Calculated value 389.1089 [M + COOH] + .

[0053] FT-IR (KBr) ν max : 3403, 2925, 1766, 1383, 1077 cm -1 .

[0054] 1 1H NMR and 13 13C NMR (DEPT) data are shown in Table 1.

[0055] Compound 1, white solid, the molecular formula was determined to be C + , according to the positive ion HR-ESI-MS (m / z 389.1099 [M+COOH] 15 H 20 O9, and the degree of unsaturation is 6. 1 The 1H NMR (Table 1) spectrum shows 1 signal of an oxygenated methylene group [δ H 4.41 (m, H-3a), 4.45 (m, H-3b)], 1 methylene group [δ H 2.57 (dd, J = 9.5, 18.1 Hz, H-9a), 2.37 (dd, J = 9.5, 18.1 Hz, H-9b)], 1 signal of a trisubstituted double bond proton [δ H 5.60 (m,H-5)], 2 methine groups [δ H3.51 (m, H-10), 3.44 (m, H-11)], two oxidized methylenes [δ H 5.24 (d, J = 4.9 Hz, H-1); 5.77 (m, H-6)], while showing an anomeric proton signal belonging to the sugar moiety [δ H 4.50 (d, J = 7.9 Hz, H-1')]; 13 The 13C NMR (Table 1) spectrum and HSQC showed 15 carbon signals (Table 1), including two methylenes [including one oxidized carbon signal δ C (65.3, C-3)], four methines [including two oxidized carbon signals, δ C 105.4 (C-1), 95.1 (C-6)], one trisubstituted double bond carbon signal [δ C 120.3 (C-5), 151.3 (C-4)], and one lactone carbonyl [δ C 179.1 (C-8)], and the remaining six carbon signals could be attributed to the signals of the sugar moiety (δ C 103.6, C-1'; 75.0, C-2'; 77.8, C-3'; 71.2, C-4'; 78.5, C-5'; 62.6, C-6'). By comparison with the literature, the data was consistent with β-D-glucopyranosyl. The lactone carbonyl, double bond, and sugar moiety accounted for three degrees of unsaturation, indicating that compound 1 is a tricyclic system compound. In the HMBC spectrum ( Figure 3 ), H-1 was correlated with C-3, C-4, C-11 (δ C 58.2) and C-1', and 1 H– 1 In the H–H COSY spectrum, H-1 and H-11 were correlated, constructing the tetrahydrofuran ring in this compound, and C-1 was attached to one β-D-glucopyranosyl. In the HMBC spectrum, H-5 was correlated with C-4, C-6, C-10 (δ C 41.3), C-11, H-1 was correlated with C-10, and 1 H– 1 In the H–H COSY spectrum, H-10 and H-11, H-6 were correlated, thus constructing the five-membered carbon ring in this compound and connecting it to the tetrahydrofuran ring through the C-4 / C-11 bond, and determining that the double bond was substituted at the C-4 and C-5 positions. However, 13 There was still one ester carbonyl signal left in the 13C NMR spectrum. Combining with the fact that in the HMBC spectrum, H-9 was correlated with C-8, C-6, C-11, C-10, and 1 H– 1In the H COSY spectrum, the correlation between H-10 and H-9 indicates the presence of a five-membered lactone ring connected by the C-6 / C-10 bond. The above information allows the planar structure of Compound 1 to be deduced as having a five-membered carbon ring, a five-membered lactone ring, and a tetrahydrofuran ring, which belongs to the structure of iridoids. In the ROESY spectrum ( Figure 3 ), the correlation between H-10 and H-6 and H-11 indicates that H-6, H-10, and H-11 are on the same side. Finally, the absolute configuration of Compound 1 was determined by calculating the ECD; as Figure 4 shown, the calculated ECD spectrum of 6R,10R,11S-1 was in good agreement with the measured ECD spectrum. Therefore, the absolute configuration of Compound 1 was determined to be 6R,10R,11S. After retrieval, Compound 1 is a novel iridoid compound named Boschnoside A.

[0056] (2) Structure data of Compound 2.

[0057]

[0058] Xylanchinoid B (2)

[0059] Molecular formula: C 16 H 20 O 10

[0060] Appearance: Amorphous powder

[0061] Optical rotation: 28.80 (c 0.150, methanol)

[0062] HRESIMS (-) m / z: Experimental value 371.0994 [M-H] – , Calculated value 371.0984 [M-H] – .

[0063] IR (KBr) ν max : 3418, 1740, 1103, 1057, 1041 cm -1 .

[0064] 1 1H NMR and 13 13C NMR (DEPT) data are shown in Table 1.

[0065] Compound 2, a white solid, has a molecular formula of C - determined by negative ion HR-ESI-MS (m / z 371.0994 [M-H] 16 H 20 O 10 , and the degree of unsaturation is 7.1 The \(^1\)H NMR (Table 1) spectrum shows one signal of an oxygen - linked methylene group [δ H 4.12 (d, J = 15.1 Hz, H - 10a), 4.21 (d, J = 15.1 Hz, H - 10b)], one signal of a trisubstituted double - bond proton [δ H 5.82 (s, H - 7)], one anomeric proton signal [δ H 4.92 (d, J = 7.2 Hz, H - 1')], and three signals of oxidized methine groups [δ H 5.53 (d, J = 1.6 Hz, H - 1), 5.32 (d, J = 2.5 Hz, H - 3), 5.59 (d, J = 7.8 Hz, H - 6)]. 13 The \(^{13}\)C NMR (Table 1) spectrum shows 16 carbon signals, including two acetal signals [δ C 92.9 (C - 1), 96.5 (C - 3)], one carbonyl carbon signal [δ C 180.6 (C - 11)], one double - bond carbon signal [δ C 126.9 (C - 7), 150.4 (C - 8)], one oxidized methylene [δ C 60.1 (C - 10)], one oxidized methine [δ C 88.7 (C - 7)], three methine signals [δ C 39.1 (C - 4), 36.1 (C - 5), 45.8 (C - 9)], and β - D - glucopyranose signals [δ C 99.2 (C - 1), 79.9 (C - 2), 75.9 (C - 3), 71.1 (C - 4), 79.3 (C - 5), 62.5 (C - 6)]. The \(^1\)H and 1 \(^{13}\)C NMR data of this compound were compared with those of the known compound Eucomoside A. The only difference is that there is no acetyl substitution at the C - 10 position of compound 2. In HMBC, H - 10b (δ 13 4.20) correlates with C - 7, C - 8 and C - 9, confirming the above speculation. In the ROESY spectrum, H - 4 (δ H 3.18) correlates with H - 5 (δ H 3.59), H - 5 correlates with H - 6 (δ H 5.59), H - 5 correlates with H - 9 (δ H 3.16), H - 1 correlates with H - 9, H - 1 correlates with H - 1', H - 4 correlates with H - 2' (δ H 3.16), H - 1 correlates with H - 1', H - 4 correlates with H - 2' (δ H3.43), indicating that H-4 and H-6 are in the β configuration; combining the above information with the coupling constants of H-1 (δ H 5.53, d, J = 1.6 Hz) and H-3 (δ H 5.32, d, J = 2.5 Hz), it is speculated that H-1 and H-3 are in the α configuration. Therefore, the structure of compound 2 was identified and named Boschnoside A, which is the second reported iridoid with a saturated bond between C-3 and C-4 and an ether bond formed between C-3 and C-2' of glucose.

[0066] Table 1. 1 1H (500 MHz) and 13 13C NMR (125 MHz) data of compounds 1 and 2

[0067]

[0068] Example 5

[0069] Activity determination of compounds 1 and 2

[0070] In vitro anti-inflammatory experiments were carried out on the prepared compounds. The cells used in the experiments were mouse RAW264.7 cells, purchased from the China Center for Type Culture Collection, Wuhan University. The CCK-8 method was used for cytotoxicity experiments to determine the drug administration concentration, and an Elisa kit was used to detect inflammatory factors IL-6, IL-1β, and TNF-α. The specific implementation methods are as follows:

[0071] Cytotoxicity experiment: Using DMEM medium, compounds 1 and 2 were prepared into solutions of 200, 100, 50, 25, and 12.5 μg / mL for standby. RAW264.7 cells were cultured routinely. When the cell growth reached 80% confluence and the cell morphology was normal under the microscope, the cells were inoculated into 96-well plates at a density of 2×10 5 cells / mL, 100 μL / well. After culturing overnight in a carbon dioxide incubator, the supernatant in the wells was discarded, and different concentrations of sample solutions were added, 100 μL per well, with 6 wells for each concentration. The blank control group was added with the same volume of DMEM and cultured in a carbon dioxide incubator. After 24 h, 10 μL of CCK-8 reagent solution was added to each well. After culturing for 4 h in a carbon dioxide incubator, the absorbance was detected at a wavelength of 450 nm using a continuous wavelength microplate reader, and the cell survival rate was statistically analyzed. Quantitative data were expressed as mean ± standard deviation. SPSS 27.0 statistical software was used for one-way analysis of variance, and pairwise comparisons were made. A p < 0.05 was considered statistically significant. The results are as Figure 1As shown, the maximum non-toxic dose of Compound 1 against RAW264.7 cells is: 200 μg / mL, and the maximum non-toxic dose of Compound 2 against RAW264.7 cells is: 100 μg / mL.

[0072] ELISA experiment: RAW264.7 cells were cultured routinely. When the cell growth reached 80% confluence and the cell morphology was normal under the microscope, the cells were seeded into 6-well plates at a density of 2×10 5 cells / mL, 2 mL per well. After culturing overnight in a carbon dioxide incubator, the supernatant in the wells was discarded. For Sample 1, sample solutions with concentrations of 200 and 100 μg / mL were added as the high- and low-dose groups; for Sample 2, sample solutions with concentrations of 100 and 50 μg / mL were added as the high- and low-dose groups (both were diluted with DMEM, and the concentrations were confirmed by the results of the cytotoxicity test). At the same time, Lipopolysaccharide (LPS) was added for modeling at a concentration of 200 ng / mL, 1 mL per well, with 3 wells for each concentration. The blank control group was added with the same volume of DMEM, and the model control group was added with DMEM solution containing the same concentration of LPS. Then they were placed in a carbon dioxide incubator for culturing. After 24 h, the cell supernatant was aspirated to detect the IL-6, IL-1β, and TNF-α kits. The experimental results are as Figure 2 shown. Considering the comprehensive effects of each sample on the 3 inflammatory mediators, the results showed that both Compound 1 and 2 could reduce the concentrations of TNF-α, IL-6, and IL-1β in the supernatant of LPS-induced RAW264.7 cells to varying degrees, indicating a certain anti-inflammatory effect.

[0073] Example 6

[0074] Preparation of pharmaceutical preparations:

[0075] In the following preparation examples, conventional reagents were selected and the preparations were prepared according to existing conventional methods. This application example only shows that at least one of the compounds 1-2 of the present invention can be prepared into different preparations, and specific reagents and operations are not specifically limited:

[0076] 1. At least one of Compounds 1-2 was dissolved in propylene glycol or polyethylene glycol, and then water for injection was added according to the conventional method, followed by fine filtration, filling, and sterilization to prepare an injection solution, and the concentration of the injection solution was 0.5 - 5 mg / mL.

[0077] 2. At least one of Compounds 1-2 was dissolved in propylene glycol, polyethylene glycol, or DMSO, and then a lyophilization protectant (such as mannitol) was added. It was dissolved in sterile water for injection, stirred to dissolve, filtered aseptically, dispensed into ampoules, freeze-dried at low temperature, and then sealed aseptically to obtain a powder injection.

[0078] 3. At least one of Compounds 1-2 was added with an appropriate excipient to prepare a powder.

[0079] 4. Add at least one of Compounds 1-2 to an appropriate amount of excipient, granulate and tableting.

[0080] 5. Prepare oral liquid with at least one of Compounds 1-2 according to the conventional preparation method of oral liquid.

[0081] 6. Add at least one of Compounds 1-2 to an appropriate amount of excipient to prepare capsules.

[0082] 7. Add at least one of Compounds 1-2 to an appropriate amount of excipient to prepare granules.

[0083] As can be seen from the above examples, the present invention provides a compound in the rhizome of Qianjinzhui, its preparation method and application, a pharmaceutical composition and its application. The present invention provides 2 novel iridoid compounds. These compounds can reduce the concentrations of TNF-α, IL-6 and IL-1β in the supernatant of LPS-induced RAW264.7 cells to varying degrees, can form a pharmaceutical composition with a pharmaceutically acceptable carrier or excipient, and can be used to prepare anti-inflammatory drugs.

[0084] The above are only the preferred embodiments of the present invention. It should be pointed out that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be regarded as the protection scope of the present invention.

Claims

1. Iridoid compounds extracted from the rhizome of Qianjinzhui, characterized in that, The structural formula is as follows:

2. The preparation method of the iridoid compound extracted from the rootstock of the Qianjinzhui, as described in claim 1, is characterized in that, The following steps are involved: Step 1: Extraction: After the dried Herba Coptidis is crushed, it is extracted with water by reflux, the amount of the extraction solvent is 8 to 15 times the mass of the raw material, the extraction times are 2 to 4 times, each time for 1 to 3 hours, the extract is filtered through a 200-mesh filter, the water extract is combined, and concentrated to a clear paste with a relative density of 0.8 to 1.5, cooled, and 95% ethanol is added to make the alcohol content reach 65% to 75%, and the mixture is allowed to stand for 24 hours, filtered, and the filtrate is concentrated to obtain a fluid extract; Step 2: Segmentation: Take the fluid extract obtained in step 1, perform silica gel column chromatography, use chloroform: methanol solvent system 15:1, 9:1, 8:2, 7:3, 6:4, 1:1 as eluent for gradient elution, collect the eluate, monitor and combine it into 11 segments Fr1 to Fr11 by TLC; Step 3 separation and purification: Take the Fr7 extract in step 2 and use water-70% methanol as the eluent for gradient elution by MCI column chromatography, then use normal phase silica gel column chromatography, use dichloromethane: methanol: water system volume ratios of 7:2.5:1, 7:3:1, and 6:4:1 as the eluent for gradient elution, use RP18 column chromatography with 15%-70% methanol gradient elution, and finally use silica gel column chromatography with ethyl acetate: ethanol volume ratio of 10:1 to elute and purify to obtain compound 1.

3. Preparation method of iridoid compounds, characterized in that, The structural formula of the iridoid compound is as follows: The preparation method comprises the following steps: Step 1: Extraction: After the dried Herba Coptidis is crushed, it is extracted with water by reflux, the amount of the extraction solvent is 8 to 15 times the mass of the raw material, the extraction times are 2 to 4 times, each time for 1 to 3 hours, the extract is filtered through a 200-mesh filter, the water extract is combined, and concentrated to a clear paste with a relative density of 0.8 to 1.5, cooled, and 95% ethanol is added to make the alcohol content reach 65% to 75%, and the mixture is allowed to stand for 24 hours, filtered, and the filtrate is concentrated to obtain a fluid extract; Step 2: Segmentation: Take the fluid extract obtained in step 1, perform silica gel column chromatography, use chloroform: methanol solvent system 15:1, 9:1, 8:2, 7:3, 6:4, 1:1 as eluent for gradient elution, collect the eluate, monitor and combine it into 11 segments Fr1 to Fr11 by TLC; Step 3 separation and purification: Take the Fr7 extract in step 2 and use water-70% methanol as the eluent for gradient elution by MCI column chromatography, then use normal phase silica gel column chromatography, use dichloromethane: methanol: water system volume ratio of 7:2.5:1, 7:3:1, 6:4:1 as the eluent for gradient elution, use RP18 column chromatography with 15%-70% methanol gradient elution, and finally use Sephadex LH-20 column chromatography, elution with methanol to purify to obtain compound 2.

4. An anti-inflammatory pharmaceutical composition, characterized in that, The pharmaceutical composition comprises: The iridoid compound extracted from the rhizome of the Herba Lycopodii as claimed in claim 1, the iridoid compound prepared by the preparation method as claimed in claim 3 or the salt of the iridoid compound and a pharmaceutically acceptable carrier.

5. The pharmaceutical composition according to claim 4, wherein The pharmaceutical composition is in the form of an edible pharmaceutical preparation.

6. The pharmaceutical composition according to claim 5, wherein The pharmaceutical preparation is selected from the form of tablets, capsules, granules, powders, oral solutions, sprays, plasters, powder injections or injection solutions.

7. Use of the iridoid compound extracted from Qianjinzhui as described in claim 1, the iridoid compound prepared by the preparation method as described in claim 3, or the pharmaceutical composition as described in any one of claims 4 to 6 in the preparation of an anti-inflammatory drug.

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