Oleanane type compound novel pentacyclic triterpenoid B extracted from radix stellariae and extraction method and application of oleanane type compound novel pentacyclic triterpenoid B

Abstract

The invention relates to an oleanane compound novel pentacyclic triterpene B extracted from radix stellariae as well as an extraction method and application of the oleanane compound novel pentacyclic triterpene B, which can effectively solve the problems of extraction and application of radix stellariae. The chemical structural formula of the oleanane type compound novel pentacyclic triterpenoid B is as follows: # imgabs0 #, compound extraction is performed on radix stellariae, the oleanane type compound novel pentacyclic triterpenoid B is separated from the radix stellariae, and related activity experiment results show that the novel pentacyclic triterpenoid B can significantly reduce C48 / 80 induced RBL-2H3 cell degranulation damage, significantly reduce the IgE level, and can be used for preparing the oleanane type compound novel pentacyclic triterpenoid B. The compound has anti-allergic activity and can be further used for preparing drugs for treating allergic diseases, and the medicinal value of radix stellariae is expanded.

Description

1. Technical Field

[0001] The invention relates to the field of drug extraction, in particular to a novel pentacyclic triterpene B of an oleanane-type compound extracted from Bupleurum chinense and an extraction method and application thereof. 2. Background Technology

[0002] Allergic disease is one of the diseases listed by the World Health Organization as a key research and prevention disease in the 21st century. It is mainly a type I hypersensitivity reaction mediated by IgE, which can affect multiple organs and systems throughout the body. It includes urticaria, atopic dermatitis, allergic conjunctivitis, allergic rhinitis, allergic asthma and other diseases. RBL-2H3 cell degranulation has been widely used in allergy and immunology research, especially in the screening of extracted and isolated compounds and the anti-allergic properties of natural extracts. At present, the long-term clinical efficacy of the treatment of allergic diseases is still not ideal. Therefore, finding novel and effective antiallergic drugs is still a technical problem that pharmaceutical researchers are eager to solve.

[0003] Bupleurum chinense is the dried root of Gypsophila oldhamiana Mip and Gypsophila licentiana Hand-Hazz, both of which belong to the Caryophyllaceae family. It belongs to the lung and stomach meridians, has the effects of clearing heat and cooling blood, and is used for asthenia, chronic malaria, infantile malnutrition, hepatitis, etc. Bupleurum chinense has a wide history of folk medicine use and has broad prospects for development and utilization. Studies have found that Gypsophila oldhamiana Mip contains many good medicinal active ingredients, but there has been no report on the research of Gypsophila licentiana Hand-Hazz. Therefore, how to prepare the required active ingredients from Gypsophila licentiana and improve the utilization value of Gypsophila licentiana is an important task, but there has been no public report so far. III. Summary of the invention

[0004] In view of the above situation, in order to solve the defects of the prior art, the purpose of the present invention is to provide a new pentacyclic triterpene B of an oleanane-type compound extracted from Bupleurum chinense and its extraction method and application, which can effectively solve the extraction and application problems of Bupleurum chinense.

[0005] The technical solution provided by the present invention is that the chemical structure of the new pentacyclic triterpene B of the oleanane type compound is as follows:

[0006]

[0007] The method for extracting the new pentacyclic triterpene B of the oleanane type compound comprises the following steps:

[0008] 1) The roots of the fine-leaved stone flower (with residual stems above the ground) were broken into coarse segments, and refluxed and extracted with 70%-80% ethanol for three times, each time for 1-3 hours, with the amount of solvent being 6-10 times, and the extract was concentrated to 40-60L by rapid centrifugation, and concentrated under reduced pressure until there was no alcohol taste, and then equal volumes of petroleum ether, ethyl acetate, and n-butanol were added in turn for repeated extraction, and after reduced pressure concentration, the extracts were transferred to an evaporating dish and dried to obtain extracts of various parts;

[0009] 2) The ethyl acetate extract of the fine-leaved stone flower obtained in step 1) is purified by macroporous resin (DM-130), activated in 95% ethanol for 24 hours before use to wash away impurities in the filler, suspended in 95% ethanol, wet-loaded on a column, 1BV=20L, and then rinsed with 95% ethanol. When the washed ethanol is mixed with an equal amount of water and does not become turbid, the ethanol is then successively transitioned from 95% ethanol to 10% ethanol, the ethyl acetate extract is dissolved in 10% ethanol, slowly added to the column bed, the lower end opening of the column is closed or reduced, and the sample is slowly adsorbed on the macroporous resin, and gradient elution is performed using 10%, 30%, 50%, 70%, and 95% ethanol solutions, each gradient washing four column volumes, and each gradient eluate is collected and concentrated under reduced pressure to obtain five components Fr1-Fr5;

[0010] 3) Purify the component Fr5 obtained in step 2) by silica gel column chromatography: wet column loading, dry loading, petroleum ether-ethyl acetate = 10:1, dichloromethane-methanol = 100:1-0:1 dual system elution, 1BV = 1500mL, 1 / 3 collected once, a total of 52 fractions were obtained, the plate was inspected under ultraviolet light, and 6 fractions Fr5-1 to Fr5-6 were obtained by merging;

[0011] 4) Purify the component Fr5-3 obtained in step 3) by silica gel column chromatography: wet column loading, dry loading, elution with dichloromethane-methanol = 100:1 -40:1 system, 1BV = 450mL, 1 / 3 collected once, a total of 38 fractions were obtained, the plate was inspected under ultraviolet light, and 5 fractions Fr5-3-1 to Fr5-3-5 were obtained by merging;

[0012] 5) Purify the component Fr5-3-3 obtained in step 4) by silica gel column chromatography: wet column packing, dry sample loading, elution with dichloromethane-methanol system, spot plate inspection under ultraviolet light, and combine to obtain two fractions Fr5-3-3-1 to Fr5-3-3-2;

[0013] 6) The component Fr5-3-3-2 obtained in step 5) was semi-preparatively purified, eluted with 90% methanol and water, flow rate 3 mL / min, dual channel wavelength: 210 nm, 254 nm, to obtain the oleanane-type compound new pentacyclic triterpene B.

[0014] The invention discloses an application of the novel pentacyclic triterpene B, an oleanane-type compound extracted from Bupleurum chinense, in the preparation of drugs for allergic diseases.

[0015] The present invention extracts compounds from Bupleurum chinense, and separates and obtains a new pentacyclic triterpene B, an oleanane-type compound, therefrom. Related activity experimental results show that the new pentacyclic triterpene B can significantly reduce C48 / 80-induced RBL-2H3 cell degranulation damage, significantly reduce IgE levels, have anti-allergic activity, and can be further used in the preparation of drugs for treating allergic diseases, thus expanding the medicinal value of Bupleurum chinense. IV. Description of the drawings

[0016] Figure 1 The present invention discloses a molecular structure of the compound Bupleurum chinense new pentacyclic triterpene B.

[0017] Figure 2 This is the HMBC correlation diagram of the compound Bupleurum chinense new pentacyclic triterpene B of the present invention.

[0018] Figure 3 It is the NOE correlation diagram of the compound of the present invention, the new pentacyclic triterpene B from Bupleurum chinense.

[0019] Figure 4 The compound of the present invention is the new pentacyclic triterpene B of Bupleurum chinense 1 H-NMR spectrum.

[0020] Figure 5 The compound of the present invention is the new pentacyclic triterpene B of Bupleurum chinense 13 C-NMR spectrum.

[0021] Figure 6 This is the DEPT spectrum of the compound Bupleurum chinense new pentacyclic triterpene B of the present invention.

[0022] Figure 7 This is the HH COSY spectrum of the compound Bupleurum chinense new pentacyclic triterpene B of the present invention.

[0023] Figure 8 This is the HSQC spectrum of the compound Bupleurum chinense new pentacyclic triterpene B of the present invention.

[0024] Fig. 9 This is the HMBC spectrum of the compound Bupleurum chinense new pentacyclic triterpene B of the present invention.

[0025] Fig.10 This is the NOE spectrum of the compound Bupleurum chinense new pentacyclic triterpene B of the present invention.

[0026] Fig.11 The IR spectrum of the compound Bupleurum chinense new pentacyclic triterpene B of the present invention.

[0027] Fig.12This is the UV spectrum of the compound Bupleurum chinense new pentacyclic triterpene B of the present invention.

[0028] Fig.13 This is the mass spectrum of the compound Bupleurum chinense new pentacyclic triterpene B of the present invention. V. Specific implementation methods

[0029] The specific implementation of the present invention is further described in detail below in conjunction with the accompanying drawings and examples.

[0030] Example 1

[0031] In the specific implementation of the present invention, the method for extracting the new pentacyclic triterpene B of the oleanane type compound comprises the following steps:

[0032] 1) 20 kg of the roots of the fine-leaved stone flower (with the residual stems above the ground) were broken into coarse segments, and refluxed and extracted with 70% ethanol for three times, each time for 2 hours, with the amount of solvent being 10, 8, and 6 times, respectively. The extract was concentrated to 50 L by rapid centrifugation, and concentrated under reduced pressure until there was no alcohol taste. Equal volumes of petroleum ether, ethyl acetate, and n-butanol were added in turn for repeated extraction. After reduced pressure concentration, the extract was transferred to an evaporating dish and dried to obtain extracts of various parts;

[0033] 2) 266.57 g of the ethyl acetate extract of the fine-leaved stone flower obtained in step 1) was purified by macroporous resin (DM-130), activated in 95% ethanol for 24 hours before use to wash away impurities in the filler, suspended in 95% ethanol, wet-loaded on a column, 1BV=20L, and then rinsed with 95% ethanol. When the washed ethanol was mixed with an equal amount of water and remained turbid, it was successively transitioned from 95% ethanol to 10% ethanol, the ethyl acetate extract was dissolved in 10% ethanol, slowly added to the column bed, the lower end opening of the column was closed or reduced, and the sample was slowly adsorbed on the macroporous resin, and gradient elution was performed using 10%, 30%, 50%, 70%, and 95% ethanol solutions, each gradient washing four column volumes, and each gradient eluate was collected and concentrated under reduced pressure to obtain five components Fr1-Fr5;

[0034] 3) Purify the component Fr5 (49.81 g) obtained in step 2) by silica gel column chromatography: wet column loading, dry loading, petroleum ether-ethyl acetate = 10:1, dichloromethane-methanol = 100:1-0:1 dual system elution, 1BV = 1500mL, 1 / 3 collected once, a total of 52 fractions were obtained, the plate was inspected under ultraviolet light, and 6 fractions Fr5-1 to Fr5-6 were obtained by merging;

[0035] 4) Purify the component Fr5-3 (11.046 g) obtained in step 3) by silica gel column chromatography: wet column loading, dry loading, elution with dichloromethane-methanol = 100:1 -40:1 system, 1BV = 450mL, 1 / 3 collected once, a total of 38 fractions were obtained, the plate was inspected under ultraviolet light, and 5 fractions Fr5-3-1 to Fr5-3-5 were obtained by merging;

[0036] 5) Purify the component Fr5-3-3 (1.51 g) obtained in step 4) by silica gel column chromatography: wet column loading, dry loading, elution with dichloromethane-methanol system, spot plate inspection under ultraviolet light, and combine to obtain two fractions Fr5-3-3-1 to Fr5-3-3-2;

[0037] 6) The component Fr5-3-3-2 (320.1 mg) obtained in step 5) was semi-preparatively purified and eluted with 90% methanol and water at a flow rate of 3 mL / min and dual channel wavelengths of 210 nm and 254 nm to obtain a new pentacyclic triterpene B of the oleanane type compound.

[0038] The compound extracted from Bupleurum chinense by the present invention is identified as a novel oleanane-type compound, a novel pentacyclic triterpene B, and an activity experimental study is conducted on the novel pentacyclic triterpene B. The experimental results show that the novel pentacyclic triterpene B can significantly reduce the degranulation injury of RBL-2H3 cells induced by C48 / 80, significantly reduce the level of IgE, have anti-allergic activity, and can be further used to prepare drugs for treating allergic diseases. The relevant specific determination and experimental data are as follows: 1 Instruments and reagents

[0039] Nuclear magnetic resonance was performed using a Bruker AVANCEⅢ500 NMR instrument (TMS internal standard) (Bruker), infrared spectroscopy was performed using a Nicolet is 10 Microscope Spectrometer (Thermo Scientific, USA), ultra-high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry was performed using an Agilent 6546 UPLC-Q-TOF-MS system (Agilent, USA), and ultraviolet spectroscopy was performed using a Shimadzu UV-2401PC apparatus, FDU-2110 freeze dryer (Shanghai Ailang Instrument Co., Ltd.), carbon dioxide incubator (Thermo Fisher Scientific Inc.), 3020 multifunctional microplate reader (Thermo Fisher Scientific Inc.), AE2000 inverted microscope (Motic Industrial Group Co., Ltd.), superconducting nuclear magnetic resonance (Bruker, Switzerland, DPX-500), semi-preparative high performance liquid chromatography (Beijing Cypress Technology Co., Ltd., LC-52), fully preparative high performance liquid chromatography (Shanghai Wufeng Science Co., Ltd., LC100), rotary evaporator (EYELA Tokyo Rika, Japan, N-1000, BSA124S-CW), condensed water circulation device (EYELA Tokyo Rika, Japan, N-1111), Agilent high performance liquid chromatograph (Agilent Technologies, USA, Agilent 1260 Infinity II).

[0040] MEM medium (Gibco Invitrogen), fetal bovine serum (Sijiqing Zhejiang Tianhang Biotechnology Co., Ltd.), trypsin digestion solution (Beijing Solaibao Technology Co., Ltd.), DMSO (McLean Biotechnology Co., Ltd.), PBS buffer (Wuhan Saiweier Biotechnology Co., Ltd.), C48 / 80 (Sigma, USA), Na2CO3, NaHCO3, NaOH (Tianjin Hengxing Chemical Reagent Manufacturing Co., Ltd.). Bupleurum chinense was collected from Tianshui County, Gansu Province in August 2022, and was identified as the underground root of Gypsophila licentiana Hand.-Mazz. by Professor Chen Suiqing of Henan University of Traditional Chinese Medicine, and was preserved in the Key Laboratory of Traditional Chinese Medicine Resources and Traditional Chinese Medicine Chemistry of Henan Province.

[0041] 2 Structure identification

[0042] Compound new pentacyclic triterpene B (3β-hydroxyolean-23-carboxyolean-28,21β-olide): white crystals, soluble in chloroform. ESI-MS m / z: 499.3413 [M+H] + , the molecular formula may be: C 31 H 46O5. UV(MeOH)λ max (logε) / nm: 208 (2.445); IR spectrum: 1770cm- 1 and 1737cm- 1 This indicates that the compound contains two carbonyl groups, of which 1770cm- 1 It is speculated that there is a lactone ring structure in the compound. 1 H-NMR (500 MHz, CDCl3) gave six methyl hydrogen signals, δ H :1.17 (3H, d) 1.08 (3H, s), 1.02 (6H, s), 0.88 (3H, d), 0.75 (3H, s), a double bond hydrogen signal 5.56 (1H, s); it is speculated to be the hydrogen signal of oleanolic acid type compound; 4.12 (1H, d, J = 6.0Hz,), 3.98 (1H, d, J = 6.2Hz) are the hydrogen signals on two oxygen-linked carbons 3.67 (1H, s) is the (-OCH3) hydrogen signal; 13 C-NMR (125MHz, CDCl3) and DEPT showed 30 carbon signals, including 6 methyl carbon signals (27.77, 27.07, 23.54, 21.60, 19.18, 16.00); 9 methylene carbon signals (36.89, 34.45, 33.39, 31.20, 28.45, 26.21, 24.01, 23.69, 19.21); 7 methine carbon signals (122.13, 84.45, 67.21, 51.88, 49.55, 40.03, 38.43); the rest were quaternary carbon signals (182.26, 175.15, 140.75, 43.54, 42.63, 39.34, 38.99, 33.71); among which δ C : 182.19, 175.15 are two carbonyl carbon signals; 140.86, 122.13 are double bond carbon signals, indicating that the compound may be a pentacyclic triterpene of olean-12-ene, 84.42, 75.12 are oxygen-linked carbon signals; combining the characteristic signals of hydrogen spectrum and carbon spectrum, it is found that the compound has a lactone ring, and the lactone ring of the compound is a lactone formed by a carbonyl group at 28 and a methylene group at 21, and the 23rd position of the compound is -COOCH3.

[0043] In summary, the chemical structure of the compound new pentacyclic triterpenoid B from Bupleurum chinense was determined to be: 3β-hydroxyolean-23-carboxyolean-28,21β-olide, methyl ester. This compound has not been reported before and is determined to be a new compound. 13 C-NMR, 1The H-NMR and HMBC related peaks are assigned as shown in Table 1. Figure 2-3 shown.

[0044] Table 1 NMR data attribution table of the new pentacyclic triterpene B of the compound of the present invention

[0045]

[0046]

[0047] 3 Activity detection

[0048] 3.1 Cell culture

[0049] Rat basophil leukemia cell line (RBL-2H3) was cultured in MEM medium containing 10% fetal bovine serum and penicillin-streptomycin solution for one week and then placed in an incubator at 37°C and 5% CO2. RBL-2H3 cells in the logarithmic growth phase were selected for the experiment. Fresh culture medium was replaced every 48 hours and the cells were passaged at a ratio of 1:2.

[0050] 3.2 Reagent preparation

[0051] C48 / 80 was taken out of the -20°C refrigerator and placed at room temperature for 0.5h, dissolved in PBS in a clean bench, and prepared into a solution with a concentration of 20mg / mL for use; in the clean bench, 100μL of Triton X-100 was taken, and 9.9mL of benchtop solution was added to dissolve it to prepare 1% Trinton X-100, which was prepared immediately before use; 4-nitrophenyl-N-acetyl-β-D-glucosamine was prepared into a substrate solution with a concentration of 1mmol / L using 1mol / L citrate buffer (pH4.5); 4.00g of NaOH particles were weighed and dissolved in 100mL of Wahaha purified water to adjust the pH value of the solution; 2.65g of Na2CO3 and 2.1g of NaHCO3 were weighed and dissolved in 250mL of Wahaha purified water, and the solution was adjusted to a pH of 10.7 using 1mol / L NaOH.

[0052] 3.3 Activity screening method

[0053] Cells in logarithmic growth phase were taken and 2×10 4200 μL of cell suspension was inoculated in a 96-well plate at a density of 1 cell / mL, with 4 replicates in each group. After 24 hours, the plates were divided into a blank control group, a model group (C48 / 80, 20 μg / mL), a group of new pentacyclic triterpenoids of Bupleurum chinense B + C48 / 80 (10 μM), and a total enzyme group (1% Trinton X-100). The drug and C48 / 80 solution were prepared with benchtop solution. In addition, 4 blank wells containing only benchtop solution without cells were set up. After 30 minutes of stimulation, 50 μL of cell supernatant was taken, 50 μL of substrate solution with a concentration of 1 mmol / L was added, and the plate was incubated at 37°C for 1 hour. 150 μL of Na2CO3 / NaHCO3 stop solution was added to terminate the reaction, and the absorbance was detected at 405 nm by a microplate reader to calculate the β-Hex release rate. The experiment was repeated three times in parallel.

[0054] The formula is: β-Hex release rate = (OD value of supernatant in experimental group - OD value of supernatant in blank well) / (OD value of supernatant in total enzyme well - OD value of supernatant in blank well) × 100%. The results are shown in Table 2. The compound can significantly inhibit the release of β-Hex (P<0.01).

[0055] Table 2 Effects of new pentacyclic triterpenoid B on C48 / 80-induced degranulation of RBL-2H3 cells ( n=4)

[0056]

[0057] Note: Compared with the blank control group, ## Indicates P < 0.01, compared with the model group, ** Indicates P<0.01.

[0058] 3.4 Immunoglobulin E (IgE) ELISA kit for determination of IgE content

[0059] RBL-2H3 cells were cultured at 2×10 4 The cells were seeded at a density of 10 cells / mL in a 96-well plate, 200 μL per well, and the IgE content was determined using an IgE kit. After 24 hours, the cells were divided into a blank control group, a model group (C48 / 80, 20 μg / mL), and a new pentacyclic triterpene B group + C48 / 80 (10 μM). After 24 hours of action, the supernatant of each group of cells was collected, centrifuged at 3000 rpm for 20 minutes at 4°C, and the supernatant was collected. The operation steps were strictly performed according to the instructions of the kit. The results are shown in Table 3. The compound new pentacyclic triterpene B in Bupleurum chinense can significantly reduce the level of IgE.

[0060] Table 3 IgE content determined by IgE kit ( n=4)

[0061]

[0062] Note: Compared with the blank control group, ## Indicates P < 0.01, compared with the model group, * Indicates P<0.05.

[0063] 4 Conclusion

[0064] In summary, it can be seen that the present invention has rich raw materials and the extraction method is easy to operate. The oleanane-type compound new pentacyclic triterpene B is separated from Bupleurum chinense. The results of relevant activity experiments show that the new pentacyclic triterpene B can significantly reduce the degranulation damage of RBL-2H3 cells induced by C48 / 80, significantly reduce the level of IgE, have anti-allergic activity, and can be further used to prepare drugs for the treatment of allergic diseases. It provides an experimental basis for the pharmacological research of Bupleurum chinense and the development of drugs for allergic diseases, expands the medicinal value of Bupleurum chinense, has huge development and application prospects, and has huge economic and social benefits.

[0065] It should be pointed out that the above is only a preferred embodiment of the present invention and does not limit the present invention in any form. Any technician familiar with the profession can make changes or modify the technical contents disclosed above into equivalent embodiments with equivalent changes without departing from the scope of the technical solution of the present invention, which all fall within the protection scope of the present invention.

Claims

1. A novel pentacyclic triterpene B of oleanane-type compound extracted from Bupleurum chinense, characterized in that: The chemical structure of the new pentacyclic triterpene B of the oleanane type compound is as follows:

2. The method for extracting the new pentacyclic triterpene B of the oleanane type compound extracted from Bupleurum chinense according to claim 1, characterized in that: The following steps are involved: 1) The roots of the fine-leaved stone flower are broken into coarse sections, and refluxed and extracted with 70%-80% ethanol for three times, each time for 1-3 hours, with the amount of solvent being 6-10 times, and the extract is concentrated by rapid centrifugation to a volume of 40-60L, and concentrated under reduced pressure until there is no alcohol taste, and equal volumes of petroleum ether, ethyl acetate, and n-butanol are added in sequence for repeated extraction, and after reduced pressure concentration, the extract is transferred to an evaporating dish and dried to obtain extracts of various parts; 2) The ethyl acetate extract of the fine-leaved stone flower obtained in step 1) is purified by macroporous resin DM-130, activated in 95% ethanol for 24 hours before use to wash away impurities in the filler, suspended in 95% ethanol, wet-loaded on a column, 1BV=20L, and then rinsed with 95% ethanol. When the washed ethanol is mixed with an equal amount of water and does not become turbid, it is then successively transitioned from 95% ethanol to 10% ethanol, the ethyl acetate extract is dissolved in 10% ethanol, slowly added to the column bed, the lower end opening of the column is closed or reduced, and the sample is slowly adsorbed on the macroporous resin, and gradient elution is performed using 10%, 30%, 50%, 70%, and 95% ethanol solutions, each gradient washing four column volumes, and each gradient eluate is collected and concentrated under reduced pressure to obtain five components Fr1-Fr5; 3) Purify the component Fr5 obtained in step 2) by silica gel column chromatography: wet column loading, dry loading, petroleum ether-ethyl acetate = 10:1, dichloromethane-methanol = 100:1-0:1 dual system elution, 1BV = 1500mL, 1 / 3 collected once, a total of 52 fractions were obtained, the plate was inspected under ultraviolet light, and 6 fractions Fr5-1 to Fr5-6 were obtained by merging; 4) Purify the component Fr5-3 obtained in step 3) by silica gel column chromatography: wet column loading, dry sample loading, elution with dichloromethane-methanol = 100:1-40:1 system, 1BV = 450mL, 1 / 3 collected once, a total of 38 fractions were obtained, the plate was inspected under ultraviolet light, and 5 fractions Fr5-3-1 to Fr5-3-5 were obtained by merging; 5) Purify the component Fr5-3-3 obtained in step 4) by silica gel column chromatography: wet column packing, dry sample loading, elution with dichloromethane-methanol system, spot plate inspection under ultraviolet light, and combine to obtain two fractions Fr5-3-3-1 to Fr5-3-3-2; 6) The component Fr5-3-3-2 obtained in step 5) was semi-preparatively purified, eluted with 90% methanol and water, flow rate 3 mL / min, dual channel wavelength: 210 nm, 254 nm, to obtain the oleanane-type compound new pentacyclic triterpene B.

3. Use of the new pentacyclic triterpene B, an oleanane-type compound extracted from Bupleurum chinense as claimed in claim 1, in the preparation of drugs for allergic diseases.

4. Use of the new pentacyclic triterpene B of the oleanane type compound prepared by the extraction method of the new pentacyclic triterpene B of the oleanane type compound extracted from Bupleurum chinense as claimed in claim 2 in the preparation of drugs for allergic diseases.

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