New pentacyclic triterpene B of oleanane type extracted from radix stellae bupleuri and extraction method and application thereof

Abstract

This invention relates to the extraction of oleanane-type compound neopentane triterpenoid B from *Bupleurum chinense*, its extraction method, and its application. It effectively solves the extraction and application problems of *Bupleurum chinense*. The technical solution is as follows: The chemical structural formula of the oleanane-type compound neopentane triterpenoid B is shown below. This invention extracts compounds from *Bupleurum chinense*, isolating the oleanane-type compound neopentane triterpenoid B. Related activity experiments show that neopentane triterpenoid B can significantly reduce C48 / 80-induced degranulation damage in RBL-2H3 cells, significantly reduce IgE levels, and has anti-allergic activity. It can be further used to prepare drugs for treating allergic diseases, thus expanding the medicinal value of *Bupleurum chinense*.

Description

I. Technical Field

[0001] This invention relates to the field of drug extraction, and in particular to a new pentacyclic triterpenoid B of oleanane type compound extracted from Bupleurum chinense, its extraction method and application. II. Background Technology

[0002] Allergic diseases are among the diseases that the World Health Organization has listed as key areas for research and prevention in the 21st century. They are mainly type I hypersensitivity reactions mediated by IgE, which can affect multiple organs and systems throughout the body. These include diseases such as urticaria, atopic dermatitis, allergic conjunctivitis, allergic rhinitis, and allergic asthma. RBL-2H3 cell degranulation has been widely used in allergy and immunology research, especially in screening the anti-allergic properties of extracted and isolated compounds and natural extracts. Currently, the long-term efficacy of clinical treatment for allergic diseases is still not ideal. Therefore, finding novel and effective anti-allergy drugs remains an urgent technical problem for pharmaceutical researchers.

[0003] *Gypsophila oldhamiana* Mip and *Gypsophila licentiana* Hand-Hazz, belonging to the Caryophyllaceae family, are dried roots that enter the lung and stomach meridians. They have the effects of clearing deficiency heat and cooling the blood, and are used for consumptive fever, chronic malaria due to yin deficiency, infantile malnutrition, and hepatitis. *Gypsophila oldhamiana* has a long history of folk medicine use and broad prospects for development and utilization. Research has found that *Gypsophila oldhamiana* contains many excellent medicinal active ingredients, but research on *Gypsophila licentiana* has not yet been reported. Therefore, how to prepare the required active ingredients from *Gypsophila licentiana* and improve its utilization value is an important task, but no public reports have been found to date. III. Summary of the Invention

[0004] In view of the above situation and to overcome the shortcomings of the existing technology, the purpose of this invention is to provide a new pentacyclic triterpenoid B of oleanane type compound extracted from Bupleurum chinense, as well as its extraction method and application, which can effectively solve the problems of extraction and application of Bupleurum chinense.

[0005] The technical solution provided by this invention is that the chemical structural formula of the new pentacyclic triterpenoid B of the oleanane type compound is shown below:

[0006]

[0007] The extraction method for this oleanane-type compound, a new pentacyclic triterpenoid B, includes the following steps:

[0008] 1) Crush the roots of the fine-leaved stone flower (with above-ground stem residues) into coarse sections, extract three times with 70%-80% ethanol under reflux for 1-3 hours each time, with a solvent volume of 6-10 times, rapidly centrifuge to concentrate the extract to 40-60L, concentrate under reduced pressure until there is no alcohol odor, add equal volumes of petroleum ether, ethyl acetate and n-butanol in sequence for repeated extraction, concentrate under reduced pressure, transfer to an evaporating dish and dry to obtain the extracts of each part;

[0009] 2) The ethyl acetate extract of *Lithops fasciata* obtained in step 1) was purified by passing it through a macroporous resin (DM-130). Before use, it was activated in 95% ethanol for 24 hours to wash away impurities in the packing material. The extract was then suspended in 95% ethanol and packed into a column using a wet method (1 BV = 20 L). The column was then washed with 95% ethanol. When the ethanol was mixed with an equal volume of water without becoming turbid, the solution was gradually changed from 95% ethanol to 10% ethanol. The ethyl acetate extract was dissolved in 10% ethanol and slowly added to the column bed. The bottom opening of the column was closed or reduced to allow the sample to be slowly adsorbed into the macroporous resin. Gradient elution was performed using 10%, 30%, 50%, 70%, and 95% ethanol solutions. Each gradient was used to wash for four column volumes. The eluents from each gradient were collected and concentrated under reduced pressure to obtain five fractions Fr1-Fr5.

[0010] 3) Purify the fraction Fr5 obtained in step 2) by silica gel column chromatography: wet packing, dry loading, elution with a two-system system of petroleum ether-ethyl acetate = 10:1 and dichloromethane-methanol = 100:1-0:1, 1 BV = 1500 mL, collect 1 / 3 at a time, and a total of 52 fractions are obtained. Spot the fractions on a plate and examine them under a UV lamp. Combine the fractions to obtain 6 fractions Fr5-1 to Fr5-6.

[0011] 4) Purify the fraction Fr5-3 obtained in step 3) by silica gel column chromatography: wet packing, dry loading, elution with dichloromethane-methanol = 100:1 - 40:1 system, 1 BV = 450 mL, collect 1 / 3 at a time, a total of 38 fractions are obtained, spot on the plate and examined under UV light, and combined to obtain 5 fractions Fr5-3-1 to Fr5-3-5;

[0012] 5) Purify the fraction Fr5-3-3 obtained in step 4) by silica gel column chromatography: wet packing, dry loading, elution with dichloromethane-methanol system, spotting on TLC and examining under UV light, and combine to obtain two fractions Fr5-3-3-1 to Fr5-3-3-2.

[0013] 6) The fraction Fr5-3-3-2 obtained in step 5) was semi-preparatively purified, eluted with 90% methanol and water, at a flow rate of 3 mL / min, with dual-channel wavelengths of 210 nm and 254 nm, to obtain the oleanane-type compound neopentane triterpenoid B.

[0014] The present invention relates to the application of the neopentane triterpenoid B, an oleanane-type compound extracted from Bupleurum chinense, in the preparation of drugs for allergic diseases.

[0015] This invention involves the extraction of compounds from *Bupleurum chinense*, from which a neopentane-type compound, neopentane triterpenoid B, was isolated. Related activity experiments showed that neopentane triterpenoid B significantly reduced C48 / 80-induced degranulation damage in RBL-2H3 cells and significantly reduced IgE levels, exhibiting anti-allergic activity. It can be further used to prepare drugs for treating allergic diseases, thus expanding the medicinal value of *Bupleurum chinense*. IV. Description of the attached drawings

[0016] Figure 1 The molecular structure of the compound *Stellaria dichotoma*, a new pentacyclic triterpenoid B, is shown below.

[0017] Figure 2 This is the HMBC correlation diagram of the new pentacyclic triterpenoid B of the compound *Stellaria dichotoma* from this invention.

[0018] Figure 3 This is a NOE correlation diagram of the compound *Stellaria dichotoma*, a new pentacyclic triterpenoid B, from the present invention.

[0019] Figure 4 The compound of this invention, *Stellaria media*, is a new pentacyclic triterpenoid B. 1 H-NMR spectrum.

[0020] Figure 5 The compound of this invention, *Stellaria media*, is a new pentacyclic triterpenoid B. 13 C-NMR spectrum.

[0021] Figure 6 This is the DEPT spectrum of the compound *Stellaria dichotoma*, a new pentacyclic triterpenoid B, which is the compound of this invention.

[0022] Figure 7 The HH COSY spectrum of the compound *Stellaria sanguisorba officinalis* B, a new pentacyclic triterpenoid of the present invention.

[0023] Figure 8 This is the HSQC spectrum of the compound *Stellaria sanguisorba officinalis* B, a new pentacyclic triterpenoid of the present invention.

[0024] Figure 9 The HMBC spectrum of the compound *Stellaria sanguisorba officinalis* B, a new pentacyclic triterpenoid of the present invention, is shown below.

[0025] Figure 10 This is the NOE spectrum of the compound *Stellaria dichotoma*, a new pentacyclic triterpenoid B, which is the compound of this invention.

[0026] Figure 11 The IR spectrum of the compound *Stellaria sanguisorba officinalis* B, a new pentacyclic triterpenoid of the present invention.

[0027] Figure 12This is the UV spectrum of the compound *Stellaria sanguisorba officinalis* B, a new pentacyclic triterpenoid of the present invention.

[0028] Figure 13 This is the mass spectrum of the compound *Stellaria sanguisorba officinalis* B, a new pentacyclic triterpenoid of the present invention. V. Detailed Implementation Methods

[0029] The specific embodiments of the present invention will be further described in detail below with reference to the accompanying drawings and examples.

[0030] Example 1

[0031] In a specific implementation of this invention, the extraction method of the oleanane-type compound neopentane triterpenoid B includes the following steps:

[0032] 1) Crush 20 kg of fine-leaved stone flower roots (with above-ground stem residues) into coarse sections, extract three times with 70% ethanol under reflux for 2 hours each time, with solvent volumes of 10, 8, and 6 times respectively. Concentrate the extract to 50 L by rapid centrifugation, concentrate under reduced pressure until no alcohol odor is detected, and add equal volumes of petroleum ether, ethyl acetate, and n-butanol for repeated extraction. After concentration under reduced pressure, transfer to an evaporating dish and dry to obtain extracts from each part.

[0033] 2) 266.57 g of the ethyl acetate extract of *Lithocarpus stenoptera* obtained in step 1) was purified through macroporous resin (DM-130). Before use, it was activated in 95% ethanol for 24 h to wash away impurities in the packing material. It was then suspended in 95% ethanol and packed into a column using a wet method (1 BV = 20 L). The column was then washed with 95% ethanol. When the ethanol was mixed with an equal volume of water without becoming turbid, the ethyl acetate extract was dissolved in 10% ethanol and slowly added to the column bed. The bottom opening of the column was closed or reduced to allow the sample to be slowly adsorbed into the macroporous resin. Gradient elution was performed using 10%, 30%, 50%, 70%, and 95% ethanol solutions. Each gradient was used to wash for four column volumes. The eluents from each gradient were collected and concentrated under reduced pressure to obtain five fractions Fr1-Fr5.

[0034] 3) Purify the fraction Fr5 (49.81 g) obtained in step 2) by silica gel column chromatography: wet packing, dry loading, elution with a two-system system of petroleum ether-ethyl acetate = 10:1 and dichloromethane-methanol = 100:1-0:1, 1 BV = 1500 mL, collect 1 / 3 at a time, and a total of 52 fractions are obtained. Spot the fractions on the TLC plate and examine them under UV light. Combine the fractions to obtain 6 fractions Fr5-1 to Fr5-6.

[0035] 4) Purify the fraction Fr5-3 (11.046 g) obtained in step 3) by silica gel column chromatography: wet packing, dry loading, elution with dichloromethane-methanol = 100:1 - 40:1 system, 1 BV = 450 mL, collect 1 / 3 at a time, a total of 38 fractions were obtained, spotted on the plate and examined under UV light, and combined to obtain 5 fractions Fr5-3-1 to Fr5-3-5;

[0036] 5) Purify the fraction Fr5-3-3 (1.51 g) obtained in step 4) by silica gel column chromatography: wet packing, dry loading, elution with dichloromethane-methanol system, TLC and UV inspection, and combine to obtain two fractions Fr5-3-3-1 to Fr5-3-3-2.

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

[0038] This invention relates to a compound extracted from *Bupleurum chinense*, identified as a neopentane-type triterpenoid B. Activity experiments were conducted on neopentane-type triterpenoid B, and the results showed that it significantly reduced C48 / 80-induced degranulation damage in RBL-2H3 cells and significantly reduced IgE levels, exhibiting anti-allergic activity. It can be further used to prepare drugs for treating allergic diseases. Specific measurements and experimental data are as follows: 1. Instruments and reagents.

[0039] Nuclear magnetic resonance (NMR) was performed using a Bruker Avance III 500 NMR spectrometer (TMS internal standard) (Bruker); infrared spectroscopy was performed using a Nicolet 10 Microscope Spectrometer (Thermo Scientific, USA); ultra-high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UHPLC-QTOF-MS) system (Agilent Technologies, USA); and ultraviolet spectroscopy was performed using a Shimadzu UV-2401PC. Apparatus, FDU-2110 freeze dryer (Shanghai Airong Instrument Co., Ltd.), CO2 incubator (Thermo Fisher Scientific), 3020 multi-functional microplate reader (Thermo Fisher Scientific), AE2000 inverted microscope (Macody Industrial Group Co., Ltd.), superconducting nuclear magnetic resonance spectrometer (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), condensate 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 Solarbio Science & Technology Co., Ltd.), DMSO (Maclean Biotechnology Co., Ltd.), PBS buffer (Wuhan Saive Biotechnology Co., Ltd.), C48 / 80 (Sigma-Aldrich, USA), Na2CO3, NaHCO3, NaOH (Tianjin Hengxing Chemical Reagent Manufacturing Co., Ltd.). *Gypsophila licentiana* root was collected in August 2022 from Tianshui County, Gansu Province. It was identified by Professor Chen Suiqing of Henan University of Traditional Chinese Medicine as the underground root of *Gypsophila licentiana* Hand.-Mazz., and is preserved in the Key Laboratory of Traditional Chinese Medicine Resources and Chemistry of Henan Province.

[0041] 2. Structural Identification

[0042] The compound neopentaneous triterpenoid B (3β-hydroxyolean-23-carboxyolean-28,21β-olide): white crystals, soluble in chloroform. ESI-MS m / z: 499.3413 [M+H] + The molecular formula is speculated to be: C 31 H 46O5. UV(MeOH)λ max (logε) / nm: 208 (2.445); IR spectrum: 1770 cm⁻¹ 1 and 1737cm- 1 The compound contains two carbonyl groups, with 1770 cm⁻¹ being the most abundant. 1 It is speculated that the compound contains an lactone ring structure. 1 The 1H-NMR (500MHz, CDCl3) spectrum yielded 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); presumably the hydrogen signal of an oleanolic acid-type compound; 4.12 (1H, d, J = 6.0 Hz), 3.98 (1H, d, J = 6.2 Hz) are hydrogen signals from two oxygen-bonded carbons; 3.67 (1H, s) is the (-OCH3) hydrogen signal; 13 A total of 30 carbon signals were observed by C-NMR (125 MHz, CDCl3) and DEPT, 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); and the remainder 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 and 175.15 are two carbonyl carbon signals; 140.86 and 122.13 are two double bond carbon signals, indicating that the compound may be a pentacyclic triterpenoid of olean-12-ene; 84.42 and 75.12 are oxygen-linked carbon signals; combining the characteristic signals of the proton and carbon spectra, it was found that the compound has a lactone ring. The lactone ring of this compound is formed by the carbonyl group at position 28 and the methylene group at position 21. The compound has a -COOCH3 group at position 23.

[0043] In summary, the chemical structure of the compound *Stellaria dichotoma*, a novel pentacyclic triterpenoid B, has been determined to be: 3β-hydroxyolean-23-carboxyolean-28,21β-olide, methyl ester. This compound has not been previously reported and is therefore identified as a new compound. Its specific... 13 C-NMR, 1The assignments of H-NMR and HMBC related peaks are shown in Table 1. Figure 2-3 As shown.

[0044] Table 1. NMR data assignment table for the novel pentacyclic triterpenoid B compound of this invention.

[0045]

[0046]

[0047] 3. Activity detection

[0048] 3.1 Cell Culture

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

[0050] 3.2 Reagent Preparation

[0051] C48 / 80 was removed from the -20℃ freezer and left at room temperature for 0.5 hours. It was then dissolved in PBS in a clean bench to prepare a 20 mg / mL solution for later use. In the clean bench, 100 μL of Triton X-100 was dissolved in 9.9 mL of Benchtop Solution to prepare 1% Triton X-100, to be prepared immediately before use. 4-Nitrophenyl-N-acetyl-β-D-glucosamine was prepared into a 1 mmol / L substrate solution using 1 mol / L citrate buffer (pH 4.5). 4.00 g of NaOH particles were weighed and dissolved in 100 mL of Wahaha purified water to adjust the pH of the solution. 2.65 g of Na₂CO₃ and 2.1 g of NaHCO₃ were weighed and dissolved in 250 mL of Wahaha purified water. The solution was then adjusted to pH 10.7 using 1 mol / L NaOH.

[0052] 3.3 Activity Screening Methods

[0053] Take cells in the logarithmic growth phase, at 2 × 10⁻⁶ 4At a density of cells / mL, 200 μL of cell suspension was seeded into 96-well plates, with 4 replicates per group. After 24 h, the cells were divided into a blank control group, a model group (C48 / 80, 20 μg / mL), a group containing *Stellaria media* var. *spinosa* (a type of terpene triterpenoid B) + C48 / 80 (10 μM), and a total enzyme group (1% Trinton X-100). Both the drugs and the C48 / 80 solution were prepared using Trussor buffer. Four blank wells containing only Trussor buffer and no cells were also included. After 30 min of stimulation, 50 μL of cell supernatant was collected, and 50 μL of a 1 mmol / L substrate solution was added. The mixture was incubated at 37°C for 1 h. The reaction was terminated by adding 150 μL of Na₂CO₃ / NaHCO₃ stop solution. The absorbance was measured at 405 nm using a microplate reader, and the β-Hex release rate was calculated. The experiment was repeated three times in parallel.

[0054] The formula is: β-Hex release rate = (OD value of experimental group supernatant - OD value of blank well supernatant) / (OD value of total enzyme well supernatant - OD value of blank well supernatant) × 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 neopentane triterpenoid B on C48 / 80-induced degranulation of RBL-2H3 cells ( n=4)

[0056]

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

[0058] 3.4 Immunoglobulin E (IgE) Enzyme-Linked Immunosorbent Assay Kit for IgE Content Determination

[0059] RBL-2H3 cells were loaded at 2×10 4 Cells were seeded at a density of 100 μL / mL in 96-well plates, with 200 μL per well. IgE levels were measured using an IgE kit. After 24 h, the cells were divided into a blank control group, a model group (C48 / 80, 20 μg / mL), and a neopentanetriterpenoid B group + C48 / 80 (10 μM). After 24 h of treatment, the supernatant of each group was collected and centrifuged at 3000 rpm for 20 minutes at 4 °C. The supernatant was collected, and the operation procedure was strictly followed according to the kit instructions. The results are shown in Table 3. The compound neopentanetriterpenoid B in Bupleurum chinense can significantly reduce IgE levels.

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

[0061]

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

[0063] 4. Conclusion

[0064] In summary, this invention utilizes abundant raw materials and employs an easy extraction method. A neopentane-type compound, neopentane triterpenoid B, was isolated from *Bupleurum chinense*. Related activity experiments showed that neopentane triterpenoid B significantly reduced C48 / 80-induced degranulation damage in RBL-2H3 cells and significantly decreased IgE levels, exhibiting anti-allergic activity. It can be further used to prepare drugs for treating allergic diseases, providing experimental evidence for pharmacological research of *Bupleurum chinense* and drug development for allergic diseases. This invention expands the medicinal value of *Bupleurum chinense*, has enormous development and application prospects, and offers significant economic and social benefits.

[0065] It should be noted that the above are merely preferred embodiments of the present invention and are not intended to limit the present invention in any way. Any modifications or alterations made by those skilled in the art without departing from the scope of the present invention to create equivalent embodiments based on the disclosed technical content shall fall within the protection scope of the present invention.

Claims

1. A neopentane triterpenoid B, an oleanane-type compound extracted from *Bupleurum chinense*, characterized in that... The chemical structural formula of the new pentacyclic triterpenoid B, an oleanane-type compound, is shown below:

2. The extraction method for the oleanane-type compound neopentane triterpenoid B from *Stellaria media* as described in claim 1, characterized in that, Includes the following steps: 1) Crush the roots of the fine-leaved stone flower into coarse sections, extract them three times by reflux with 70%-80% ethanol for 1-3 hours each time, with a solvent volume of 6-10 times, and concentrate the extract by rapid centrifugation to a volume of 40-60L. Concentrate under reduced pressure until there is no alcohol odor, and add equal volumes of petroleum ether, ethyl acetate and n-butanol in sequence for repeated extraction. After concentration under reduced pressure, transfer to an evaporating dish and dry to obtain the extracts of each part. 2) The ethyl acetate extract of *Gynostemma pentaphyllum* obtained in step 1) was purified by passing it through macroporous resin DM-130. Before use, it was activated in 95% ethanol for 24 hours to wash away impurities in the packing material. It was then suspended in 95% ethanol and packed into a column using a wet method (1 BV = 20 L). The column was then washed with 95% ethanol. When the ethanol was mixed with an equal volume of water without becoming turbid, the ethyl acetate extract was gradually dissolved in 10% ethanol and slowly added to the column bed. The bottom opening of the column was closed or reduced to allow the sample to be slowly adsorbed into the macroporous resin. Gradient elution was performed using 10%, 30%, 50%, 70%, and 95% ethanol solutions, with each gradient wash consisting of four column volumes. The eluents from each gradient were collected and concentrated under reduced pressure to obtain five fractions Fr1-Fr5. 3) Purify the fraction Fr5 obtained in step 2) by silica gel column chromatography: wet packing, dry loading, elution with a two-system system of petroleum ether-ethyl acetate = 10:1 and dichloromethane-methanol = 100:1-0:1, 1 BV = 1500 mL, collect 1 / 3 at a time, and a total of 52 fractions are obtained. Spot the fractions on a plate and examine them under a UV lamp. Combine the fractions to obtain 6 fractions Fr5-1 to Fr5-6. 4) Purify the fraction Fr5-3 obtained in step 3) by silica gel column chromatography: wet packing, dry loading, elution with dichloromethane-methanol = 100:1-40:1 system, 1 BV = 450 mL, collect 1 / 3 at a time, a total of 38 fractions are obtained, spot on the plate and examined under UV light, and combined to obtain 5 fractions Fr5-3-1 to Fr5-3-5; 5) Purify the fraction Fr5-3-3 obtained in step 4) by silica gel column chromatography: wet packing, dry loading, elution with dichloromethane-methanol system, spotting on TLC and examining under UV light, and combine to obtain two fractions Fr5-3-3-1 to Fr5-3-3-2. 6) The fraction Fr5-3-3-2 obtained in step 5) was semi-preparatively purified, eluted with 90% methanol and water, at a flow rate of 3 mL / min, with dual-channel wavelengths of 210 nm and 254 nm, to obtain the oleanane-type compound neopentane triterpenoid B.

3. The use of the neopentane triterpenoid B, an oleanane-type compound extracted from Bupleurum chinense as described in claim 1, in the preparation of drugs for allergic diseases.

4. The application of the oleanane-type compound neopentane triterpenoid B prepared by the extraction method of oleanane-type compound neopentane triterpenoid B extracted from Bupleurum chinense as described in claim 2 in the preparation of drugs for allergic diseases.

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