Compound with hypoglycemic effect in pieris fortunei as well as separation method and application of compound

Abstract

The invention relates to a compound with a hypoglycemic effect in leaves of drynaria fortunei, and a separation preparation method and application thereof, and belongs to the technical field of medicines. The invention discloses a structural general formula of triterpenoids in leaves of drynaria fortunei, and structures and activities of compounds 1-37. The invention particularly discloses a separation and preparation method and application of a compound which is separated from leaves of drynaria fortunei and has a hypoglycemic effect. Researches prove that the compounds 1-37 achieve the effect of reducing blood sugar by inhibiting the activity of alpha-glucosidase; the triterpenoids can be used as lead compounds for development of hypoglycemic drugs, and have important significance on comprehensive utilization and development of pieris fortunei.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and more specifically, relates to compounds with hypoglycemic effects in the leaves of Pieris wilsonii, a separation method, and an application thereof. Background Art

[0002] Plants of the genus Pieris were called "Qin" in ancient times. "Er Ya" once recorded: "Qin, Osmanthus-like tree". Because the branches and leaves of this genus of plants are similar to those of osmanthus trees, they were regarded as a kind of osmanthus tree by the ancients. In modern plant taxonomy, the genus Pieris belongs to the Ericaceae family. There are seven species in the world, distributed in eastern Asia, eastern North America, and the West Indies. There are three species in China, namely Pieris wilsonii, Pieris swinhoei, and Pieris japonica. Plants of the genus Pieris have iconic urn-shaped flowers. Because horses eat their leaves and become lethargic as if drunk, they got this name.

[0003] Pieris japonica and Pieris wilsonii in the genus Pieris are used as traditional folk medicines. According to "Chinese Herbal Medicine", the roots and leaves of Pieris japonica can be used as medicine to treat heatstroke, vomiting, diarrhea, and scabies. According to "Illustrated Guide to Zhuang Medicines in China", the whole plant of Pieris wilsonii can reduce inflammation, relieve pain, and activate the meridians and collaterals, and is mainly used to treat rheumatic arthralgia, etc. Externally, it is mainly used to treat scabies and poisonous sores.

[0004] Diabetes is a chronic metabolic disease, mainly characterized by abnormally elevated blood glucose levels, and is divided into types such as type 1 diabetes (T1DM), type 2 diabetes (T2DM), and gestational diabetes. Among them, T2DM accounts for more than 90% of diabetes cases and is mainly caused by insulin resistance and reduced pancreatic beta-cell function. With the increase in the global obesity rate and population aging, the prevalence of diabetes has increased year by year and has become a major challenge to global public health. According to data from the International Diabetes Federation (IDF), the number of global diabetes patients exceeded 537 million in 2021 and is expected to increase to 783 million by 2045. Diabetes not only seriously affects the quality of life of patients but may also lead to serious complications such as cardiovascular diseases, kidney diseases, retinopathy, and neuropathy, bringing a huge burden to the medical system. Currently, diabetes treatment drugs mainly include insulin and its analogs, oral hypoglycemic drugs (such as metformin, sulfonylureas, DPP-4 inhibitors, SGLT-2 inhibitors), and new drugs (such as GLP-1 receptor agonists). As a first-line treatment drug, metformin reduces blood glucose by inhibiting hepatic gluconeogenesis and improving insulin sensitivity, but its long-term use may cause gastrointestinal side effects and vitamin B12 deficiency. Although insulin treatment can effectively control blood glucose, there are problems such as hypoglycemia risk and weight gain. In recent years, GLP-1 receptor agonists and SGLT-2 inhibitors have received extensive attention because of their significant hypoglycemic effects and cardiovascular protection effects. However, existing drugs still cannot fully meet the needs of patients, and some patients have problems such as poor drug tolerance or obvious side effects. Therefore, the development of new diabetes drugs has become a research hotspot. SUMMARY OF THE INVENTION

[0005] The object of the present invention is to provide triterpenoid compounds with hypoglycemic effects in Pieris formosa var. wilsonii and a method for separating and preparing the same. Another object of the present invention is to clarify the application value of the triterpenoid compounds in the preparation of hypoglycemic drugs. Thus, the technical problems of gastrointestinal side effects, hypoglycemia risk, weight gain, poor drug tolerance or obvious side effects existing in hypoglycemic drugs in the prior art are solved.

[0006] According to the first aspect of the present invention, a triterpenoid compound is provided, and the structural formula of the triterpenoid compound is shown as follows:

[0007]

[0008] R1-R4 in Formula I, R1-R6 in Formula II, R1-R6 in Formula III, R1-R8 in Formula IV, R1-R4 in Formula V, R1-R3 in Formula VI, R1-R4 in Formula VII, R1-R7 in Formula VIII, R1-R4 in Formula IX, R1-R4 in Formula X, R1-R6 in Formula XI, and R1-R4 in Formula XII are each independently selected from hydrogen, hydroxyl, methylene, oxygen-containing methylene, methyl, ethyl, allyl, carbonyl, halogen, acyl, acyl halide, nitro, cyano, glucosyl, arabinopyranosyl, rhamnosyl, cis-feruloyloxy, trans-feruloyloxy, cis-p-coumaroyloxy, trans-p-coumaroyloxy, benzene ring, and indole ring.

[0009] Preferably, Formula I is Compound 1 or Compound 2, Formula II is Compound 3, Compound 4, Compound 5, or Compound 6, Formula III is Compound 7, Formula IV is one of Compound 8-10, Compound 13-17, Compound 20-22, Formula V is Compound 18, Formula VI is Compound 19, Formula VII is Compound 11 or Compound 12, Formula VIII is one of Compound 23-26, Compound 28, Compound 30-32, Formula IX is Compound 27, Formula X is Compound 29, Formula XI is one of Compound 33-35, Compound 37, and Formula XII is Compound 36;

[0010]

[0011]

[0012]

[0013] According to another aspect of the present invention, a method for preparing the triterpenoid compound is provided, including the following steps:

[0014] (1) Dry and powder the leaves of Pieris formosa (Wall.) D. Don, extract with ethanol, and concentrate under reduced pressure to obtain the total extract.

[0015] (2) Suspend the total extract obtained in step (1) in water, extract successively with petroleum ether and chloroform, and concentrate under reduced pressure to obtain the extraction extract.

[0016] (3) Mix the extraction extract obtained in step (2) with silica gel, perform normal-phase silica gel column chromatography, with the eluent being a mixed solution of dichloromethane and methanol, gradient elution, combine with TLC spotting detection, and combine to obtain 7 fractions A1–A7 with increasing polarity.

[0017] When fraction A3 is separated by ODS column chromatography to obtain 15 fractions A3a–A3o with decreasing polarity; fraction A3h is separated by Sephadex LH-20 column chromatography and silica gel column chromatography to obtain compound 17 and compound 25 with increasing polarity.

[0018] When fraction A3i is separated by Sephadex LH-20 gel column chromatography and silica gel column chromatography to obtain compound 33 and 5 fractions A3i1–A3i5 with increasing polarity; fraction A3i4 is further purified by semi-preparative HPLC to obtain compound 15 and compound 16 with increasing retention time.

[0019] When fraction A3j is separated by Sephadex LH-20 gel column chromatography and silica gel column chromatography to obtain compound 19 with increasing retention time.

[0020] When fraction A3k is separated by Sephadex LH-20 gel column chromatography and silica gel column chromatography to obtain compound 21 and compound 27.

[0021] When fraction A3n is separated by Sephadex LH-20 gel column chromatography and silica gel column chromatography to obtain 7 fractions A3n1–A3n7 with increasing polarity, and fraction A3n2 is purified by semi-preparative HPLC to obtain compound 28.

[0022] When fraction A3n5 is purified by semi-preparative HPLC to obtain compound 37.

[0023] When fraction A4 is subjected to reverse-phase C 18 Silica gel column chromatography, with the eluent being a mixed solution of methanol and water, gradient elution, to obtain 10 fractions A4a–A4j with decreasing polarity; fraction A4b is separated by Sephadex LH-20 gel column chromatography and silica gel column chromatography and finally purified by semi-preparative HPLC to obtain compound 31, compound 20, and compound 34 with increasing retention time.

[0024] When component A4c was subjected to Sephadex LH-20 gel column chromatography, component A4c3 was obtained; component A4c3 was separated by semi-preparative high performance liquid chromatography column C 18 to obtain compounds 5, 6, and 7 with increasing retention times;

[0025] When component A4d was subjected to Sephadex LH-20 gel column chromatography, four components A4d1–A4d4 with decreasing molecular weights were obtained; component A4d4 was separated by semi-preparative high performance liquid chromatography phenyl column to obtain compounds 9 and 10 with increasing retention times;

[0026] When component A4h was obtained by Sephadex LH-20 gel column chromatography and silica gel column chromatography, compounds 26 and 13 with increasing polarities were obtained;

[0027] When component A5 was subjected to reversed-phase C 18 silica gel column chromatography with gradient elution to obtain 17 components A5a–A5q with decreasing polarities; when component A5h was separated by Sephadex LH-20 gel column chromatography and silica gel column chromatography, compounds 35 and five components A5h1–A5h5 with increasing polarities were obtained;

[0028] When component A5h3 was separated by semi-preparative HPLC to obtain four components A5h3a–A5h3d with decreasing polarities, component A5h3b was purified by semi-preparative HPLC to obtain compounds 22 and 23 with increasing retention times, and component A5h3c was purified by semi-preparative HPLC to obtain compounds 30 and 36 with increasing retention times;

[0029] When component A5j was subjected to Sephadex LH-20 gel column chromatography and reversed-phase C 18 silica gel column chromatography, component A5j2 was obtained; component A5j2 was separated by semi-preparative high performance liquid chromatography column C 18 to obtain compounds 3 and 4 with increasing retention times;

[0030] When component A5k was separated by Sephadex LH-20 gel column chromatography and silica gel column chromatography, compounds 18, 24, 14 with increasing polarities and seven components A5k1–A5k7 with increasing polarities were obtained;

[0031] When component A5k7 was purified by semi-preparative HPLC to obtain compounds 29 and 32 with increasing retention times;

[0032] When component A5m was subjected to Sephadex LH-20 gel column chromatography and reversed-phase C 18Silica gel column chromatography was performed to obtain fraction A5m2. When fraction A5m2 was subjected to semi-preparative high performance liquid chromatography C 18 column separation, compounds 11 and 12 with increasing retention times were obtained;

[0033] When fraction A5o was further subjected to Sephadex LH-20 gel column chromatography and reverse-phase C 18 silica gel column chromatography, fraction A5o3 was obtained. Fraction A5o3 was subjected to reverse-phase phenyl silica gel column chromatography to obtain four fractions A5o3a–A5o3d with decreasing polarity; fraction A5o3b was subjected to semi-preparative high performance liquid chromatography C 18 column separation to obtain compound 8;

[0034] When fraction A5p was further subjected to Sephadex LH-20 gel column chromatography and reverse-phase C 18 silica gel column chromatography, fraction A5p3 was obtained. Fraction A5p3 was subjected to semi-preparative high performance liquid chromatography C 18 column separation to obtain compounds 1 and 2 with increasing retention times.

[0035] According to another aspect of the present invention, there is provided the use of the triterpenoid compound, and the use of the triterpenoid compound in the preparation of a hypoglycemic drug.

[0036] Preferably, the triterpenoid compound is used to inhibit α-glucosidase activity.

[0037] Preferably, the hypoglycemic drug is a drug for treating diabetes.

[0038] Preferably, the diabetes is type 2 diabetes.

[0039] Generally speaking, compared with the prior art by the above technical solutions conceived by the present invention, the following technical advantages are mainly possessed:

[0040] (1) Compounds 1–37 provided in the present invention are triterpenoid compounds and are all first prepared and discovered from Pieris formosa (Wall.) D. Don var. wilsonii (Rehd.) W. W. Smith. Compounds 1–7 are taraxastane-type triterpenoids with a 6 / 6 / 6 / 6 / 6 pentacyclic carbon ring system, compounds 8–10, 13–22 are ursane-type triterpenoids with a 6 / 6 / 6 / 6 / 6 pentacyclic carbon ring system, compounds 11–12, 23–32 are oleanane-type triterpenoids with a 6 / 6 / 6 / 6 / 6 pentacyclic carbon ring system, and compounds 33–37 are lupane-type triterpenoids with a 6 / 6 / 6 / 6 / 5 pentacyclic carbon ring system.

[0041] (2) The triterpenoid compounds 1–37 provided in the present invention have α-glucosidase inhibitory activity. In particular, compounds 1, 2, 4, 9–12, 23, and 26 have significant inhibitory effects on α-glucosidase, stronger than the positive drug acarbose. Thus, the technical problems of gastrointestinal side effects, hypoglycemia risk, weight gain, poor drug tolerance, or obvious side effects existing in hypoglycemic drugs in the prior art are solved. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1 is the X-ray single crystal diffraction pattern of compound 1.

[0043] Figure 2 is the X-ray single crystal diffraction pattern of compound 7.

[0044] Figure 3 is the X-ray single crystal diffraction pattern of compound 11.

[0045] Figure 4 is the dose-response curve of the half inhibitory concentration values of the α-glucosidase inhibitory effects of compounds 1–7, 9–12. DETAILED DESCRIPTION OF THE INVENTION

[0046] In order to make the objectives, technical solutions, and advantages of the present invention clearer and more understandable, the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention. In addition, the technical features involved in the various embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0047] Pieris formosa (Wall.) D. Don, also known as Pieris formosa and Pieris formosa var. wilsonii, is an evergreen shrub or small tree, mainly distributed in provinces and regions such as Hubei, Zhejiang, Jiangxi, Hunan, Guangxi, Guangdong, Guizhou, and Sichuan in China, and is a plant of the genus Pieris in the family Ericaceae.

[0048] The present invention specifically relates to the separation and purification process, structure determination, and α-glucosidase inhibitory effect of triterpenoid compounds in Pieris formosa.

[0049] The structural formulas of the triterpenoid compounds of the present invention are shown as follows:

[0050]

[0051]

[0052] R1-R4, R1-R6 in Formula II, R1-R6 in Formula III, R1-R8 in Formula IV, R1-R4 in Formula V, R1-R3 in Formula VI, R1-R4 in Formula VII, R1-R7 in Formula VIII, R1-R4 in Formula IX, R1-R4 in Formula X, R1-R6 in Formula XI, and R1-R4 in Formula XII are each independently selected from hydrogen, hydroxyl, methylene, oxygenated methylene, methyl, ethyl, allyl, carbonyl, halogen, acyl, acyl halide, nitro, cyano, glucosyl, arabinopyranosyl, allosyl, rhamnosyl, benzene ring and indole ring, cis-feruloyloxy, trans-feruloyloxy, cis-p-coumaroyloxy, trans-p-coumaroyloxy.

[0053] Preferably, Formula I is Compound 1 or Compound 2, Formula II is Compound 3, Compound 4, Compound 5 or Compound 6, Formula III is Compound 7, Formula IV is one of Compound 8-10, Compound 13-17, Compound 20-22, Formula V is Compound 18, Formula VI is Compound 19, Formula VII is Compound 11 or Compound 12, Formula VIII is one of Compound 23-26, Compound 28, Compound 30-32, Formula IX is Compound 27, Formula X is Compound 29, Formula XI is one of Compound 33-35, Compound 37, and Formula XII is Compound 36;

[0054]

[0055]

[0056]

[0057] The compounds of the present invention are isolated and purified from the medicinal plant Pieris formosa (Wall.) D. Don of the genus Pieris in the family Ericaceae. The present invention obtains the above triterpenoids 1-37 by repeatedly subjecting the chloroform fraction extract of the 95% ethanol extract of Pieris formosa to column chromatography separation and purification, among which Compounds 1-12 are new compounds. Compounds 1-7 are taraxastane-type triterpenoids with a 6 / 6 / 6 / 6 / 6 pentacyclic carbon ring system, Compounds 8-10, 13-22 are ursane-type triterpenoids with a 6 / 6 / 6 / 6 / 6 pentacyclic carbon ring system, Compounds 11-12, 23-32 are oleanane-type triterpenoids with a 6 / 6 / 6 / 6 / 6 pentacyclic carbon ring system, and Compounds 33-37 are lupane-type triterpenoids with a 6 / 6 / 6 / 6 / 5 pentacyclic carbon ring system.

[0058] The present invention determines its structure by using various spectroscopic analysis methods, quantum chemical calculation methods, and X-ray single crystal diffraction methods.

[0059] The above-mentioned compounds 1-37 provided by the present invention have significant α-glucosidase inhibitory effects and can be used to prepare hypoglycemic drugs.

[0060] The present invention provides a study on the α-glucosidase inhibitory effects of compounds 1-37. The α-glucosidase inhibitory activity was determined by the pNPG method. The results showed that compounds 1-37 all exhibited varying degrees of α-glucosidase inhibitory activity. Compounds 1, 2, 4, 9-12, 23, and 26 had significant inhibitory effects on α-glucosidase, which were stronger than the positive drug acarbose.

[0061] The following are specific examples:

[0062] Example 1: The separation and preparation process of the compounds of the present invention

[0063] After drying and pulverizing the leaves of Pieris formosa var. wilsonii (31.9 kg), it was soaked and extracted 5 times (5 days each time) with industrial-grade 95% ethanol (120.0 L) at room temperature. After removing ethanol by reduced pressure concentration, 12.0 kg of crude extract was obtained. After suspending and dispersing in an appropriate amount of warm water, it was extracted successively with petroleum ether and chloroform to obtain the petroleum ether fraction (445.5 g) and chloroform fraction (1386.5 g) of the leaves of Pieris formosa var. wilsonii. The chloroform fraction was mixed with silica gel of 100-200 mesh and subjected to normal-phase silica gel column chromatography. The eluent was gradient elution with dichloromethane / methanol (200:1 to 2:1, V / V). Combining TLC spotting detection, 7 fractions (A1-A7) with increasing polarity were obtained.

[0064] The A3 fraction was separated by ODS column chromatography (methanol / water, 40:60 → pure methanol, V / V) to obtain 15 sub-fractions (A3a-A3o) with decreasing polarity. A3h was separated by Sephadex LH-20 gel column chromatography (methanol) and silica gel column chromatography (dichloromethane / methanol, 500:1 → pure methanol, V / V) to obtain compound 17 (14.5 mg) and compound 25 (11.1 mg) with increasing polarity. A3i was separated by Sephadex LH-20 gel column chromatography (methanol) and silica gel column chromatography (dichloromethane / methanol, 500:1 → pure methanol, V / V) to obtain compound 33 (7.8 mg) and 5 sub-fractions (A3i1-A3i5) with increasing polarity. A3i4 was further purified by semi-preparative HPLC (methanol / water, 80:20, V / V, 1.5 mL / min, C 18 column) to obtain compound 15 (t R = 35.0 min, 4.2 mg) and compound 16 (t R= 38.8 min, 7.3 mg). A3j was separated by Sephadex LH-20 gel column chromatography (methanol) and silica gel column chromatography (dichloromethane / methanol, 200:1 → pure methanol, V / V) to obtain compound 19 (23.0 mg). A3k was separated by Sephadex LH-20 gel column chromatography (methanol) and silica gel column chromatography (dichloromethane / methanol, 200:1 → pure methanol, V / V) to obtain compound 21 (3.0 mg) and compound 27 (11.0 mg) with increasing retention time. A3n was separated by Sephadex LH-20 gel column chromatography (methanol) and silica gel column chromatography (dichloromethane / methanol, 150:1 → pure methanol, V / V) to obtain seven sub-components (A3n1–A3n7) with increasing polarity. A3n2 was purified by semi-preparative HPLC (methanol / water, 73:27, V / V, 1.5 mL / min, C 18 column) to obtain compound 28 (t R = 27.7 min, 18.9 mg). A3n5 was purified by semi-preparative HPLC (methanol / water, 66:34, V / V, 1.5 mL / min, phenyl column) to obtain compound 37 (t R = 18.2 min, 2.5 mg).

[0065] Component A4 was subjected to reversed-phase C 18 silica gel column chromatography, and the eluent was gradient eluted with methanol / water (40:60 to 100:1, V / V) to obtain sub-components A4a–A4j with decreasing polarity. A4b was purified by Sephadex LH-20 gel column chromatography (methanol), silica gel column chromatography (dichloromethane / methanol, 100:1 → pure methanol, V / V), and finally by semi-preparative HPLC (methanol / water, 78:22, 1.5 mL / min, phenyl column) to obtain compound 31 (t R = 27.9 min, 3.0 mg), compound 20 (t R = 31.1 min, 8.0 mg), and compound 34 (t R = 42.5 min, 5.3 mg) with increasing retention time. When component A4c was subjected to Sephadex LH-20 gel column chromatography with methanol as the eluent, component A4c3 was obtained. Component A4c3 was separated by semi-preparative high-performance liquid chromatography C 18 column with acetonitrile / water (95:5, V / V) as the eluent and a flow rate of 2.0 mL / min to obtain compound 5 (t R = 30.5 min, 14.5 mg), compound 6 (t R = 50.1 min, 7.5 mg), and compound 7 (t R= 52.2 min, 6.6 mg). When fraction A4d was subjected to Sephadex LH-20 gel column chromatography with methanol as the eluent, four sub-fractions A4d1–A4d4 with decreasing molecular weights were obtained. Fraction A4d4 was separated by semi-preparative high performance liquid chromatography on a phenyl column with acetonitrile / water (70:30, V / V) as the eluent at a flow rate of 2.0 mL / min, and compounds 9 (t R = 31.3 min, 3.5 mg) and compound 10 (t R = 33.5 min, 4.5 mg) with increasing retention times were obtained. A4h was separated by Sephadex LH-20 gel column chromatography (methanol) and silica gel column chromatography (dichloromethane / methanol, 200:1 → pure methanol) to obtain compound 26 (8.4 mg) and compound 13 (13.3 mg) with increasing polarity.

[0066] Fraction A5 was subjected to reversed-phase C 18 silica gel column chromatography with a gradient elution of methanol / water (40:60 to 100:1, V / V) to obtain sub-fractions A5a–A5q with decreasing polarity. A5h was separated by Sephadex LH-20 gel column chromatography (methanol) and silica gel column chromatography (dichloromethane / methanol, 100:1 → pure methanol, V / V) to obtain compound 35 (4.8 mg) and five sub-fractions (A5h1–A5h5) with increasing polarity. A5h3 was separated by semi-preparative HPLC (methanol / water, 70:30, V / V, 1.5 mL / min, C18 column) to obtain four sub-fractions (A5h3a–A5h3d). A5h3b was purified by semi-preparative HPLC (methanol / water, 72.5:27.5, V / V, 1.5 mL / min, C18 column) to obtain compounds 22 (t R = 21.3 min, 7.3 mg) and compound 23 (t R = 27.0 min, 14.0 mg) with increasing retention times. A5h3c was purified by semi-preparative HPLC (methanol / water, 69:31, V / V, 1.5 mL / min, C18 column) to obtain compounds 30 (t R = 26.9 min, 7.3 mg) and compound 36 (t R = 33.1 min, 13.7 mg) with increasing retention times. When fraction A5j was further subjected to Sephadex LH-20 gel column chromatography (eluent: methanol) and reversed-phase C 18 silica gel column chromatography (eluent: methanol / water, 75:25 to 90:10, V / V), fraction A5j2 was obtained. Fraction A5j2 was subjected to semi-preparative high performance liquid chromatography C 18Column separation was performed with methanol / water (80:20, V / V) as the eluent at a flow rate of 2.0 mL / min, yielding compounds 3 (t R = 45.5 min, 15.2 mg) and 4 (t R = 47.3 min, 4.5 mg) with retention times increasing from short to long. A5k was separated by Sephadex LH-20 column chromatography (methanol) and silica gel column chromatography (dichloromethane / methanol, 100:1 → pure methanol, V / V) to obtain compounds 18 (3.8 mg), 24 (17.3 mg), 14 (4.5 mg) with increasing polarity and 7 sub-components (A5k1 - A5k7) with increasing polarity. A5k7 was purified by semi-preparative HPLC (acetonitrile / water, 70:30, V / V, 1.5 ml / min, C 18 column) to obtain compounds 29 (t R = 23.9 min, 4.3 mg) and 32 (t R = 32.3 min, 6.5 mg) with retention times increasing from short to long. When fraction A5m was further subjected to Sephadex LH-20 column chromatography (eluent: methanol) and reversed-phase C 18 silica gel column chromatography (eluent: methanol / water, 75:25 to 90:10, V / V), fraction A5m2 was obtained. Fraction A5m2 was separated by semi-preparative high-performance liquid chromatography C 18 column with methanol / water (60:40, V / V) as the eluent at a flow rate of 2.0 mL / min, yielding compounds 11 (t R = 27.7 min, 7.2 mg) and 12 (t R = 31.5 min, 4.0 mg) with retention times increasing from short to long. When fraction A5o was further subjected to Sephadex LH-20 column chromatography (eluent: methanol) and reversed-phase C 18 silica gel column chromatography (eluent: methanol / water, 60:40 to 90:10, V / V), fraction A5o3 was obtained. Fraction A5o3 was subjected to reversed-phase phenyl silica gel column chromatography with gradient elution using methanol / water (55:45 to 100:1, V / V) to obtain 4 sub-components A5o3a - A5o3d with decreasing polarity. Fraction A5o3b was separated by semi-preparative high-performance liquid chromatography C 18 column with methanol / water (65:35, V / V) as the eluent at a flow rate of 2.0 mL / min, yielding compound 8 (t R = 37.5 min, 6.0 mg). When fraction A5p was further subjected to Sephadex LH-20 column chromatography (eluent: methanol) and reversed-phase C 18Silica gel column chromatography (eluent: methanol / water, 55:45 to 90:10, V / V) gave fraction A5p3. Fraction A5p3 was separated by semi-preparative high performance liquid chromatography C 18 column with eluent methanol / water (62:38, V / V) at a flow rate of 2.0 mL / min to give compounds 1 (t R = 25.6 min, 4.5 mg) and 2 (t R = 28.9 min, 3.2 mg) with retention times increasing from short to long.

[0067] Example 2: Separation and preparation process of the compounds of the present invention

[0068] Combined with a variety of spectroscopic analysis methods (high resolution mass spectrometry HRESIMS, ultraviolet spectroscopy UV, infrared spectroscopy IR and nuclear magnetic resonance spectroscopy NMR), quantum chemical ECD calculation method and X-ray single crystal diffraction and other methods for comprehensive analysis to determine the structures of new compounds 1–12. The structures of known compounds 13–37 were determined by comparing literature data. The absolute configurations of compounds 1, 7 and 11 were determined by X-ray single crystal diffraction, as shown in Figure 1 , Figure 2 , Figure 3 .

[0069] Compound 1 (Pieristriterpenin A): Colorless crystal, m.p. 225–227 °C. Optical rotation value (c 0.1, methanol); UV (methanol) λ max (logε): 199 (2.88), 218 (2.79), 237 (2.73), 298 (2.80), 328 (2.98) nm; IR (KBr) ν max 3489, 2972, 2935, 2869, 1738, 1697, 1631, 1594, 1519, 1457, 1375, 1259, 1161, 1042, 994 cm -1 ; CD (methanol) λ max (Δε): 210 (-2.5), 230 (-1.3), 250 (+8.8), 330 (+15.0) nm; 1 1H and 13 13C NMR spectral data are shown in Tables 1 and 2; high resolution mass spectrometry m / z 671.3912 [M+Na] + (calculated for C 40 41H 56 37O7Na, 697.3924).

[0070] Compound 2 (Pieristriterpenin B): White powder; optical rotation value (c0.1, methanol); UV (methanol) λ max (logε): 199 (3.03), 212 (2.86), 232 (2.53), 298 (2.70), 310 (2.73) nm; IR (KBr) ν max 3456, 2974, 2928, 2871, 1756, 1694, 1608, 1514, 1451, 1375, 1270, 1175, 1093, 849 cm -1 ; CD (methanol) λ max (Δε): 215 (+8.5), 240 (+8.3), 310 (19.6) nm; 1 H and 13 C NMR spectral data are shown in Tables 2 and 3; HRMS m / z 671.3933 [M+Na] + (calcd for C 40 H 56 O7Na, 671.3924).

[0071] Compound 3 (Pieristriterpenin C): white powder; optical rotation value (c 0.1, methanol); UV (methanol) λ max (logε): 199 (2.88), 211 (2.79), 229 (2.69), 293 (2.93) and 314 (3.02) nm; IR (KBr) 3462, 2967, 2928, 2864, 1679, 1628, 1601, 1161, 983, 828 cm -1 ; CD (methanol) λ max (Δε): 202 (-3.1), 220 (+12.3), 272 (+6.5), 307 (+19.4) nm; 1 H and 13 C NMR spectral data are shown in Tables 1 and 2; HRMS HRESIMS m / z 611.4073 [M+Na] + (calcd for C 39 H 56 O4Na, 611.4076) and 1199.8166 [2M+Na] + (calcd for C 78 H 112 O8Na, 1199.8255).

[0072] Compound 4 (Pieristriterpenin D): white powder; optical rotation value (c 0.05, methanol); UV (methanol) λmax (logε): 199(2.97), 211(2.77), 225(2.59), 288(2.63), 312(2.67) nm; infrared (KBr) ν max 3473, 2971, 2937, 2862, 1686, 1640, 1608, 1517, 1446, 1250, 1170, 992, 890, 837 cm -1 ; CD (methanol) λ max (Δε): 220(+7.5), 260(+6.9), 307(+7.3) nm; 1 H and 13 C NMR spectral data are shown in Table 1 and Table 2; high-resolution mass spectrometry m / z 611.4070 [M+Na] + (calculated for C 39 H 56 O4Na, 611.4076).

[0073] Compound 5 (Pieristriterpenin E): white powder; optical rotation value (c 0.1, methanol); ultraviolet (methanol) λ max (logε): 201(2.83) nm; infrared (KBr) ν max 3482, 2963, 2931, 2889, 2564, 1642, 1462, 1385, 1031, 997, 876 cm -1 ; CD (methanol) λ max (Δε): 206(+6.9) nm; 1 H and 13 C NMR spectral data are shown in Table 1 and Table 2; high-resolution mass spectrometry m / z 465.3690 [M+Na] + (calculated for C 30 H 50 O2Na, 465.3709).

[0074] Compound 6 (Pieristriterpenin F): white powder; optical rotation value (c 0.05, methanol); ultraviolet (methanol) λ max (logε): 201(2.83) nm; infrared (KBr) ν max 3483, 2937, 2869, 1449, 1380, 1035, 1052, 908 cm -1 ; CD (methanol) λ max (Δε): 207(+5.8) nm; 1 H and 1313C NMR spectral data are shown in Table 1 and Table 2; high-resolution mass spectrometry m / z 465.3707 [M+Na] + (calcd for C 30 H 50 O2Na, 465.3709) and 907.7620 [2M+Na] + (calcd for C 78 H 112 O8Na, 907.7519).

[0075] Compound 7 (Pieristriterpenin G): colorless crystals, m.p. 231–232 °C. Optical rotation (c 0.05, MeOH); UV (MeOH) λ max (logε): 272 (4.03) nm; IR (KBr) ν max 3414, 2961, 2929, 2872, 1644, 1601, 1461, 1377, 1318, 1246, 1077, 1012, and 757 cm -1 ; CD (MeOH) λ max (Δε): 212 (+13.7) nm; 1 1H and 13 13C NMR spectral data are shown in Table 1 and Table 2; high-resolution mass spectrometry m / z 465.3705 [M+Na] + (calcd for C 30 H 50 O2Na, 465.3709).

[0076] Compound 8 (Pieristriterpenin H): white powder; optical rotation (c 0.1, MeOH); UV (MeOH) λ max (logε): 198 (2.65), 250 (2.65) nm; IR (KBr) ν max 3418, 2972, 2949, 2928, 2869, 1654, 1455, 1383, 1221, 1195, 1038, 995 cm -1 ; CD (MeOH) λ max (Δε): 248 (+9.7), 325 (-0.9) nm; 1 1H and 13 13C NMR spectral data are shown in Table 2 and Table 3; high-resolution mass spectrometry m / z 479.3487 [M+Na] + (calcd for C 30 H 48 O3Na, 479.3501).

[0077] Compound 9 (Pieristriterpenin I): white powder; optical rotation value (c 0.1, methanol); UV (methanol) λ max (logε): 199 (2.98), 210 (2.81), 228 (2.73), 290 (2.92), 312 (2.97) nm; IR (KBr) ν max 3411, 2931, 2880, 1686, 1631, 1604, 1512, 1453, 1270, 1163, 1013, 967, 830 cm -1 ; 1 H and 13 13C NMR spectral data are shown in Tables 2 and 3; high-resolution mass spectrometry m / z 641.3814 [M+Na] + (calcd for C 39 H 54 O6Na, 641.3818).

[0078] Compound 10 (Pieristriterpenin J): white powder; optical rotation value (c 0.05, methanol); UV (methanol) λ max (logε): 198 (2.93), 215 (2.65), 234 (2.54), 300 (2.55), 326 (2.68) nm; IR (KBr) ν max 3475, 2972, 2939, 2871, 1743, 1704, 1633, 1592, 1515, 1464, 1373, 1168, 1040, 817 cm -1 ; CD (methanol) λ max (Δε): 218 (-12.5), 310 (+4.2) nm; 1 H and 13 13C NMR spectral data are shown in Tables 2 and 3; high-resolution mass spectrometry m / z 641.3818 [M+Na] + (calcd for C 39 H 54 O6Na, 641.3818).

[0079] Compound 11 (Pieristriterpenin K): colorless crystals, m.p. 266–267 °C. Optical rotation value (c 0.05, methanol); UV (methanol) λ max(logε): 199(2.91), 213(2.77), 221(2.92), 234(2.98), 241(3.02), 255(2.89), 292(2.67), 310(2.70) nm; Infrared (KBr) ν max 3387, 3036, 2951, 2864, 1697, 1603, 1509, 1198, 1162, 1141 cm -1 ; 1 H and 13 C NMR spectral data are shown in Tables 2 and 3; High-resolution mass spectrometry m / z 623.3702 [M+Na] + (Calculated for C 39 H 54 O5Na, 623.3712).

[0080] Compound 12 (Pieristriterpenin L): White powder; Optical rotation value (c 0.05, methanol); Ultraviolet (methanol) λ max (logε): 199(2.90), 216(2.76), 234(2.95), 242(2.96), 250(2.97), 261(2.83), 299(2.88), 312(2.93) nm; Infrared (KBr) ν max 3416, 2947, 2865, 1685, 1607, 1509, 1272, 1199, 1164, 836 cm -1 ; 1 H and 13 C NMR spectral data are shown in Tables 2 and 3; High-resolution mass spectrometry m / z 623.3714 [M+Na] + (Calculated for C 39 H 54 O5Na, 623.3712).

[0081] Table 1 1 H NMR data of Compounds 1–6 (400 MHz)

[0082]

[0083] a Pyridine-d5; b Chloroform-d; c Methanol-d4

[0084] Table 2 13 C NMR data of Compounds 1–12 (100 MHz)

[0085]

[0086] a deuterated pyridine; b deuterated chloroform; c deuterated methanol

[0087] 1H NMR data (400 MHz) of compounds 7–12 1 1H NMR data (400 MHz)

[0088]

[0089]

[0090] a deuterated pyridine; b deuterated chloroform; c deuterated methanol

[0091] Compound 13 (3β,12-dihydroxyurs-12-en-11-one): white powder; optical rotation value (c 0.2, methanol); ESIMS m / z: 479.4 [M+Na] + , with the molecular formula C 30 H 48 O3, and the hydrogen deficiency index is 7. 1 1H NMR (400 MHz, deuterated chloroform) δ H : 0.86 (3H, s, H3-23), 0.87 (3H, s, H3-24), 1.10 (3H, s, H3-25), 1.11 (3H, s, H3-26), 1.33 (3H, s, H3-27), 0.97 (3H, s, H3-28), 0.77 (3H, d, J = 6.3 Hz, H3-29), 0.90 (3H, d, J = 6.5 Hz, H3-30), 2.07 (1H, td, J = 13.6, 4.7 Hz, H-16b), 3.20 (1H, dd, J = 10.8, 5.5 Hz, H-3α), 5.56 (1H, s, 12-OH); 13 13C NMR (100 MHz, deuterated chloroform) δ C: 29.0 (C-28), 134.5 (C-13), 144.7 (C-12), 78.9 (C-3), 41.0 (C-19), 28.3 (C-23), 49.1 (C-18), 55.1 (C-5), 33.6 (C-17), 59.9 (C-9), 45.7 (C-8), 39.5 (C-20), 41.9 (C-14), 39.4 (C-4), 39.3 (C-1), 41.4 (C-22), 37.4 (C-10), 33.2 (C-7), 27.5 (C-15), 31.4 (C-21), 27.7 (C-2), 16.7 (C-29), 27.5 (C-16), 21.2 (C-27), 195.6 (C-11), 17.7 (C-6), 18.8 (C-26), 16.8 (C-25), 21.2 (C-30), 15.8 (C-24).

[0092] Compound 14 (3β-hydroxy-11-oxo-ursan-12-ene): white powder; optical rotation value (c 0.1, methanol); ESI MS m / z: 463.4 [M+Na] + , molecular formula C 30 H 48 O2, hydrogen deficiency index is 7. 1 1H NMR (400 MHz, chloroform-d) δ H : 0.98 (3H, s, H3-23), 0.91 (3H, s, H3-24), 1.13 (3H, s, H3-25), 1.14 (3H, s, H3-26), 1.27 (3H, s, H3-27), 0.79 (3H, s, H3-28), 0.77 (3H, d, J = 6.3 Hz, H3-29), 1.10 (3H, d, J = 6.5 Hz, H3-30), 2.10 (1H, td, J = 12.5, 4.2 Hz, H-16b), 3.20 (1H, dd, J = 11.0, 5.4 Hz, H-3α), 5.51 (1H, s, H-12), 0.69 (1H, dd, J = 12.1, 1.1 Hz, H-5), 2.30 (1H, s, H-9); 13 13C NMR (100 MHz, chloroform-d) δ C: 29.0 (C-28), 130.7 (C-13), 165.1 (C-12), 79.0 (C-3), 39.5 (C-19), 28.3 (C-23), 61.7 (C-18), 55.2 (C-5), 34.1 (C-17), 59.2 (C-9), 45.3 (C-8), 39.5 (C-20), 41.1 (C-14), 39.4 (C-4), 39.3 (C-1), 41.1 (C-22), 37.2 (C-10), 33.1 (C-7), 27.5 (C-15), 31.1 (C-21), 27.7 (C-2), 16.7 (C-29), 27.5 (C-16), 21.3 (C-27), 200.0 (C-11), 17.8 (C-6), 18.7 (C-26), 17.6 (C-25), 21.3 (C-30), 15.8 (C-24).

[0093] Compound 15 (3β,28-dihydroxy-11-oxo-ursan-12-ene): white powder; optical rotation value (c 0.1, methanol); ESIMS m / z: 456.3 [M+Na] + , molecular formula C 30 H 48 O3, hydrogen deficiency index is 7. 1 1H NMR (400 MHz, chloroform-d) δ H : 0.98 (3H, s, H3-23), 0.91 (3H, s, H3-24), 1.13 (3H, s, H3-25), 1.14 (3H, s, H3-26), 1.27 (3H, s, H3-27), 3.14 (1H, d, J = 10.9 Hz, H-28a), 3.43 (1H, d, J = 10.9 Hz, H-28b), 0.77 (3H, d, J = 6.3 Hz, H3-29), 1.10 (3H, d, J = 6.5 Hz, H3-30), 2.10 (1H, td, J = 12.5, 4.2 Hz, H-16b), 3.20 (1H, dd, J = 11.0, 5.4 Hz, H-3α), 5.51 (1H, s, H-12), 0.69 (1H, dd, J = 12.1, 1.1 Hz, H-5), 2.30 (1H, s, H-9); 13 13C NMR (100 MHz, chloroform-d) δ C: 69.9 (C-28), 130.7 (C-13), 165.1 (C-12), 79.0 (C-3), 39.5 (C-19), 28.3 (C-23), 61.7 (C-18), 55.2 (C-5), 36.1 (C-17), 59.2 (C-9), 45.3 (C-8), 39.5 (C-20), 41.1 (C-14), 39.4 (C-4), 39.3 (C-1), 41.1 (C-22), 37.2 (C-10), 33.1 (C-7), 27.5 (C-15), 31.1 (C-21), 27.7 (C-2), 16.7 (C-29), 27.5 (C-16), 21.3 (C-27), 200.0 (C-11), 17.8 (C-6), 18.7 (C-26), 17.6 (C-25), 21.3 (C-30), 15.8 (C-24).

[0094] Compound 16 (3β,12-dihydroxyurs-12-en-11-one): white powder; optical rotation value (c0.2, methanol); ESIMS m / z: 472.4 [M+Na] + , molecular formula is C 31 H 52 O3, the hydrogen deficiency index is 6. 1 1H NMR (400 MHz, chloroform-d) δ H : 1.04 (3H, s, H3-23), 0.85 (3H, s, H3-24), 1.14 (3H, s, H3-25), 1.11 (3H, s, H3-26), 1.20 (3H, s, H3-27), 0.81 (3H, s, H3-28), 0.92 (3H, d, J = 6.6 Hz, H3-29), 0.94 (3H, d, J = 6.6 Hz, H3-30), 2.01 (1H, td, J = 13.6, 4.7 Hz, H-16b), 3.25 (1H, dd, J = 8.9, 7.5 Hz, H-3α), 4.49 (1H, s, 12-OH), 3.17 (1H, s, 11-OCH3); 13 13C NMR (100 MHz, chloroform-d) δ C: 28.5 (C-28), 118.4 (C-13), 142.7 (C-12), 78.9 (C-3), 41.2 (C-19), 28.5 (C-23), 48.3 (C-18), 55.1 (C-5), 33.6 (C-17), 46.9 (C-9), 40.6 (C-8), 39.9 (C-20), 42.8 (C-14), 39.4 (C-4), 39.3 (C-1), 41.4 (C-22), 37.4 (C-10), 34.8 (C-7), 27.5 (C-15), 31.4 (C-21), 27.7 (C-2), 16.7 (C-29), 27.5 (C-16), 23.5 (C-27), 76.9 (C-11), 17.7 (C-6), 18.8 (C-26), 16.8 (C-25), 21.2 (C-30), 15.8 (C-24), 51.4 (C-OCH3).

[0095] Compound 17 (3β,11α-Methoxyurs-12-en-3-ol): white powder; optical rotation value (c 0.1, methanol); ESIMS m / z: 456.4 [M+Na] + , with the molecular formula C 31 H 52 O2, and the hydrogen deficiency index is 6. 1 1H NMR (400 MHz, chloroform-d) δ H : 0.97 (3H, s, H3-23), 0.77 (3H, s, H3-24), 1.01 (3H, s, H3-25), 1.03 (3H, s, H3-26), 1.11 (3H, s, H3-27), 0.78 (3H, s, H3-28), 0.84 (3H, d, J = 5.8 Hz, H3-29), 0.90 (3H, d, J = 6.4 Hz, H3-30), 1.90 (1H, dt, J = 13.8, 3.5 Hz, H-16b), 3.20 (1H, dd, J = 10.3, 6.0 Hz, H-3α), 3.25 (3H, s, -OCH3), 3.77 (1H, dd, J = 8.9, 3.1 Hz, H-11); 13 13C NMR (100 MHz, chloroform-d) δ C: 28.9 (C-28), 143.2 (C-13), 124.2 (C-12), 79.0 (C-3), 39.4 (C-19), 28.1 (C-23), 58.8 (C-18), 55.0 (C-5), 33.9 (C-17), 54.5 (C-9), 43.6 (C-8), 39.2 (C-20), 41.3 (C-14), 39.4 (C-4), 40.2 (C-1), 41.7 (C-22), 38.3 (C-10), 33.8 (C-7), 27.0 (C-15), 31.8 (C-21), 27.5 (C-2), 17.8 (C-29), 28.3 (C-16), 22.7 (C-27), 77.0 (C-11), 18.2 (C-6), 18.8 (C-26), 17.4 (C-25), 21.1 (C-30), 15.9 (C-24).

[0096] Compound 18 (3β-hydroxy-12-oxours-11-ene): white powder; optical rotation value (c 0.1, methanol); ESIMS m / z: 463.4 [M+Na] + , with the molecular formula C 30 H 48 O2, and the hydrogen deficiency index is 7. 1 1H NMR (400 MHz, chloroform-d) δ H : 1.05 (3H, s, H3-23), 0.84 (3H, s, H3-24), 1.21 (3H, s, H3-25), 1.29 (3H, s, H3-26), 1.09 (3H, s, H3-27), 0.99 (3H, s, H3-28), 0.69 (3H, d, J = 6.7 Hz, H3-29), 0.87 (3H, d, J = 6.4 Hz, H3-30), 1.95 (1H, dt, J = 13.5, 4.6 Hz, H-16b), 3.17 (1H, dd, J = 11.8, 5.4 Hz, H-3α), 5.92 (1H, s, H-11), 2.01 (1H, dt, J = 13.3, 3.5 Hz, H-1b); 13 13C NMR (100 MHz, chloroform-d) δ C: 28.5 (C-28), 48.2 (C-13), 203.2 (C-12), 78.3 (C-3), 39.4 (C-19), 28.1 (C-23), 47.9 (C-18), 50.0 (C-5), 34.9 (C-17), 179.5 (C-9), 45.6 (C-8), 39.2 (C-20), 41.3 (C-14), 39.4 (C-4), 36.4 (C-1), 41.7 (C-22), 40.1 (C-10), 33.2 (C-7), 27.0 (C-15), 31.8 (C-21), 27.9 (C-2), 20.2 (C-29), 27.3 (C-16), 20.0 (C-27), 123.4 (C-11), 18.2 (C-6), 24.8 (C-26), 24.4 (C-25), 21.1 (C-30), 15.9 (C-24).

[0097] Compound 19 (3β-hydroxy-urs-11-en-28(13)-olide): white powder; optical rotation value (c 0.1, methanol); ESIMS m / z: 477.3 [M+Na] + , with the molecular formula C 30 H 46 O3, and the hydrogen deficiency index is 8. 1 1H NMR (400 MHz, chloroform-d) δ H : 0.99 (3H, s, H3-23), 0.78 (3H, s, H3-24), 0.93 (3H, s, H3-25), 0.94 (3H, s, H3-26), 1.16 (3H, s, H3-27), 1.05 (3H, d, J = 6.5 Hz, H3-29), 1.00 (3H, d, J = 6.5 Hz, H3-30), 2.12 (1H, td, J = 11.5, 4.7 Hz, H-16b), 3.20 (1H, dd, J = 11.4, 4.8 Hz, H-3α), 5.53 (1H, dd, J = 10.4, 3.0 Hz, H-12), 0.74 (1H, dd, J = 11.8, 2.1 Hz, H-5), 1.95 (1H, s, H-9); 13 13C NMR (100 MHz, chloroform-d) δ C: 179.8 (C-28), 89.6 (C-13), 128.9 (C-12), 78.9 (C-3), 38.2 (C-19), 27.8 (C-23), 60.6 (C-18), 54.8 (C-5), 45.1 (C-17), 53.1 (C-9), 42.0 (C-8), 40.3 (C-20), 42.1 (C-14), 38.9 (C-4), 39.4 (C-1), 31.4 (C-22), 36.4 (C-10), 31.3 (C-7), 25.6 (C-15), 30.9 (C-21), 27.1 (C-2), 17.8 (C-29), 22.9 (C-16), 16.1 (C-27), 133.4 (C-11), 17.7 (C-6), 18.9 (C-26), 9.2 (C-25), 17.9 (C-30), 14.9 (C-24).

[0098] Compound 20 (pomolic acid): white powder; optical rotation value (c 0.1, methanol); ESIMS m / z: 486.3 [M+Na] + , molecular formula is C 30 H 46 O4, hydrogen deficiency index is 8. 1 1H NMR (400 MHz, chloroform-d) δ H : 1.01 (3H, s, H3-24), 3.26 (1H, d, J = 11.7 Hz, H-23a), 3.61 (1H, d, J = 11.7 Hz, H-23b), 1.10 (3H, s, H3-25), 1.06 (3H, s, H3-26), 1.25 (3H, s, H3-27), 1.19 (3H, s, H3-29), 0.92 (3H, d, J = 6.6 Hz, H3-30), 1.90 (1H, ddd, J = 15.5, 7.3, 3.4 Hz, H-16b), 2.01 (1H, dd, J = 9.0, 3.6 Hz, H-9), 5.36 (1H, t, J = 3.5 Hz, H-12), 2.50 (1H, s, H-18); 13 13C NMR (100 MHz, chloroform-d) δ C: 182.5 (C-28), 137.3 (C-13), 128.2 (C-12), 216.9 (C-3), 72.1 (C-19), 66.6 (C-23), 52.0 (C-18), 55.9 (C-5), 47.7 (C-17), 46.0 (C-9), 39.1 (C-8), 41.8 (C-20), 41.1 (C-14), 47.5 (C-4), 40.1 (C-1), 37.4 (C-22), 37.8 (C-10), 34.2 (C-7), 29.2 (C-15), 25.9 (C-21), 32.2 (C-2), 26.6 (C-29), 25.3 (C-16), 24.6 (C-27), 23.7 (C-11), 19.6 (C-6), 17.0 (C-26), 14.9 (C-25), 16.8 (C-30), 22.7 (C-24).

[0099] Compound 21 (19,24-dihydroxy-urs-12-en-3-one-28-oic acid): white powder; optical rotation value (c 0.1, methanol); ESIMS m / z: 493.3 [M+Na] + , with the molecular formula C 30 H 46 O4, and the index of hydrogen deficiency is 8. 1 1H NMR (400 MHz, chloroform-d) δ H : 1.01 (3H, s, H3-23), 0.76 (3H, s, H3-24), 1.04 (3H, s, H3-25), 1.06 (3H, s, H3-26), 1.24 (3H, s, H3-27), 1.19 (3H, s, H3-29), 0.92 (3H, d, J = 6.6 Hz, H3-30), 1.86 (1H, ddd, J = 15.8, 7.3, 3.4 Hz, H-16b), 2.01 (1H, dd, J = 9.0, 3.6 Hz, H-9), 5.36 (1H, t, J = 3.5 Hz, H-12), 2.50 (1H, s, H-18); 13 13C NMR (100 MHz, chloroform-d) δ C: 184.1 (C-28), 138.3 (C-13), 129.2 (C-12), 217.9 (C-3), 73.1 (C-19), 27.6 (C-23), 52.9 (C-18), 55.3 (C-5), 47.8 (C-17), 46.5 (C-9), 39.9 (C-8), 41.2 (C-20), 41.1 (C-14), 47.5 (C-4), 39.1 (C-1), 37.4 (C-22), 36.8 (C-10), 34.2 (C-7), 28.2 (C-15), 25.9 (C-21), 32.2 (C-2), 26.6 (C-29), 25.3 (C-16), 24.6 (C-27), 23.7 (C-11), 19.6 (C-6), 17.0 (C-26), 15.1 (C-25), 16.3 (C-30), 21.7 (C-24).

[0100] Compound 22 (rotundanoic acid): white powder; optical rotation value (c 0.1, methanol); ESIMS m / z: 486.3 [M+Na] + , molecular formula is C 30 H 46 O4, the hydrogen deficiency index is 8. 1 1H NMR (400 MHz, chloroform-d) δ H : 0.99 (3H, s, H3-23), 3.42 (1H, d, J = 11.7 Hz, H-24a), 3.71 (1H, d, J = 11.7 Hz, H-24b), 1.10 (3H, s, H3-25), 1.06 (3H, s, H3-26), 1.25 (3H, s, H3-27), 1.19 (3H, s, H3-29), 0.92 (3H, d, J = 6.6 Hz, H3-30), 1.90 (1H, ddd, J = 15.5, 7.3, 3.4 Hz, H-16b), 2.01 (1H, dd, J = 9.0, 3.6 Hz, H-9), 5.36 (1H, t, J = 3.5 Hz, H-12), 2.50 (1H, s, H-18); 13 13C NMR (100 MHz, chloroform-d) δ C: 182.5 (C-28), 137.3 (C-13), 128.2 (C-12), 216.9 (C-3), 72.1 (C-19), 26.6 (C-23), 52.0 (C-18), 55.9 (C-5), 47.7 (C-17), 46.0 (C-9), 39.1 (C-8), 41.8 (C-20), 41.1 (C-14), 47.5 (C-4), 40.1 (C-1), 37.4 (C-22), 37.8 (C-10), 34.2 (C-7), 29.2 (C-15), 25.9 (C-21), 32.2 (C-2), 26.6 (C-29), 25.3 (C-16), 24.6 (C-27), 23.7 (C-11), 19.6 (C-6), 17.0 (C-26), 14.9 (C-25), 16.8 (C-30), 69.7 (C-24).

[0101] Compound 23 (3β-tran-p-coumaroyl maslinic acid): white powder; optical rotation value (c 0.1, methanol); ESIMS m / z 641.4 [M+Na] + , molecular formula is C 39 H 54 O6, the index of hydrogen deficiency is 13. 1 1H NMR (400 MHz, deuterated methanol) δ H : 5.33 (1H, t, J = 3.0 Hz, H-12), 1.10 (3H, H-23), 0.89 (3H, s, H3-24), 1.23 (3H, s, H3-25), 0.89 (3H, s, H3-26), 1.17 (3H, s, H3-27), 1.29 (3H, s, H3-29), 4.64 (1H, d, J = 9.9 Hz, H-3), 3.85 (1H, ddd, J = 10.4, 9.8, 4.5 Hz, H-2), 6.39 (1H, d, J = 15.9 Hz, H-8′), 7.63 (1H, d, J = 15.9 Hz, H-7′), 6.81 (2H, d, J = 8.6 Hz, H-2′, 6′), 7.47 (2H, d, J = 8.6 Hz, H-3′, 5′); 13 13C NMR (100 MHz, deuterated methanol) δ C: 181.1 (C-28), 136.2 (C-13), 127.1 (C-12), 78.9 (C-3), 74.0 (C-19), 32.3 (C-30), 71.4 (C-23), 67.3 (C-2), 51.2 (C-20), 51.2 (C-18), 48.8 (C-9), 45.1 (C-1), 44.7 (C-5), 43.2 (C-14), 42.6 (C-17), 41.9 (C-8), (C-14), 25.3 (C-15), 26.3 (C-16), 40.9 (C-17), 50.4 (C-18), 73.1 (C-19), 87.4 (C-20), 41.2 (C-4), 39.6 (C-10), 34.0 (C-7), 26.8 (C-21), 26.0 (C-22), 51.2 (C-26), 23.6 (C-26), 23.6 (C-27), 26.0 (C-29), 25.8 (C-16), 25.3 (C-15), 19.4 (C-6), 17.6 (C-24), 17.5 (C-25), 166.6 (C-1′), 117.8 (C-2′), 144.3 (C-3′), 127.5 (C-4′), 132.2 (C-5′), 115 (C-6′), 156.7 (C-7′), 115.0 (C-8′), 132.3 (C-9′).

[0102] Compound 24 (xiongterpene): white powder; optical rotation value (c 0.1, methanol); ESIMS m / z: 602.3 [M+Na] + , molecular formula is C 39 H 54 O5, and the hydrogen deficiency index is 13. 1 1H NMR (400 MHz, chloroform-d) δ H: 0.80 (3H, s, H3-23), 0.72 (3H, s, H3-24), 0.86 (3H, s, H3-25), 0.91 (3H, s, H3-26), 0.88 (1H, s, H-27), 1.11 (1H, s, H-28), 0.90 (1H, s, H-29), 4.52 (1H, dd, J = 11.0, 5.2 Hz, H-3α), 2.80 (1H, dd, J = 13.8, 4.5 Hz, H-18), 5.26 (1H, t, J = 3.5 Hz, H-12), 5.18 (1H, d, J = 12.7 Hz, H-8′), 6.82 (1H, d, J = 12.7 Hz, H-7′), 6.75 (2H, d, J = 8.6 Hz, H-2′, 6′), 7.57 (2H, d, J = 8.6 Hz, H-3′, 5′); 13 13C NMR (100 MHz, chloroform-d) δ C : 28.5 (C-28), 145.7 (C-13), 121.9 (C-12), 81.0 (C-3), 46.9 (C-19), 27.8 (C-23), 47.5 (C-18), 55.7 (C-5), 32.8 (C-17), 47.9 (C-9), 41.4 (C-8), 31.4 (C-20), 40.0 (C-14), 38.1 (C-4), 38.1 (C-1), 37.9 (C-22), 36.4 (C-10), 32.8 (C-7), 26.5 (C-15), 33.9 (C-21), 23.6 (C-2), 33.7 (C-29), 27.5 (C-16), 25.6 (C-27), 23.2 (C-11), 18.4 (C-6), 16.3 (C-26), 15.9 (C-25), 23.1 (C-30), 14.9 (C-24), 166.6 (C-1′), 117.8 (C-2′), 144.3 (C-3′), 127.5 (C-4′), 132.2 (C-5′), 115 (C-6′), 156.7 (C-7′), 115.0 (C-8′), 132.3 (C-9′).

[0103] Compound 25 (2α,23-dihydroxy-3β-cis-p-coumaroyloxy-olean-12-en-28-oic acid): white powder; optical rotation (c 0.1, methanol); ESIMS m / z: 613.3 [M+Na] + , molecular formula C 39 H 54 O5, degree of unsaturation is 13. 11H NMR (400 MHz, chloroform-d) δ H : δ H : 0.86 (3H, s, H3-26), 1.12 (3H, s, H3-25), 1.12 (3H, s, H3-24), 1.17 (3H, s, H3-27), 1.64 (3H, s, H3-29), 2.56 (1H, s, H-18), 2.64 (1H, dt, J = 13.0, 12.4, 6.8 Hz, H-16a), 2.73 (1H, m, H-27a), 3.76 (1H, d, J = 10.5 Hz, H-23a), 4.26 (1H, m, H-3), 4.27 (1H, m, H-23b), 4.31 (1H, m, H-2), 6.19 (1H, KBr s, H-12); 13 13C NMR (100 MHz, pyridine-d5) δ C : 47.3 (C-1), 68.8 (C-2), 78.0 (C-3), 43.6 (C-4), 48.0 (C-5), 18.6 (C-6), 32.9 (C-7), 40.2 (C-8), 47.8 (C-9), 38.5 (C-10), 24.4 (C-11), 125.8 (C-12), 135.6 (C-13), 42.0 (C-14), 25.3 (C-15), 26.3 (C-16), 40.9 (C-17), 50.4 (C-18), 73.1 (C-19), 87.4 (C-20), 25.8 (C-21), 25.9 (C-22), 66.2 (C-23), 14.2 (C-24), 17.4 (C-25), 16.1 (C-26), 23.4 (C-27), 178.8 (C-28), 25.8 (C-29), 63.1 (C-30).

[0104] Compound 26 (23Z-coumaroyl hederagenin): white powder; optical rotation (c 0.1, methanol); ESIMS m / z 472.4 [M+Na] + , with the molecular formula C 30 H 48 O4 and a hydrogen deficiency index of 7. 1 1H NMR (400 MHz, methanol-d4) δ H: 5.48 (1H, t, J = 3.3 Hz, H-12), 3.96 (1H, d, J = 10.9 Hz, H-23a), 4.17 (1H, d, J = 10.9 Hz, H-23b), 1.16 (3H, s, H3-24), 1.27 (3H, s, H3-25), 1.10 (3H, s, H3-26), 1.26 (3H, s, H3-27), 1.23 (3H, s, H3-29), 3.62 (1H, dd, J = 10.9, 4.0 Hz, H-3), 5.18 (1H, d, J = 12.7 Hz, H-8′), 6.82 (1H, d, J = 12.7 Hz, H-7′), 6.75 (2H, d, J = 8.6 Hz, H-2′, 6′), 7.57 (2H, d, J = 8.6 Hz, H-3′, 5′); 13 13C NMR (100 MHz, methanol-d4) δ C : 182.5 (C-28), 140.0 (C-13), 129.2 (C-12), 78.2 (C-3), 73.3 (C-19), 33.0 (C-30), 70.9 (C-23), 26.5 (C-2), 42.9 (C-20), 55.8 (C-18), 47.9 (C-9), 34.2 (C-1), 55.9 (C-5), 42.5 (C-14), 47.9 (C-17), 40.9 (C-8), 42.2 (C-14), 28.9 (C-15), 25.9 (C-16), 48.1 (C-17), 39.4 (C-19), 42.9 (C-20), 43.9 (C-4), 37.2 (C-10), 34.9 (C-7), 26.9 (C-21), 38.9 (C-22), 16.6 (C-26), 24.9 (C-27), 27.0 (C-29), 25.8 (C-16), 29.9 (C-15), 19.9 (C-6), 14.4 (C-24), 17.5 (C-25), 166.6 (C-1′), 117.8 (C-2′), 144.3 (C-3′), 127.5 (C-4′), 132.2 (C-5′), 115 (C-6′), 156.7 (C-7′), 115.0 (C-8′), 132.3 (C-9′).

[0105] Compound 27 (3β,28-dihydroxy-olean-18-ene): colorless crystal; optical rotation (c 0.1, methanol); ESIMS m / z: 442.4 [M+Na] + , with the molecular formula C 30 H 50 O2 and a hydrogen deficiency index of 6.1 H NMR (400 MHz, chloroform-d) δ H : 0.94 (3H, s, H3-23), 0.75 (3H, s, H3-24), 0.85 (3H, s, H3-25), 1.02 (3H, s, H3-26), 0.74 (1H, s, H-27), 3.61 (1H, d, J = 10.8 Hz, H-28a), 3.44 (1H, d, J = 10.8 Hz, H-28b), 3.18 (1H, dd, J = 11.2, 5.1 Hz, H-3α), 0.66 (1H, dd, J = 11.0, 1.1 Hz, H-5); 13 C NMR (100 MHz, chloroform-d) δ C : 65.7 (C-28), 38.8 (C-13), 26.4 (C-12), 79.2 (C-3), 134.7 (C-19), 28.2 (C-23), 138.8 (C-18), 55.7 (C-5), 39.8 (C-17), 51.4 (C-9), 43.4 (C-8), 32.4 (C-20), 41.0 (C-14), 39.1 (C-4), 39.1 (C-1), 31.9 (C-22), 37.4 (C-10), 34.8 (C-7), 27.5 (C-15), 33.5 (C-21), 27.6 (C-2), 30.7 (C-29), 31.5 (C-16), 15.6 (C-27), 21.2 (C-11), 18.4 (C-6), 16.3 (C-26), 16.9 (C-25), 30.1 (C-30), 14.9 (C-24).

[0106] Compound 28 (23,24-dihydroxy-3-oxo-olean-12-en-28-oic acid): white powder; optical rotation value (c 0.1, methanol); ESIMS m / z: 486.3 [M+Na] + ,with the molecular formula C 30 H 46 O5 and the hydrogen deficiency index of 8. 1 HNMR (400 MHz, chloroform-d) δ H: 4.08 (1H, d, J = 11.1 Hz, H-23a), 3.44 (1H, d, J = 11.1 Hz, H-23b), 3.90 (1H, d, J = 11.7 Hz, H-24a), 3.57 (1H, d, J = 11.7 Hz, H-24b), 1.14 (3H, s, H3-25), 0.88 (3H, s, H3-26), 1.19 (1H, s, H-27), 0.91 (1H, s, H-29), 0.95 (1H, s, H-30), 2.88 (1H, dd, J = 13.7, 4.1 Hz, H-18), 5.28 (1H, dd, J = 3.7, 3.7 Hz, H-12); 13 13C NMR (100 MHz, chloroform-d) δ C : 181.9 (C-28), 145.5 (C-13), 123.6 (C-12), 217.3 (C-3), 47.4 (C-19), 64.2 (C-23), 133.8 (C-18), 49.9 (C-5), 47.8 (C-17), 48.4 (C-9), 40.4 (C-8), 31.4 (C-20), 43.0 (C-14), 59.1 (C-4), 39.1 (C-1), 33.9 (C-22), 37.4 (C-10), 34.8 (C-7), 28.5 (C-15), 35.5 (C-21), 37.6 (C-2), 33.7 (C-29), 24.5 (C-16), 26.6 (C-27), 25.2 (C-11), 20.4 (C-6), 17.3 (C-26), 15.9 (C-25), 24.1 (C-30), 63.9 (C-24).

[0107] Compound 29 (oleanderolide): white powder; optical rotation (c 0.1, methanol); ESIMS m / z: 495.4 [M+Na] + , molecular formula C 30 H 48 O3, degree of unsaturation is 7. 1 1H NMR (400 MHz, chloroform-d) δ H : 0.96 (3H, s, H3-23), 0.75 (3H, s, H3-24), 0.88 (3H, s, H3-25), 1.12 (3H, s, H3-26), 1.28 (3H, s, H3-27), 0.96 (3H, s, H3-29), 0.88 (3H, s, H3-30), 3.20 (1H, dd, J = 11.4, 4.9 Hz, H-3α); 13 13C NMR (100 MHz, chloroform-d) δC : 180.2 (C-28), 90.8 (C-13), 76.6 (C-12), 79.1 (C-3), 39.6 (C-19), 28.2 (C-23), 51.3 (C-18), 55.4 (C-5), 44.8 (C-17), 44.8 (C-9), 42.3 (C-8), 31.8 (C-20), 42.3 (C-14), 39.1 (C-4), 39.1 (C-1), 27.4 (C-22), 36.6 (C-10), 34.2 (C-7), 28.2 (C-15), 34.2 (C-21), 27.4 (C-2), 33.5 (C-29), 21.4 (C-16), 18.8 (C-27), 29.0 (C-11), 17.9 (C-6), 18.7 (C-26), 16.5 (C-25), 24.1 (C-30), 15.6 (C-24).

[0108] Compound 30 (11-oxo-β-amyrin): white powder; optical rotation value (c 0.1, methanol); ESIMS m / z: 463.4 [M+Na] + , with the molecular formula C 30 H 48 O2, and the hydrogen deficiency index is 7. 1 1H NMR (400 MHz, chloroform-d) δ H : 0.86 (3H, s, H3-23), 0.87 (3H, s, H3-24), 1.10 (3H, s, H3-25), 1.11 (3H, s, H3-26), 1.33 (3H, s, H3-27), 0.97 (3H, s, H3-28), 0.78 (3H, s, H3-29), 0.83 (3H, s, H3-30), 2.07 (1H, td, J = 13.6, 4.7 Hz, H-16b), 3.20 (1H, dd, J = 11.0, 5.4 Hz, H-3α), 5.56 (1H, s, H-12), 0.69 (1H, dd, J = 12.1, 1.1 Hz, H-5), 2.30 (1H, s, H-9); 13 13C NMR (100 MHz, chloroform-d) δ C: 33.3 (C-28), 128.3 (C-13), 170.8 (C-12), 79.0 (C-3), 45.4 (C-19), 28.3 (C-23), 47.8 (C-18), 55.2 (C-5), 32.6 (C-17), 62.0 (C-9), 43.6 (C-8), 31.2 (C-20), 45.6 (C-14), 37.3 (C-4), 39.3 (C-1), 36.7 (C-22), 39.3 (C-10), 33.0 (C-7), 26.7 (C-15), 34.7 (C-21), 27.5 (C-2), 33.3 (C-29), 26.6 (C-16), 23.6 (C-27), 200.5 (C-11), 17.7 (C-6), 18.9 (C-26), 16.6 (C-25), 23.7 (C-30), 15.8 (C-24).

[0109] Compound 31 (11-oxo-erythrodiol): white powder; optical rotation value (c 0.1, methanol); ESIMS m / z: 465.4 [M+Na] + , with the molecular formula C 30 H 48 O3, and the hydrogen deficiency index is 7. 1 1H NMR (400 MHz, chloroform-d) δ H : 0.97 (3H, s, H3-23), 0.98 (3H, s, H3-24), 1.11 (3H, s, H3-25), 1.11 (3H, s, H3-26), 3.44 (1H, d, J = 12.5 Hz, H-28a), 3.62 (1H, d, J = 12.5 Hz, H-28b), 3.22 (1H, dd, J = 11.5, 4.8 Hz, H-3α), 5.54 (1H, s, H-12), 0.72 (1H, dd, J = 11.8, 1.9 Hz, H-5), 2.28 (1H, s, H-9); 13 13C NMR (100 MHz, chloroform-d) δ C: 69.8 (C-28), 169.8 (C-13), 128.4 (C-12), 79.0 (C-3), 45.1 (C-19), 15.7 (C-23), 42.9 (C-18), 55.1 (C-5), 37.3 (C-17), 62.0 (C-9), 45.6 (C-8), 31.1 (C-20), 43.6 (C-14), 39.3 (C-4), 39.3 (C-1), 30.8 (C-22), 37.2 (C-10), 32.9 (C-7), 26.0 (C-15), 33.1 (C-21), 27.2 (C-2), 33.1 (C-29), 21.7 (C-16), 23.6 (C-27), 200.1 (C-11), 17.7 (C-6), 18.8 (C-26), 16.6 (C-25), 23.6 (C-30), 28.3 (C-24).

[0110] Compound 32 (3β-hydroxy-11α-methoxy-olean-12-ene): white powder; optical rotation value (c 0.1, methanol); ESIMS m / z: 465.4 [M+Na] + , with the molecular formula C 31 H 52 O2, and the hydrogen deficiency index is 6. 1 1H NMR (400 MHz, chloroform-d) δ H : 0.77 (3H, s, H3-23), 0.85 (3H, s, H3-24), 0.81 (3H, s, H3-25), 0.97 (3H, s, H3-26), 0.86 (3H, s, H-27), 0.98 (3H, s, H-28), 1.01 (3H, s, H-29), 1.18 (3H, s, H-30), 1.90 (1H, dt, J = 13.8, 3.5 Hz, H-16b), 3.20 (1H, dd, J = 10.3, 6.0 Hz, H-3α), 3.25 (3H, s, -OCH3), 3.77 (1H, dd, J = 8.9, 3.1 Hz, H-11); 13 13C NMR (100 MHz, chloroform-d) δ C: 28.7 (C-28), 149.4 (C-13), 121.6 (C-12), 78.2 (C-3), 46.7 (C-19), 28.3 (C-23), 47.8 (C-18), 55.4 (C-5), 32.1 (C-17), 52.0 (C-9), 42.0 (C-8), 31.1 (C-20), 43.9 (C-14), 39.0 (C-4), 39.8 (C-1), 38.2 (C-22), 38.1 (C-10), 33.5 (C-7), 26.7 (C-15), 34.3 (C-21), 33.4 (C-2), 33.4 (C-29), 27.5 (C-16), 26.1 (C-27), 76.4 (C-11), 18.5 (C-6), 18.5 (C-26), 17.1 (C-25), 23.2 (C-30), 15.7 (C-24).

[0111] Compound 33 (alphitolic acid): White powder; optical rotation value (c 0.1, methanol); ESIMS m / z 541.3 [M+Na] + , with the molecular formula C 30 H 46 O7, and the hydrogen deficiency index is 7. 1 1H NMR (400 MHz, deuterated methanol) δ H : 5.42 (1H, t, J = 4.0 Hz, H-12), 1.11 (1H, s, H-23), 0.80 (3H, s, H3-24), 1.02 (3H, s, H3-25), 0.85 (3H, s, H3-26), 1.20 (3H, s, H3-27), 1.39 (3H, s, H3-29), 3.76 (1H, s H-30), 2.91 (1H, d, J = 9.6 Hz, H-3α), 3.62 (1H, ddd, J = 11.4, 9.6, 4.6 Hz, H-2β).

[0112] Compound 34 (3β,30-dihydroxy-lup-20(29)-en-28-oic acid): White powder; optical rotation value (c 0.1, methanol); ESIMS m / z: 464.4 [M+Na] + , with the molecular formula C 30 H 44 O2, and the hydrogen deficiency index is 7. 1 1H NMR (400 MHz, deuterated chloroform) δ H: 4.96 (1H, s, H-28a), 4.92 (1H, s, H-28b), 3.18 (1H, dd, J = 11.5, 5.1 Hz, H-3), 4.12 (3H, s, H3-29), 0.71 (3H, s, H3-27), 0.95 (3H, s, H3-24), 0.81 (3H, s, H3-26), 0.90 (3H, s, H3-25), 0.96 (3H, s, H3-23); 13 13C NMR (100 MHz, chloroform-d) δ C : 149.9 (C-20), 138.9 (C-17), 120.0 (C-22), 107.6 (C-28), 78.9 (C-3), 54.4 (C-5), 53.5 (C-19), 51.2 (C-9), 49.8 (C-18), 47.8 (C-13), 41.9 (C-14), 40.9 (C-8), 39.5 (C-4), 39.1 (C-1), 37.8 (C-21), 36.5 (C-10), 34.4 (C-21), 32.1 (C-15), 28.2 (C-24), 27.6 (C-2), 26.6 (C-12), 25.9 (C-16), 21.3 (C-11), 68.5 (C-29), 18.5 (C-6), 15.9 (C-25), 15.4 (C-26), 15.5 (C-23), 13.7 (C-27).

[0113] Compound 35 (betulin): white powder; optical rotation value (c 0.1, methanol); ESIMS m / z: 465.4 [M + Na] + , molecular formula C 30 H 50 O2, degree of unsaturation is 6. 1 1H NMR (400 MHz, chloroform-d) δ H : 0.94 (3H, s, H3-23), 0.73 (3H, s, H3-24), 0.80 (3H, s, H3-25), 0.99 (3H, s, H3-26), 3.78 (1H, d, J = 10.8 Hz, H-28a), 3.31 (1H, d, J = 10.8 Hz, H-28b), 4.55 (1H, s, H-29a), 4.65 (1H, s, H-29b), 1.65 (3H, s, H3-30), 3.17 (1H, dd, J = 11.4, 4.8 Hz, H-3α), 0.66 (1H, d, J = 9.6 Hz, H-5); 13 13C NMR (100 MHz, chloroform-d) δ C: 60.8 (C-28), 37.5 (C-13), 25.4 (C-12), 79.2 (C-3), 48.0 (C-19), 28.2 (C-23), 48.9 (C-18), 55.5 (C-5), 48.0 (C-17), 50.6 (C-9), 41.1 (C-8), 150.7 (C-20), 42.9 (C-14), 39.1 (C-4), 38.9 (C-1), 34.4 (C-22), 37.4 (C-10), 34.2 (C-7), 27.6 (C-15), 29.9 (C-21), 27.2 (C-2), 109.9 (C-29), 29.4 (C-16), 14.9 (C-27), 21.1 (C-11), 18.5 (C-6), 16.2 (C-26), 16.3 (C-25), 19.3 (C-30), 15.5 (C-24).

[0114] Compound 36 (3β-hydroxy-28-nor-lup-17(22),20(28)-diene): white powder; optical rotation value (c 0.1, methanol); ESIMS m / z: 433.4 [M+Na] + , molecular formula is C 29 H 46 O, hydrogen deficiency index is 7. 1 1H NMR (400 MHz, chloroform-d) δ H : 5.13 (1H, s, H-22), 4.71 (1H, s, H-28a), 4.60 (1H, s, H-28b), 3.18 (1H, dd, J = 11.2, 5.1 Hz, H-3), 1.66 (3H, s, H3-29), 0.96 (3H, s, H3-27), 0.95 (3H, s, H3-24), 0.92 (3H, s, H3-26), 0.81 (3H, s, H3-25), 0.73 (3H, s, H3-23); 13 13C NMR (100 MHz, chloroform-d) δ C: 150.3 (C-20), 145.6 (C-17), 119.1 (C-22), 109.6 (C-28), 79.2 (C-3), 55.7 (C-5), 53.5 (C-19), 51.2 (C-9), 49.8 (C-18), 47.8 (C-13), 41.9 (C-14), 40.9 (C-8), 39.1 (C-4), 39.1 (C-1), 38.5 (C-21), 37.5 (C-10), 34.4 (C-21), 32.1 (C-15), 28.2 (C-24), 27.6 (C-2), 26.6 (C-12), 25.3 (C-16), 21.3 (C-11), 20.2 (C-29), 18.5 (C-6), 16.7 (C-25), 15.9 (C-26), 15.6 (C-23), 14.1 (C-27).

[0115] Compound 37 (jughopenoid C): white powder; optical rotation value (c 0.1, methanol); ESIMS m / z: 430.4 [M+Na] + , molecular formula is C 28 H 46 O3, hydrogen deficiency index is 6. 1 1H NMR (400 MHz, chloroform-d) δ H : 3.19 (1H, dd, J = 11.3, 5.0 Hz, H-3), 2.21 (3H, s, H3-30), 0.94 (3H, s, H3-27), 0.75 (3H, s, H3-24), 0.94 (3H, s, H3-26), 0.83 (3H, s, H3-25), 0.96 (3H, s, H3-23), 1.66 (1H, d, J = 10.9 Hz, H-18); 13 13C NMR (100 MHz, chloroform-d) δ C : 214.7 (C-20), 80.9 (C-17), 36.9 (C-22), 78.9 (C-3), 55.5 (C-5), 56.0 (C-19), 51.2 (C-9), 52.4 (C-18), 41.9 (C-13), 41.4 (C-14), 40.9 (C-8), 39.1 (C-4), 39.1 (C-1), 25.4 (C-21), 37.5 (C-10), 28.2 (C-15), 15.6 (C-24), 27.6 (C-2), 26.2 (C-12), 30.7 (C-16), 21.4 (C-11), 18.5 (C-6), 16.7 (C-25), 15.9 (C-26), 28.1 (C-23), 14.1 (C-27).

[0116] Example 3

[0117] The inhibitory effect of Compounds 1-37 on α-glucosidase was determined by the PNPG method. The results are shown in Table 4 and Figure 4 as follows.

[0118] Table 4 Inhibitory Activity of Compounds 1-37 on α-Glucosidase

[0119]

[0120] a IC 50 values are expressed as mean ± deviation, n = 3; b positive drug acarbose.

[0121] Conclusion: Compounds 1-37 all showed varying degrees of α-glucosidase inhibitory activity. Among them, Compounds 1, 2, 4, 9-12, 23 and 26 had significant inhibitory effects on α-glucosidase, and the IC 50 values ranged from 31.99 ± 1.15 to 189.70 ± 2.08 μM, which was about 1 to 7 times the efficacy of the positive drug acarbose.

[0122] It is easy for those skilled in the art to understand that the above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, and improvements made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A triterpenoid compound, characterized in that, The structural formula of the triterpenoid compound is as follows: R1-R4 in Formula I, R1-R6 in Formula II, R1-R6 in Formula III, R1-R8 in Formula IV, R1-R4 in Formula V, R1-R3 in Formula VI, R1-R4 in Formula VII, R1-R7 in Formula VIII, R1-R4 in Formula IX, R1-R4 in Formula X, R1-R6 in Formula XI, and R1-R4 in Formula XII are each independently selected from hydrogen, hydroxyl, methylene, oxygen-containing methylene, methyl, ethyl, allyl, carbonyl, halogen, acyl, acyl halide, nitro, cyano, glucosyl, arabinopyranosyl, rhamnosyl, cis-feruloyloxy, trans-feruloyloxy, cis-p-coumaroyloxy, trans-p-coumaroyloxy, benzene ring, and indole ring.

2. The triterpenoid compound according to claim 1, wherein Formula I is Compound 1 or Compound 2, Formula II is Compound 3, Compound 4, Compound 5, or Compound 6, Formula III is Compound 7, Formula IV is one of Compound 8-10, Compound 13-17, Compound 20-22, Formula V is Compound 18, Formula VI is Compound 19, Formula VII is Compound 11 or Compound 12, Formula VIII is one of Compound 23-26, Compound 28, Compound 30-32, Formula IX is Compound 27, Formula X is Compound 29, Formula XI is one of Compound 33-35, Compound 37, and Formula XII is Compound 36; 3. The preparation method of the triterpenoid compound according to claim 2, characterized in that, It includes the following steps: (1) Dry and powder the leaves of (2) Suspend the total extract obtained in step (1) in water, extract successively with petroleum ether and chloroform, and concentrate under reduced pressure to obtain an extraction extract; (3) Mix the extraction extract obtained in step (2) with silica gel, perform normal-phase silica gel column chromatography, the eluent is a mixed solution of dichloromethane and methanol, gradient elution, combine TLC spotting detection, and combine to obtain 7 components A1–A7 with increasing polarity; When component A3 is separated by ODS column chromatography to obtain 15 components A3a–A3o with decreasing polarity; Component A3h is separated by Sephadex LH-20 column chromatography and silica gel column chromatography to obtain Compound 17 and Compound 25 with increasing polarity; When component A3i is separated by Sephadex LH-20 gel column chromatography and silica gel column chromatography to obtain Compound 33 and 5 components A3i1–A3i5 with increasing polarity; Component A3i4 is further purified by semi-preparative HPLC to obtain Compound 15 and Compound 16 with increasing retention time; When component A3j is separated by Sephadex LH-20 gel column chromatography and silica gel column chromatography to obtain Compound 19; When component A3k is separated by Sephadex LH-20 gel column chromatography and silica gel column chromatography to obtain Compound 21 and Compound 27 with increasing retention time; When component A3n was separated by Sephadex LH-20 gel column chromatography and silica gel column chromatography to obtain 7 components A3n1–A3n7 with increasing polarity from small to large, component A3n2 was purified by semi-preparative HPLC to obtain compound 28; When component A3n5 was purified by semi-preparative HPLC to obtain compound 37; When component A4 passed through a reversed-phase C 18 silica gel column chromatography, the eluent was a mixed solution of methanol and water, and gradient elution was carried out to obtain 10 components A4a–A4j with decreasing polarity; component A4b was purified by Sephadex LH-20 gel column chromatography, silica gel column chromatography, and finally by semi-preparative HPLC to obtain compounds 31, 20, and 34 with increasing retention time; When component A4c was subjected to Sephadex LH-20 gel column chromatography, component A4c3 was obtained; Component A4c3 was separated by semi-preparative high performance liquid chromatography C 18 column chromatography to obtain Compounds 5, 6 and 7 with increasing retention times; When component A4d was subjected to Sephadex LH-20 gel column chromatography, 4 components A4d1–A4d4 with decreasing molecular weight from large to small were obtained; Component A4d4 was separated by semi-preparative high performance liquid chromatography phenyl column to obtain compound 9 and compound 10 with increasing retention time from short to long; When component A4h was obtained by Sephadex LH-20 gel column chromatography and silica gel column chromatography, compound 26 and compound 13 with increasing polarity from small to large were obtained; When component A5 passes through a reversed-phase C 18 silica gel column chromatography with gradient elution, 17 components A5a–A5q with decreasing polarity are obtained; when component A5h is separated by Sephadex LH-20 gel column chromatography and silica gel column chromatography, compounds 35 and 5 components A5h1–A5h5 with increasing polarity are obtained; When component A5h3 was separated by semi-preparative HPLC to obtain 4 components A5h3a–A5h3d with decreasing polarity from large to small, component A5h3b was purified by semi-preparative HPLC to obtain compound 22 and compound 23 with increasing retention time from short to long, and component A5h3c was purified by semi-preparative HPLC to obtain compound 30 and compound 36 with increasing retention time from short to long; When component A5j was subjected to Sephadex LH-20 gel column chromatography and reverse-phase C 18 silica gel column chromatography, component A5j2 was obtained; component A5j2 was separated by semi-preparative high performance liquid chromatography C 18 column to obtain compounds 3 and 4 with increasing retention times; When component A5k was separated by Sephadex LH-20 gel column chromatography and silica gel column chromatography, compound 18, compound 24, compound 14 and 7 components A5k1–A5k7 with increasing polarity from small to large were obtained; When component A5k7 was purified by semi-preparative HPLC to obtain compound 29 and compound 32 with increasing retention time from short to long; When component A5m was subjected to Sephadex LH-20 gel column chromatography and reverse-phase C 18 silica gel column chromatography, component A5m2 was obtained; When component A 5m2 was separated by semi-preparative high performance liquid chromatography C 18 column, compounds 11 and 12 with retention times from short to long were obtained; When component A5o was further subjected to Sephadex LH-20 gel column chromatography and reverse-phase C 18 silica gel column chromatography, component A5o3 was obtained; component A5o3 was subjected to reverse-phase phenyl silica gel column chromatography to obtain four components A5o3a–A5o3d with decreasing polarity; component A5o3b was separated by semi-preparative high-performance liquid chromatography C 18 column to obtain compound 8; When component A5p was further subjected to Sephadex LH-20 gel column chromatography and reverse-phase C 18 silica gel column chromatography, component A5p3 was obtained; component A5p3 was separated by semi-preparative high performance liquid chromatography C 18 column to obtain compounds 1 and 2 with increasing retention times.

4. Use of the triterpenoid compound according to claim 1 or 2, characterized in that, Application of the triterpenoid compound in the preparation of antidiabetic drugs.

5. The application according to claim 4, characterized in that The triterpenoid compound is used to inhibit α-glucosidase activity.

6. The application according to claim 4 or 5, characterized in that, The antidiabetic drug is a drug for treating diabetes.

7. The application according to claim 6, wherein The diabetes is type 2 diabetes.

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