Use of a class of pentacyclic triterpenoids in the treatment of neurodegenerative diseases

Abstract

The application discloses functions and uses of a kind of pentacyclic triterpenoid compounds in preventing and / or treating neurodegenerative diseases, and the compounds are obtained by isolating and extracting from Cynomorium songaricum Rupr medicinal materials.The inventors found that the compounds have significant protective effects on glutamic acid-induced nerve cell injury and oxygen-glucose deprivation-induced nerve cell injury during pharmacological activity evaluation of the compounds, and thus can be used for treating and preventing neurodegenerative diseases such as stroke, brain injury, amyotrophic lateral sclerosis, Huntington's disease, Parkinson's disease and Alzheimer's disease, and can be developed into drugs.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically the use of a class of pentacyclic triterpenoid compounds in the preparation of medicaments for the prevention and / or treatment of neurodegenerative diseases. Background Technology

[0002] Neurodegenerative diseases, characterized by the massive loss of specific neurons, are a group of progressive, severe, and potentially fatal complex illnesses. They can be divided into acute and chronic neurodegenerative diseases. Acute neurodegenerative diseases mainly include stroke and brain injury; chronic neurodegenerative diseases mainly include amyotrophic lateral sclerosis (ALS), Huntington's disease (HD), Parkinson's disease (PD), and Alzheimer's disease (AD). With an aging population, neurodegenerative diseases have become a serious medical problem in modern society. The pathogenic factors of neurodegenerative diseases are diverse and their pathogenesis is complex; therefore, there are many types of drugs used to treat them. Among these, neuroprotective drugs, including antioxidants, glutamate receptor antagonists, calcium channel blockers, anti-inflammatory drugs, and anti-apoptotic drugs, are an important treatment approach.

[0003] Traditional Chinese medicine believes that the kidneys are connected to the brain, and tonifying the kidneys can prevent and treat diseases of the central nervous system. Numerous modern medical studies also suggest that tonifying the kidneys is a very promising approach to treating neurodegenerative diseases.

[0004] The traditional Chinese medicine *Cynomorium songaricum* Rupr., a plant in the Cynomoraceae family, is known as Suoyang. It is used to tonify kidney yang, replenish essence and blood, and moisten the intestines to relieve constipation. It is indicated for kidney yang deficiency, deficiency of essence and blood, soreness and weakness of the lower back and knees, impotence, premature ejaculation, and constipation. As a commonly used tonic, Suoyang is mainly distributed in arid regions of Xinjiang, Qinghai, Gansu, Ningxia, Inner Mongolia, and Shaanxi, and is known as "desert ginseng." As an important kidney-yang tonifying herb in traditional Chinese medicine, recent years have seen significant progress in the pharmacological activities of Suoyang extract, including anti-aging, neuroprotective, and antioxidant effects. In particular, studies have shown that the ethyl acetate extract of Suoyang (Zheng Lingyan & Han Ruilan, 2016) can significantly improve learning, memory, and memory retention in D-galactose-induced and scopolamine-induced aged mice. However, research on the active components of Suoyang is very limited, and there are currently no reports on neuroprotective active components in Suoyang.

[0005] Based on previous research on the chemical components of Cynomorium songaricum, the inventors discovered that most of the pentacyclic triterpenoids in Cynomorium songaricum, including the unique malonate type triterpenoids such as compound 16 and p-hydroxycinnamate type triterpenoids such as compounds 19-23, have significant neuroprotective activity and therefore have good application prospects in the treatment of neurodegenerative diseases. Summary of the Invention

[0006] The technical problem to be solved by the present invention is to provide a class of pentacyclic triterpenoid compounds, the use of pharmaceutical compositions containing the same in the preparation of products for the prevention and / or treatment of neurodegenerative diseases, and further to provide a method for preparing such compounds.

[0007] To solve the technical problem of this invention, the present invention adopts the following technical solution:

[0008] The first aspect of this invention provides the use of a class of pentacyclic triterpenoid compounds derived from Cynomorium songaricum in the preparation of products for the prevention and / or treatment of neurodegenerative diseases, the structure of which is as follows:

[0009]

[0010] A second aspect of this invention provides the use of a pharmaceutical composition containing a pentacyclic triterpenoid compound as described in the first aspect in the preparation of a product for the prevention and / or treatment of neurodegenerative diseases, the pharmaceutical composition being prepared according to methods known in the art. The compounds of this invention can be formulated into any dosage form suitable for human or animal use by combining them with one or more pharmaceutically acceptable solid or liquid excipients and / or adjuvants. The content of the compounds of this invention in its pharmaceutical composition is typically 0.1-95% by weight.

[0011] The compounds of this invention or pharmaceutical compositions containing them can be administered in unit dose form via enteral or non-enteric routes, such as oral, intravenous, intramuscular, subcutaneous, nasal, oral mucosa, eye, lung and respiratory tract, skin, vagina, rectum, etc.

[0012] Dosage forms can be liquid, solid, or semi-solid. Liquid dosage forms can include solutions (including true solutions and colloidal solutions), emulsions (including o / w, w / o, double emulsions, and liposomes), suspensions, injections (including aqueous injections, powder injections, and infusions), eye drops, nasal drops, lotions, and liniments, etc.; solid dosage forms can include tablets (including regular tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, and orally disintegrating tablets), capsules (including hard capsules, soft capsules, and enteric-coated capsules), granules, powders, microcapsules, pellets, suppositories, films, patches, aerosols, and sprays, etc.; semi-solid dosage forms can include ointments, gels, and pastes, etc.

[0013] The compounds of this invention or pharmaceutical compositions containing them can be formulated into conventional formulations, sustained-release formulations, controlled-release formulations, targeted formulations, and various microparticle delivery systems.

[0014] To formulate the compounds of the present invention or pharmaceutical compositions containing them into tablets, a wide variety of excipients known in the art can be used, including diluents, binders, wetting agents, disintegrants, lubricants, and flow aids. Diluents can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, dicalcium phosphate, calcium carbonate, etc.; wetting agents can be water, ethanol, isopropanol, etc.; binders can be starch paste, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, gum arabic paste, gelatin paste, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; disintegrants can be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfonate, etc.; lubricants and flow aids can be talc, silica, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.

[0015] Tablets can also be further processed into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or bilayer and multilayer tablets.

[0016] To formulate the drug delivery unit into capsules, the compound of the present invention or a pharmaceutical composition containing it can be mixed with a diluent and a dispersant, and the mixture can be placed directly into hard or soft capsules. Alternatively, the compound of the present invention or a pharmaceutical composition containing it can be first formed into granules or microspheres with a diluent, a binder, and a disintegrant, and then placed into hard or soft capsules. The diluents, binders, wetting agents, disintegrants, and dispersants used to prepare tablets of the compound of the present invention or a pharmaceutical composition containing it can also be used to prepare capsules of the compound of the present invention or a pharmaceutical composition containing it.

[0017] To prepare an injectable formulation of the compound of this invention or a pharmaceutical composition containing it, water, ethanol, isopropanol, propylene glycol, or mixtures thereof can be used as solvents, and appropriate amounts of commonly used solubilizers, co-solvents, pH adjusters, and osmotic pressure regulators can be added. Solubilizers or co-solvents can be poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc.; pH adjusters can be phosphates, acetates, hydrochloric acid, sodium hydroxide, etc.; osmotic pressure regulators can be sodium chloride, mannitol, glucose, phosphates, acetates, etc. If preparing a lyophilized powder for injection, mannitol, glucose, etc., can also be added as a support agent.

[0018] To formulate the compounds of the present invention or pharmaceutical compositions containing them into liposomes, phospholipids and cholesterol can be used as carrier materials. Phospholipids can be phosphatidylcholine, dipalmitoylphosphatidylcholine, distearylphosphatidylcholine, dipalmitoylethanolamine, dioleoylphosphatidylcholine, dioleoylphosphatidylglycerol, phosphatidylethanolamine, sphingomyelin, phosphatidic acid, phosphatidylinositol, acylserine, phosphatidylglycerol, and octadecylamine.

[0019] In addition, colorants, preservatives, flavorings, tasters, or other additives may be added to the pharmaceutical preparation if necessary. To achieve the intended therapeutic effect, the medicine or pharmaceutical composition of the present invention can be administered using any known method of administration.

[0020] The dosage of the compounds of the present invention or pharmaceutical compositions containing them can vary widely depending on the nature and severity of the disease to be prevented or treated, the individual circumstances of the patient or animal, the route of administration, and the dosage form. Generally, the suitable daily dosage range of the compounds of the present invention is 0.001-150 mg / kg body weight, preferably 0.1-100 mg / kg body weight, more preferably 1-60 mg / kg body weight, and most preferably 2-30 mg / kg body weight. The above dosages can be administered as a single dose unit or divided into several dose units, depending on the physician's clinical experience and the administration regimen, including the use of other treatment methods.

[0021] The compounds or pharmaceutical compositions of the present invention can be taken alone or in combination with other therapeutic or symptomatic drugs. When the compounds of the present invention have a synergistic effect with other therapeutic drugs, their dosage should be adjusted according to the actual situation.

[0022] The aforementioned neurodegenerative diseases include, but are not limited to, stroke, brain injury, amyotrophic lateral sclerosis (ALS), Huntington's disease, Parkinson's disease, and Alzheimer's disease.

[0023] The products described in the first and second aspects of this invention include, but are not limited to, pharmaceuticals.

[0024] The third aspect of this invention is to provide a method for preparing the pentacyclic triterpenoid compounds described in the first and second aspects, comprising the following steps: extracting Cynomorium songaricum with 95% ethanol under reflux, concentrating the extract and then separating and purifying it by organic solvent extraction, silica gel column chromatography, gel column chromatography, reversed-phase silica gel column chromatography and preparative HPLC to obtain the above-mentioned compounds, whose structures are identified by various spectroscopic methods as pentacyclic triterpenoid compounds, mainly of the ursane type and oleanane type.

[0025] The organic solvent extraction method described herein is a well-known method in the art. The extract is suspended in water and can be extracted with ethyl acetate, chloroform, dichloromethane, petroleum ether, or chloroform-methanol (1:1); the number of extractions is 1-3 times, preferably 3 times.

[0026] The macroporous adsorption resin mentioned therein is a non-polar, weakly polar, moderately polar, polar, or strongly polar macroporous adsorption resin known in the art; preferably, a non-polar, weakly polar, or moderately polar macroporous adsorption resin is used.

[0027] The macroporous adsorption resin, in terms of its structure, is a styrene-type, styrene-divinylbenzene-type, cross-linked polystyrene-type, styrene-nitrile-type, methylstyrene-type, methacrylate-type, methacrylic acid-type, acrylic acid-type, acrylamide-type, vinylpyrrolidone-type, sulfoxide-type, phenol-formaldehyde-type, or nitrogen oxide-type macroporous adsorption resin known in the art; preferably, a styrene-type, acrylate-type, or methacrylate-type macroporous adsorption resin is used.

[0028] The specific models of the macroporous adsorption resins mentioned include, but are not limited to, D101, DA201, D301, D3520, D4006, D4020, H103, H107, H30, H60, AB-8, X-5, NKA, Diaion HP-20, Sepabedads SP-700, HP2MGL, HPD100, HPD722, HPD-600, HPD-826, ADS-17, Amberlite XAD-4, XAD-1600, etc.

[0029] The elution solvent for the macroporous adsorption resin is 50%–95% ethanol, eluting for 2–4 column volumes; preferably, 70%–95% ethanol is used for elution.

[0030] The silica gel column chromatography method described herein is well-known in the art. After evaporating the extract to dryness, it is dissolved in chloroform or methanol, an appropriate amount of silica gel is added, mixed well, dried, and then packed onto a silica gel column. Gradient elution is performed using elution solvent systems such as chloroform-methanol, chloroform-acetone, petroleum ether-acetone, dichloromethane-methanol, and dichloromethane-acetone; preferably, chloroform-methanol or dichloromethane-methanol is used as the elution solvent.

[0031] The gel column chromatography described herein can use gels of type Sephdex LH-20, HW-40, etc., and methanol, chloroform-methanol (2:1), or petroleum ether-chloroform-methanol (5:5:1) as elution solvents.

[0032] The reversed-phase silica gel column chromatography mentioned above typically uses reversed-phase C64S silica gel column chromatography. 18 C8 and other packing materials use methanol-water or acetonitrile-water as the mobile phase; preferably, C8 is used. 18 The packing material is methanol-water, and the mobile phase is methanol-water.

[0033] The preparative HPLC described above typically uses reversed-phase C240 ​​HPLC. 18C8 and other packing materials use methanol-water or acetonitrile-water as the mobile phase. If necessary, 0.1-1% of formic acid, acetic acid, trifluoroacetic acid, etc., can be added to the mobile phase.

[0034] Beneficial technical effects:

[0035] 1. This invention provides a novel use of a class of pentacyclic triterpenoid compounds in the prevention or treatment of neurodegenerative diseases.

[0036] 2. Experiments have shown that the compounds described in this invention have the following two significant pharmacological activities: First, these compounds can protect against glutamate-induced neuronal damage; second, these compounds can significantly protect against oxygen-glucose deprivation-induced neuronal damage.

[0037] 3. The preparation methods for this type of compound are mature, simple and easy to obtain. Detailed Implementation

[0038] The following examples and pharmacological activity experiments are provided to further illustrate the present invention, but do not imply any limitation thereof. Experimental methods not specifically described in the following examples were performed according to conventional methods and conditions, or as selected in the product instructions.

[0039] In the following embodiments, the full names or corresponding Chinese names of some substances are as follows:

[0040] UV: Ultraviolet-Visible Spectrum

[0041] IR: Infrared Spectrum

[0042] FT-IR: Fourier Transform Infrared Spectroscopy

[0043] ODS: Carbon-18 bonded silica gel

[0044] ESIHRMS: Electrospray High-Resolution Mass Spectrometry

[0045] ESIMS: Electrospray mass spectrometry

[0046] 1 H-NMR: 1H NMR spectrum

[0047] 13 C-NMR: Carbon NMR spectrum

[0048] The room temperature mentioned in the following examples is as is commonly known in the art, generally referring to 15–25°C.

[0049] Example 1. Preparation method of pentacyclic triterpenoids

[0050] 60 kg of dried Cynomorium songaricum was pulverized and extracted using an extraction tank. The extraction was performed three times under reflux with 95% ethanol, each time for 2 hours. After filtration, the supernatant was collected, and the ethanol was recovered under reduced pressure to obtain 8.4 kg of crude extract. The Cynomorium songaricum extract was then dispersed in water and extracted with chloroform. The solvent was recovered from the chloroform fraction to obtain 1.5 kg of extract. The chloroform fraction was mixed with silica gel at a 1:1 ratio and loaded onto a silica gel column. A chloroform-methanol gradient elution (1:0 → 1:1) was used, and the triterpenoid components were detected by TLC. The triterpenoid-enriched fraction was collected. After solvent recovery, compounds 1 (11.5 mg), 2 (1.95 mg), 3 (7.3 mg), 4 (3.42 mg), 5 (7 mg), 6 (2.6 mg), 7 (5.1 mg), 8 (5 g), 9 (1.2 mg), 10 (2.5 mg), 11 (5.51 mg), 12 (4.1 mg), 13 (3.3 mg), 14 (3.0 mg), 15 (2.9 mg), 16 (13.8 mg), 18 (4 mg), 19 (14.1 mg), 20 (3.41 mg), 21 (3.06 mg), 22 (30.8 mg), and 23 (8.1 mg) were purified repeatedly by silica gel column chromatography, ODS reversed-phase silica gel column chromatography, Sephadex LH-20 gel column chromatography, and preparative HPLC to obtain compounds 2 (11.5 mg), 3 (1.95 mg), 4 (7.3 mg), 5 (7 mg), 6 (2.6 mg), 7 (5.1 mg), 8 (5 g), 9 (1.2 mg), 10 (2.5 mg), 11 (5.51 mg), 12 (4.1 mg), 13 (3.3 mg), 14 (3.0 mg), 15 (2.9 mg), 16 (13.8 mg), 18 (4 mg), 19 (14.1 mg), 20 (3.41 mg), 21 (3.06 mg), 22 (30.8 mg), and 23 (8.1 mg). Their structures were identified by various spectroscopic methods and were found to be pentacyclic triterpenoids.

[0051] The spectral information and NMR signal assignments of the above compounds are as follows:

[0052] Compound 1: White powder, +33.1(c 0.07,CH3CN).UV(CH3CN)λ max nm(logε):191(3.95),228(3.86).CD(c 0.07,CH3CN)Δε 209 -3.72,Δε 235 9.88, Δε 343 -1.95.IRν max 3453,2951,1699,1662cm -1 .(-)-ESIHRMS m / z 467.3159[MH] - (calcd 467.3167 for C) 30 H 43 O4-). 1 H-NMR and 13 C-NMR data are shown in Tables 1 and 2.

[0053] Compound 2: white powder, -183.7(c 0.02,CH3CN).CD(c 0.02,CH3CN)Δε 215-2.00.IRν max 3276,2959,1693cm -1 .(-)-ESIHRMS m / z 455.3506[MH] - (calcd 455.3531forC 30 H 47 O3-). 1 H-NMR and 13 C-NMR data are shown in Tables 1 and 2.

[0054] Compound 3: white powder. +28.5(c 0.1,CH3CN).CD(c 0.1,CH3CN)Δε 222 -1.39.IRν max 2925, 1735, 1690cm -1 .(-)-ESIHRMS m / z 555.3682[MH] - (calcd 555.3691forC 34 H 51 O6-). 1 H-NMR (500MHz, CDCl3) δ3.75(3H,s,H-4'),3.38(2H,s,H-2'). 13 C-NMR (125MHz, CDCl3) δ 167.2 (C-3'), 166.3 (C-1'), 52.5 (C-4'), 41.8 (C-2'). Other 1 H-NMR and 13 C-NMR data are shown in Tables 1 and 2.

[0055] Compound 4: white powder. +27.8(c 0.07,CH3CN).CD(c 0.07,CH3CN)Δε 221 -1.22.IRν max 3230, 2924, 1757, 1730, 1692cm -1 .(-)-ESIHRMS m / z 556.3699[MH] - (calcd555.3691for C 34 H 51 O6-). 1 H-NMR (500MHz, CDCl3) δ3.68(3H,s,H-4'),3.31(2H,s,H-2'). 13C-NMR (125MHz, CDCl3) δ 166.2 (C-3'), 165.2 (C-1'), 51.4 (C-4'), 40.6 (C-2'). Other 1 H-NMR and 13 C-NMR data are shown in Tables 1 and 2.

[0056] Table 1. Compounds 1-4 1 H-NMR data (δin ppm)

[0057]

[0058] # Coupling constants were read from the HMQC spectrum.

[0059] Determined at a 500MHz in CDCl3 or b 600 MHz in Pyridine-d5.

[0060] Table 2 1-4 13 C-NMR data (δin ppm)

[0061]

[0062] Determined at a 125 MHz in CDCl3 or b 150 MHz in Pyridine-d5.

[0063] Compound 5: White powder, (-)-ESIMS m / z 467[MH]-. 1 H-NMR (500MHz, CDCl3) δ7.02 (1H, d, J = 10.1 Hz, H-1), 5.81 (1H, d, J = 10.1 Hz, H-2 ),5.42(1H,t,J=3.7Hz,H-12),4.20(1H,dd,J=11.5,5.1Hz,H-15),2.82(1H,dd ,J=13.7,4.0Hz,H-18),1.17(3H,s,H-27),1.16(3H,s,H-23),1.14(3H,s,H-26 ),1.07(3H,s,H-24),0.94(3H,s,H-25),0.92(3H,s,H-30),0.90(3H,s,H-29); 13C-NMR (125MHz, CDCl3) δ205.3(C-3),183.0(C-28),158.8(C-1),144.7(C-13),125.4(C-2),124.0(C-1 2),68.2(C-15),53.2(C-5),47.8(C-14),46.0(C-17),45.1(C-19),44.6(C-4),41.9(C-18),41.8(C-9) ,41.5(C-8),39.6(C-10),35.9(C-7),33.6(C-16),33.2(C-21),33.1(C-29),32.2(C-22),30.8(C-20), 27.9(C-23),23.7(C-11),23.6(C-30),21.7(C-24),20.0(C-27),19.2(C-6),18.8(C-25),18.2(C-26).

[0064] Compound 6: White powder, (-)-ESIMS m / z 513 [MH] - . 1 H-NMR (500MHz, CDCl3) δ5.27 (1H, t, J = 3.7 Hz, H-12), 4.65 (1H, d, J = 3.3 Hz, H-3) ,4.12(1H,dd,J=6.6,3.3Hz,H-2),2.19(1H,d,J=11.4Hz,H-18),2.15(3H,s,H- 2'),1.30(3H,s,H-25),1.08(3H,s,H-23),1.07(3H,s,H-27),0.95(3H,d,J=6. 4Hz,H-30),0.88(3H,s,H-24),0.86(3H,d,J=6.4Hz,H-29),0.81(3H,s,H-26); 13C-NMR (125MHz, CDCl3) δ181.7(C-28),170.5(C-1'),138.0(C-13),125.8(C-12),80.8(C-3),69.4(C-2),55. 4(C-5),52.6(C-18),48.0(C-9),47.9(C-17),43.6(C-1),42.1(C-14),39.6(C-8),39.0(C-20),39.0(C-19), 38.8(C-22),37.5(C-4),36.7(C-10),36.7(C-7),30.6(C-21),29.3(C-24),27.9(C-15),24.1(C-16),23.6(C -11),23.4(C-6),21.2(C-30),21.2(C-2'),18.1(C-27),18.0(C-23),17.1(C-29),17.0(C-26),16.5(C-25).

[0065] Compound 7: White powder, (+)-ESIMS m / z 465 [M+Na] + . 1 H-NMR (500MHz, CDCl3) δ5.14(1H,t,J=3.5Hz,H-12),3.53(1H,d,J=10.8Hz,H-27),3.22(1H,dd,J=11.0,4.6Hz,H-3),3.20(1H,d,J=10.8Hz,H-27),1. 10(3H,s,H-28),1.00(3H,s,H-23),0.99(3H,s,H-24),0.95(3H,s,H-26),0 .94(3H,d,J=6.3Hz,H-29),0.81(3H,d,J=5.8Hz,H-30),0.79(3H,s,H-25); 13C-NMR (150MHz, CDCl3) δ138.7(C-13),125.0(C-12),79.0(C-3),69.9(C-27),55.2(C-5),54.0(C-18) ,47.7(C-9),42.1(C-14),40.0(C-8),39.4(C-19),39.4(C-20),38.8(C-1),38.8(C-17),38.1(C-4),3 6.9(C-10),35.2(C-22),32.8(C-7),30.6(C-21),28.1(C-12),27.3(C-2),26.0(C-16),23.4(C-11),2 3.3(C-12),23.3(C-28),21.3(C-29),18.3(C-6),17.4(C-30),16.8(C-24),15.7(C-26),15.6(C-25).

[0066] Compound 8: White powder, (-)-ESIMS m / z 455 [MH] - . 1 H-NMR (500MHz, Pyridine-d5) δ5.51(1H,t,J=3.3Hz,H-12),3.47(1H,dd,J=11.2,4.5Hz,H-3),2.65(1H,d,J=11.3Hz,H-18),1.26(3H,s,H- 23),1.25(3H,s,H-27),1.07(3H,s,H-26),1.04(3H,s,H-24),1.02(3H,d,J=6.2Hz,H-29),0.97(3H,d,J=5.9Hz,H-30),0.91(3H,s,H-25); 13C-NMR(125MHz,Pyridine-d5)δ179.62(C-28),138.98(C-13),125.4(C-12),77.9(C-3),55.5(C-5),53.3 (C-18),49.4(C-17),47.8(C-9),42.2(C-14),39.7(C-8),39.2(C-19),39.1(C-20),39.1(C-1),38.8(C- 4),37.2(C-22),37.0(C-10),33.3(C-7),30.8(C-21),28.5(C-23),28.4(C-15),27.8(C-2),24.6(C-16) ,23.6(C-27),23.4(C-11),21.1(C-30),18.5(C-6),17.2(C-29),17.2(C-26),16.3(C-24),15.4(C-25).

[0067] Compound 9: White powder, (-)-ESIMS m / z 453 [MH] - . 1 H-NMR(600MHz,Pyridine-d5)δ6.70(1H,dd,J=10.5,2.8Hz,H-11),5.81(1H,d,J=10.5Hz,H-12),3.51(1H,dd,J=11.2,4.6Hz,H-3),1 .27(3H,s,H-23),1.12(3H,s,H-26),1.12(3H,s,H-25),1.05(3H,s,H-27),1.00(3H,s,H-29),0.95(3H,s,H-24),0.93(3H,s,H-30); 13C-NMR(150MHz,Pyridine-d5)δ178.92(C-28),136.58(C-13),133.8(C-18,overlapped with solvent)127.0(C-12),126.0(C-11),78.1(C-3),55.3(C-5),54.9(C-9),48.7(C-17),4 2.5(C-14),41.1(C-8),41.0(C-19),39.6(C-4),38.5(C-1),37.5(C-21),37.1(C-10),36 .3(C-22),33.3(C-15),32.1(C-7),32.8(C-20),32.4(C-29),28.5(C-23),28.1(C-2),2 5.7(C-16),24.4(C-30),20.1(C-27),18.8(C-6),18.4(C-25),17.1(C-26),16.1(C-24).

[0068] Compound 10: White powder, (-)-ESIMS m / z 453 [MH] - . 1 H-NMR (500MHz, CDCl3) δ5.28(1H,t,J=3.3Hz,H-12),3.22(1H,dd,J=11.3,4.3Hz,H-3),2.82(1H,dd,J=13.7,4.0Hz,H-18),1.13 (3H,s,H-27),0.99(3H,s,H-23),0.93(3H,s,H-26),0.91(3H,s,H-24),0.90(3H,s,H-30),0.77(3H,s,H-29),0.75(3H,s,H-25); 13C-NMR (125MHz, CDCl3) δ182.8(C-28),143.6(C-13),122.6(C-12),79.0(C-3),55.2(C-5),47.6(C-9), 46.5(C-17),45.9(C-19),41.6(C-14),41.0(C-18),39.3(C-8),38.7(C-4),38.4(C-1),37.1(C-10),3 3.8(C-21),33.0(C-7),32.6(C-29),32.4(C-22),30.7(C-20),28.1(C-23),27.7(C-15),27.2(C-2),2 5.9(C-27),23.6(C-11),23.4(C-16),23.0(C-30),18.3(C-6),17.1(C-26),15.5(C-24),15.3(C-25).

[0069] Compound 11: White powder, (-)-ESIMS m / z 453 [MH] - . 1 H-NMR(500MHz,Pyridine-d5)δ5.49(1H,brs,H-21),3.50(1H,t,J=8.0Hz,H-3),1.75(3H,s,H-30),1.26(3H,s, H-24),1.14(3H,d,J=6.0Hz,H-29),1.09(3H,s,H-26),1.06(3H,s,H-27),1.05(3H,s,H-23),0.89(3H,s,H-25); 13 C-NMR(150MHz,Pyridine-d5)δ178.5(C-28),143.5(C-20),118.3(C-21),78.5(C-3),56.3(C-5),51.42 (C-9),50.0(C-18),49.7(C-17),42.7(C-14),41.6(C-8),39.9(C-13),39.8(C-1),39.7(C-4),38.8(C-2 2),38.2(C-19),37.8(C-10),35.1(C-7),34.0(C-12),30.0(C-16),29.0(C-24),28.7(C-15),28.3(C-2) ,24.1(C-29),22.6(C-30),22.3(C-11),19.1(C-6),17.0(C-25),16.8(C-23),16.7(C-26),15.4(C-27).

[0070] Compound 12: White powder, (-)-ESIMS m / z 455.5 [MH] - . 1 H-NMR(500MHz,Pyridine-d5)δ3.51(1H,dd,J=10.2,5.8Hz,H-3),1.28(s,3H,H-23),1.28(s,3H,H- 27),1.19(s,3H,H-26),1.05(s,3H,H-24),0.98(s,3H,H-25),0.93(s,3H,H-29),0.88(s,3H,H-30); 13 C-NMR(150MHz,Pyridine-d5)δ179.4(C-28),138.3(C-13),129.7(C-18),78.5(C-3),56.3(C-5),51.6( C-9),50.0(C-17),45.2(C-14),42.2(C-8),42.0(C-19),39.9(C-4),39.7(C-1),38.1(C-21),37.8(C-10 ),36.9(C-12),35.9(C-7),34.1(C-16),33.3(C-20),32.7(C-30),29.2(C-11),28.7(C-23),28.2(C-15) ,26.00(C-2),24.8(C-29),22.5(C-22),21.7(C-27),19.2(C-6),18.6(C-26),17.1(C-24),16.9(C-25).

[0071] Compound 13: White powder, (-)-ESIMS m / z 507 [M+Cl] - . 1 H-NMR(500MHz,CDCl3)δ5.36(1H,t,J=3.2Hz,H-12),3.22(1H,dd,J=11.2,4.4Hz,H-3),2.60(1H,s,H-18),1.26(3H,s,H-27), 1.21(3H,s,H-29),0.99(3H,s,H-23),0.95(3H,d,J=6.7Hz,H-30),0.91(3H,s,H-25),0.79(3H,s,H-24),0.69(3H,s,H-26).; 13C-NMR (125MHz, CDCl3) δ178.5(C-28),138.2(C-13),129.4(C-12),79.2(C-3),73.3(C-19),55.3(C-5) ,53.4(C-18),51.7(C-17),48.0(C-9),47.4(C-20),41.3(C-14),40.1(C-8),38.9(C-4),38.6(C-1),3 7.6(C-22),37.1(C-10),32.9(C-7),28.4(C-15),28.3(C-23),27.6(C-2),27.4(C-21),26.2(C-29),2 5.7(C-16),24.7(C-27),23.8(C-11),18.6(C-6),16.8(C-26),16.3(C-24),15.7(C-30),15.4(C-25).

[0072] Compound 14: White powder, (+)-ESIMS m / z 429 [M+H] + . 1 H-NMR (600MHz, CDCl3) δ5.31(1H,t,J=3.7Hz,H-12),3.22(1H,dd,J=11.5,4.4Hz,H-3),2.23(1H,dd,J=13.3,3.7Hz,H-18),1.14 (3H,s,H-27),1.00(3H,s,H-23),0.96(3H,s,H-28),0.95(3H,s,H-26),0.93(3H,s,H-29),0.90(3H,s,H-24),0.79(3H,s,H-25); 13 C-NMR (150MHz, CDCl3) δ143.2(C-13),124.4(C-12),79.0(C-3),72.2(C-17),55.2(C-5),48.8(C-1 8),48.3(C-19),47.6(C-9),41.5(C-14),39.6(C-8),38.8(C-1),38.5(C-4),37.1(C-10),36.8(C- 22),36.5(C-21),32.8(C-28),32.8(C-7),31.0(C-20),28.1(C-23),27.3(C-16),27.2(C-15),25. 5(C-27),25.5(C-29),24.0(C-11),23.6(C-2),18.4(C-6),17.1(C-26),15.6(C-24),15.3(C-25).

[0073] Compound 15: White powder, (+)-ESIMS m / z 443 [M+H] + . 1 H-NMR (500MHz, Pyridine-d5) δ5.25 (2H, t, J = 3.3 Hz, H-12), 3.95 (1H, d, J = 10.5 Hz, H-28), 3.51 (1H, d, J = 10.5 Hz, H-28), 3.49 (1H, dd, J = 10.8, 5. 4Hz,H-3),1.28(3H,s,H-27),1.25(3H,s,H-25),1.08(3H,s,H-23),1.0 5(3H,s,H-26),0.99(3H,m,H-30),0.97(3H,s,H-29),0.96(3H,s,H-24); 13 C-NMR(150MHz,Pyridine-d5)δ140.0(C-13),125.4(C-12),78.5(C-3),69.5(C-18),56.1(C-5),55.0(C -18),48.5(C-9),42.8(C-14),40.7(C-8),40.3(C-20),40.2(C-19),39.8(C-1),39.6(C-4),39.1(C-10 ),37.5(C-17),36.6(C-7),33.7(C-21),31.6(C-15),29.2(C-22),28.6(C-2),27.0(C-23),24.2(C-11) ,24.1(C-16),24.0(C-27),22.0(C-30),19.2(C-6),18.2(C-29),17.4(C-26),17.0(C-25),16.3(C-24).

[0074] Compound 16: Colorless needle-like crystals (methanol), (-)-ESIMS m / z 541 [MH] - . 1H-NMR(500MHz, Pyridine-d5) δ 5.48 (1H, t, J = 3.4 Hz, H-12), 4.85 (1H, dd, J = 11.6, 4.3 Hz, H-3), 3.82 (2H, s, H-2'), 2.64 (1H, d, J = 11.3 Hz, H-18), 1.23 (3H, s, H-27), 1.06 (3H, s, H-25), 1.02 (3H, s, H-23), 1.02 (3H, d, J = 6.2 Hz, H-30), 0.97 (3H, d, J = 6.2 Hz, H-29), 0.93 (3H, s, H-26), 0.82 (3H, s, H-24); 13 C-NMR(125MHz, Pyridine-d5) δ 179.9 (C-28), 170.3 (C-3'), 167.9 (C-1'), 139.3 (C-13), 125.4 (C-12), 81.6 (C-3), 55.6 (C-5), 53.6 (C-18), 48.1 (C-17), 47.8 (C-9), 42.5 (C-14), 39.9 (C-2'), 39.9 (C-8), 39.5 (C-19), 39.4 (C-20), 38.3 (C-1), 38.1 (C-4), 37.5 (C-22), 37.0 (C-10), 33.4 (C-7), 31.1 (C-21), 28.7 (C-23), 28.7 (C-15), 28.2 (C-2), 24.9 (C-16), 23.9 (C-27), 23.6 (C-11), 21.4 (C-30), 18.5 (C-6), 17.5 (C-29), 17.4 (C-26), 17.0 (C-24), 15.5 (C-25).

[0075] Compound 18: White powder, (-)-ESIMS m / z 507 [M + Cl] - . 1 H-NMR(500MHz, Pyridine-d5) δ 5.49 (1H, t, J = 3.5, H-12), 4.12 (1H, td, J = 10.5, 4.3 Hz, H-2), 3.42 (1H, d, J = 10.5 Hz, H-3), 2.65 (1H, d, J = 11.3 Hz, H-18), 1.30 (3H, s, H-27), 1.23 (3H, s, H-25), 1.10 (3H, s, H-23), 1.07 (3H, s, H-26), 1.00 (3H, s, H-24), 1.00 (3H, d, J = 5.9 Hz, H-30), 0.97 (3H, d, J = 6.2 Hz, H-29); 13C-NMR(150MHz,Pyridine-d5)δ180.3(C-28),139.7(C-13),125.9(C-12),84.2(C-3),69.0(C-2),56.3( C-5),53.9(C-18),48.5(C-17),48.4(C-1),48.4(C-9),42.9(C-14),40.4(C-8),40.2(C-4),39.8(C-19) ,39.8(C-20),38.8(C-10),37.8(C-22),33.9(C-7),31.4(C-21),29.8(C-23),29.0(C-15),25.3(C-16) ,24.3(C-11),24.1(C-27),21.8(C-30),19.2(C-6),18.1(C-29),17.9(C-26),17.9(C-24),17.4(C-25).

[0076] Compound 19: White powder, (-)-ESIMS m / z 617 [MH] - . 1 H-NMR(500MHz,Pyridine-d5)δ8.01(1H,d,J=15.9Hz,H-7'),7.57(2H,d,J=8.5Hz,H-2',6'),7.16(2 H,d,J=8.5Hz,H-3',5'),6.62(1H,d,J=15.9Hz,H-8'),5.64(1H,td,J=10.9,4.4Hz,H-2),5.48(1H,t ,J=3.3Hz,H-12),2.66(1H,d,J=11.2Hz,H-18),1.32(3H,s,H-27),1.26(3H,s,H-23),1.13(3H,s,H- 24),1.09(3H,s,H-26),1.04(3H,s,H-25),1.03(3H,d,J=6.3Hz,H-29),0.97(3H,d,J=6.2Hz,H-30). 13C-NMR(125MHz,Pyridine-d5)δ180.1(C-28),167.5(C-9'),161.4(C-4'),144.71(C-7'),139.4(C-13),130.6(C-2',6'),126.2(C- 1'),125.4(C-12),116.9(C-3',5'),116.00(C-7'),79.9(C-3),73.4(C-2),55.6(C-5),53.6(C-18),48.1(C-17),48.0(C-9),44.9( C-1),42.6(C-14),40.5(C-8),40.0(C-19),39.5(C-4),39.4(C-20),38.6(C-10),37.5(C-22),33.5(C-7),31.2(C-21),29.3(C-15 ),28.7(C-23),25.0(C-16),23.9(C-27),23.7(C-11),21.5(C-30),18.8(C-6),17.7(C-24),17.5(C-25),17.4(C-26),16.8(C-29).

[0077] Compound 20: White powder, (-)-ESIMS m / z 617 [MH] - . 1 H-NMR(500MHz,Pyridine-d5)δ7.99(1H,d,J=15.9Hz,H=7'),7.55(2H,d,J=8.6Hz,H-2',6'),7.17(2H,d ,J=8.6Hz,H-3',5'),6.58(1H,d,J=15.9Hz,H-8'),5.67(1H,td,J=10.5,4.5Hz,H-2),5.53(1H,t,J=3.8 Hz,H-12),3.66(1H,d,J=10.5Hz,H-3),3.34(1H,dd,J=14.0,4.5Hz,H-18),1.34(3H,s,H-27),1.33(3H, s,H-25),1.15(3H,s,H-23),1.11(3H,s,H-30),1.03(3H,s,H-29),1.01(3H,s,H-24),0.96(3H,s,H-26); 1313C-NMR (150 MHz, Pyridine-d5) δ 180.0 (C-28), 167.2 (C-9'), 161.1 (C-4'), 144.7 (C-7'), 144.4 (C-13), 130.3 (C-2',6'), 125.9 (C-1'), 122.1 (C-12), 116.6 (C-3',5'), 115.8 (C-8'), 79.6 (C-3), 73.1 (C-2), 55.4 (C-5), 47.8 (C-9), 46.2 (C-17,19), 44.4 (C-1), 42.0 (C-8), 41.7 (C-18), 40.2 (C-4), 39.6 (C-14), 38.4 (C-10), 34.0 (C-21), 33.0 (C-7), 33.3 (C-22), 32.9 (C-29), 30.7 (C-20), 29.0 (C-23), 28.1 (C-15), 26.0 (C-27), 23.7 (C-11), 23.5 (C-16,30), 18.6 (C-6), 17.4 (C-26), 17.2 (C-24), 16.4 (C-25).

[0078] Compound 21: White powder, (-)-ESIMS m / z 617 [M-H] - . 1 1H-NMR (500 MHz, Pyridine-d5) δ 8.02 (1H, d, J = 15.9 Hz, H-7'), 7.57 (2H, overlapped with solvent), 7.18 (2H, d, J = 8.6 Hz, H-3',5'), 6.71 (1H, d, J = 15.9 Hz, H-8'), 5.48 (1H, t, J = 3.2 Hz, H-12), 5.27 (1H, d, J = 10.5 Hz, H-3), 4.31 (1H, td, J = 10.5, 4.4 Hz, H-2), 3.32 (1H, dd, J = 13.5, 3.4 Hz, H-18), 1.28 (3H, s, H-27), 1.07 (3H, s, H-25), 1.05 (3H, s, H-26), 1.02 (3H, s, H-30), 1.02 (3H, s, H-29), 1.01 (3H, s, H-23), 0.97 (3H, s, H-24); 13C-NMR(150MHz, Pyridine-d5) δ 180.3 (C-28), 168.0 (C-9'), 161.4 (C-4'), 145.0 (C-6'), 145.0 (C-13), 130.6 (C-2', 6'), 126.3 (C-1'), 122.3 (C-12), 116.8 (C-3', 5'), 116.1 (C-8'), 85.1 (C-3), 66.4 (C-2), 55.5 (C-5), 48.4 (C-9), 48.1 (C-19), 46.7 (C-1), 46.5 (C-17), 42.2 (C-14), 42.0 (C-18), 39.9 (C-8), 39.8 (C-4), 38.4 (C-10), 34.3 (C-21), 33.3 (C-7), 33.2 (C-29), 33.0 (C-22), 31.0 (C-20), 29.0 (C-23), 28.3 (C-15), 26.2 (C-27), 23.9 (C-16), 23.8 (C-11), 23.7 (C-30), 18.7 (C-6), 18.3 (C-26), 17.4 (C-24), 16.8 (C-25).

[0079] Compound 22: white powder, (-)-ESIMS m / z 617 [M-H] - . 1 H-NMR(600MHz, pyridine-d5) δ 8.02 (1H, d, J = 15.9 Hz, H-7'), 7.57 (2H, overlapped with solvent), 7.16 (2H, d, J = 8.6 Hz, H-3', 5'), 6.69 (1H, d, J = 15.9 Hz, H-2'), 5.48 (1H, t, J = 3.4 Hz, H-12), 5.28 (1H, d, J = 10.5 Hz, H-3), 4.31 (1H, td, J = 10.5, 4.5 Hz, H-2), 2.65 (1H, d, J = 11.3 Hz, H-18), 1.24 (3H, s, H-27), 1.08 (3H, s, H-23), 1.06 (3H, s, H-24), 1.05 (3H, s, H-26), 1.01 (3H, s, H-25), 1.00 (3H, d, J = 6.3 Hz, H-29), 0.97 (3H, d, J = 6.3 Hz, H-30); 13C-NMR(150MHz,Pyridine-d5)δ179.9(C-28),167.9(C-9'),161.3(C-4'),144.8(C-7'),139.4(C-13),130.6(C-2',6'),126.3(C-1 '),125.4(C-12),116.8(C-3',5'),116.1(C-8'),85.1(C-3),66.4(C-2),55.6(C-5),53.5(C-18),48.6(C-9),48.1(C-17),48.0(C -1),42.6(C-14),40.0(C-8),39.8(C-4),39.5(C-19),39.4(C-20),38.3(C-10),37.5(C-22),33.4(C-7),31.1(C-21),29.1(C-23) ,28.7(C-15),24.9(C-16),24.0(C-27),23.7(C-11),21.4(C-30),18.7(C-6),18.3(C-24),17.5(C-26),17.4(C-29),16.9(C-25).

[0080] Compound 23: White powder, (-)-ESIMS m / z 617 [MH] - . 1 H-NMR(500MHz,Pyridine-d5)δ8.16(1H,d,J=8.6Hz,H-3',5'),7.16(1H,d,J=8.6Hz,H-2',6'),6. 94(1H,d,J=12.9Hz,H-3”),6.10(1H,d,J=12.9Hz,H-2”),5.47(1H,t,J=3.2Hz,H-12),5.22(1H,d, J=9.8Hz,H-3),4.27(1H,m,H-2),2.65(1H,d,J=11.2Hz,H-18),1.20(3H,s,H-27),1.06(3H,s,H-2 3),1.04(3H,s,H-24),0.99(3H,s,H-29),0.99(3H,s,H-26),0.98(3H,s,H-25),0.97(3H,s,H-30). 13C-NMR(150MHz,Pyridine-d5)δ167.7(C-1'),161.0(C-7'),144.1(C-3'),139.9(C-13),134.3(C-5',9'),127.2(C-4'),125.9( C-12),117.6(C-2'),116.4(C-6',8'),85.5(C-3),66.8(C-2),56.1(C-5),54.1(C-18),49.2(C-9),48.6(C-17),48.5(C-1),43 .1(C-14),40.53(C-4),40.2(C-8),40.0(C-19),38.8(C-20),38.0(C-10),33.8(C-7),31.6(C-22),30.5(C-21),29.6(C-23),2 9.2(C-15),25.4(C-16),24.5(C-11),24.2(C-27),21.9(C-30),19.2(C-6),18.7(C-24),18.1(C-29),18.0(C-26),17.4(C-25).

[0081] Pharmacological experiments on the neuroprotective activity of pentacyclic triterpenoids

[0082] Experimental Example 1. Glutamate-induced SK-N-SH neuroblastoma cell damage model

[0083] Experimental methods: SK-N-SH cells were cultured routinely, and then co-incubated with donepezil (3 μmol / L) and screening compound (10 μmol / L) for 4 hours respectively. A glutamate injury model was then established by damaging SK-N-SH cells with glutamate. Cell viability was assessed using the MTT assay 4 hours after injury.

[0084] Experimental results: As shown in Table 3, among the compounds isolated from Cynomorium songaricum, 12 pentacyclic triterpenoids exhibited significant neuroprotective activity against glutamate-induced neuronal damage.

[0085] Experimental Example 2. Sodium dithionite-induced oxygen-glucose deprivation model in SK-N-SH neuroblastoma cells

[0086] Experimental Methods: SK-N-SH cells were cultured routinely, and then co-incubated with donepezil (3 μmol / L), PHPB (10 μmol / L), and screening compound (10 μmol / L) for 4 hours, respectively. An oxygen-glucose deprivation model was then established by damaging SK-N-SH cells with sodium dithionite. Cell viability was assessed using the MTT assay 24 hours after injury.

[0087] Experimental results: As shown in Table 3, among the compounds isolated from Cynomorium songaricum, 12 pentacyclic triterpenoids have significant protective activity against nerve cells damaged by oxygen-glucose deprivation.

[0088] Table 3. Neuroprotective activity of Cynomorium isolated compounds (10 μmol / ml)

[0089]

[0090] ** P<0.01 vs Control; # P < 0.05 vs Model, ## P<0.01 vs Model, ### P<0.001 vs Model

[0091] References:

[0092] Zheng Lingyan, Han Ruilan, 2016. Study on the active components and mechanisms of the effects of Cynomorium songaricum on Alpinia oxyphylla. Vol. Master's Thesis. Inner Mongolia Medical University, Hohhot, p.47.

Claims

1. The use of the pentacyclic triterpenoids and their pharmaceutically acceptable salts as shown below in the preparation of medicaments for the prevention and / or treatment of neurodegenerative diseases. 。 2. The use of a pharmaceutical composition in the preparation of a medicament for the prevention and / or treatment of neurodegenerative diseases, characterized in that, The pharmaceutical composition contains an effective dose of the pentacyclic triterpenoid compound as described in claim 1 and a pharmaceutically acceptable carrier.

3. The application according to claim 2, characterized in that: The dosage form of the pharmaceutical composition is selected from tablets, capsules, pills, granules, oral liquids, injections, and suspensions.

4. In the application of claim 1, the neurodegenerative disease is selected from stroke, brain injury, amyotrophic lateral sclerosis, Huntington's disease, Parkinson's disease, or Alzheimer's disease.

5. The method for preparing the pentacyclic triterpenoid compound according to claim 1 includes the following steps: 95% ethanol reflux extraction of Cynomorium songaricum, concentration of the extract followed by separation and purification by organic solvent extraction, macroporous adsorption resin, silica gel column chromatography, gel column chromatography, reversed-phase silica gel column chromatography, and preparative HPLC to obtain the target compound.

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