CN105503988B - Natural antiepileptic activity compound and its purposes in pharmaceutical preparation - Google Patents

Abstract

The present invention relates to natural antiepileptic activity compound and its purposes in pharmaceutical preparation.The invention discloses the different monomers compound in Cynanchum otophyllum Schneid to have the function that the rush epilepsy completely contradicted and anti-epileptic.Specifically, the present inventor extraction one-component from Apocynaceae Cynanchum (Cynanchum) plant cynanchum auriculatum Royle ex Wight (C.auriculatum Royle ex Wight) and cynanchum wilfordii (C.wilfordii Hemsl.), and experiments prove that the antiepileptic activity of each compound.

Description

Natural antiepileptic active compounds and their use in pharmaceutical preparations

technical field

The invention belongs to the field of biomedicine, in particular, the invention relates to natural antiepileptic active compounds and their application in pharmaceutical preparations.

Background technique

Epilepsy, also known as cerebral epilepsy or sheep epilepsy, is a syndrome of chronic recurrent transient brain dysfunction. According to statistics, the incidence of epilepsy in my country is about 23/100,000 per year, and the prevalence rate is 3.5‰-4.8‰. Seizures can cause a series of symptoms, and convulsions are one of the typical symptoms of epilepsy, including repetitive movements of limbs, abnormal behavior, and general convulsions with loss of consciousness, etc., and even cause body damage or death. The treatment of epilepsy includes various methods such as drug therapy, surgical therapy, physical therapy and psychotherapy, among which drug therapy is the most commonly used and most important means at present. Currently clinically used drugs for the treatment of epilepsy include Carbamazepine (Carbamazepine), Sodium Valproate (Depakene), Sodium Phenytoin (Dilantin), etc., but these drugs usually have very obvious side effects, and long-term use is harmful to the body. Large effects, and may even trigger suicidal thoughts or behaviors in a small proportion of patients taking the drug. Traditional Chinese medicine compound and Chinese patent medicine preparation products also occupy a lot of markets, but due to the complex ingredients of traditional Chinese medicine prescriptions, and the different plant origins, changing climates, and different harvest periods affect the content of the main ingredients of the medicine, so the quality of traditional Chinese medicine prescriptions is limited. It is quite difficult to control the photosynthesis, which also limits the promotion of traditional Chinese medicine preparations. Therefore, finding safe, non-toxic, effective and quality-controllable new antiepileptic drugs is of great significance in the prevention and treatment of epilepsy.

Cynanchum is a genus of Apocynum, about 200 species are distributed in tropical and temperate regions, and about 53 species are produced in my country. C.auriculatum, also known as Baishouwu, is a unique plant species in my country. About 95% of it grows in Binhai County, Jiangsu Province. It has the effects of nourishing liver and kidney, nourishing essence and blood, and anti-aging. Geshanxiao (C. wilfordii) is produced in Liaoning, Henan, Hubei and other places. It is often used to strengthen the stomach, relieve fullness, and treat choking. Qingyang ginseng (C. otophyllum) is produced in Hunan, Guangxi, Guizhou, Yunnan, Sichuan, and Tibet provinces and regions. It has detoxification, antispasmodic, and rheumatism effects. It is clinically used to treat rheumatic bone pain, epilepsy, and snake bites.

Qingyang ginseng extract Qingyang ginseng total glycosides is an adjuvant drug for clinical treatment of epilepsy. However, the composition of Qingyangshen tablets is complex, which brings certain difficulties to quality control, and the target of action is unknown. Clinically, it is mainly used in combination with antiepileptic drugs such as phenytoin or phenobarbital.

Contents of the invention

The object of the present invention is to provide natural antiepileptic active compounds and their use in pharmaceutical preparations.

The first aspect of the present invention provides a compound with a structure as shown in formula A or a pharmaceutically acceptable salt thereof, characterized in that:

In _{formula A} , R1 is sugar group, R2 is substituted or unsubstituted C1～C6 alkyl or substituted or unsubstituted C1～C6 alkenyl, _R3 is substituted or unsubstituted C1～C6 alkyl;

The substitution means that one or more hydrogen atoms are substituted by a substituent selected from the following group: C1～C10 alkyl, C3～C10 cycloalkyl, C1～C10 alkoxy, hydroxyl, carboxyl, C1～C10 carbonyl, C1-C10 amide group, C2-C10 ester group, C6-C30 aryl group and oxygen (=O).

In another preferred example, the sugar base is composed of cymarosyl, oleandrosyl, diginosyl, thevetosyl, digitoxin sugar One or more of (digitoxosyl), canarosyl (canarosyl), rhamnosyl (rhamnosyl) and glucosyl (glucosyl).

In another preferred example, the sugar group includes 1 to 6 identical or different monosaccharide units, preferably, includes 2 to 4 identical or different monosaccharide units.

_In another preferred embodiment, R does not contain sugar units

In another preferred example, when R ₂ is hydroxyphenyl, the number of sugar units in R ₁ is not 3.

In another preferred example, the number of sugar units may be 0, 1, 2, 3 or 4.

In another preferred embodiment, the _R3 is Wherein R ₄ is a substituted or unsubstituted C1～C6 alkyl group; R ₅ is a substituted or unsubstituted C1～C6 alkyl group.

In another preferred embodiment, the R ₁ is n is a positive integer from 1 to 4, R ₇ is H or methyl or C1-C10 ester group, R ₈ is H or C2-C10 ester group.

In another preferred example, the R1 is selected from the following group:

_In another preferred example, the R2 is selected from the following group:

In another preference, the _R3 is selected from the following group:

In another preferred example, the structure of the compound is shown in Formula I:

In another preferred example, the structure of the compound is shown in Formula II:

In another preferred example, the structure of the compound is shown in formula III:

In another preferred example, the structure of the compound is shown in formula IV:

In another preferred example, the structure of the compound is shown in formula V:

In another preferred example, the structure of the compound is shown in formula VI:

In another preferred example, the structure of the compound is shown in formula VII:

In another preferred example, the structure of the compound is shown in formula VIII:

In another preferred example, the structure of the compound is shown in formula IX:

In another preferred example, the structure of the compound is shown in formula X:

In another preferred example, the structure of the compound is shown in formula XI:

In another preferred example, the structure of the compound is shown in formula XII:

In another preferred example, the structure of the compound is shown in formula XIII:

In another preferred example, the structure of the compound is shown in formula XV:

In another preferred example, the structure of the compound is shown in formula XVI:

The first aspect of the present invention provides a pharmaceutical composition, which comprises the compound as described in the first aspect of the present invention or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable carrier or excipient .

In another preferred example, the pharmaceutical composition further includes an optional drug selected from the group consisting of Carbamazepine, Depakene and Dilantin.

The third aspect of the present invention provides the compound as described in the first aspect of the present invention or a pharmaceutically acceptable salt thereof, wilfoside G or a pharmaceutically acceptable salt thereof, or the drug as described in the second aspect of the present invention Application of composition in preparation of medicine for treating or preventing epilepsy

In another preferred example, the wilfoside G structure is shown in formula XIV:

The fourth aspect of the present invention provides Apocynaceae Cynanchum plant C. auriculatum Royle ex Wight or C. wilfordii Hemsl. or its extract in the preparation of treatment or Use of antiepileptic drugs.

The fifth aspect of the present invention provides a total phytoglycoside or a traditional Chinese medicine extract containing the phytoglycoside, the phytoglycoside or the extract contains anti-epileptic active ingredients, and the extract contains The content C1 of the antiepileptic active ingredient satisfies the following formula B:

C1/(C1+C2)≥50％ Formula B

In formula B, C1 is the content of anti-epileptic active ingredients; C2 is the content of promoting epilepsy active ingredients.

In another preferred example, C1/(C1+C2)≥80%; more preferably, ≥90%; most preferably, ≥95%, such as 96%, 97%, 98%, 99%, 99.5%.

In another preferred example, the anti-epileptic active ingredients are selected from one or more of the following groups:

Compounds represented by formulas I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV, XVI.

In another preferred example, the epilepsy-promoting active ingredients are selected from one or more of the following groups:

Compounds shown in formula a, b, c and d

In another preferred example, the plants include: C. auriculatum Royle ex Wight and C. wilfordii Hemsl.

In another preferred example, the preparation method of the extract is as follows:

Dipping and extracting the medicinal material with 95% ethanol three times, concentrating the extract under reduced pressure to obtain ethanol extract, adding water to suspend, extracting with ethyl acetate, recovering ethyl acetate, dissolving the ethyl acetate extract in 70% ethanol solution, filtering Or after centrifuging to remove insoluble matter, perform macroporous resin column chromatographic separation, collect 75% ethanol eluted fractions, and concentrate to obtain the final product.

The sixth aspect of the present invention provides a pharmaceutical composition, comprising the traditional Chinese medicine extract as described in the fifth aspect of the present invention, and optional pharmaceutically acceptable carriers or excipients.

The seventh aspect of the present invention provides a method for quality control of anti-epileptic products, including steps:

(a) Determining the content C2 of the epilepsy-promoting active ingredient in the product or its raw materials, and comparing it with the predetermined value C0;

(b) When the content C2 is higher than C0, it is judged that the quality of the product is unqualified; when the content C2 is lower than or equal to C0, it is judged that the quality of the product is qualified.

In another preferred example, the epilepsy-promoting active ingredients are selected from one or more of the following groups: compounds represented by formulas a, b, c and d.

In another preferred example, the anti-epileptic products include anti-epileptic drugs containing Qingyangshen extract.

In another preferred example, the C2 is the weight percentage of the epilepsy-promoting active ingredient in the product or raw material.

In another preferred example, the C0≤10% (w/w), preferably, the C0≤5% (w/w), more preferably, the C0≤1%, the C0 can be 0.01%, 0.05%, 0.1%, 0.2%, 0.5%, 1.0%, 5%, 10%.

The eighth aspect of the present invention provides the use of a compound selected from the following group in the preparation of a medicament or reagent for constructing an animal epilepsy model: compounds represented by formulas a, b, c and d.

It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described in the following (such as embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, we will not repeat them here.

Description of drawings

Figure 1A shows the fast movement trajectory of zebrafish in each concentration group of cynawilfoside A and the epilepsy model group; Figure 1B shows the bar graph drawn according to the mean ± SE of the fast movement distance of zebrafish.

Figure 2A shows the fast movement trajectory of zebrafish in each concentration group of wilfoside C1N and the epilepsy model group; Figure 2B shows the bar graph drawn according to the mean ± SE of the fast movement distance of zebrafish.

Figure 3A shows the fast movement trajectory of zebrafish in each concentration group of wilfoside K1N and the epilepsy model group; Figure 3B shows the bar graph drawn according to the mean ± SE of the fast movement distance of zebrafish.

Figure 4A shows the fast movement trajectory of zebrafish in each concentration group of taiwanoside C and the epilepsy model group; Figure 4B shows the bar graph drawn according to the mean ± SE of the fast movement distance of zebrafish.

Figure 5A shows the fast movement trajectory of zebrafish in different concentration groups of qingyangshenside and epilepsy model group; Figure 5B shows the bar graph drawn according to the mean ± SE of the fast movement distance of zebrafish.

Figure 6A shows the 3-O-β-D-oleandyranosyl-(1→4)-β-D-canadaranyl-(1→4)-β-D -Zebrafish of various concentration groups and epilepsy model group Fast movement trajectories; Figure 6B shows a histogram plotted against the mean ± SE of fast movement distance in zebrafish.

Figure 7A shows the fast movement trajectory of zebrafish in each concentration group of wilfoside C3N and the epilepsy model group; Figure 7B shows the bar graph drawn according to the mean ± SE of the fast movement distance of zebrafish.

Figure 8A shows the fast movement trajectory of zebrafish in each concentration group of cynauricoside A and the epilepsy model group; Figure 8B shows the bar graph drawn according to the mean ± SE of the fast movement distance of zebrafish.

Figure 9A shows the fast movement trajectory of zebrafish in each concentration group and epilepsy model group of the deactivated part of psoriasis in the ear lobe; Figure 9B shows the bar graph drawn according to the mean±SE of the mean ± SE of the fast movement distance of zebrafish.

Figure 10 shows Qingyangshengenin (qingyangshengenin), Caudatin (caudatin), otophylloside A (otophylloside A), otophylloside C (otophylloside C), and otophylloside F (otophylloside F, formula I), otophylloside M (otophylloside M, formula b), Caudatin 3-O-β-D-Canadian sesame sugar [caudatin3-O-β-D-cymaropyranoside, formula IX], otophylloside aglycon 3-O-β-D-Cymaropyranosyl-(1→4)-β-D-Digoxigenin [qinyangshengenin 3-O-β-D-cymaropyranosyl-(1→4)-β-D -digitoxopyranoside, formula XVII], Caudatin 3-O-β-D-Canadian sesame sugar pyranyl-(1 → 4)-β-D-Canada sesame sugar [caudatin3-O-β-D-cymaropyranosyl-( 1→4)-β-D-cymaropyranoside, formula X], rusutamine 3-O-β-D-oleandopyranosyl-(1→4)-β-D-Canada hempyranose pyranoside base-(1→4)-β-D-Canada sesame sugar Formula II], rusutamine 3-O-α-L-Canada hempyranosyl-(1→4)-β-D-Canada hempyranosyl-(1→4)-α-L- Digi sugar pyranosyl-(1→4)-β-D-Canada hemp sugar α-L-diginopyranosyl-β-D-cymaropyranoside, formula XIII], Qingyangshen aglycone 3-O-β-D-oleandyl pyranosyl-(1→4)-β-D-Canada hemp Glycopyranyl-(1→4)-β-D-digitoxin [qinyangshengenin3-O-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β- D-digitoxopyranoside, formula a], Kodatine 3-O-β-D-glucopyranosyl-(1→4)-β-D-oleandyl-(1→4)-β -D-Canadian sesame sugar pyranyl-(1→4)-β-D-Canadian sesame sugar [caudatin3-O-β-D -glucopyranoside-(1→4)-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-cymaropyranoside, formula c], tell the court 3-O- β-D-Canadian sugar pyranosyl-(1→4)-β-D-Canada hempyranosyl-(1→4)-β-D-Canada hemp sugar [caudatin 3-O-β- D-thevetopyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-cymaropyranoside, formula XII], Qingyangshen aglycone 3-O-β-D-oleander Panosyl-(1→4)-β-D-Canada hemp sugar [qinyangshengenin3-O-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyranoside, formula XI], taiwanoside C (formula III) , wilfoside K1N (formula V), cynawilfoside A (formula VI), wilfoside M1N (formula d), cynauricoside E (formula XV), cynauricuoside A (formula XVI), wilfoside G (formula XIV) and other compounds in the maximum non-toxic dose Effects on PTZ-induced zebrafish.

Detailed ways

Through extensive and in-depth research, the present inventor unexpectedly found that different monomeric compounds in Qingyang ginseng have diametrically opposite epilepsy-promoting and anti-epileptic effects. Specifically, the inventors extracted a single component from C. auriculatum Royle ex Wight and C. wilfordii Hemsl. of the genus Cynanchum of the family Apocynaceae, and confirmed its anti- epileptic activity.

the term

As used herein, the term "substituted" means that one or more hydrogen atoms on the group are replaced by a substituent selected from the group consisting of: C1~C10 alkyl, C3~C10 cycloalkyl, C1~C10 alkoxy, hydroxyl , carboxyl group (-COOH), C1~C10 carbonyl group, C1~C10 amide group, C2~C20 ester group, C6~C30 aryl group, fluorine group and sulfide group.

The term "C1～C10 alkyl" refers to a straight chain or branched chain alkyl group having 1 to 10 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or similar groups.

The term "C3-C10 cycloalkyl" refers to a cycloalkyl group having 3-10 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cycloheptyl, or the like.

The term "C1-C10 alkoxy" refers to a straight-chain or branched alkoxy group with 1-10 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, iso Butoxy, sec-butoxy, tert-butoxy, or the like.

The term "carbonyl" refers to a A group; for example, "C1-10 carbonyl" refers to a carbonyl group having 1-10 carbon atoms.

The term "amido" refers to a The group; for example, "C1-10 amido group" refers to an amido group having 1-10 carbon atoms.

The term "C2-C20 ester group" refers to having an alkyl-COO- structure or having a -COO-alkyl structure, wherein the alkyl group can be straight or branched. It is preferably a group having a C1-C10 alkyl-COO- structure, such as CH ₃ COO-, C ₂ H ₅ COO-, C ₃ H ₈ COO-, (CH ₃ ) ₂ CHCOO-, nC ₄ H ₉ COO- , tC ₄ H ₉ COO-, or similar groups. Or have -COO-C1~C19 alkyl structure, such as -COOCH ₃ , -COOC ₂ H ₅ , -COOC ₃ H ₈ or -COO-(CH ₂ )n-CH ₃ , n is an integer of 0-18.

The term "aryl", preferably having 6 to 30 carbon atoms, refers to a monocyclic or bicyclic aromatic group with 6 to 12 carbon atoms in the ring, such as: phenyl, biphenyl, naphthyl , or similar groups in which each carbon atom may be optionally substituted. It preferably has 1-3 substituents, and the substituents are selected from: halogen, C1-C10 alkyl, cyano, OH, nitro, C3-C10 cycloalkyl, C1-C10 alkoxy, amino.

The term "thioether group" refers to a group having the structure -hydrocarbyl-S-hydrocarbyl.

The term "halogenated compound" refers to a compound in which at least one H atom in an organic compound is replaced by a halogen.

The term "halogen" refers to fluorine, chlorine, bromine, iodine. The term "halo" refers to fluoro, chloro, bromo, iodo.

Each group in the present invention may be unsubstituted or substituted, and the substituted refers to being substituted by a substituent selected from the following group: hydroxyl, halogen, C1～C6 alkyl, halogenated C1～C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, cyano, tert-butylcarbamoyl, -O-(CH ₂ ) _n -O-; wherein, n is an integer between 1 and 3. The substituents may be substituted at various positions of each group.

In the embodiment of the present invention, relate to otophylloside F), rostratamine3-O-triglycoside (rostratamine 3-O-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyra-nosyl-( 1→4)-β-D-cymaropyranoside), taiwanoside C, wilfoside C1N, wilfoside C3N, wilfoside K1N, cynauricoside A, cynawilfoside A and other monomer compounds in the preparation of drugs for the treatment or prevention of epilepsy and the application of Apocynaceae goose down Application of the extracts of C. auriculatum Royleex Wight and C. wilfordii Hemsl. from plants of the genus Cynanchum in the preparation or prevention of epilepsy drugs. And the anti-epileptic effects of these monomeric compounds are better than otophylloside A, qingyangshengenin and caudatin, which are the main components of Qingyangshen Total Glycosides Tablets.

About otophylloside F, rusutamine 3-O-β-D-oleandylopyranosyl-(1→4)-β-D-canadinopyranosyl-(1 →4)-β-D-Canada sesame sugar [rostratamine3-O-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-cymaropyranoside], taiwanoside C, The application of wilfoside C1N, wilfoside C3N, wilfoside K1N, cynauricoside A, cynauricoside A, and the antiepileptic effects of the extracts of Niupixiao and Geshanxiao in related products has not been reported in the literature, and cynawilfoside A is isolated and identified from natural products for the first time compound.

In the present invention, the anti-epileptic effect of the monomeric compounds in Qingyang ginseng has been thoroughly studied, and it is unexpectedly found that different monomeric compounds in Qingyang ginseng have diametrically opposite epilepsy-promoting and anti-epileptic effects. Qingyangshen glycoside (otophylloside F), one of the main components of Qingyang ginseng, and rusutamine 3-O-β-D-oleandopyranosyl-(1→4)-β-D-Canada Rostratamine3-O-β-D-oleandropyranosyl-(1→4)-β-D-Canada sesame sugar[rostratamine3-O-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β- D-cymaropyranoside] has significant anti-clonic convulsions induced by pentylenetetrazol in the zebrafish model; Oleandropyranosyl-(1→4)-β-D-Canada hempyranosyl-(1→4)-β-D-digitoxin [qinyangshengenin3-O-β-D-oleandropyranosyl- (1→4)-β-D-cymaropyranosyl-(1→4)-β-D-digitoxopyranoside] and otophylloside M (otophylloside M) have a significant role in promoting convulsions.

The present invention provides compounds with significant anti-epileptic effects. There is no relevant report on the anti-epileptic or anti-convulsant effects of these compounds. Cynawilfoside A is a new natural product isolated and identified for the first time. Since these compounds have a large content in crude drugs, extraction , The preparation is relatively easy, and the quality is controllable, so it has great development prospects in the development of antiepileptic drugs.

Through animal experiments, the present invention finds that extracts derived from C. auriculatum and C. wilfordii of the genus Cynanchum of the family Apocynaceae have certain antiepileptic activity, and the monomeric compound Otophylloside F, rusutamine 3-O-β-D-oleandylopyranosyl-(1→4)-β-D-canadamosylpyranosyl-(1→ 4)-β-D-Canada sesame sugar [rostratamine3-O-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-cymaropyranoside], taiwanoside C, wilfosideC1N , wilfoside C3N, wilfoside K1N, cynauricoside A, cynawilfoside A, etc. have significant antiepileptic activity, and their structures are represented by the following formulas Ⅰ to XVI respectively:

The structure of the compound otophylloside F (compound I) is shown in formula I:

Compound rusutamine 3-O-β-D-oleandopyranosyl-(1→4)-β-D-canapyranosyl-(1→4)-β-D-canapyranosyl The structure of the sugar [rostratamine 3-O-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-cymaropyranoside, Compound II] is shown in Formula II:

The structure of compound taiwanoside C (compound III) is shown in formula III:

The structure of compound wilfoside C1N (compound IV) is shown in formula IV:

The structure of compound wilfoside K1N (compound V) is shown in formula V:

The structure of compound cynawilfoside A (compound VI) is shown in formula VI:

The structure of compound wilfoside C3N (compound VII) is shown in formula VII:

The structure of compound cynauricoside A (compound VIII) is shown in formula VIII:

Compound IX:

Compound X:

Compound XI:

Compound XII:

Compound XIII:

Compound XV:

Compound XVI:

However, some monomeric compounds derived from Cynanchum of the family Apocynaceae have significant epilepsy-promoting activity.

Qingyang ginseng aglycon 3-O-β-D-oleandyl-(1→4)-β-D-canadamosyl-(1→4)-β-D-pyranosyl The structural formula of rehmanthin (compound a) is shown in formula a:

The structural formula of qingyangshenside M (compound b) is shown in formula b:

Koldatin 3-O-β-D-glucopyranosyl-(1→4)-β-D-oleandopyranyl-(1→4)-β-D-canapyranosepyranosyl The structural formula of base-(1→4)-β-D-Canada sesame sugar (compound c) is shown in formula c:

The structural formula of wilfoside M1N (compound d) is shown in formula d:

Pharmaceutical compositions and methods of administration

The compound of the present invention has excellent antiepileptic effect. The compounds of the present invention can be administered to mammals (eg, humans), and can be administered orally, rectally, parenterally (intravenously, intramuscularly or subcutaneously), topically, and the like. The compounds can be administered alone or in combination with other pharmaceutically acceptable compounds. It should be noted that the compounds of the present invention may be administered in admixture.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is admixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with (a) fillers or extenders, for example, Starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders such as hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, For example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow agents, such as paraffin; (f) Absorption accelerators such as quaternary ammonium compounds; (g) wetting agents such as cetyl alcohol and glyceryl monostearate; (h) adsorbents such as kaolin; and (i) lubricants such as talc, hard Calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage form may also contain buffering agents.

Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shell materials, such as enteric coatings and others well known in the art. They may contain opacifying agents and, in such compositions, the release of the active compound or compounds may be in a certain part of the alimentary canal in a delayed manner. Examples of usable embedding components are polymeric substances and waxy substances. The active compounds can also be in microencapsulated form, if desired, with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1 , 3-butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances, etc.

Besides such inert diluents, the compositions can also contain adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspending agents, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.

Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols, and suitable mixtures thereof.

Dosage forms for topical administration of a compound of this invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants which may be required, if necessary.

The compounds of the present invention may be administered alone or in combination with other active ingredients such as other clinically available antiepileptic drugs.

When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage is a pharmaceutically effective dosage when administered. For an individual with a body weight of 60 kg, The daily dosage is usually 1-1000 mg, preferably 20-500 mg. Of course, factors such as the route of administration and individual health conditions should also be considered for the specific dosage, which are within the skill of skilled physicians.

The main advantages of the present invention are:

(1) For the first time, the antiepileptic activity of the single-component extracts of C. auriculatum and C. wilfordii from the plants of the genus Cynanchum of the family Apocynaceae was evaluated, and it was found that part of the extracts Extracts have anti-epileptic function, and some extracts have epilepsy-promoting effect;

(2) The antiepileptic activity of the involved monomeric compounds with antiepileptic activity is superior to that of Qingyangshen glycoside A, Qingyangshen aglycone, and Gaodating, which are the main components of Qingyangshen Tablets in clinical use;

(3) The involved compounds with antiepileptic activity have a large content in plants and a wide range of sources, and have great research value and development prospects;

(4) The compound cynawilfoside A is a new compound isolated and identified for the first time, and its antiepileptic activity is better than that of the positive drug phenytoin;

(5) The procedure for obtaining effective parts rich in active compounds such as wilfoside C1N, wilfoside C3N, wilfoside K1N, and cynauricoside A from the traditional Chinese medicine Erye Niupixiao is simple and convenient for production and application.

Below in conjunction with specific embodiment, further state the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. Percentages and parts are by weight unless otherwise indicated.

Example 1

Evaluation of antiepileptic effect of compound cynawilfoside A (formula VI) in vivo:

The preparation method of the compound cynawilfoside A: 10kg of fresh Geshanxiao roots, peeled and sliced to obtain 2.7kg of dry product, crushed and extracted three times with 95% ethanol percolation, each time for 2-3 days, combined extracts, concentrated under reduced pressure to obtain Geshan Disinfected alcoholic extract, suspend the extract in water, extract with ethyl acetate to obtain ethyl acetate extract, mix the sample with 100 mesh silica gel, separate with silica gel column chromatography (200-300 mesh silica gel), and use chloroform- Gradient elution with methanol (10:1→5:1) yielded components Fr1-Fr9. Fr6 was further purified repeatedly with petroleum ether-acetone and preparative HPLC to obtain pure compound cynawilfoside A (1.4 g).

The physical and chemical data of the compound cynawilfoside A are as follows: white amorphous powder, easily soluble in methanol, ethanol, acetone, chloroform and other organic solvents, the molecular formula is C ₆₃ H ₉₆ O ₂₀ , ¹ H NMR (CDCl ₃ , 500 MHz): δ _H 1.49 (s,3H,H-18),1.20,1.24,1.26,1.26(d,each 3H,J＝6.3 Hz,H-6 of sugar moiety),1.15(s,3H,19-CH ₃ ),1.16( d,3H,J=6.5 Hz,H-21),3.39,3.40,3.41,3.45(s,each 3H,3-OCH ₃ of sugarmoieties),4.76(d,1H,J=9.8,anomeric H),4.77 (brs,1H,anomeric H),4.84(1H,d,J＝9.6 Hz,anomeric H),4.98(brs,anomeric H),5.37(brs,1H,H-6),6.31(d,1H,J ＝16.0Hz,H _Cin-2 ),7.37(m,3H,H _Cin-6,7,8 ),7.51(m,2H,H _Cin-5,9 ),7.63(d,1H,J＝16.0 Hz ,H _Cin-3 ),2.33(m,1H,H _{2′-methyl-butyryl} -2),1.44(m,1H,H _{2′-methyl-butyryl} -3),0.95(t,1H,J=6.5 Hz, H _{2′-methyl-butyryl} -4), 1.17 (d, 1H, J=6.5 Hz, H _{2′-methyl-butyryl} -5); ¹³ C NMR (CDCl ₃ , 125MHz): δ _C 38.9(t ,C-1),29.1(t,C-2),78.0(d,C-3),38.8(t,C-4),139.3(s,C-5),118.6(d,C-6) ,34.6(t,C-7),74.2(s,C-8),43.5(d,C-9),37.1(s,C-10),25.2(t,C-11),73.9(d, C-12),56.3(s,C-13),87.8(s,C-14),34.4(t,C-15),34.6(t,C-16),88.1(s,C-17), 10.4(q,C-18),18.4(q,C-19),73.6(d,C-20),15.1(q,C-21),166.9(s,C _Cin-1 ),119.1(d, C _Cin-2 ), 144.7(d, C _Cin-3 ),134.7(s,C _Cin-4 ),128.2(d,C _Cin-5,9 ),130.3(d,C _Cin-6,8 ),128.9(d,C _Cin-7 ), 174.5(s,C _{2′-methyl-butyryl} -1),41.5(d,C _{2′-methyl-butyryl} -2),26.4(t,C _{2′-methyl-butyryl} -3),11.9(q,C _{2′-methyl-butyryl} -4),16.7(q,C _{2′-methyl-butyryl-} 5),95.8(d,C _cym1-1 ),34.2(t,C _cym1-2 ),77.2(d,C _cym1-3 ),81.9(d,C _cym1-4 ),68.7(d,C _cym1-5 ),18.3(q,C _cym1-6 ),57.6(q,C _cym1-OMe ),101.0(d,C _digin-1 ),31.7(t,C _digin-2 ),73.9(d,C _digin-3 ),74.5(d,C _digin-4 ),66.9(d,C _digin-5 ),18.2(q,C _digin-6 ),55.7(q,C _digin-OMe ),99.5(d,C _cym2-1 ),34.4(t,C _cym2-2 ),77.3(d,C _cym2-3 ),81.9(d,C _cym2-4 ),69.2(d,C _cym2-5 ),18.4(q,C _cym2-6 ),57.1(q,C _cym2-OMe ),98.8(d,C _cym3-1 ),31.2(t,C _cym3-2 ),74.9(d,C _cym3-3 ),72.1(d,C _cym3-4 ),65.7(d,C _cym3-5 ),17.6(q,C _cym3-6 ),56.5(q,C _cym3-OMe ).

The protective effect of compound cynawilfoside A on pentylenetetrazole (Pentylenetetrazole)-induced zebrafish seizure model:

The breeding of zebrafish embryos is carried out in the way of natural pair mating. For each mating, 4 to 5 pairs of adult zebrafish that can produce an average of 200 to 300 embryos are prepared. The dead embryos are removed 6 hours and 24 hours after fertilization, and Select the appropriate embryo according to the developmental stage of the embryo, and incubate the embryo with fish culture water at 28°C (water quality for fish culture: add 200 mg of instant sea salt per 1 L of reverse osmosis water, the conductivity is 480-510 μS/cm; pH is 6.9-7.2 ; The hardness is 53.7～71.6mg/LCaCO ₃ ). After the experiment was completed, the zebrafish at various developmental stages were overexposed to tricaine methanesulfonic acid, so that the zebrafish were anesthetized and killed.

Epilepsy model group: zebrafish treated with Pentylenetetrazole; positive control group: Phenytoin Sodium (Phenytoin Sodium) 300 μM; blank group: blank group (1% DMSO), and the rest of the treatments were the same as the model group. After adding the compound to be tested into the microwell plate, use the movement/behavior analyzer to record the movement of the zebrafish within 60 minutes, quantitatively analyze the distance (D) of the fast movement (V>20mm/sec) of the zebrafish, and calculate the Efficiency of anti-epileptic therapy, according to the zebrafish trajectory map, the qualitative evaluation of the anti-epileptic therapeutic effect of the drug. It can be seen from Figure 1A that, compared with the epilepsy model group, the rapid movement trajectory of zebrafish in each concentration group of cynawilfoside A was significantly reduced. Figure 1B is a histogram drawn according to mean ± SE of the average distance of fast movement of zebrafish. It can be seen from Figure 1B that the compound cynawilfoside A has anti-epileptic effects at concentrations of 30, 100, and 300 μM, and the therapeutic efficiencies are 34%, 44.6%, respectively. %, 61.6% (P<0.05, P<0.01, P<0.01), the dose-effect relationship is clear. At the same time, the antiepileptic effect of the positive control drug phenytoin at a concentration of 300 μM was only 57.2% (P<0.01).

Example 2

Evaluation of antiepileptic effect of compound wilfoside C1N (formula IV) in vivo:

The preparation method of Wilfoside C1N: a. 10kg of fresh Auricularia saponis root tuber, sliced and dried to obtain 3Kg dry product, pulverized and extracted three times with 95% ethanol, the extracts were combined, concentrated under reduced pressure to obtain ethanol extract, mixed with silica gel, passed 200-300 mesh silica gel, gradient elution with petroleum ether-acetone (3:1→2:1), and then gradient elution with chloroform-methanol (10:1→5:1) to obtain components Fr1-Fr6. Fr3 was further purified repeatedly by petroleum ether-acetone silica gel column chromatography to obtain pure compound wilfoside C1N (1.5g); b. 10kg of fresh Geshanxiao root, peeled, sliced and dried to obtain 2.7kg dry product, pulverized and diafiltered with 95% ethanol Extract three times, each time for 2-3 days, combine the extracts, concentrate under reduced pressure to obtain the alcohol extract of Geshanxiao, suspend the extract in water, extract with ethyl acetate to obtain the ethyl acetate extract, mix with 100 mesh silica gel The sample was separated by silica gel column chromatography (200-300 mesh silica gel), and chloroform-methanol (10:1→5:1) gradient elution was used to obtain components Fr1-Fr9. Fr6 was further purified repeatedly with petroleum ether-acetone and preparative HPLC to obtain pure compound wilfoside C1N (2.5 g).

The physical and chemical data of compound wilfoside C1N are as follows: white amorphous powder, easily soluble in methanol, ethanol, acetone, chloroform and other organic solvents, molecular formula is C ₅₆ H ₉₀ O ₁₉ , ESIMS m/z 1089[M+Na] ⁺ ; ¹ H NMR(pyridine-d ₅ ,300MHz):δ _H 0.95,0.97(d,each 3H,J＝6.5Hz,H _Ikem-5,6 ),1.32(s,3H,H-19),1.36,1.37,1.40 ,1.48(d,each 3H,J＝6.0Hz,H-6 of sugar moieties),1.96(s,3H,H-18),2.29(s,3H,H _Ikem-7 ),2.49(s,3H, H-21),3.45,3.48,3.52,3.53(s,each 3H,3-OCH ₃ of sugarmoieties),5.02(brd,1H,J＝3.0 Hz,anomeric H),5.09(d,1H,J＝9.3 Hz, anomeric H), 5.15 (brd, 1H, J＝3.0 Hz, anomeric H), 5.22 (d, 1H, J＝9.0 Hz, anomeric H), 5.30 (brs, 1H, H-6), 5.84 (br s,1H,H _Ikem-2 ); ¹³ C NMR (Pyridine-d ₅ , 100 MHz): δ _C 39.2(t,C-1), 29.9(t,C-2), 77.6(d,C-3 ),38.9(t,C-4),139.3(s,C-5),119.2(d,C-6),33.8(t,C-7),74.2(s,C-8),44.5(d ,C-9),37.4(s,C-10),25.0(t,C-11),72.5(d,C-12),57.9(s,C-13),89.4(s,C-14) ,34.8(t,C-15),32.9(t,C-16),92.4(s,C-17),10.7(q,C-18),18.4(q,C-19),209.4(s, C-20),27.5(q,C-21),166.0(s,C _Ikem-1 ),114.1(d,C _Ikem-2 ),165.5(s,C _Ikem-3 ),38.1(s,C _Ikem -3 _-4 ),20.8(q,C _Ikem-5 ),20.9(q,C _Ikem-6 ),16.4(q,C _Ikem-7 ),96.1(d,C _cym1-1 ),35.3(t,C _{cym1 -2} ),77.6 (d,C _cym1-3 ),82.4(d,C _cym1-4 ),69.2(d,C _cym1-5 ),18.7(q,C _cym1-6 ),57.3(q,C _cym1-OMe ),100.9 (d,C _digin-1 ),32.5(t,C _digin-2 ),73.8(d,C _digin-3 ),74.6(d,C _digin-4 ),67.4(d,C _digin-5 ),17.9 (q,C _digin-6 ),55.3(q,C _digin-OMe ),99.4(d,C _cym2-1 ),36.3(t,C _cym2-2 ),77.8(d,C _cym2-3 ),82.3 (d,C _cym2-4 ),69.4(d,C _cym2-5 ),18.6(q,C _cym2-6 ),58.2(q,C _cym2-OMe ),99.0(d,C _cym3-1 ),32.1 (t,C _cym3-2 ),76.4(d,C _cym3-3 ),73.3(d,C _cym3-4 ),66.4(d,C _cym3-5 ),18.1(q,C _cym3-6 ),56.6 (q,C _cym3-OMe ).

The protective effect of the compound wilfoside C1N on the zebrafish seizure model induced by Pentylenetetrazole:

According to the method in Example 1, the antiepileptic therapeutic effect of the compound was qualitatively evaluated. It can be seen from Figure 2A that when wilfoside C1N was administered at a dose of 300 μM, compared with the epilepsy model group, the fast movement trajectory of zebrafish was significantly reduced. Figure 2B is a histogram drawn according to mean ± SE of the average distance of fast movement of zebrafish. It can be seen from Figure 2B that the compound wilfoside C1N has a tendency to treat epilepsy at concentrations of 30 and 100 μM, but there is no statistical difference; at a concentration of 300 μM , the epilepsy treatment efficiency was 46.4%, (P<0.01), and the dose-effect relationship was clear.

Example 3

Evaluation of compound wilfoside K1N (formula V) antiepileptic effect in vivo:

The preparation method of Wilfoside K1N: a. 10kg of fresh cowhide tuber, sliced and dried to obtain 3kg of dry product, pulverized and extracted three times with 95% ethanol, the extracts were combined, concentrated under reduced pressure to obtain ethanol extract, mixed with silica gel, passed 200- 300 mesh silica gel, gradient elution with petroleum ether-acetone (3:1→2:1), and then gradient elution with chloroform-methanol (10:1→5:1) to obtain fractions Fr1-Fr6. Fr3 was further purified repeatedly by petroleum ether-acetone silica gel column chromatography to obtain pure compound wilfoside K1N (2.5g); b. 10kg of fresh Geshanxiao root, peeled and sliced to obtain 2.7kg of dry product, crushed and percolated with 95% ethanol Extract three times, each time for 2-3 days, combine the extracts, concentrate under reduced pressure to obtain the alcohol extract of Geshanxiao, suspend the extract in water, extract with ethyl acetate to obtain the ethyl acetate extract, mix with 100 mesh silica gel The sample was separated by silica gel column chromatography (200-300 mesh silica gel), and chloroform-methanol (10:1→5:1) gradient elution was used to obtain components Fr1-Fr9. Fr7 was further purified repeatedly with petroleum ether-acetone and preparative HPLC to obtain pure compound wilfoside K1N (1.7 g).

The physical and chemical data of compound wilfoside K1N are as follows: white amorphous powder, easily soluble in methanol, ethanol, acetone, chloroform and other organic solvents, molecular formula is C ₅₈ H ₈₆ O ₁₉ , ESIMS m/z 1109[M+Na] ⁺ ; ¹ H NMR(pyridine-d ₅ ,300MHz):1.36(s,3H,H-19),1.37,1.38,1.42,1.49(d,each 3H,J＝6.0Hz,H-6 of sugarmoieties),2.02(s, 3H,H-18),2.54(s,3H,H-21),3.42,3.44,3.53,3.54(s,each 3H,3-OCH ₃ of sugar moieties),4.98(brd,1H,J＝3.0 Hz ,anomeric H),5.09(d,1H,J＝8.0Hz,anomeric H),5.15(brd,1H,J＝3.0 Hz,anomeric H),5.23(d,1H,J＝9.0Hz,anomericH),5.32 (brs,1H,H-6),6.81(d,1H,J＝16.0 Hz,H _Cin-2 )7.38(m,3H,H _Cin-6,7,8 ),7.65(m,2H,H _{Cin -5,9} ),7.99(d,1H,J＝16.0 Hz,H _Cin-3 ); ¹³ C NMR(Pyridine-d ₅ ,100 MHz):δ _C 39.2(t,C-1),29.9(t ,C-2),77.6(d,C-3),38.9(t,C-4),139.3(s,C-5),119.2(d,C-6),33.8(t,C-7) ,74.5(s,C-8),44.5(d,C-9),37.4(s,C-10),25.0(t,C-11),73.6(d,C-12),58.1(s, C-13),89.5(s,C-14),34.7(t,C-15),33.0(t,C-16),92.4(s,C-17),10.7(q,C-18), 18.6(q,C-19),209.9(s,C-20),27.5(q,C-21),165.8(s,C _Cin-1 ),119.2(d,C _Cin-2 ),145.0(d ,C _Cin-3 ),135.0(s,C _Cin-4 ),128.6(d,C _Cin-5,9 ),129.3(d,C _Cin-6,8 ),130.6(d,C _Cin-7 ) ,96.1(d,C _cym1-1 ),35.3(t,C _cym1-2 ),77.5(d,C _cym1-3 ),82.5(d,C _cym1-4 ),69.2(d,C _cym1-5 ),18.7(q,C _cym1-6 ),57.3(q,C _cym1-OMe ), 100.9(d,C _digin-1 ),32.5(t,C _digin-2 ),73.8(d,C _digin-3 ),74.6(d,C _digin-4 ),67.4(d,C _digin-5 ),17.9(q,C _digin-6 ),55.3(q,C _digin-OMe ),99.4(d,C _cym2-1 ),36.3(t,C _cym2-2 ),77.8(d,C _cym2-3 ),82.3(d,C _cym2-4 ),69.4(d,C _cym2-5 ),18.7(q,C _cym2-6 ),58.3(q,C _cym2-OMe ),99.0(d,C _cym3-1 ),32.1(t,C _cym3-2 ),76.4(d,C _cym3-3 ),73.2(d,C _cym3-4 ),66.4(d,C _cym3-5 ),18.2(q,C _cym3-6 ), 56.6 (q, C _cym3-OMe ).

The protective effect of the compound wilfoside K1N on the zebrafish seizure model induced by Pentylenetetrazole:

According to the method in Example 1, the antiepileptic therapeutic effect of the compound was qualitatively evaluated. It can be seen from Figure 3A that when wilfoside K1N was administered at a dose of 100 μM, compared with the epilepsy model group, the fast movement trajectory of zebrafish was significantly reduced. Figure 3B is a histogram based on mean ± SE of the average distance of zebrafish fast movement. It can be seen from Figure 3B that the compound wilfoside K1N has a tendency to treat epilepsy at concentrations of 10 and 30 μM, but there is no statistical difference; at a concentration of 100 μM It has anti-epileptic effect, and the epilepsy treatment efficiency is 49% (P<0.01), and the dose-effect relationship is clear.

Example 4

Evaluation of antiepileptic effect of compound taiwanoside C (formula III) in vivo:

The preparation method of Taiwanoside C: a. 10kg of fresh cowhide tuber root, sliced and dried to obtain 3kg of dry product, pulverized and extracted three times with 95% ethanol, the extracts were combined, concentrated under reduced pressure to obtain ethanol extract, mixed with silica gel, passed 200- 300 mesh silica gel, gradient elution with petroleum ether-acetone (3:1→2:1), and then gradient elution with chloroform-methanol (10:1→5:1) to obtain fractions Fr1-Fr6. Fr4 was further purified repeatedly by chloroform-methanol silica gel column chromatography to obtain the pure compound taiwanoside C (80.4 mg); b. 10 kg of fresh Geshanxiao root, peeled, sliced and dried to obtain 2.7 kg of dry product, crushed and extracted by diafiltration with 95% ethanol Three times, 2 to 3 days each time, combine the extracts, concentrate under reduced pressure to obtain the alcohol extract of Geshanxiao, suspend the extract in water, extract with ethyl acetate to obtain ethyl acetate extract, mix the sample with 100 mesh silica gel , separated by silica gel column chromatography (200-300 mesh silica gel), and eluted with a gradient of chloroform-methanol (10:1→5:1) to obtain components Fr1-Fr9. Fr6 was further purified repeatedly with petroleum ether-acetone and preparative HPLC to obtain pure compound taiwanoside C (640 mg).

The physical and chemical data of compound taiwanoside C are as follows: white amorphous powder, easily soluble in methanol, ethanol, acetone, chloroform and other organic solvents, molecular formula is C ₄₉ H ₈₀ O ₁₈ , ESIMS m/z 979[M+Na] ⁺ ; ¹ H NMR(CDCl ₃ ,300 MHz):δ _H 1.14(s,3H,H-18),1.20,1.21,1.22,1.24(d,each 3H,J＝6.3 Hz,H-6 of sugarmoieties),1.27(s ,3H,H-19),2.34(s,3H,H-21),3.25,3.30(dd,each 1H,J＝9.6,2.7 Hz,H _cym-4 ),3.37,3.40,3.41,3.45(s ,each 3H,3-OCH ₃ of sugar moieties),3.70,3.77(q,each 1H,J＝3.0 Hz,H _cym-3 ),3.83,3.86,4.04(dq,each 1H,J＝8.9,6.4 Hz ,H _cym-5 ),3.85(brs,1H,H _digin-4 ),3.96(brq,1H,J＝6.3 Hz,H _digin-5 ),4.75(d,1H,J＝9.8,anomeric H), 4.77 (brs, 1H, anomeric H), 4.81 (1H, d, J = 9.6 Hz, anomeric H), 4.97 (brd, J = 3.0 Hz, anomeric H), 5.36 (brs, 1H, H-6). ¹³ C NMR (Pyridine-d ₅ , 125 MHz): δC 39.4(t, _C -1), 30.3(t,C-2), 78.3(d,C-3), 39.4(t,C-4) ,139.7(s,C-5),119.7(d,C-6),34.2(t,C-7),74.7(s,C-8),44.9(d,C-9),37.8(s, C-10),25.3(t,C-11),74.0(d,C-12),58.7(s,C-13),89.9(s,C-14),35.2(t,C-15), 33.3(t,C-16),92.8(s,C-17),10.9(q,C-18),18.6(q,C-19),210.7(s,C-20),28.0(q,C -21),170.4(s,C _Ac-1 ),21.2(q,C _Ac-2 ),96.6(d,C _cym1-1 ),35.7(t,C _cym1-2 ),78.0(d,C _{cym1 -3} ),82.8(d,C _cym1-4 ),69.7(d,C _cym1-5 ),19.2(q,C _cym1-6 ),57.7(q,C _cym1-OMe ),101.4(d,C _{digin -1} ),32.9(t,C _digin-2 ),74.3(d,C _digin-3 ),75.0(d,C _digin-4 ),67.9(d,C _digin-5 ),18.3(q,C _{digin -6} ),55.8(q,C _digin-OMe ),99.8(d,C _cym2-1 ),36.8(t,C _cym2-2 ),78.1(d,C _cym2-3 ),82.8(d,C _{cym2 -4} ),69.9(d,C _cym2-5 ),18.9(q,C _cym2-6 ),58.4(q,C _cym2-OMe ),99.5(d,C _cym3-1 ),32.6(t,C _{cym3 -2} ),76.9(d,C _cym3-3 ),73.7(d,C _cym3-4 ),66.9(d,C _cym3-5 ),19.1(q,C _cym3-6 ),57.0(q,C _{cym3 -OMe} ).

The protective effect of compound taiwanoside C on pentylenetetrazole (Pentylenetetrazole)-induced zebrafish seizure model:

According to the method in Example 1, the antiepileptic therapeutic effect of the compound was qualitatively evaluated. It can be seen from Figure 4A that compared with the epilepsy model group, the rapid movement trajectory of zebrafish was significantly reduced at each dose of taiwanoside C. Figure 4B is a histogram drawn according to mean ± SE of the average distance of fast movement of zebrafish. It can be seen from Figure 4B that the compound taiwanoside C has an epilepsy treatment efficiency of 28.1% at a concentration of 100 μM, and there is no statistical difference; at concentrations of 300 and 1000 μM Both of them have antiepileptic effect, and the therapeutic efficiency is 48.5% and 84.8% respectively (P<0.01, P<0.01), and the dose-effect relationship is clear.

Example 5

Evaluation of antiepileptic effect of compound otophylloside F (formula I) in vivo:

The preparation method of otophylloside F: dry 5 kg of otophylloside F, pulverize and extract three times with 95% ethanol percolation, each time for 2 to 3 days, combine the extracts, concentrate under reduced pressure to obtain otophylloside F Alcoholic extract, suspending the extract in water, extracting with ethyl acetate to obtain ethyl acetate extract, C-18 reverse phase column chromatography with methanol aqueous solution (10% ~ 100%) gradient elution, to obtain the component Fr1 - Fr5, Fr4 were further purified repeatedly by preparative HPLC to obtain the pure compound otophylloside F (1.1 g).

The physical and chemical data of the compound qingyang shenxi is as follows: white amorphous powder, easily soluble in methanol, ethanol, acetone, chloroform and other organic solvents, the molecular formula is C ₆₃ H ₉₆ O ₂₀ , ¹ H NMR (CDCl ₃ , 400 MHz): δ _H 1.04,1.06(d,each 3H,J=6.5Hz,H _Ikem-5,6 ),1.11(s,3H,H-19),1.24,1.26,1.33(d,each 3H,J=6.0Hz ,H-6 of sugarmoieties),1.40(s,3H,H-18),2.11(s,3H,H _Ikem-7 ),2.16(s,3H,H-21),3.38,3.45(s,each3H, 3-OCH ₃ of sugar moieties), 4.49 (dd, 1H, J＝2.0, 9.6 Hz, anomeric H), 4.81 (dd, 1H, J＝2.2, 9.3 Hz, anomeric H), 4.92 (dd, 1H, J =2.2,9.6 Hz, anomeric H),5.35(brs,1H,H-6),5.51(br s,1H,H _Ikem-2 ); ¹³ C NMR(CDCl ₃ ,125 MHz):δ _C 38.9(t ,C-1),29.0(t,C-2),78.0(d,C-3),38.9(t,C-4),140.8(s,C-5),117.7(d,C-6) ,34.3(t,C-7),74.4(s,C-8),43.8(d,C-9),37.3(s,C-10),24.4(t,C-11),71.7(d, C-12),58.0(s,C-13),88.1(s,C-14),33.2(t,C-15),32.0(t,C-16),91.6(s,C-17), 9.5(q,C-18),18.7(q,C-19),209.0(s,C-20),27.3(q,C-21),166.0(s,C _Ikem-1 ),113.1(d, C _Ikem-2 ),166.1(s,C _Ikem-3 ),38.3(s,C _Ikem-4 ),20.9(q,C _Ikem-5 ),21.0(q,C _Ikem-6 ),16.7(q, C _Ikem-7 ),95.9(d,C _digit-1 ),37.2(t,C _digit-2 ),75.5(d,C _digit-3 ),82.6(d,C _digit-4 ),68.2(d, C _digit-5 ),18.3(q,C _digit-6 ),98.5(d ,C _cym-1 ),35.7(t,C _cym-2 ),77.0(d,C _cym-3 ),82.5(d,C _cym-4 ),68.8(d,C _cym-5 ),18.1(q ,C _cym-6 ),58.5(q,C _cym-OMe ),101.6(d,C _ole-1 ),35.4(t,C _ole-2 ),80.7(d,C _ole-3 ),75.5(d , C _ole-4 ), 71.6 (d, C _ole-5 ), 18.1 (q, C _ole-6 ), 56.4 (q, C _ole-OMe ).

The protective effect of the compound qingyangshenxi on the zebrafish seizure model induced by Pentylenetetrazole:

According to the method in Example 1, the antiepileptic therapeutic effect of the compound was qualitatively evaluated. It can be seen from Fig. 5A that, compared with the epilepsy model group, the rapid movement trajectory of zebrafish was significantly reduced at each dose of Qingyangshenside. Figure 5B is a histogram based on mean ± SE of the average distance of zebrafish fast movement. It can be seen from Figure 5B that the compound qingyangshenside has a tendency to treat epilepsy at concentrations of 30 and 100 μM, but there is no statistical difference; It has anti-epileptic effect at the concentration of 300μM, and the epilepsy treatment efficiency is 55% (P<0.01), and the dose-effect relationship is clear.

Example 6

Compound rusutamine 3-O-β-D-oleandopyranosyl-(1→4)-β-D-canapyranosyl-(1→4)-β-D-canapyranosyl Sugar [rostratamine 3-O-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-cymaropyranoside] (formula II) (hereinafter referred to as "compound II") in vivo Evaluation of antiepileptic effect:

The preparation method of compound II: dry 5 kg of Qingyang ginseng tuber, pulverize and extract three times with 95% ethanol percolation, each time for 2 to 3 days, combine the extracts, concentrate under reduced pressure to obtain the alcoholic extract of Qingyang ginseng, and extract Suspended in water, extracted with ethyl acetate to obtain ethyl acetate extract, C-18 reverse phase column chromatography was eluted with methanol aqueous solution (10% ~ 100%) gradient to obtain components Fr1-Fr5, Fr4 was further prepared by Repeated purification by HPLC gave 100 mg of pure compound II.

rostratamine3-O-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-cymaropyranoside

The physical and chemical data of Compound II are as follows: white needle-like crystals, easily soluble in methanol, ethanol, acetone, chloroform and other organic solvents, the molecular formula is C ₆₃ H ₉₆ O ₂₀ , ¹ H NMR (CDCl ₃ , 400 MHz): δ _H 1.46( s,3H,H-18),1.20,1.22,1.31(d,each 3H,J＝6.3 Hz,H-6 of sugar moiety),1.12(s,3H,19-CH ₃ ),2.19(s,3H ,H-21),3.38,3.44,3.44(s,each 3H,3-OCH ₃ of sugar moieties),4.48(dd,1H,J＝1.9,9.8,anomeric H),4.74(dd,J＝1.9, 9.8, 1H, anomeric H), 4.84 (dd, 1H, J＝1.8, 9.6 Hz, anomeric H), 5.37 (brs, 1H, H-6), 6.30 (d, 1H, J＝16.0 Hz, H _{Cin- 2} ),7.38(m,3H,H _Cin-6,7,8 ),7.51(m,2H,H _Cin-5,9 ),7.62(d,1H,J=16.0 Hz,H _Cin-3 ); ¹³ C NMR (Pyridine-d ₅ , 125 MHz): δC 39.9(t, _C -1), 29.8(t,C-2), 77.6(d,C-3), 38.8(t,C-4) ,139.3(s,C-5),119.2(d,C-6),34.6(t,C-7),74.2(s,C-8),44.5(d,C-9),37.1(s, C-10),25.2(t,C-11),73.9(d,C-12),58.3(s,C-13),89.5(s,C-14),34.7(t,C-15), 33.6(t,C-16),92.1(s,C-17),10.7(q,C-18),18.1(q,C-19),209.9(s,C-20),27.7(q,C -21),165.9(s,C _Cin-1 ),119.1(d,C _Cin-2 ),144.7(d,C _Cin-3 ),135.0(s,C _Cin-4 ),128.6(d,C _Cin -4 _-5,9 ),129.3(d,C _Cin-6,8 ),130.6(d,C _Cin-7 ),96.3(d,C _cym1-1 ),37.9(t,C _cym1-2 ),77.8( d,C _cym1-3 ),83.4(d,C _cym1-4 ),68.7(d,C _cym1-5 ),18 .3(q,C _cym1-6 ),58.6(q,C),100.5(d,C _cym2-1 ),37.2(t,C _cym2-2 ),78.0(d,C _cym2-3 ),83.2( d,C _cym2-4 ),68.9(d,C _cym2-5 ),18.5(q,C _cym2-6 ),58.8(q,C _cym2-OMe ),102.2(d,C _ole-1 ),37.2( d,C _ole-2 ),81.3(d,C _ole-3 ),76.2(d,C _ole-4 ),72.8(d,C _ole-5 ),18.6(d,C _ole-6 ),57.1( d, C _ole-OMe ),

Protective effect of compound II on pentylenetetrazole (Pentylenetetrazole)-induced zebrafish seizure model:

According to the method in Example 1, the antiepileptic therapeutic effect of the compound was qualitatively evaluated. It can be seen from Fig. 6A that compared with the epilepsy model group, the rapid movement trajectory of zebrafish was significantly reduced at each dose of compound II. Figure 6B is a histogram drawn according to mean ± SE of the average distance of fast movement of zebrafish. It can be seen from Figure 6B that Compound II has a tendency to treat epilepsy at a concentration of 30 μM, but there is no statistical difference; It has antiepileptic effect, and the treatment efficiency is 65% and 52% respectively (P<0.01, P<0.01).

Example 7

Evaluation of compound wilfoside C3N (formula VII) antiepileptic effect in vivo:

The preparation method of Wilfoside C3N: dry 10kg of Pseudoides auricularia root, pulverize and extract three times by percolation with 95% ethanol for 2 to 3 days each time, combine the extracts, concentrate under reduced pressure to obtain the alcoholic extract of Pseudosides auricularis, and Suspend the extract in water, extract with ethyl acetate to obtain ethyl acetate extract, dissolve the ethyl acetate extract in 70% ethanol solution, filter or centrifuge to remove insoluble matter, then conduct macroporous resin column chromatography, collect The fractions were eluted with 75% ethanol and concentrated to obtain the active site, which was further purified repeatedly by preparative HPLC to obtain pure Wilfoside C3N.

The physical and chemical data of Wilfoside C3N are as follows: white amorphous powder, easily soluble in methanol, ethanol, acetone, chloroform and other organic solvents, the molecular formula is C ₄₉ H ₇₈ O ₁₆ , ¹ H NMR (Pyridine-d ₅ , 400 MHz): δ _H 0.92,0.94(d,each 3H,J＝6.5Hz,H _Ikem-5,6 ),1.32(s,3H,H-19),1.36,1.34,1.51(d,each 3H,J＝6.0Hz,H -6 ofsugar moieties),1.97(s,3H,H-18),2.26(s,3H,H _Ikem-7 ),2.50(s,3H,H-21),3.42,3.44,3.51(s,each 3H ,3-OCH ₃ of sugar moieties),5.24(dd,1H,J＝1.9,9.4,anomeric H),5.15(d,J＝3.3,1H,anomeric H),5.10(dd,1H,J＝1.8, 9.8 Hz, anomeric H),5.86(br s,1H,H _Ikem-2 ); ¹³ C NMR(Pyridine-d ₅ ,125 MHz):δ _C 39.0(t,C-1),29.9(t,C- 2),77.5(d,C-3),39.3(t,C-4),139.3(s,C-5),119.2(d,C-6),34.8(t,C-7),74.3( s,C-8),44.6(d,C-9),37.4(s,C-10),25.1(t,C-11),72.6(d,C-12),58.0(s,C-13 ),89.5(s,C-14),33.9(t,C-15),33.0(t,C-16),92.4(s,C-17),10.8(q,C-18),18.2(q ,C-19),209.5(s,C-20),27.6(q,C-21),166.0(s,C _Ikem-1 ),114.1(d,C _Ikem-2 ),165.5(s,C _Ikem -2 _-3 ),38.2(s,C _Ikem-4 ),20.9(q,C _Ikem-5 ),20.9(q,C _Ikem-6 ),16.5(q,C _Ikem-7 ),96.1(d,C _{cym1 -1} ),35.3(t,C _cym1-2 ),77.5(d,C _cym1-3 ),82.5(d,C _cym1-4 ),69.2(d,C _cym1-5 ),18.8(q,C _{cym1 -6} ),57.2(q,C),101.0( d,C _digin-1 ),32.5(t,C _digin-2 ),74.5(d,C _digin-3 ),73.8(d,C _digin-4 ),67.5(d,C _digin-5 ),17.9( q,C _digin-6 ),55.3(q,C _digin-OMe ),99.5(d,C _cym2-1 ),35.3(d,C _cym2-2 ),79.9(d,C _cym2-3 ),74.1( d,C _cym2-4 ),71.0(d,C _cym2-5 ),18.7(d,C _cym2-6 ),58.0(d,C _cym2-OMe ).

The protective effect of Wilfoside C3N on the zebrafish seizure model induced by Pentylenetetrazole:

According to the method in Example 1, the antiepileptic therapeutic effect of the compound was qualitatively evaluated. It can be seen from Figure 7A that compared with the epilepsy model group, the rapid movement trajectory of zebrafish was significantly reduced at each dose of wilfoside C3N. Figure 7B is a histogram drawn according to the mean ± SE of the fast moving distance of zebrafish. It can be seen from Figure 7B that wilfoside C3N has a tendency to treat epilepsy at a concentration of 100 μM, but there is no statistical difference; at a concentration of 200 μM, it has antiepileptic effect effect, the treatment efficiency was 55.2% (P<0.001).

Example 8

Evaluation of antiepileptic effect of compound cynauricoside A (formula VIII) in vivo:

The preparation method of Cynauricoside A: dry 10kg of Auricularia pachyrhiza roots, pulverize and extract three times with 95% ethanol percolation, each time for 2 to 3 days, combine the extracts, concentrate under reduced pressure to obtain the alcoholic extract of Cynauricoside, extract Suspended in water, extracted with ethyl acetate to obtain ethyl acetate extract, dissolved ethyl acetate extract in 70% ethanol solution, filtered or centrifuged to remove insoluble matter, then separated by macroporous resin column chromatography, collected 75 The fractions were eluted with % ethanol, concentrated to obtain the active site, and the active site was further purified repeatedly by preparative HPLC to obtain pure cynauricoside A.

The physical and chemical data of Cynauricoside A are as follows: white amorphous powder, easily soluble in methanol, ethanol, acetone, chloroform and other organic solvents, the molecular formula is C ₅₁ H ₇₄ O ₁₆ , ¹ H NMR (Pyridine-d ₅ , 400 MHz): δ _H 2.03(s,3H,H-18),1.37,1.45,1.52(d,each 3H,J＝6.3 Hz,H-6 of sugar moiety),1.35(s,3H,19-CH ₃ ),2.49(s ,3H,H-21),3.52,3.42,3.45(s,each 3H,3-OCH ₃ of sugar moieties),5.24(dd,1H,J＝1.9,9.4,anomeric H),5.15(d,J＝ 3.3, 1H, anomeric H), 5.10 (dd, 1H, J＝1.8, 9.8Hz, anomeric H), 5.30 (brs, 1H, H-6), 6.81 (d, 1H, J＝16.0 Hz, H _{Cin- 2} ),7.34(m,3H,H _Cin-6,7,8 ),7.63(m,2H,H _Cin-5,9 ),8.00(d,1H,J＝16.0 Hz,H _Cin-3 ); ¹³ C NMR(Pyridine-d ₅ ,125MHz): δC 38.9(t, _C -1),29.8(t,C-2),77.6(d,C-3),39.2(t,C-4), 139.2(s,C-5),119.1(d,C-6),34.7(t,C-7),74.2(s,C-8),44.5(d,C-9),37.4(s,C -10),25.0(t,C-11),73.6(d,C-12),58.1(s,C-13),89.5(s,C-14),33.8(t,C-15),33.0 (t, C-16), 92.4 (s, C-17), 10.7 (q, C-18), 18.1 (q, C-19), 209.9 (s, C-20), 27.7 (q, C- 21),165.8(s,C _Cin-1 ),119.2(d,C _Cin-2 ),144.9(d,C _Cin-3 ),135.0(s,C _Cin-4 ),128.6(d,C _Cin -4 _5,9 ),129.3(d,C _Cin-6,8 ),130.6(d,C _Cin-7 ),96.1(d,C _cym1-1 ),35.3(t,C _cym1-2 ),77.5(d ,C _cym1-3 ),82.5(d,C _cym1-4 ),69.2(d,C _{c ym1-5} ),18.8(q,C _cym1-6 ),57.2(q,C),101.0(d,C _digin-1 ),32.5(t,C _digin-2 ),74.5(d,C _digin-3 ),73.8(d,C _digin-4 ),67.5(d,C _digin-5 ),17.9(q,C _digin-6 ),55.3(q,C _digin-OMe ),99.5(d,C digin-OMe ),99.5(d,C _cym2-1 ),35.3(d,C _cym2-2 ),79.9(d,C _cym2-3 ),74.1(d,C _cym2-4 ),71.0(d,C _cym2-5 ),18.7(d,C _cym2-6 ),58.0(d,C _cym2-OMe ).

The protective effect of Cynauricoside A on the zebrafish seizure model induced by Pentylenetetrazole:

According to the method in Example 1, the antiepileptic therapeutic effect of the compound was qualitatively evaluated. It can be seen from Fig. 8A that compared with the epilepsy model group, the rapid movement trajectory of zebrafish was significantly reduced at each dose of cynauricoside A. Fig. 8B is a histogram drawn according to mean±SE of the mean ± SE of the fast movement distance of zebrafish. It can be seen from Fig. 8B that cynauricoside A has a tendency to treat epilepsy at concentrations of 30 and 100 μM, but there is no statistical difference; It has antiepileptic effect, and the treatment efficiency is 57.5% (P<0.01).

Comparative example 1

The compounds qingyangshengenin, caudatin, otophylloside A, otophylloside C, which are isolated and extracted from three plants of the genus A. C), otophylloside F (formula I), otophylloside M (otophylloside M, formula b), caudatin 3-O-β-D-Canadian sesame sugar [caudatin 3-O-β -D-cymaropyranoside, formula IX], Qingyangshen aglycone 3-O-β-D-Canada hemp sugar pyranyl-(1→4)-β-D-digoxigenin [qinyangshengenin3-O-β- D-cymaropyranosyl-(1→4)-β-D-digitoxopyranoside, formula XVII], Kodatine 3-O-β-D-Canada hemp sugar pyranyl-(1→4)-β-D-Canada hemp Sugar [caudatin 3-O-β-D-cymaropyranosyl-(1→4)-β-D-cymaropyranoside, formula X], rusutamine 3-O-β-D-oleander glycopyranosyl-( 1→4)-β-D-Oleandropyranosyl-(1→4)-β-D-Canada [rostratamine 3-O-β-D-oleandropyranosyl-(1→4)-β-D -cymaropyranosyl-(1→4)-β-D-cymaropyranoside, formula II], rusutamine 3-O-α-L-Canada hemp sugar pyranosyl-(1→4)-β-D-Canada hemp Glycopyranosyl-(1→4)-α-L-digi sugarpyranosyl-(1→4)-β-D-Canada hemp sugar [rostratamine3-O-α-L-cymaropyranosyl-(1→ 4)-β-D-cymaropyranosyl-(1→4)-α-L-diginopyranosyl-β-D-cymaropyranoside, formula XIII], Qingyangshen aglycone 3-O-β-D-oleander glycopyran Glycosyl-(1→4)-β-D-canadacanose pyranyl-(1→4)-β-D-digitoxin [qinyangshengenin3-O-β-D-oleandropyranosyl-(1→4) -β-D-cymaropyranosyl-(1→4)-β-D-digitoxopyranoside, formula a], Koudatine 3-O-β-D-glucopyranosyl-(1→4)-β-D- Oleandopyranyl-(1→4)-β-D-Canadasapyranyl-(1→4)-β-D- Canadian sesame sugar [caudatin3-O-β-D-glucopyranoside-(1→4)-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-cymaropyranoside, Formula c], Kodatin 3-O-β-D-xanthopyranosyl-(1→4)-β-D-Canada hempyranosyl-(1→4)-β-D- Canadian sesame sugar [caudatin 3-O-β-D-thevetopyranosyl-(1→4)-β-D-cymaropyranosyl-(1→4)-β-D-cymaropyranoside, formula XII], qingyangshen aglycone 3- O-β-D-oleandropyranosyl-(1→4)-β-D-Canada sesame sugar [qinyangshengenin3-O-β-D-oleandropyranosyl-(1→4)-β-D-cymaropyranoside , Formula XI], taiwanoside C (Formula III), wilfoside K1N (Formula V), cynawilfoside A (Formula VI), wilfoside M1N (Formula d), cynauricoside E (Formula XV), cynauricuoside A (Formula XVI), wilfoside G (Formula XIV) Effects on PTZ-induced zebrafish at the maximum non-toxic dose:

According to the method in Example 1, the antiepileptic therapeutic effect of the compound was qualitatively evaluated. It can be seen from Figure 10 that the anti-epileptic effects of monomeric compounds such as formulas I, II, III, V, and VI are better than those of Qingyang ginseng, which are the main components of Qingyang ginseng (qingyangshengenin), caudatin, and Qingyang ginseng. Otophylloside A (otophylloside A), otophylloside C (otophylloside C), formula b compound, formula c compound, formula IX compound, and Qingyang ginseng main components formula a compound, formula b compound, formula c compound and Geshan Xiaozhong The compound of component formula d has the effect of significantly enhancing the epileptic seizures induced by pentylenetetrazol.

The preparation of the extract of embodiment 9 auricularia bougainvillea extract

Add 10 kg of Pseudomonas auricularis medicinal material, add 5 L of 95% ethanol, soak and extract three times, concentrate the extract under reduced pressure to obtain ethanol extract, add water to suspend, extract with 5 times the volume of ethyl acetate, recover ethyl acetate, and dilute the ethyl acetate The extract was dissolved in 3L of 70% ethanol solution, filtered to remove insoluble matter, and subjected to macroporous resin column (resin model ZH801) chromatographic separation, and 75% ethanol eluted fractions were collected and concentrated to obtain 15 g of extract.

After testing, the contents of the anti-epileptic active ingredients wilfoside C1N, wilfoside C3N, wilfoside K1N, and cynauricoside A in the extract were 17.4%, 42.1%, 7.1%, and 14.2%, respectively, and the total content of anti-epileptic active ingredients reached more than 80%. In terms of epilepsy-promoting ingredients, qingyangshenside M, qingyangshen aglycone 3-O-β-D-oleandopyranosyl-(1→4)-β-D-canadasaminopyranosyl- (1→4)-β-D-Digoxigenin was not detected.

The method in Example 1 was used to evaluate the antiepileptic activity of Pseudomonas auricularia extract, and the results showed that at a concentration of 50 μg/mL, there was a tendency for epilepsy treatment, but there was no statistical difference; The treatment efficiency was 61.4% (p<0.001).

The preparation of embodiment 10 Qingyang ginseng extract

Dipping and extracting Qingyang ginseng with 95% ethanol three times, concentrating the extract under reduced pressure to obtain ethanol extract, adding water to suspend, extracting with ethyl acetate, recovering ethyl acetate, dissolving the ethyl acetate extract in 70% ethanol solution , after filtering or centrifuging to remove insoluble matter, perform macroporous resin column chromatography, collect 75% ethanol eluted fractions, and concentrate to obtain the product.

All documents mentioned in this application are incorporated by reference in this application as if each were individually incorporated by reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims (17)

1. A total plant glycoside or a plant extract containing the total plant glycoside, characterized in that, the total plant glycoside or the extract contains anti-epileptic active ingredients and epilepsy-promoting active ingredients, and the extract The content C1 of the anti-epileptic active ingredient in the formula satisfies the following formula B: C1/(C1+C2)≥50％ Formula B In formula B, C1 is the content of antiepileptic active ingredients; C2 is the content of promoting epilepsy active ingredients; Wherein, the anti-epileptic active ingredient is selected from one or more of the following groups: Compounds shown in formulas I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV, XVI The epilepsy-promoting active ingredients are selected from one or more of the following groups: Compounds shown in formula a, b, c and d 2. The phytoglycoside or the plant extract containing the phytoglycoside according to claim 1, wherein C1/(C1+C2)≥80%. 3. The phytoglycoside or the plant extract containing the phytoglycoside according to claim 2, wherein C1/(C1+C2)≥90%. 4. The phytoglycoside or the plant extract containing the phytoglycoside according to claim 2, wherein C1/(C1+C2)≥95%. 5. The phytoglycoside or the plant extract containing the phytoglycoside according to claim 2, wherein C1/(C1+C2) is 96% to 99.5%. 6. The phytoglycoside or the plant extract containing the phytoglycoside according to claim 2, wherein C1/(C1+C2) is 96%, 97%, 98%, 99%, or 99.5%. 7. The total plant glycosides as claimed in claim 1 or the plant extract containing the total plant glycosides, wherein the plant is C. auriculatum Royle ex Wight or C. wilfordii Hemsl.). 8. A pharmaceutical composition, characterized in that it comprises the phytoglycosides or the plant extract containing the phytoglycosides according to claim 1, and optionally a pharmaceutically acceptable carrier or excipient. 9. A method for quality control of anti-epileptic products, comprising the steps of: (a) Determining the content C2 of the epilepsy-promoting active ingredient in the product or its raw materials, and comparing it with the predetermined value C0; (b) When the content C2 is higher than C0, it is determined that the quality of the product is unqualified; when the content C2 is lower than or equal to C0, it is determined that the quality of the product is qualified; The epilepsy-promoting active ingredients are selected from one or more of the following groups: Compounds shown in formula a, b, c and d 10. The method of claim 9, wherein the anti-epileptic product comprises an anti-epileptic drug containing an extract of Cyan ginseng. 11. The method according to claim 9, wherein the C2 is the weight percentage of the epilepsy-promoting active ingredient in the product or raw material. 12. The method of claim 9, wherein said C0 < 10% (w/w). 13. The method of claim 9, wherein said C0 < 5% (w/w). 14. The method of claim 9, wherein said C0 < 1% (w/w). 15. The method of claim 9, wherein the CO is 0.01% to 10% (w/w). 16. The method of claim 9, wherein the CO is 0.01%, 0.05%, 0.1%, 0.2%, 0.5%, 1.0%, 5%, or 10%. 17. The use of a compound selected from the following group in the preparation of a medicament or a reagent, which is used to construct an animal epilepsy model: compounds shown in formulas a, b, c and d: