CN103183719B - Saponin derivative and uses thereof - Google Patents

Abstract

A saponin derivative, its structure is shown in the following formula. Animal experiments have proved that the saponin derivatives have significant antidepressant pharmacological activity, and can be used as active ingredients to make medicines, foods and health products, and can be used alone or in combination with other substances to prevent and treat depression.

Description

Saponin derivatives and uses thereof

technical field

The invention relates to a saponin derivative, in particular to a furostane-type saponin derivative, and the application of the derivative in preventing and treating depression.

Background technique

Anemarrhena (also known as rabbit oil grass, Chuandilong) is the dry rhizome of Anemarrhena asphodeloides, a plant of the Liliaceae family. According to "Flora", it is bitter and cold in nature, and has the effects of nourishing yin and reducing fire, moistening dryness and smoothing intestines, and facilitating defecation.

The main chemical components of Chinese herbal medicine Anemarrhena include steroidal saponins, diphenylpyrone, polysaccharides and lignin, such as: timosaponin (timosaponin) A-Ⅰ, A-Ⅱ, A- Ⅲ, A-Ⅳ, B-Ⅰ, B-Ⅱ and B-Ⅲ, among them, the structures of timosaponin A-Ⅱ and A-Ⅳ are still unknown. And timosaponin (amemarsaponin) A2, that is, marcosapogenin-3-O-β-D-glucopyranosyl (1→2)-β-D-galactopyranoside B (marlogenin-3-O- β-D-glucopyranosy (1→2)-β-D-galactopyranoside B), desgalactotigonin, F-gitonin and smilageninoside, etc. In addition, it also contains anemaran A/B/C/D, cis-hinokiresinol, monomethyl-cis-hinokiresinol, oxidized-cis- Oxy-cis-himokiresinol, 2,6,4'-trihydroxy-4-methoxybenzophenone (2,6,4'-trihydroxy-4-methoxybenzophenone), p-hydroxyphenyl p-hydroxyphenyl crotonic acid, pentacosyl vinyl ester, β-sitosterol, mangiferin, nicotinic acid, nicotinamide ) and pantothenic acid (pantothenic acid) and so on.

Timosaponin B-II can improve learning and memory impairment caused by cerebral ischemia (Neuroscience letter., 2007, 421, 147-151). Timosaponin A-Ⅲ can relieve memory loss mainly by inhibiting acetylcholinesterase and the expression of inflammatory factors (Pharmacol. Biochem. Be., 2009, 93, 121-127).

Many studies have proved that these substances contained in Anemarrhena have therapeutic effects on various diseases, such as: Anemarrhena total saponins can be used to treat myocardial ischemia (CN1682873A), mainly timosaponins A-Ⅲ and B-Ⅱ Medicaments made from the composition have also been used in the treatment of thrombotic disorders (WO2011/026259). Timosaponin A-Ⅲ can induce the apoptosis of human cervical cancer cells, and it has been proved to have the effect of treating colon and rectal cancer in vivo and in vitro (Cancer Res., 2008, 68, 10229-10237), and has the function of inhibiting blood vessels. It has a remarkable effect on the proliferation of endothelial cells and has a therapeutic effect on the treatment of diseases such as tumors and rheumatoid arthritis (CN102030812A).

Recent related studies have also shown that Anemarrhena total saponin has antidepressant effects in various depression models, which may be related to its enhancement of noradrenaline and serotonergic nervous systems (New Drugs and Clinical Pharmacology of Traditional Chinese Medicine, 2007, 18, 29) . Through animal experiments, the smilagenin contained in Anemarrhena has certain effects on experimental depression in mice, and can affect the activity of dopamine and monoamine oxidase in the mouse brain, so that this type of saponin has antidepressant activity (Biol. Pharm. Bull., 2006, 29, 2304-2306). Yijia et al. found that timosaponin B-II has antidepressant activity, and its mechanism of action may be related to the enhancement of 5-HT and DA nervous system in the brain (CN101214253A; Journal of Pharmaceutical Practice, 2010, 28, 283-287 ).

Traditional Chinese medicine believes that depression is mostly caused by excessive worry and fatigue, and the treatment should focus on calming the mind and calming the mind, nourishing blood and nourishing yin. Therefore, in most of the prescriptions used in traditional Chinese medicine to treat depression, Anemarrhena is also a commonly used Chinese herbal medicine. The commonly used antidepressants in clinical practice include tricyclic and tetracyclic antidepressants, monoamine oxidase inhibitors, selective serotonin reuptake inhibitors (SSRIs), atypical antidepressants and lithium salts.

In recent years, fluoxetine, the most widely used SSRI drug in clinical practice, has many adverse reactions, such as: systemic or local allergy, gastrointestinal dysfunction, etc. (Chinese Modern Applied Pharmacy, 2008, 25, 763-765; China Journal of New Drugs, 2001, 10, 20; Journal of Practical Medicine, 2004, 20, 318-319). Studies have pointed out that young children who take fluoxetine may increase suicidal tendencies, and it has also been reported that taking fluoxetine may cause increased violent tendencies and congenital cardiovascular birth defects.

SSRI drugs——Sertraline (Sertraline) is a brand-name antidepressant drug with a large number of prescriptions in the United States, but it is easy to cause sinus tachycardia and ST-T changes in patients (Chinese Journal of New Drugs and Clinics, 2002, 21, 711 -713) adverse reactions, in the process of treating depressive episodes, it may cause manic episodes or turn manic episodes to a certain extent (Foreign Medical Psychiatry Volume, 2004, 3l, 104).

Although venlafaxine and clomipramine are equally well tolerated, about 1/3 of the drug users will experience nausea, especially in the first few weeks of starting the drug. The adverse reactions caused by venlafaxine have been found to include malignant syndrome, induced mania, purpura, hallucinations, hallucinations caused by sudden arrest, withdrawal reactions, 5-HT syndrome, chest tightness, dizziness, hypotension, menopause and ocular Elevated blood pressure, etc., have a higher rate of dysphoria in children (Journal of Clinical Psychiatry, 2002, 12, 382).

According to the forecast of the World Health Organization experts, the total social burden of depression will rise to the first place among all disease burdens by 2020, and the active molecules contained in the active drugs used to treat depression are mainly obtained by chemical synthesis , Most of them have problems such as high toxicity and side effects, narrow antidepressant spectrum and high price. Only 2% of patients with depression are currently receiving treatment in our country, which has the potential to cause great harm to families and society. Therefore, it is particularly necessary to develop safe, effective and inexpensive innovative antidepressant drugs.

Traditional Chinese medicine and natural medicine are one of the important sources of innovative drug research and development. In recent years, international and domestic research and development of natural medicines such as Hypericum perforatum have achieved a series of results (Modern Chinese Medicine, 2009, 11, 6-9; Chinese Patent Medicine, 2006, 28, 713-716; Guangdong Medicine, 2005, 859-860; Journal of Nanjing University of Traditional Chinese Medicine, 2001, 17, 294-298; Journal of Pharmaceutical Sciences of Jiefang Bureau, 2003, 19, 426-428; Journal of Pharmaceutical Practice, 2004, 22, 104-105; Pharm.Biol ., 2006, 44, 503-510). But so far no new traditional Chinese medicine for the treatment of depression has been listed, and there are few research reports on the relationship between chemical structure and pharmacodynamic mechanism in this regard.

Contents of the invention

An object of the present invention is to provide a steroidal saponin derivative, which has an antidepressant pharmacological effect, and can be used as an active ingredient for preparing medicines for treating and preventing depression.

Another object of the present invention is to provide a furostane-type saponin derivative, which has an antidepressant pharmacological effect and can be used as an active ingredient to prepare medicines for treating and preventing depression.

Another object of the present invention is to provide a furostane-type saponin derivative prepared from timosaponin B-III or from the Chinese medicinal material Anemarrhena, which has antidepressant pharmacological effects.

Another object of the present invention is to provide a composition, which uses the provided saponin derivatives as active ingredients to make medicines, foods and health products, which are used alone or in combination with other substances for the prevention and treatment of depression.

The term "saponin derivatives" referred to in the present invention refers to saponin derivatives and their pharmaceutically acceptable salts, such as: but not limited to hydrochloride, sulfate, carbonate, bicarbonate, acetate, Potassium salt, sodium salt, calcium salt, magnesium salt, barium salt, manganese salt, zinc salt, iron salt and quaternary ammonium salt, etc.

The "furostane-type saponin derivatives" referred to in the present invention refers to furostane-type saponin derivatives and their pharmaceutically acceptable salts, such as: but not limited to hydrochloride, sulfate, carbonate, Bicarbonate, acetate, potassium salt, sodium salt, calcium salt, magnesium salt, barium salt, manganese salt, zinc salt, iron salt and quaternary ammonium salt, etc.

The "furostane-type saponin derivatives obtained from the Chinese herbal medicine Zhimu" in the present invention refers to compounds formed from the derivatives and various inorganic acids, inorganic bases, organic acids or organic bases, including In the process of preparing furostane-type saponin derivatives from the medicinal material Anemarrhena, the furostane-type saponin derivatives and their direct or indirect precursor compounds (such as: timosaponin B-Ⅲ) are related to the preparation Take the pharmaceutically acceptable salts formed by the various reagents used, such as: hydrochloride, sulfate, carbonate, bicarbonate, acetate, sodium salt, potassium salt or quaternary ammonium salt, etc.

The term "Anemarrhena asphodeloides" in the present invention refers to the various parts of Anemarrhena asphodeloides, such as roots, stems, leaves, and fruits. Since the rhizomes of this plant can produce the most target Saponin derivatives, so the present invention preferably selects its dried rhizomes. Whether the material selection and processing process of these Anemarrhena medicinal materials conform to the processing technology and standards of Chinese medicinal materials shall not limit the present invention.

The "depression" referred to in the present invention belongs to mood disorders, and is a syndrome characterized by significant and persistent depression. Depression is usually referred to as clinical major depression (Chinese Journal of Experimental Formulas, 2005, 11, 46; Journal of Pharmaceutical Practice, 2008, 26, 282). Patients with depression are often accompanied by anxiety symptoms, and anxiety can aggravate the development of depression (China Clinical Rehabilitation, 2006, 10, 58). The onset of depression is mainly characterized by low mood, which is not commensurate with the situation, and can range from gloomy to distraught, and even stupor. In severe cases, psychotic symptoms such as hallucinations and delusions may appear. Anxiety and motor agitation were prominent in some cases. Its diagnostic criteria are also mainly depressed mood, accompanied by at least 4 of the following: (1) loss of interest, no sense of pleasure; (2) loss of energy or fatigue; (3) psychomotor retardation or agitation; (4) Low self-evaluation, self-blame, or guilt; (5) Difficulty associating or decline in conscious thinking ability; (6) Recurring thoughts of death or suicide or self-injury behavior; (7) Sleep disturbance, Such as insomnia, early awakening, or excessive sleep; (8) decreased appetite or significant weight loss; and (9) decreased libido. Impairment of social functions, causing pain or adverse consequences to the person is a serious standard for the development of the disease. From the perspective of its course of disease, the patient has met the symptom criteria and severe criteria for at least 2 weeks; some schizoid symptoms may be present, but the diagnosis of schizophrenia is not met. If the symptoms of schizophrenia are met at the same time, after the symptoms of schizophrenia are relieved, the criteria for depressive episodes should be met for at least 2 weeks (Chinese Classification and Diagnostic Criteria of Mental Disorders [M], Jinan: Shandong Science and Technology Press, 2001, 28-35).

"Anti-depression" in the present invention refers to the purpose of treating, improving or curing the symptoms of patients with depression, so that the patients' conditions can be improved so as to return to normal.

The "forced swimming test (Forced Swimming Test, FST)" or "forced swimming animal model of depression" referred to in the present invention refers to putting experimental animals (such as: mice) in a limited environment (such as: water ), the animal struggled desperately in this environment and tried to escape but could not escape, thus providing an unavoidable oppressive environment. After a period of experiment, the animal showed a typical "immobility state", in fact, the animal gave up the escape. Hope, which reflects a so-called "behavioral hopelessness state", records a series of parameters in the process of producing hopeless immobility in animals in this environment. The system uses a camera to record the experimental process and uses corresponding software to track and analyze the images, reducing human operations and making the data objective. The forced-swimming animal model of depression is a reliable experimental model for the study of human depression pharmacology and its pathogenesis, and for screening and observation of antidepressant drugs. Its main feature is the high specificity of drug action. Antidepressants are distinguished from strong tranquillizers and anxiolytics, and the effects produced by most antidepressants are significantly related to clinical potency (Arch. Int. Pharmacodyn. Ther., 1977, 229, 327-336; Neurosei. Biobehav. Rev., 1995, 19, 377-395).

The "Tail Suspension Test (TST)" referred to in the present invention is a classic method that can quickly evaluate the efficacy of antidepressant drugs. The principle is to use the mouse to try to escape after suspending its tail but cannot escape, so that it gives up struggling and enters a unique state of depression and immobility. During the experiment, the immobility time of the animals is recorded to reflect the state of depression. state.

The "organism", "animal" or "patient" referred to in the present invention refers to human beings, wild animals and livestock (Livestock). Wild animals are animals that have not been domesticated in their natural state. Livestock are animals raised in captivity to provide a source of food, such as, but not limited to, dogs, cats, rats, rats, hamsters, pigs, rabbits, cows, buffaloes, bulls, sheep, goats, geese, and chickens. The "patient", "animal" or "organism" to be treated is preferably a mammal, especially a human.

The "prevention" referred to in the present invention refers to various means or measures used to prevent the occurrence or development of diseases before they are identified by clinical standards, including medical, physical or chemical methods, in order to prevent and reduce various diseases. Onset or progression of symptoms.

"Treatment" referred to in the present invention means to prevent and reduce the occurrence or development of the disease, so that the development or aggravation of the disease course can be inhibited, curbed, alleviated, improved, slowed down, stopped, delayed or reversed, the described maintenance and Various indicators of a disease, disorder or pathological state when administered include alleviation or reduction of symptoms or complications, or cure or elimination of the disease, disorder or condition, and/or administration.

"Food" in the present invention refers to an edible single compound or composition including various saponin derivatives provided by the present invention. The production and manufacture of the single compound or composition should comply with relevant food safety standards, but these food safety standards should not limit the present invention.

The "health product" referred to in the present invention refers to the preparation of various saponin derivatives provided by the present invention into edible single compounds or compositions to be administered to patients for the purpose of preventing and treating diseases. It belongs to the food mentioned in the present invention, but its production, manufacture and sales should also meet various relevant requirements, standards and norms.

The term "medicine" in the present invention refers to a single compound, a composition formed by multiple compounds, Chinese medicinal materials and their extracts, or a composition with a single compound as the main active ingredient, which can be used to prevent or treat a certain disease Or preparation (formulation), also refers to the composition or preparation with multiple compounds as active ingredients. "Drug" should be understood not only as a product that is approved and approved for production by the administrative agency established by it according to the laws and regulations of a country, but also refers to a product containing a single compound as the active ingredient formed in the process of obtaining approval and approval for production. various material forms. "Formation" should be understood as obtaining through chemical synthesis, biological transformation or purchase.

The "preparation" referred to in the present invention refers to a dosage form containing isoflavone compounds of the present invention that is beneficial to drug delivery, such as: but not limited to, aqueous solution injection, powder injection, pill, powder, tablet, patch, Suppositories, emulsions, creams, gels, granules, capsules, aerosols, sprays, powder sprays, sustained-release and controlled-release formulations, etc. These pharmaceutical excipients can be conventionally used in various preparations, such as: but not limited to, isotonic agents, buffers, flavoring agents, excipients, fillers, binders, disintegrants and lubricants, etc. ; It can also be selected for use in order to be compatible with the substance, such as: emulsifier, solubilizer, bacteriostat, analgesic and antioxidant, etc., this type of adjuvant can effectively improve the stability of the compound contained in the composition and solubility or change the release rate and absorption rate of the compound, etc., thereby improving the metabolism of the compound of the present invention in vivo, thereby enhancing the administration effect. In addition, excipients that can be used to achieve specific administration purposes or methods, such as sustained-release administration, controlled-release administration, and pulse administration, such as, but not limited to, gelatin, albumin, chitosan , polyether and polyester polymer materials, such as: but not limited to, polyethylene glycol, polyurethane, polycarbonate and its copolymers, etc. The main manifestations of the so-called "beneficial to drug administration" include, but are not limited to, improving therapeutic effect, improving bioavailability, reducing toxic and side effects, and improving patient compliance.

The "effective therapeutic dose" referred to in the present invention refers to the amount that can alleviate various pathological symptoms and use the compound of the present invention as an active ingredient. The given amount is generally determined according to the body weight of the organism, such as: the total amount given every day 0.05mg/kg-50mg/kg. According to the actual conditions of the organism and its disease, the amount and frequency of use can also be adjusted, such as: give a total amount of 0.05-0.5mg/kg, 0.6-1mg/kg, 1-10mg/kg, 11 times a day or more. - 25 mg/kg, 26-40 mg/kg or 41-50 mg/kg. Depending on the dosage form of the drug, the dosage will also change. For example, 10 mg/kg is required for ordinary tablets, while the dosage required for slow and controlled release preparations is lower. The specific dose of the steroidal saponin derivatives of the present invention needs to be determined according to specific conditions, such as: the way of administration, the route of administration, the state of the patient at the time of administration, and the pathological condition during treatment.

The "basic compound" referred to in the present invention is a compound obtained by replacing all variable substituents on the structural formula of the compound with "hydrogen". For example, the compound obtained by substituting "hydrogen" for the substituents R ₁ , R ₂ and R ₃ in the structure shown in the following formula I at the same time is the basic compound.

The "substituent" referred to in the present invention refers to the general designation of one or more inorganic or organic groups that replace hydrogen atoms at specific positions on the basic compound, which can be atoms, molecules, ions, compounds and polymers.

The "halogen" referred to in the present invention includes fluorine, chlorine, bromine and iodine, and a combination of -F, -Cl, -Br and -I is applied to the basic compound as a substituent.

The "sulfate group" referred to in the present invention is -OSO ₃ H, -OSO ₃ Na, -OSO ₃ K and -OSO ₃ NH ₄ .

The "phosphate group" referred to in the present invention is -OPO(HO) ₂ , and the pharmaceutically acceptable salts formed by substituting hydrogen on it.

The "trifluoroacetoxy group" referred to in the present invention is -OCOCF ₃ .

The "substituted amino" referred to in the present invention refers to the amino group whose hydrogen on the amino group is partially or completely substituted by C1-C3 saturated alkane, such as but not limited to methylamino and dimethylamino.

The "C1-C3 saturated alkanes" referred to in the present invention are straight-chain or branched-chain alkanes with 1-3 carbon atoms. Among them, the letter C represents a carbon atom, and the subsequent numbers are positive integers, such as: 1, 2 or 3, etc., indicating the number of carbon atoms contained in the group, such as: methane, ethane and propane.

The "sugar" referred to in the present invention, also known as "sugar", is a kind of basic organic matter in chemistry or biochemistry, and is composed of polyhydroxyl (more than 2) aldehydes and polyhydric ketones composed of C, H and O elements (more than 2) and organic compounds that can be hydrolyzed to form polyhydroxy aldehydes or polyhydroxy ketones. According to its molecular structure, it can be divided into "monosaccharide", "disaccharide" and "polysaccharide". An organic compound composed of several monosaccharide molecules condensed through glycosidic bonds, also known as "sugar chain". In the present invention, monosaccharides or disaccharides are preferably used as substituents in the base compound.

The "monosaccharide" referred to in the present invention refers to an organic compound composed of C, H and O elements, which is also the basic unit of other sugar substances, generally a polyhydroxy group containing 3-6 carbon atoms Organics of aldehydes or polyhydroxyketones. Preference is given to organic polyhydroxy aldehydes or polyhydroxy ketones with 5 or 6 carbon atoms, also known as "pentose" and "hexose" such as: but not limited to ribose, xylose, arabinose, glucose, mannose, galactose, Allulose, fructose, sorbose, tagatose, gulose, idose, talose, allose and altrose etc. In the monosaccharide structure of the present invention, the O element can be partially or completely replaced by the S element.

The "disaccharide" referred to in the present invention refers to an organic compound composed of C, H and O elements, which is generally formed by connecting two monosaccharide molecules through condensation and dehydration to form a glycosidic bond. Well-known disaccharides such as: sucrose, lactose, cellobiose, trehalose and maltose, etc. Glycosidic bonds formed between monosaccharide molecules such as but not limited to α-1,1-, α-1,2-, α-1,3-, α-1,4-, β-1,4- and α-1,6- and several other forms of glycosidic bonds. In the disaccharide structure of the present invention, the O element can be partially or completely replaced by the S element.

The "polysaccharide" referred to in the present invention also includes oligosaccharide, which refers to an organic compound composed of C, H and O elements, generally composed of more than three monosaccharide molecules through condensation and dehydration. A linear or branched polymer linked by glycosidic bonds. Preference is given to linear or branched polymers with 3-15 monosaccharide molecules connected by glycosidic bonds. Well-known polysaccharides such as: gentiobiose, melezitose, acacitriose, verbascose, stachyose and raffinose wait. The glycosidic bonds formed between monosaccharide molecules include but not limited to α-1,4-, β-1,4- and α-1,6- and other forms of glycosidic bonds. In the polysaccharide structure of the present invention, the O element can be partially or completely replaced by the S element.

The "C1-C6 alcohol" referred to in the present invention is a linear or branched saturated or unsaturated hydrocarbon substituted by one or more hydroxyl groups. Among them, the letter C represents a carbon atom, and the subsequent numbers are positive integers, such as: 1, 2, 3, 4 or 5, etc., indicating the number of carbon atoms contained in the group, such as: methanol, ethanol, propanol, isopropanol Alcohol and n-butanol etc.

A saponin derivative provided by the present invention is specifically a furostane-type saponin derivative, and its structure is shown in formula I. Among them, R ₁ represents a substituent selected from one of -OH, -SH, halogen and C1-C3 saturated alkane substituted amino groups; R ₂ represents a substituent selected from -OH, - One of SH, halogen, sulfate ester group, phosphate ester group, trifluoroacetate group, C1-C3 saturated alkane substituted amino group, sugar linked by O-glycosidic bond and sugar linked by S-glycosidic bond.

Formula Ⅰ

Another saponin derivative provided by the present invention has a structure as shown in formula I, wherein R represents a substituent selected from _one of -OH, -SH, halogen and C1-C3 saturated alkane substituted amino Group; R Represents a substituent selected from _-OH , -SH, halogen, sulfate ester, phosphate, trifluoroacetate, C1-C3 saturated alkane substituted amino, linked by O-glycosidic bond A group of monosaccharides and monosaccharides linked by S-glycosidic bonds.

Another saponin derivative provided by the present invention has a structure as shown in formula I, wherein R ₁ represents a substituent selected from -OH or -SH; R ₂ represents a substituent selected from -OH One of , -SH, halogen, sulfate group, phosphate group, trifluoroacetate group, amino group substituted by C1-C3 saturated alkane, monosaccharide linked by O-glycosidic bond and monosaccharide linked by S-glycosidic bond group.

Another saponin derivative provided by the present invention has a structure as shown in formula _II , wherein R represents a substituent selected from -OH, -SH, halogen, sulfate ester group, phosphate ester group, trifluoroacetic acid A group, one of C1-C3 saturated alkane-substituted amino groups, monosaccharides linked by O-glycosidic bonds and monosaccharides linked by S-glycosidic bonds.

Formula II

Another saponin derivative provided by the present invention has a structure shown in formula III-I.

Formula III-I

Another saponin derivative provided by the present invention has a structure shown in formula III-II.

Formula III-II

The various steroidal saponin derivatives provided by the present invention can be obtained in various ways, such as dissolving timosaponin B-Ⅲ or timosaponin B-Ⅲ saponin in a solvent, and adding various necessary reagents, React under the protection of an inert gas (such as: but not limited to nitrogen, hydrogen or helium). After the reaction, use a quencher such as trimethylamine, triethylamine, sodium bicarbonate or sodium thiosulfate to terminate the reaction. The resulting product is separated by column chromatography with fillers such as silica gel, alumina, ODS or macroporous resin. Solvents used for separation such as but not limited to tetrahydrofuran, petroleum ether, ethyl acetate, DMF, pyridine, dichloromethane, ethanol, One or more of methanol or water.

The present invention provides a relatively direct method for obtaining various saponin derivatives of the present invention, which is obtained by separating and hydrolyzing timosaponin B-III as a raw material. The specific method is as follows: according to the weight of timosaponin B-III Solvent (solvent can be aqueous solution of 10v/v%-100v/v%C1-C6 alcohol and aqueous solution of other common organic solvents, or add co-solvent in water, such as: surfactant) volume ratio 1:0.2-10, will know Dissolve timosaponin B-Ⅲ in a container, add mineral acid according to the volume ratio of timosaponin B-Ⅲ to inorganic acid (such as hydrochloric acid and sulfuric acid, etc.) in a volume ratio of 1:1-4, and react at 60°C-100°C for 1 -10 hours, then stop the reaction, add an appropriate amount of lye (such as sodium bicarbonate and sodium hydroxide) to the reaction solution to neutralize to pH 6-8, according to the volume ratio of timosaponin B-Ⅲ to n-butanol 1 : 1-10 extractions 1-3 times, combined n-butanol extraction fractions, recovered the solvent under reduced pressure to obtain a solid. The solid is dissolved in a common organic solvent (such as acetonitrile and methanol, etc.), separated by analytical, semi-preparative or preparative HPLC or an open ODS column and the main chromatographic peaks are collected. After the solvent is recovered from the solution containing the target product of the present invention, A slightly yellow or white product is obtained, which is the furostane-type saponin derivative of the present invention.

Various direct or indirect methods can obtain various saponin derivatives, pharmaceutically acceptable salts or compositions thereof provided by the present invention. Those skilled in the art can foresee that, extracting timosaponin B-Ⅲ (such as: CN101307090A) from Anemarrhena or its extracts, and then indirectly producing various furostanols provided by the present invention through the acid hydrolysis method disclosed above Alkane-type saponin derivatives, for example: after hydrolysis of timosaponin B-III obtained from the Chinese medicinal material Anemarrhena chinensis, the furostane-type saponin derivatives represented by formula I are obtained through separation. In addition, the target derivatives of the present invention can also be obtained through timosaponin B-Ⅲ sapogenin, and other methods such as: organic synthesis, extraction from biological metabolites, separation and purification, chemical total synthesis, biocatalysis and transformation, etc. These derivatives can also be obtained. The various preparation methods listed in the present invention should be understood as necessary disclosures for implementing the technical solutions of the present invention. Those skilled in the art can obtain the saponin derivatives of the present invention according to the teachings of textbooks and experimental manuals, and through necessary experiments, and the methods for obtaining the saponin derivatives described here should not be regarded as limitations of the present invention.

Various saponin derivatives provided by the present invention can be used as the sole or main active ingredient in the preparation of drugs for the prevention or treatment of viral diseases, or administered together with one or more other chemical substances or drugs with antidepressant effects organism. These chemical substances such as: but not limited to Anemarrhena extract, timosaponin and its hydrolyzate, timosaponin B-Ⅱ and its hydrolyzate, timosaponin B-Ⅲ saponin, timosaponin B-Ⅲ and its hydrolyzate Hydrolyzates, tricyclic and tetracyclic antidepressants, monoamine oxidase inhibitors, selective 5-HT reuptake inhibitors (SSRIs), atypical antidepressants and lithium salts, etc. The so-called "administration to organisms together" means that various saponin derivatives of the present invention are administered alone or mixed with one or more other compounds or drugs with antidepressant effects through a single route of administration, such as: but not limited to , oral (Oral), nasal (Nasal), (buccal), transdermal (Transdermal), pulmonary (Pulmonal), vaginal (Vaginal), subcutaneous (Subcutaneous) or intravenous (Intravenous) administration of organisms; or And one or more other chemical substances or drugs with antidepressant effect are given to organisms through multiple administration routes, such as: but not limited to immediate-release oral preparations and slow-release implant preparations.

It is proved by forced swimming test and tail suspension test that administering an effective therapeutic dose of the furostane-type saponin derivative represented by formula I of the present invention to the organism can improve or eliminate the depression symptoms of the organism. This shows that the furostane-type saponin derivatives provided by the present invention have significant antidepressant activity, and they are used as active ingredients to make medicines, foods and health products, and to be used in the prevention and treatment of depression.

The pharmaceutical composition provided by the invention uses various furostane-type saponin derivatives as active ingredients for preventing and treating depression.

The food provided by the invention uses various furostane-type saponin derivatives as active ingredients for preventing and treating depression.

The health care product provided by the invention uses various furostane-type saponin derivatives as active ingredients to prevent and treat depression.

The beneficial effect that technical solution of the present invention realizes:

The furostane-type saponin derivatives represented by the formula I of the present invention, especially the furostane-type saponin derivatives with the structural conformations shown in the formulas III-I and III-II, have significant antidepressant pharmacological activity and can be used as The active ingredients are made into medicines, foods and health products, and are used alone or in combination with other substances for the prevention and treatment of depression.

Description of drawings

Figure 1 is the ¹ H NMR spectrum of formula III-I (400 MHz, Pyr-d ₅ );

Figure 2 is the ¹³ C-NMR spectrum and DEPT135 spectrum (100MHz, Pyr-d ₅ ) of formula III-I;

Figure 3 is the HMBC spectrum of formula III-II (Pyr-d ₅ );

Figure 4 is the formula III-II ROESY spectrum (Pyr-d ₅ );

Figure 5 is the HMBC correlation diagram of formula III-II, in which "→" represents the long-range correlation between hydrogen and carbon;

Figure 6 is the ROESY correlation diagram of formula III-II, in which "←→" represents the hydrogen-hydrogen NOE correlation.

Detailed ways

The technical solution of the present invention will be described in detail below in conjunction with the accompanying drawings. The embodiments of the present invention are only used to illustrate the technical solutions of the present invention without limitation. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the invention can be modified or equivalently replaced , without departing from the spirit and scope of the technical solution of the present invention, all of which shall be covered by the claims of the present invention.

All reagents used in the present invention were purchased from Sigma-Aldrich unless otherwise specified.

The preparation of embodiment 1 furostane type saponin monosaccharide derivative

Using timosaponin B-Ⅲ as the reaction starting material to prepare furostane-type saponin monosaccharide derivatives, the specific steps are as follows:

(1) Dissolve 0.05mol timosaponin B-Ⅲ in 300ml methanol solution (water or 30v/v% ethanol solution or 1w/v% Tween solution) with a volume ratio of 10v/v%, and add 0.5ml-1ml Concentrated sulfuric acid, react at 80°C-150°C for 1 hour-6 hours.

(2) Adjust the pH of the above reaction solution to 6-7 with sodium hydroxide.

(3) Use 900ml-1200ml n-butanol to extract the solution obtained after neutralization in step (2) three times, and concentrate the extract under reduced pressure to evaporate to dryness.

(4) Dissolve the above solid with water, make the saponin adsorb on the ODS column, elute with 40v/v%-60v/v% methanol, and collect the eluate.

(5) The solution was recovered under reduced pressure and evaporated to dryness to obtain furostane-type saponin monosaccharide derivative 2 (NMR: see Table 1 for details).

Preparation of embodiment 2 furostane type saponin derivatives

(1) Dissolve 0.05mol timosaponin B-III in 300mL methanol solution (water or 30% ethanol solution or 1% Tween solution) with a volume ratio of 10%, add 2mL-4mL concentrated hydrochloric acid, 80-150℃ React for 1 hour to 6 hours.

(2) Adjust the pH of the above reaction solution to 6-7 with sodium hydroxide.

(3) Use 900mL-1200mL n-butanol to extract the solution obtained after neutralization in step (2) three times, and concentrate the extract under reduced pressure until evaporated to dryness.

(4) Dissolve the above solid with water, make the saponin adsorb on the ODS column, elute with 40v/v%~60v/v% methanol, and collect the eluate.

(5) The solution was recovered under reduced pressure and evaporated to dryness to obtain furostane-type saponin monosaccharide derivative 2 (NMR: see Table 1 for details).

Preparation of embodiment 3 furostane type saponin monosaccharide derivatives

Using timosaponin B-Ⅲ as the reaction starting material to prepare furostane-type saponin monosaccharide derivatives, the specific steps are as follows:

(1) Dissolve 0.1mol timosaponin B-III in 600ml methanol solution with a volume ratio of 10v/v%, add 30ml-50ml concentrated hydrochloric acid, and react at 80°C-100°C for 2 hours.

(2) Adjust to pH=6-7 with sodium hydroxide.

(3) Use 1.8L-2.4L n-butanol to extract the solution obtained after neutralization in step (2) three times, and concentrate the extract under reduced pressure until evaporated to dryness.

(4) Dissolve the above solid with water, make the saponin adsorb on the ODS column, elute with 40v/v%-60v/v% methanol, and collect the eluate.

(5) The solution was recovered under reduced pressure and evaporated to dryness to obtain aglycone 3 of mother saponin B-Ⅲ (NMR: 30.63(1-C); 27.14(2-C); 66.07(3-C); 34.41(4 -C);37.04(5-C);26.97(6-C);28.65(7-C);35.27(8-C);40.02(9-C);35.57(10-C);21.38(11-C); C);40.15(12-C);43.88(13-C);54.83(14-C);31.39(15-C);84.59(16-C);64.68(17-C);14.45(18-C );24.26(19-C);103.59(20-C);11.83(21-C);152.38(22-C);34.41(23-C);23.62(24-C);33.68(25-C) ;68.76(26-C);17.16(27-C)).

(6) Dissolve 0.05mol timosaponin B-Ⅲ aglycone and 0.05mol-0.07mol reagent T1 in dichloromethane, cool down to -20°C-0°C under nitrogen protection, add 0.91ml-1.4ml CF ₃ SO ₃ Si(CH ₃ ) ₃ , the reaction stopped after 40 minutes to 1 hour.

(7) Triethylamine was added to quench the reaction, and the solvent was spin-dried under reduced pressure.

(8) Add the precipitate to 200mL CH ₂ Cl ₂ -MeOH (2:1, v/v) solution, then add 50ml of 1mol/L-1.5mol/L sodium methoxide methanol solution, react for 3-5 hours, and evaporate to dryness liquid

(9) Dissolve the above solid with water, make the saponin adsorb on the ODS column, elute with 40v/v%-60v/v% methanol, and collect the eluate. The solution was recovered under reduced pressure and evaporated to dryness to obtain the furostane-type saponin monosaccharide derivative 2.

The preparation of embodiment 4 furostane type saponin iodo derivatives

Using the aglycone 3 of timosaponin B-Ⅲ prepared in Example 3 as the reaction starting material to prepare furostane-type saponin iodo derivatives, the specific steps are as follows:

(1) Dissolve 0.05 mol of saponin aglycon and 0.05 mol of sodium iodide obtained in step (5) of Example 3 in 150ml-200ml of 2-butanone, and react at 25°C-50°C for 1-3 hours.

(2) After the reaction, pour the reaction liquid into 1.25L-2L water, filter, and wash with water twice.

(3) Dry the filter cake and recrystallize it with methanol.

(4) Filter and dry the product to obtain furostane-type saponin iodo derivative 4 (NMR: 30.60(1-C); 27.27(2-C); 66.05(3-C); 37.37(4-C) ;40.68(5-C);27.22(6-C);29.64(7-C);34.92(8-C);40.02(9-C);35.57(10-C);21.38(11-C); 40.15(12-C);43.88(13-C);54.83(14-C);31.39(15-C);84.59(16-C);64.68(17-C);14.45(18-C);24.26 (19-C);103.59(20-C);11.83(21-C);152.38(22-C);17.14(23-C);32.62(24-C);35.82(25-C);32.87( 26-C); 20.23(27-C)).

Preparation of embodiment 5 furostane type saponin brominated derivatives

Using the aglycon 3 of timosaponin B-Ⅲ prepared in Example 3 as the reaction starting material to prepare furostane-type saponin brominated derivatives, the specific steps are as follows:

(1) Dissolve 0.05 mol of the saponin aglycone obtained in step (5) of Example 3 in 250ml-300ml CH ₂ Cl ₂ and add 25ml-30ml boron trifluoride-ether complexing reagent (CAS: 109-63- 7) Add 0.05mol-0.1mol of acetic anhydride dropwise, react at 20°C-60°C for 30 minutes-1 hour, wash the reaction solution with 1L of saturated NaCl aqueous solution, then wash the reaction solution with 750ml-1L water, separate the organic phase, wash with NaCl ₂ SO ₄ drying; the organic phase was filtered, the filtrate was spin-dried, and the product was purified by silica gel column chromatography, and the developing solvent was petroleum ether and ethyl acetate at a volume ratio of 6:1.

(2) Dissolve the product obtained in step (1) in 250ml-300ml tetrahydrofuran solution, add 0.05mol-0.1mol NBS and 0.005mol-0.01mol (Ph) ₃ P, and react at room temperature for 5-7 hours.

(3) Filter the reaction solution, evaporate the solvent under reduced pressure, add 200mL CH ₂ Cl ₂ -MeOH (2:1, v/v) solution for extraction, add 1mol/L-1.5mol/L sodium methoxide methanol solution 50ml-100ml , 20°C-50°C for 5-8 hours.

(4) Distill under reduced pressure, concentrate the reaction, and recrystallize from a mixture of ethanol and ethyl acetate to obtain the final product furostane-type saponin brominated derivative 7 (NMR: 31.52 (1-C); 26.44 (2-C) ;33.85(3-C);38.32(4-C);40.68(5-C);27.22(6-C);29.64(7-C);34.92(8-C);40.02(9-C); 35.57(10-C);21.38(11-C);40.15(12-C);43.88(13-C);54.83(14-C);31.39(15-C);84.59(16-C);64.68 (17-C);14.45(18-C);24.26(19-C);103.59(20-C);11.83(21-C);152.38(22-C);34.41(23-C);23.63( 24-C);35.43(25-C);68.07(26-C);17.04(27-C)).

Preparation of embodiment 6 furostane type saponin phosphate derivatives

Using the aglycon 3 of timosaponin B-Ⅲ prepared in Example 3 as the reaction starting material, the furostane-type saponin phosphate derivatives were prepared, and the specific steps were as follows:

(1) Dissolve 0.05 mol of saponin aglycon obtained in step (5) of Example 3 in 200 ml of tetrahydrofuran and cool down to -50°C-20°C, slowly add 0.25mol-0.3mol of pyrophosphoryl chloride, Keep the temperature constant, and react for 1 hour to 3 hours.

(2) Dilute the reaction solution with water, raise the temperature to about 40°C-60°C, add a certain amount of Na ₂ CO ₃ to it until no bubbles are released, stir and react at 25°C-35°C for 0.5-1.5 hours, filter, and separate layers. The aqueous solution of the lower layer was rotary evaporated at 50°C until no obvious bubbles were found. Add hydrochloric acid to it, adjust the pH=1-2, and precipitate a white precipitate.

(3) Filtrate the precipitate, wash the precipitate three times with acid, and dry to obtain the furostane-type saponin phosphate derivative 8 (NMR: 30.60(1-C); 27.27(2-C); 66.05(3-C );37.37(4-C);40.68(5-C);27.22(6-C);29.64(7-C);34.92(8-C);40.02(9-C);35.57(10-C) ;21.38(11-C);40.15(12-C);43.88(13-C);54.83(14-C);31.39(15-C);84.59(16-C);64.68(17-C); 14.45(18-C);24.26(19-C);103.59(20-C);11.83(21-C);152.38(22-C);33.43(23-C);24.12(24-C);34.82 (25-C);71.13(26-C);16.21(27-C)).

Preparation of embodiment 7 furostane type saponin sulfate derivatives

Using the aglycon 3 of timosaponin B-Ⅲ prepared in Example 3 as the reaction starting material to prepare the furostane-type saponin sulfate derivatives, the specific steps are as follows:

(1) Dissolve 0.05 mol of saponin aglycone obtained in step (5) of Example 3 in 300 mL of DMF, pass through nitrogen protection, and cool down to -15°C-0°C. Add 0.05mol-0.1mol Et ₃ N·SO ₃ and 0.4mol-0.6mol p-toluenesulfonic acid into the reaction liquid, and react for 3-5 hours.

(2) Add 0.05mol-0.07mol triethylamine, and then add methanol with a volume ratio of 1:1 to the solution. Continue to stir and react for 3 hours to 5 hours; raise the temperature to 20°C to 30°C and continue to stir for 2 hours to 4 hours.

(3) Spin the solvent in vacuum, dissolve the above solid with water, and adsorb the saponin on the ODS column, elute with 40v/v%-70v/v% methanol, and collect the eluate.

(4) The solution was recovered under reduced pressure and evaporated to dryness to obtain a solid.

(5) Dissolve the solid with 50ml-60ml of 1mol/L NaOH solution, heat to 40°C-70°C, and stir for 1-3 hours.

(6) Evaporate the solvent to dryness, recrystallize with acetone, and filter to obtain the furostane-type saponin sulfate derivative 9 (NMR: 30.60(1-C); 27.27(2-C); 66.05(3-C) ;37.37(4-C);40.68(5-C);27.22(6-C);29.64(7-C);34.92(8-C);40.02(9-C);35.57(10-C); 21.38(11-C);40.15(12-C);43.88(13-C);54.83(14-C);31.39(15-C);84.59(16-C);64.68(17-C);14.45 (18-C);24.26(19-C);103.59(20-C);11.83(21-C);152.38(22-C);33.43(23-C);24.12(24-C);32.82( 25-C); 84.13(26-C); 17.41(27-C)).

Example 8 Preparation of furostane-type saponin trifluoroacetate-substituted derivatives

Using the aglycone 3 of timosaponin B-Ⅲ prepared in Example 3 as the reaction starting material, the furostane-type saponin trifluoroacetate-substituted derivatives were prepared, and the specific steps were as follows:

(1) Dissolve 0.05mol of the saponin aglycon obtained in step (5) of Example 3 in 250ml CH ₂ Cl ₂ , add 50ml-60ml of boron trifluoride-diethyl ether complexing reagent, and dropwise add acetic anhydride 0.1mol-0.13 mol, react at 20°C-40°C for 1 hour-2 hours, wash the reaction solution with 1L of saturated NaCl aqueous solution, then wash the reaction solution with 750ml of water, separate the organic phase, and dry it with Na ₂ SO ₄ ; filter the organic phase, and spin the filtrate , the product was purified by silica gel column chromatography, and the developing solvent was petroleum ether and ethyl acetate at a volume ratio of 6:1.

(2) Dissolve the product in (1) in 250ml CH ₂ Cl ₂ , cool down to -40°C--15°C, add 0.075mol-0.1mol trifluoroacetyl trifluoromethanesulfonate (TFAT), and react for 2 hours- 4 hours.

(3) After the reaction is completed, add sodium bicarbonate to quench the reaction, continue to stir for 30 minutes to 1 hour, raise the temperature to 20°C to 30°C, and continue to stir for 30 minutes to 1 hour.

(4) Spin the solvent in vacuum, dissolve the above solid with water, and adsorb the saponin on the ODS column, elute with 40v/v%-70v/v% methanol, and collect the eluate.

(5) The solution was recovered under reduced pressure and evaporated to dryness to obtain furostane-type saponin trifluoroacetate derivative 11 (NMR: 30.60(1-C); 27.27(2-C); 66.05(3-C); 37.37 (4-C);40.68(5-C);27.22(6-C);29.64(7-C);34.92(8-C);40.02(9-C);35.57(10-C);21.38( 11-C);40.15(12-C);43.88(13-C);54.83(14-C);31.39(15-C);84.59(16-C);64.68(17-C);14.45(18 -C);24.26(19-C);103.59(20-C);11.83(21-C);152.38(22-C);33.43(23-C);24.12(24-C);32.82(25-C); C);68.73(26-C);17.41(27-C);156.60(28-C);113.19(29-C)).

Preparation of embodiment 9 furostane type saponin alkylamino derivatives

Using the aglycone 3 of timosaponin B-Ⅲ prepared in Example 3 as the reaction starting material, the furostane-type saponin alkylamino-substituted derivatives were prepared, and the specific steps were as follows:

(1) Dissolve 0.05 mol of the saponin aglycone obtained in step (5) of Example 3 in 300 mL of tetrahydrofuran, and lower the temperature to -30°C-0°C; Add 0.2mol diethyl azodicarboxylate and 0.12mol-0.22mol PPh ₃ into the solution, pass through nitrogen protection, and react for 2-3 hours.

(2) After the reaction, filter and spin the filtrate to dryness.

(3) Spin the solvent in vacuum, dissolve the above solid with water, and adsorb the saponin on the ODS column, elute with 50v/v%-70v/v% methanol, and collect the eluate.

(4) The solution was recovered under reduced pressure and evaporated to dryness to obtain the furostane-type saponin alkylamino-substituted derivative 12 (NMR: 30.60(1-C); 27.27(2-C); 66.05(3-C); 37.37(4 -C);40.68(5-C);27.22(6-C);29.64(7-C);34.92(8-C);40.02(9-C);35.57(10-C);21.38(11-C); C);40.15(12-C);43.88(13-C);54.83(14-C);31.39(15-C);84.59(16-C);64.68(17-C);14.45(18-C );24.26(19-C);103.59(20-C);11.83(21-C);152.38(22-C);23.43(23-C);28.12(24-C);30.82(25-C) ;68.73(26-C);18.41(27-C);41.49(3-N-CH ₃ );47.13(26-N-CH ₃ )).

Embodiment 10 structure confirmation

The solid obtained in Example 1 or Example 2 was tested positive by Molish reaction and Liebermann-Burchard reaction, and its nuclear magnetic resonance spectrum (Nuclear Magnetic Resonance, NMR) data is shown in Table 1. Then it is analyzed as the derivative represented by formula III-II through comprehensive analysis of HMBC, ROESY, MS and NMR and other spectral results.

Derivatives of formula Ⅲ-Ⅱ, white amorphous powder in appearance, easily soluble in pyridine, soluble in acetone-water, acetonitrile-water and aqueous solution. The Liebermann-Burchard and Molish reactions were positive, proving that the compound was timosaponin. (c0.05mg/ml, pyridine).

HR-ESI-MS (positive ion mode) showed [M+Na] ⁺ m/z 601.3713 (calculated as C ₃₃ H ₅₆ O ₉ Na: 601.3711), and its molecular formula was determined to be C ₃₃ H ₅₅ O ₈ .

The nuclear magnetic resonance spectrum suggested that the structure contained a molecule of glucose, and the δ4.87 (d, J=7.7Hz) in the hydrogen spectrum ( ¹ H-MNR, see Figure 1) was the signal of the glucose end group proton, which was determined to be the β configuration. Comprehensive analysis of ¹³ C-NMR spectrum and DEPT spectrum (see Figure 2), it is confirmed that there are 4 methyl carbon signals, 12 methylene carbon signals, 13 methine carbon signals and 4 quaternary carbon signals. The carbon spectrum and hydrogen spectrum data of the aglycone part of the compound are similar to timosaponin B3, indicating that it has a similar mother nucleus with timosaponin B3. In the HMBC spectrum (see Figure 3), H21 is remotely correlated with C20 and C22, indicating that the position of the double bond is consistent with that of timosaponin B3. In addition, the δ _H 4.87 (H-1') glucose terminal proton has a long-range correlation with the δ _C 75.22 (C-26), which determines the position of glycosylation, that is, the glucose is connected with 26-OH to form a glycoside. The relative stereo configuration was determined by the ROESY spectrum (see Figure 4), that is, there is a NOE correlation between the H-5 proton and the H-19 proton, and the A ring and the B ring were determined to be cis configurations. See Table 1, Figure 5 and Figure 6 for other NMR data and related relationships. This compound has not been reported in the literature and is a new compound named timosaponin BⅢ-4 (or abbreviated as: BⅢ-4).

The NMR data of derivative shown in table 1 formula Ⅲ-Ⅱ

Example 11 Antidepressant Activity Verification

The FST and TST two behavioral hopelessness models are sensitive to most antidepressants, and are simple and quick to operate, and are widely used in the screening of such drugs. This example is also prepared by using this model in Example 1-Example 9 The antidepressant pharmacological effects of the derivatives were verified.

Male ICR mice, weighing (20±2) g, were used in the experiment, purchased from the Experimental Animal Center of Shanghai Institute of Materia Medica, Chinese Academy of Sciences, free to eat and drink, room temperature (23±2)°C, and natural light. All mice were randomly divided into 10 mice/cage, and the experiment started after 3 days of adaptation in the feeding environment. Before the experiment, they were fasted for 12 hours and had free access to water. The specific administration method is as follows: a blank control group was given an equal volume of normal saline.

FST verification experiment

1) Specific operation: Continuous administration for 6 days, test 1 hour after the last administration. Firstly, the autonomous activity of the mice was measured by the open-field method, that is, the mice were placed alone in a cylindrical glass jar, timed for 4 minutes, and the number of arm lifts within 2 minutes after that was recorded. Then the mice were individually placed in a cylindrical glass jar with a height of 20 cm and a diameter of 14 cm. The water depth in the jar was 10 cm, and the water temperature was 23-25°C. Time the mouse for 6 minutes after entering the water, and record the cumulative immobility time within 4 minutes (the immobility criterion: the mouse stops struggling in the water, or is in a floating state, and only has small limb movements to keep the head floating on the water). Each group of mice was operated in parallel.

2) Experimental data processing: The experimental results are expressed as mean ± standard error (x ± SE). The t-test was used for statistical analysis to determine whether it was significant. Firstly, the t-test was carried out on the autonomic activity index, and its P>0.05 indicated that the autonomic hindrance of the mice had no effect, so as to avoid the interference of central stimulant drugs. Then the t test was carried out on the indicators of the forced swimming test, and the blank control group was compared with the three experimental groups of high, middle and low doses to judge whether it had antidepressant effect and the relationship between the intensity of antidepressant effect and the dose.

TST verification experiment

1) Specific operation: Continuous administration for 6 days, test 1 hour after the last administration. Firstly, the autonomous activity of the mice was measured by the open-field method, that is, the mice were placed alone in a cylindrical glass jar, timed for 4 minutes, and the number of arm lifts within 2 minutes after that was recorded. Then stick the tail of the mouse on the horizontal bar at 2 cm from the tip of the tail with adhesive tape, and isolate the sight of the animal with a board around it. The horizontal bar is about 25 cm away from the ground, so that the mouse is about 10 cm away from the ground, and the time is 6 minutes. The mice in each group were operated in parallel.

2) Experimental data processing: The experimental results are expressed as mean ± standard error (x ± SE). The t-test was used for statistical analysis to determine whether it was significant. Firstly, the t-test was carried out on the autonomic activity index, and its P>0.05 indicated that the autonomic hindrance of the mice had no effect, so as to avoid the interference of central stimulant drugs. Then the t test was carried out on the indicators of the tail suspension test, and the blank control group was compared with the three experimental groups of high, middle and low doses to judge whether it had antidepressant effect and the relationship between the intensity of antidepressant effect and the dose.

The antidepressant effect of the derivative prepared by the present invention was verified through animal experiments, and the results and data are shown in Table 2 and Table 3. The mice were given 10 mg/Kg of various compounds by intraperitoneal injection, and a blank control group (normal saline) was set up separately. The t test was carried out on the indicators of the autonomous activities of the mice, and there was no significant difference, indicating that these compounds had no effect on the autonomous activities of the mice. Carry out t-test to the immobility time of mouse forced swimming and tail suspension test respectively, the result shows, the various derivatives that embodiment 1-embodiment 9 make all can obviously shorten the immobility of mouse forced swimming and tail suspension test. time, suggesting that they have significant antidepressant activity.

Table 2 The effect of intraperitoneal administration on the immobility time of forced swimming in ICR mice

"*" means P<0.05; "**" means P<0.01.

Table 3 Effect of intraperitoneal administration on tail suspension time of ICR mice

"*" means P<0.05; "**" means P<0.01.

Claims (4)

1. A saponin derivative as shown in formula III-I, 2. The saponin derivative according to claim 1, characterized in that the conformation of the formula III-I is shown in the formula III-II . 3. The saponin derivative according to claim 1 or 2 is used as an active ingredient to prepare medicines, food or health care products for preventing and treating depression. 4. A composition for antidepressant, the active ingredient of which contains the saponin derivative according to claim 1 or 2.