CN115368426B

CN115368426B - Triterpene compound, preparation method and antiasthmatic application thereof

Info

Publication number: CN115368426B
Application number: CN202110942150.4A
Authority: CN
Inventors: 谭道鹏; 何芋岐; 鲁艳柳; 秦琳; 张倩茹; 陆安静
Original assignee: Zunyi Medical University
Current assignee: Zunyi Medical University
Priority date: 2021-08-17
Filing date: 2021-08-17
Publication date: 2023-07-18
Anticipated expiration: 2041-08-17
Also published as: CN115368426A

Abstract

The invention discloses a triterpene compound with the following formula I or pharmaceutically acceptable salts, esters, stereoisomers, tautomers, deuterated compounds, ¹⁴ The C marker, solvate, metabolite or prodrug has the advantages of simple preparation method, good reproducibility, high purity and better asthma relieving effect.

Description

Triterpene compound, preparation method and antiasthmatic application thereof

Technical Field

The invention relates to the technical field of medicines, relates to a triterpene compound and a preparation method thereof, and also relates to application of the triterpene compound in the aspect of treating respiratory diseases.

Background

Asthma is a common and frequently occurring disease, and currently, about 3 hundred million people are worldwide asthmatics, and about 3000 ten thousand people are Chinese asthmatics. Asthma prevalence has increased at a striking rate worldwide, particularly in developing countries, for the last 20 years, and has increased by nearly a factor of 1 in many areas between 10 and 20 years.

Asthma is also known as bronchial asthma. Bronchial asthma is a chronic airway inflammation involving a variety of cells and cellular components, which is often accompanied by increased airway responsiveness, resulting in recurrent wheezing, shortness of breath, chest distress and/or cough, and the like, which often occurs at night and/or in the early morning, with extensive and variable airflow obstruction, which can be reversed by itself or by therapy.

At present, western medicines are mainly taken orally, atomized and administrated in clinic for treating asthma, and have quick response, but difficult radical treatment of asthma, and the traditional Chinese medicines have general and slow response.

The principle of western medicine treatment is generally that the symptoms of inflammation diminishing, asthma relieving, spasmolysis and antiallergic, hormone control, heart strengthening and diuretic are temporarily controlled, the development of the symptoms is only inhibited, and the symptoms of patients are aggravated year by year, so that the symptoms of patients are not radically cured. The persistent and repeated attacks of asthma cause great pain in the spirit and flesh of patients, and some patients with serious illness are pessimistic disappointment and self-abandonment of the patients are caused. Western medicine mainly uses bronchodilators, hormones and other chemicals to temporarily relieve symptoms in the period of onset, but the symptoms usually recur after stopping taking the medicine when encountering external stimulus, and the long-term taking of the medicine can cause series of toxic and side effects such as hypophysis function reduction and the like. Inhalation of glucocorticoids is one of the methods of using hormones. However, long-term intake of glucocorticoid can cause a patient to have hormone dependency, cause endocrine dyscrasia and metabolic disturbance in the patient, and cause trouble in subsequent treatments, for example, osteoporosis may occur in patients who have been treated for more than 1 year by glucocorticoid inhalation. Oral hormone therapy can be used as maintenance therapy of severe asthma, is used for symptom acute onset, but is harmful and useless for a small number of patients with hormone resistance to the treatment, and has high treatment cost. Anticholinergic bronchodilators have weak effect on treating asthma, have a plurality of adverse reactions, and may cause adverse reactions such as liver and kidney function damage.

The traditional Chinese medicine mainly regulates the whole body (mainly the immune system), increases the immunity of the organism, improves the capability of the organism for resisting attack of diseases, reduces the attack of asthma and even achieves the aim of long-term alleviation; the traditional Chinese medicine for treating asthma has long treatment time, obvious effect and slow effect, and brings much pain to patients; moreover, most of traditional Chinese medicines are decoction, and are required to be decocted first and then taken, and the decoction is long in time and inconvenient to carry; the oral administration needs to pass through the gastrointestinal circulation, has great stimulation to the intestines and the stomach, and further affects the physical health of patients.

Gynostemma pentaphyllum (Gynostemma pentaphyllum Makino.) is also known as Gynostemma pentaphyllum, herba Gynostemmatis, etc. Wherein the gypenosides are the main active ingredients in gynostemma pentaphyllum. Pharmacological experiments and clinical application show that the composition has the effects of reducing blood fat, resisting tumors, protecting liver, enhancing the immune function of a human body and the like, is mainly used for treating proliferation of cancer cells such as liver cancer, lung cancer, uterine cancer, skin cancer and the like, and has the effects of nourishing, calming, hypnotizing, resisting stress, stimulating appetite, reducing cholesterol, reducing transaminase and the like on a human body.

The inventor discovers that the triterpene compounds with brand new chemical skeleton structures in gynostemma pentaphylla have better asthma relieving effect and possibly have wide prospects in preparation of prevention or/and treatment of respiratory diseases. The triterpene compounds in gynostemma pentaphylla have wide pharmacological activity, are derived from natural plants, are safe and reliable, and are not reported in the related report of the triterpene compounds for treating or/and preventing respiratory diseases at present.

Disclosure of Invention

The invention mainly aims to provide a natural and safe medicament for treating respiratory diseases, especially asthma, with definite curative effect and few side effects, so as to solve the problems of easy generation of drug resistance, poor curative effect, easy repeated attack, easy generation of dependence, easy initiation of complications, high price and larger toxic and side effects of common medicaments in the prior art.

In order to achieve the above object, according to one aspect of the present invention, there is provided a triterpene compound of the following formula I or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, deuterate, ¹⁴ A C-tag, solvate, metabolite or prodrug,

wherein R is ₁ 、R ₂ 、R ₃ And R is ₄ Are identical or different and are each independently selected from hydrogen, halogen, hydroxy, C _1-12 Alkyl, C _1-12 Haloalkyl, C _1-12 Alkoxy, C _2-12 Alkenyl, C _2-12 Haloalkenyl, C _2-12 Alkynyl, C _2-12 Haloalkynyl, cyano, amino or carboxyl.

Further, R ₁ 、R ₂ 、R ₃ And R is ₄ Are identical or different and are each independently selected from hydrogen, halogen, hydroxy, C _1-8 Alkyl, C _1-8 Haloalkyl, C _1-8 Alkoxy, C _2-8 Alkenyl, C _2-8 Haloalkenyl, C _2-8 Alkynyl or C _2-8 Haloalkynyl.

Further, R ₁ 、R ₂ 、R ₃ And R is ₄ Are identical or different and are each independently selected from hydrogen, halogen, hydroxy, C _1-5 Alkyl, C _1-5 Haloalkyl, C _1-5 Alkoxy, C _2-5 Alkenyl, C _2-5 Haloalkenyl, C _2-5 Alkynyl or C _2-5 Haloalkynyl.

Further, R ₁ 、R ₂ 、R ₃ And R is ₄ Are identical or different and are each independently selected from hydrogen, halogen, hydroxy or methyl.

Further, the halogen is fluorine, chlorine or bromine.

Further, the halogen is chlorine.

Further, the triterpene compound has the following structural formula:

further, the triterpene compound is obtained by hydrolyzing the aqueous extract of gynostemma pentaphylla by an acid hydrolysis method.

According to another aspect of the present invention, there is provided a pharmaceutical composition comprising the above triterpene compound and pharmaceutically acceptable excipients.

Further, the auxiliary material is selected from one or more of the following: preservatives, lubricants, diluents, suspending agents, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, flavoring agents, coloring agents, buffers, disintegrants, thickening agents, masking agents and antioxidants.

Further, the pharmaceutical composition further comprises one or more of the following drugs: antiinfective, antitussive, antiasthmatic, expectorant and antihistamine.

Further, the pharmaceutical composition further comprises an antiasthmatic agent.

Further, the antiasthmatic is selected from one or more of the following: bronchodilators, anti-inflammatory antiasthmatics and antiallergic antiasthmatics.

Further, the antiasthmatic is one or more of the following drugs: salbutamol sulphate, aminophylline, diprophylline, ipratropium bromide, oxitropium bromide, sodium tryptophanate, beclomethasone dipropionate, epinephrine, isoprenaline, ephedrine, albuterol and amicin.

According to another aspect of the present invention there is provided a process for the preparation of the above triterpenes, wherein the process comprises the steps of:

(1) Taking a proper amount of gynostemma pentaphylla dried whole herb as a raw material, extracting with water by ultrasonic or reflux for 1 to 3 times, 1 to 3 hours each time, combining the extracting solutions, and concentrating under reduced pressure to obtain a water extract;

(2) Adding 0.5-3% acid solution to the water extract, extracting with ultrasound or reflux for 0.5-3 hr, filtering, and washing with water to obtain neutral precipitate; and

(3) The sediment is subjected to silica gel column chromatography to obtain the triterpene compound shown in the formula I.

Further, the water used for extraction in the step (1) is 1 to 5 times by mass of the dry whole herb of gynostemma pentaphylla.

Further, the acid in step (2) is selected from one or more of hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, acetic acid, propionic acid, malonic acid and butyric acid.

Further, the column chromatography in the step (3) is performed by using silica gel having a particle size of 100 to 400 mesh.

Further, the eluting mobile phase of the column chromatography is a mixed solvent with the volume ratio of n-hexane to ethyl acetate of 25:1 to 15:1.

Further, the volume ratio of the acid solution to the aqueous extract is from 1:3 to 1:10.

Further, the acid in the step (2) is hydrochloric acid.

According to another aspect of the present invention there is provided the use of a triterpene compound as defined above or a pharmaceutical composition as defined above in the manufacture of a medicament for the treatment of respiratory diseases.

Further, the respiratory disease is selected from one or more of the following: asthma, tracheitis, bronchitis, chronic obstructive pulmonary disease, pulmonary heart disease and tuberculosis.

Further, the asthma is selected from one or more of the following: allergic asthma, drug-induced asthma, senile asthma, cough variant asthma, chronic asthma, athletic asthma and childhood asthma.

Further, the drug is acetylcholine chloride and/or histamine phosphate.

Further, the medicine is a volume mixed solution of 2% of acetylcholine chloride and 0.1% of histamine phosphate.

The invention has the beneficial effects that:

the Gynostemma pentaphyllum is a dried whole herb of Gynostemma pentaphyllum (Thunb.) Makino of Cucurbitaceae, and is a perennial climbing herb, and is mainly distributed in China, japan, korean, etc. Modern pharmaceutical research shows that the main active ingredient is gynostemma pentaphylla saponin (Gypenoside) which has the same or very similar structure as that of ginsenoside. The research shows that the triterpene compounds with new chemical structures, which are extracted and separated from the dried whole herb of gynostemma pentaphylla, have obvious asthma relieving effect, and the application range of gynostemma pentaphylla can be expanded and the utilization value of gynostemma pentaphylla plant resources can be improved by developing the triterpene compounds into medicaments for treating respiratory diseases.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings that are needed in the description of the embodiments will be briefly described below, and it is apparent that the drawings in the following description are only some embodiments of the present application, and that other drawings can be obtained by those skilled in the art from these drawings without departing from the scope of protection of the present application.

Fig. 1 is a chemical structural formula of Gypensapogenin III.

Fig. 2 is a hydrogen spectrum of Gypensapogenin III.

Fig. 3 is a carbon spectrum of Gypensapogenin III.

Detailed Description

The following description of the embodiments of the present application will be made clearly and fully with reference to the accompanying drawings, in which it is evident that the embodiments described are some, but not all, of the embodiments of the present application. All other embodiments, which can be made by those skilled in the art based on the embodiments herein without making any inventive effort, are intended to be within the scope of the present application.

The invention is intended to cover all alternatives, modifications and equivalents, which may be included within the art of the invention as defined by the appended claims. Those skilled in the art will recognize many methods and materials similar or equivalent to those described herein that can be used in the practice of the present invention. The invention is in no way limited to the description of methods and materials.

The invention will apply to the following definitions unless otherwise indicated. According to the object of the invention, the chemical elements are according to the periodic Table of the elements, CAS version and handbook of chemistry and Physics, 75 ^th Ed., 1994. In addition, the general principles of organic chemistry are found in "Organic Chemistry", thomas Sorrell, university Science Books, sausalato: 1999, and "March's Advanced Organic Chemistry", michael B.Smith and Jerry March, john Wiley&Sons, new York:2007, thus allIs incorporated by reference.

The articles "a," "an," and "the" are intended to include "at least one" or "one or more" unless the context clearly dictates otherwise or otherwise. Thus, as used herein, the articles refer to articles of manufacture that include one or more than one (i.e., at least one) object.

As described in the background section, the existing common medicines for treating respiratory diseases, especially asthma, are easy to generate drug resistance, have poor curative effect, are easy to repeatedly attack, are easy to generate dependence, are easy to cause complications, are high in price and have the problems of large toxic and side effects. In order to solve the problems, the invention provides a triterpene compound of the following formula I or pharmaceutically acceptable salts, esters, stereoisomers, tautomers, deuterated compounds thereof, ¹⁴ A C-tag, solvate, metabolite or prodrug,

The term "salt" as used herein refers to salts of the compounds of the present invention that are pharmaceutically acceptable and have the desired pharmacological activity of the parent compound. Suitable salts include all known pharmaceutically acceptable salts, including those formed with organic and inorganic acids. Thus, suitable salts include those formed from the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, citric acid, tartaric acid, lactic acid, valeric acid, acetic acid, succinic acid, fumaric acid, maleic acid, methanesulfonic acid, and benzenesulfonic acid.

The term "ester" as used herein refers to esters of the compounds of the present invention that hydrolyze in vivo (in vivo) and include esters that readily decompose in the human body leaving the parent compound or salt thereof. Suitable ester groups include, for example, those derived from pharmaceutically acceptable aliphatic carboxylic acids (especially alkanoic, alkenoic, cyclic alkanoic and alkanedioic acids), wherein each alkyl or alkenyl moiety preferably has less than 6 carbon atoms. Representative examples of specific esters include, but are not limited to, formates, acetates, propionates, butyrates, acrylates and ethylsuccinates.

The term "stereoisomer" as used herein refers to a compound having the same chemical structure but different arrangements of atoms or groups in space. Stereoisomers include enantiomers, diastereomers, conformational isomers (rotamers), geometric (cis/trans) isomers, atropisomers, and the like.

The term "tautomer" or "tautomeric form" as used herein refers to structural isomers having different energies that can be interconverted by a low energy barrier (lowerergybarrier). If tautomerism is possible (e.g., in solution), chemical equilibrium of the tautomers can be achieved. For example, proton tautomers (also known as proton transfer tautomers (prototropic tautomer)) include interconversions by proton transfer, such as keto-enol isomerisation and imine-enamine isomerisation.

When tautomerism (e.g., keto-enol tautomerism) of the compounds of the invention or prodrugs thereof is present, both protection of their individual forms (e.g., keto or enol forms) and mixtures thereof in any ratio are required. The same applies to their stereoisomers, e.g., enantiomers, diastereomers, conformational isomers (rotamers), geometric (cis/trans) isomers, atropisomers, and the like.

If desired, the tautomers can be separated according to methods known in the art (e.g., liquid chromatography). The same applies to their enantiomers, for example, by separation using a chiral stationary phase. Alternatively, the enantiomers may be separated by conversion to diastereomers, i.e. coupling with enantiomerically pure auxiliary compounds, followed by separation of the resulting diastereomers and cleavage of the auxiliary residues. Alternatively, any enantiomer of a compound of the invention may be obtained by stereoselective synthesis using optically pure starting materials.

The term "deuterated" as used herein refers to a deuterium-containing compound produced by substitution of one or more hydrogens in the group of the compound of formula I with deuterium. The disclosed deuterium labeled or alternative therapeutic compounds may improve DMPK (drug metabolism and pharmacokinetics) properties, related to distribution, metabolism and excretion (ADME). Substitution with heavier isotopes (such as deuterium) may be of some therapeutic advantage because of greater metabolic stability, for example increased in vivo half-life, reduced dosage requirements and/or improved therapeutic guidelines.

The term "substituted" as used herein refers to any one or more hydrogen atoms on a specified atom or group, substituted with one or more substituents other than hydrogen, so long as the normal valence of the specified atom is not exceeded.

The term "as used herein" ¹⁴ C-tag "means that in the compounds of the present invention, the radionuclide carbon-14 (i.e ¹⁴ C) Substituted compound of its stable isotope carbon-12 (i.e ¹² C) And with carbon-14 (i.e ¹⁴ C) Radiolabeled compounds as labels.

Certain isotopically-labeled compounds of the present invention (e.g., with ³ H and ¹⁴ c-labeled) are used in compound and/or substrate tissue distribution assays. Tritium (i.e. tritium) ³ H) And carbon-14 (i.e ¹⁴ C) Isotopes are particularly preferred for their ease of preparation and detection. Furthermore, substituents containing heavier isotopes such as deuterium (i.e ² H) May have certain therapeutic advantages because of their better metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and thus may be preferred for use in certain circumstances. Isotopically-labeled compounds of the present invention can generally be prepared by those methods well known in the art by substituting a suitable isotopically-labeled reagent for a non-isotopically-labeled reagent.

The term "solvate" as used herein refers to an association of one or more solvent molecules with a compound of the present invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethylsulfoxide, ethyl acetate, acetic acid, ethanolamine, or mixtures thereof. The term "hydrate" refers to an association of solvent molecules that are water. When the solvent is water, the term "hydrate" may be used. In one embodiment, a molecule of a compound of the invention may be associated with a water molecule, such as a monohydrate; in another embodiment, one molecule of the compounds of the present invention may be combined with more than one water molecule, such as dihydrate; in yet another embodiment, one molecule of the compounds of the present invention may be associated with less than one water molecule, such as a hemihydrate. It should be noted that the hydrates described in the present invention retain the biological effectiveness of the compounds in a non-hydrated form.

The term "metabolite" as used herein refers to a product obtained by metabolizing a specific compound or salt thereof in vivo. The metabolites of a compound may be identified by techniques well known in the art and their activity may be characterized by employing the assay methods as described herein. Such products may be obtained by oxidation, reduction, hydrolysis, amidization, deamination, esterification, degreasing, enzymatic cleavage, etc. of the administered compound. Accordingly, the present invention includes metabolites of compounds, including metabolites produced by contacting a compound of the present invention with a mammal for a period of time sufficient.

The term "prodrug" as used herein, means a compound that is converted in vivo to a compound of formula I. Such conversion is effected by hydrolysis of the prodrug in the blood or enzymatic conversion to the parent structure in the blood or tissue. The prodrug of the invention can be ester, and in the prior invention, the ester can be phenyl ester, aliphatic (C _1-24 ) Esters, acyloxymethyl esters, carbonates, carbamates and amino acid esters. For example, one compound of the invention may contain a hydroxyl group, i.e., it may be acylated to provide the compound in a prodrug form. Other prodrugs Forms include phosphates, such as those obtained by phosphorylation of a hydroxyl group on the parent.

Unless otherwise indicated, all suitable pharmaceutically acceptable salts, esters, stereoisomers, tautomers, deuterides, and, ¹⁴ The C-tag, solvate, metabolite or prodrug are included within the scope of the invention.

In the various parts of the present specification, substituents of the presently disclosed compounds are disclosed in terms of the type or scope of groups. It is specifically noted that the present invention includes each individual subcombination of the individual members of these group classes and ranges. For example, the term "C _1-12 Alkyl "means in particular methyl, ethyl, C independently disclosed ₃ Alkyl, C ₄ Alkyl, C ₅ Alkyl, C ₆ Alkyl, C ₇ Alkyl, C ₈ Alkyl, C ₉ Alkyl, C ₁₀ Alkyl, C ₁₁ Alkyl and C ₁₂ An alkyl group.

The term "alkyl" or "alkyl group" as used herein means a saturated, straight or branched, monovalent hydrocarbon group containing 1 to 12 carbon atoms, wherein the alkyl group may be optionally substituted with one or more substituents as described herein. In some embodiments, the alkyl group contains 1 to 6 carbon atoms; in other embodiments, the alkyl group contains 1 to 4 carbon atoms; in some embodiments, the alkyl group contains 1 to 3 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl (Me, -CH ₃ ) Ethyl (Et, -CH) ₂ CH ₃ ) N-propyl (n-Pr, -CH) ₂ CH ₂ CH ₃ ) Isopropyl (i-Pr, -CH (CH) ₃ ) ₂ ) N-butyl (n-Bu, -CH) ₂ CH ₂ CH ₂ CH ₃ ) Isobutyl (i-Bu, -CH) ₂ CH(CH ₃ ) ₂ (2-methylpropyl)) sec-butyl (s-Bu, -CH (CH) ₃ )CH ₂ CH ₃ (1-methylpropyl)) tert-butyl (t-Bu, -C (CH) ₃ ) ₃ ) Etc.

The term "halo" as used herein refers to one or more halogen substituents, i.e., one or more chlorine, fluorine, bromine or iodine atoms.

As used herein, the term "alkenyl" or "alkenyl group" means a straight or branched chain monovalent hydrocarbon radical containing 2 to 12 carbon atoms having at least one site of unsaturation, i.e., having one carbon-carbon sp ² A double bond, wherein the alkenyl group may be optionally substituted with one or more substituents described herein, including "cis" and "trans" positioning, or "E" and "Z" positioning. In one embodiment, the alkenyl group contains 2 to 8 carbon atoms; in another embodiment, the alkenyl group comprises 2 to 6 carbon atoms; in yet another embodiment, the alkenyl group contains 2 to 4 carbon atoms. Examples of alkenyl groups include, but are not limited to, vinyl (-ch=ch) ₂ ) Allyl (-CH) ₂ CH＝CH ₂ ) 1-propenyl (i.e., propenyl, -ch=ch-CH) ₃ ) Etc.

The term "alkynyl" or "alkynyl group" as used herein means a straight or branched chain monovalent hydrocarbon radical containing from 2 to 12 carbon atoms, wherein there is at least one site of unsaturation, i.e., one carbon-carbon sp triple bond, wherein the alkynyl group may be optionally substituted with one or more substituents as described herein. In one embodiment, the alkynyl group contains 2 to 8 carbon atoms; in another embodiment, the alkynyl group contains 2 to 6 carbon atoms; in yet another embodiment, the alkynyl group contains 2 to 4 carbon atoms. Examples of alkynyl groups include, but are not limited to, ethynyl (-C≡CH), propargyl (-CH) ₂ C.ident.CH), 1-propynyl (i.e., propynyl, -C.ident.C-CH ₃ ) And so on.

In a preferred embodiment, R ₁ 、R ₂ 、R ₃ And R is ₄ Are identical or different and are each independently selected from hydrogen, halogen, hydroxy, C _1-8 Alkyl, C _1-8 Haloalkyl, C _1-8 Alkoxy, C _2-8 Alkenyl, C _2-8 Haloalkenyl, C _2-8 Alkynyl or C _2-8 Haloalkynyl.

In a preferred embodiment, R ₁ 、R ₂ 、R ₃ And R is ₄ Is the same asOr different and each independently selected from hydrogen, halogen, hydroxy, C _1-5 Alkyl, C _1-5 Haloalkyl, C _1-5 Alkoxy, C _2-5 Alkenyl, C _2-5 Haloalkenyl, C _2-5 Alkynyl or C _2-5 Haloalkynyl.

In a preferred embodiment, R ₁ 、R ₂ 、R ₃ And R is ₄ Are identical or different and are each independently selected from hydrogen, halogen, hydroxy or methyl.

In a preferred embodiment, the halogen is fluorine, chlorine or bromine.

In a preferred embodiment, the halogen is chlorine.

In a preferred embodiment, the triterpene compound has the following structural formula:

in a preferred embodiment, the triterpenes are obtained by hydrolysis of aqueous extracts of gynostemma pentaphylla by acid hydrolysis.

An effective dose of the triterpene compound or the pharmaceutically acceptable salt, ester, stereoisomer, tautomer and deuterate thereof, ¹⁴ C markers, solvates, metabolites or prodrugs and pharmaceutically acceptable excipients are used for the production of the pharmaceutical composition. The dosage of the active compound can vary with the route of administration, the age and weight of the patient, the nature and severity of the condition to be treated, and the like.

The pharmaceutical compositions of the compounds of the present invention may be administered in any of the following ways: oral, spray inhalation, rectal, nasal, buccal, topical, parenteral, e.g., subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal and intracranial injection or infusion, or by means of an explanted reservoir. Among them, oral, topical, intraperitoneal or intravenous administration is preferred.

In a preferred embodiment, the adjuvant is selected from one or more of the following: preservatives, lubricants, diluents, suspending agents, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, flavoring agents, coloring agents, buffers, disintegrants, thickening agents, masking agents and antioxidants.

When administered orally, the compounds of the present invention may be formulated in any orally acceptable formulation, including, but not limited to, tablets, capsules, aqueous solutions or suspensions. Wherein the tablet comprises lactose and corn starch, and optionally lubricant such as magnesium stearate. Diluents used in capsule formulations generally include lactose and dried corn starch. Aqueous suspension formulations are usually prepared by mixing the active ingredient with suitable emulsifying and suspending agents. If desired, some sweetener, flavoring agent or coloring agent may be added to the above oral preparation.

Aqueous solutions for oral administration can be prepared as follows: the active ingredient is dissolved in water and appropriate colorants, flavors, stabilizers and thickeners are added as desired. Aqueous suspensions for oral administration can be prepared as follows: finely divided active component is dispersed in water together with viscous materials such as natural/synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose and other suspending agents known in the art of pharmaceutical formulations.

When topically applied, the compounds of the present invention can be formulated into skin external preparations which can include any general term for application to the outside of the skin, and various dosage forms of pharmaceuticals and cosmetics can be included therein. The selection may be made appropriately according to the intended purpose. For example, one or more dosage forms selected from the group consisting of skin lotions, skin softeners, skin lotions, astringent lotions, milk lotions, moisturizing lotions, nutritional lotions, massage creams, nutritional creams, moisturizing creams, hand creams, foundations, essences, nutritional essences, facial masks, soaps, foam facial washes, makeup removers, cleansing creams, body milks, and bath milks may be prepared, but are not limited thereto.

Within the understanding of the person skilled in the art, the auxiliary materials mentioned are only illustrative and are not intended to be limiting.

Non-limiting examples of preservatives include, but are not limited to, benzoic acid, EDTA, phenolic acid, sorbic acid, benzyl alcohol, isopropyl alcohol, benzethonium chloride, bronopol, butyl paraben, cetyltrimethylammonium bromide, chlorhexidine, chlorobutanol, chlorocresol, cresol, ethylparaben, glycerol, imidurea, methyl paraben, phenol, phenoxyethanol, phenylmercuric acetate, phenylmercuric borate, phenylmercuric nitrate, potassium sorbate, propylene glycol, propylparaben, sodium benzoate, sodium propionate, sorbic acid, sodium ethyl mercuric sulfosalicylate, and mixtures thereof.

Non-limiting examples of lubricants include, but are not limited to, calcium stearate, low mustard seed oil, glyceryl palmitostearate, hydrogenated vegetable oil, magnesium oxide, mineral oil, poloxamer, polyethylene glycol, polyvinyl alcohol, sodium benzoate, sodium lauryl sulfate, sodium stearyl fumarate, stearic acid, sterilized corn starch, talc, zinc stearate, and mixtures thereof.

Non-limiting examples of diluents include, among others, calcium phosphate, calcium sulfate, carboxymethylcellulose calcium, cellulose acetate, dextrose binder (dextran), dextrin, dextrose, fructose, glyceryl palmitostearate (glycerolpalmitoleate), kaolin, lactitol, lactose, magnesium carbonate, magnesium oxide, maltitol, maltodextrin, maltose, microcrystalline cellulose, polymethacrylates, powdered cellulose, pregelatinized starch, silicified microcrystalline cellulose, sodium chloride, sorbitol, starch, sucrose, sugar, talc, hydrogenated vegetable oils, and mixtures thereof.

Non-limiting examples of suspending agents include, among others, alginic acid, bentonite, carbomer, carboxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, microcrystalline cellulose, dextrin, gelatin, guar gum, xanthan gum, kaolin, magnesium aluminum silicate, maltitol, methylcellulose, polysorbate, povidone, propylene glycol alginate, sodium alginate, sorbitan esters, tragacanth, and mixtures thereof.

Non-limiting examples of the solubilizing agent include, but are not limited to, benzalkonium chloride, beaver oil, cyclodextrin, polyoxyethylene ether, glyceryl monostearate, lecithin, poloxamer, polysorbate, polyoxyethylene stearate, sorbitan ester, stearic acid, and mixtures thereof.

Non-limiting examples of stabilizers include, among others, acacia, albumin, polyvinyl alcohol, alginic acid, bentonite, carboxymethylcellulose, hydroxypropyl cellulose, silica gel, cyclodextrin, glyceryl monostearate, hydroxypropyl methylcellulose, magnesium aluminum silicate, propylene glycol alginate, sodium alginate, waxes, xanthan gum and mixtures thereof.

Non-limiting examples of humectants include, but are not limited to, glycerin, propylene glycol, sorbitol, triacetin, and mixtures thereof.

Non-limiting examples of disintegrants include, among others, alginic acid, sodium carboxymethyl cellulose, hydroxypropyl cellulose, microcrystalline cellulose, silica gel, croscarmellose sodium (croscarmellose sodium), crospovidone, magnesium aluminum silicate, methylcellulose, polacrilin, povidone, sodium alginate, sodium starch glycolate, starch, and mixtures thereof.

Non-limiting examples of emulsifiers include, among others, pectin, soy lecithin, glycerol monostearate, castor oil, and poloxamer.

Non-limiting examples of sweeteners include, among others, sugar alcohols, such as sorbitol, xylitol; monosaccharides, including sucrose, glucose, fructose, and lactose; sorbitol, mannitol, glycerol, xylitol, erythritol, maltitol syrup, isomalt, lactitol; and mixtures of sugars, including glucose syrup; invert syrup, high sugar syrup; malt or malt extract.

Non-limiting examples of colorants include, among others, titanium dioxide, flavone dyes, isoquinoline dyes, polyene colorants, pyran colorants, naphthoquinone dyes, quinone and anthraquinone dyes, benzopyran dyes, benzopyrone dyes, and indigoid dyes and indole colorants. Examples thereof are caramel coloring, carmine, chlorophyllin, cochineal, betanin, turmeric, saffron, red pepper, lycopene, paphiopedilum and pterocarpus marsupium.

Non-limiting examples of fragrances include, but are not limited to, bergamot oil, coriander oil, dimethyl heptanol, dimethylbenzyl orthoacetate, geranyl acetate, citronellyl acetate, rose composition, phyllostachys pratensis, methyl dihydrojasmonate, isoeugenol, methyl eugenol threonate acetate, elemene, l-rose oxide, C-11 cycloundecalaldehyde, derivatives of 2, 6-dimethyl-2-alkoxyoct-7-ol, miscanthus oil, vetiverol acetate, guaiac wood oil, anthranilate, benzyl salicylate, benzyl benzoate, oak, eugenol, p-tert-butylcyclohexyl acetate, and coumarin.

Non-limiting examples of antioxidants include, among others, vitamin E (i.e., tocopherol), ascorbic acid, sodium metabisulfite, butylhydroxytoluene, butylated hydroxyanisole, edetic acid, and edetate.

Among these, it may be advantageous to use thickeners that are resistant to the lower pH range. For example, neutralized thickeners do not impart desirable thickening or sensory properties to the composition formulation at low pH. On the other hand, fatty alcohols such as cetyl alcohol and stearyl alcohol are generally stable at low pH.

Non-limiting examples of masking agents include, among others, essential oils, aroma chemicals, fragrances, and the like, such as ambergris, borneol and its esters, carvone, castoreum, civet, cinnamaldehyde, citral, clove oil, white rosin, jasmine yellow, camphol and its esters, pinene, rosemary oil, sandalwood, terpineol, terpinene, benzaldehyde, benzoin, and isoamyl acetate.

In a preferred embodiment, the pharmaceutical composition further comprises one or more of the following drugs: antiinfective, antitussive, antiasthmatic, expectorant and antihistamine.

In a preferred embodiment, the pharmaceutical composition further comprises an antiasthmatic agent.

In a preferred embodiment, the anti-asthmatic is selected from one or more of the following: bronchodilators, anti-inflammatory antiasthmatics and antiallergic antiasthmatics.

Among them, bronchodilators include β2 receptor agonists, theophyllines, and M receptor blockers.

Among them, anti-inflammatory antiasthmatic drugs, which can achieve the effect of preventing asthma attacks for a long period of time by inhibiting airway inflammatory responses, have become the first-line drugs among antiasthmatic drugs, for example, glucocorticoids are the drugs having the strongest anti-inflammatory effect among anti-inflammatory antiasthmatic drugs and antiallergic effect.

Among them, antiallergic antiasthmatic agents exert antiallergic and slightly antiinflammatory effects by inhibiting the immunoglobulin E-mediated mast cell release mediator and inhibiting the activity of inflammatory cells such as macrophages, eosinophils, monocytes, etc. It has slow effect of relieving asthma, is not suitable for treating asthma in acute attack stage, and is clinically mainly used for preventing and controlling asthma attack for a long time.

In a preferred embodiment, the anti-asthmatic is one or more of the following: salbutamol sulphate, aminophylline, diprophylline, ipratropium bromide, oxitropium bromide, sodium tryptophanate, beclomethasone dipropionate, epinephrine, isoprenaline, ephedrine, albuterol and amicin.

The compounds of the invention may be administered simultaneously with, or before or after, one or more additional therapeutic agents for the treatment of asthma. The compounds of the present invention may be administered separately from other additional therapeutic agents by the same or different routes of administration, or in the same pharmaceutical compositions as they are.

Wherein salbutamol sulfate is used for treating bronchospasm of patients suffering from bronchial asthma, asthmatic bronchitis and emphysema.

Among them, ipratropium bromide is a bronchodilator, clinically used: (1) relieving bronchospasm and wheezing symptoms caused by Chronic Obstructive Pulmonary Disease (COPD); (2) preventing and treating asthma, especially for patients who cannot tolerate such drugs due to muscle tremor and tachycardia caused by beta receptor agonists.

Among them, aminophylline has strongest relaxation effect on smooth muscle of bronchus, can expand bronchus, increase vital capacity, and has lasting effect, especially has remarkable effect on bronchus in spasticity.

Among them, diprophylline is a smooth muscle relaxant, has the effect of dilating bronchi and coronary arteries, and has diuretic effect. It has similar antiasthmatic effect to aminophylline, and is especially suitable for tachycardia.

Among them, oxitropium bromide is useful for the treatment of chronic obstructive respiratory disease, chronic obstructive bronchitis, bronchial asthma and pulmonary edema asthma accompanied by reversible tone elevation of bronchial smooth muscle.

Wherein, cromolyn sodium inhibits degranulation caused by various stimulus of mast cells, inhibits release of allergic medium after antigen-antibody combination, and can prevent asthma attack.

Among them, beclomethasone dipropionate is a powerful external glucocorticoid medicine, has the functions of anti-inflammatory, antiallergic and antipruritic, and can inhibit bronchial exudates, eliminate bronchial mucosa swelling and relieve bronchospasm.

Wherein ephedrine is used for preventing bronchial asthma attacks and relieving mild asthma attacks.

Wherein albuterol is used for treating asthmatic bronchitis, bronchial asthma or bronchial asthma of emphysema patients, wherein aerosol type is used for treating the attack, and oral administration is used for preventing the attack.

Wherein, the ammonia wheezing is used for preventing and treating bronchial spasm caused by respiratory diseases such as bronchial asthma, asthma type chronic bronchitis, emphysema and the like.

In a preferred embodiment, the amount of water used for the extraction in step (1) is 1 to 5 times by mass, respectively, the mass of the dried whole herb of gynostemma pentaphylla.

In a preferred embodiment, the acid in step (2) is selected from one or more of hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, acetic acid, propionic acid, malonic acid and butyric acid.

In a preferred embodiment, the column chromatography in step (3) is performed with silica gel having a particle size of 100 to 400 mesh.

In a preferred embodiment, the eluting mobile phase of the column chromatography is a mixed solvent with a volume ratio of n-hexane to ethyl acetate of 25:1 to 15:1.

In a preferred embodiment, the volume ratio of the acid solution to the aqueous extract is from 1:3 to 1:10.

In a preferred embodiment, the acid in step (2) is hydrochloric acid.

In a preferred embodiment, the respiratory disease is selected from one or more of the following: asthma, tracheitis, bronchitis, chronic obstructive pulmonary disease, pulmonary heart disease and tuberculosis.

In a preferred embodiment, the asthma is selected from one or more of the following: allergic asthma, drug-induced asthma, senile asthma, cough variant asthma, chronic asthma, athletic asthma and childhood asthma.

Among them, asthma is also called bronchial asthma, which is a heterogeneous disease characterized by chronic airway inflammation involving various cells (such as eosinophils, mast cells, T lymphocytes, neutrophils, airway epithelial cells, etc.) and cellular components, which is associated with airway hyperreactivity, and which usually presents with extensive and variable reversible expiratory airflow limitation, resulting in recurrent wheezing, shortness of breath, chest distress, and/or cough, and the like, with intensity varying with time. Most patients can self-relieve or treated relieve, usually at night and/or in the early morning. Bronchial asthma, such as untimely diagnosis and treatment, can produce irreversible airway constriction and airway remodeling with prolonged course of disease.

Among these, all asthma attacks caused by drugs are collectively referred to as drug-induced asthma, including asthma caused by the administration of certain drugs to asthmatics or to patients who have exacerbated asthma attacks and have no history of asthma, of which aspirin-based drugs are most common.

In a preferred embodiment, the drug is acetylcholine chloride and/or histamine phosphate.

In a preferred embodiment, the drug is an isovolumetric mixture of 2% acetylcholine chloride and 0.1% histamine phosphate.

In the present invention, the term "treatment" also includes "prophylaxis" unless there is a specific description of the contrary. The terms "therapeutic" and "therapeutic" should be understood accordingly.

The term "treating" as used herein refers in some embodiments to ameliorating a disease or disorder (i.e., slowing or preventing or alleviating the progression of the disease or at least one clinical symptom thereof). In other embodiments, "treating" refers to moderating or improving at least one physical parameter, including physical parameters that may not be perceived by the patient. In other embodiments, "treating" refers to modulating a disease or disorder physically (e.g., stabilizing a perceived symptom) or physiologically (e.g., stabilizing a parameter of the body) or both. In other embodiments, "treating" refers to preventing or delaying the onset, or exacerbation of a disease or disorder.

The term "preventing" as used herein refers to a reduction in the risk of a disease or disorder (i.e., stopping the progression of at least one clinical symptom of a disease in a subject who may or may not have been predisposed to such a disease, but who has not experienced or exhibited symptoms of the disease).

The present invention also provides a method for preventing and/or treating respiratory diseases, which comprises administering to said patient a therapeutically effective amount of the above triterpene compound or the above pharmaceutical composition.

The term "therapeutically effective amount" as used herein refers to the total amount of each active ingredient sufficient to exhibit a meaningful patient benefit. When separate active ingredients are used for separate administration, the term refers only to the ingredient. When applied in combination, the term refers to the combined amounts of the active ingredients that result in a therapeutic effect, whether administered in combination, sequentially or simultaneously. This means that any amount of the compound, when administered appropriately, is sufficient to achieve the desired prophylactic or therapeutic effect in the patient to whom it is administered. Generally, depending on the condition to be prevented or treated and the route of administration. Substantially satisfactory results are obtained when a daily dose of about 0.01mg to about 100mg per kg of animal body weight is administered, preferably a single daily dose. For large mammals, the total daily dose is from about 0.1mg to about 1000mg, preferably from about 0.2mg to about 50mg. In the case of a 70kg adult human, the total daily dose is approximately from about 0.2mg to 200mg. The dosage regimen can be adjusted to provide an optimal therapeutic response. The amount administered, the route of administration and further treatment regimen may be determined by the treating clinician, depending on, for example: age, sex, general condition of the patient, nature and severity of the disease/condition to be treated, and the like.

The term "patient" as used herein refers to a human (including adults and children) or other animals. In some embodiments, "patient" refers to a human.

It should be noted that, in the case of no conflict, the embodiments and features in the embodiments may be combined with each other. The present invention will be described in detail with reference to examples.

The invention is described in further detail below in connection with specific embodiments, which should not be construed as limiting the scope of the invention as claimed.

Examples

Preparation example one

50Kg of gynostemma pentaphylla medicinal material is added with 100L of water for reflux extraction for 2 hours, reduced pressure concentration is carried out until the concentration reaches 5L, 1L of 1% hydrochloric acid aqueous solution is added, reflux is carried out for 1 hour, filtration is carried out, sediment is collected, and is eluted by n-hexane-ethyl acetate (15:1) through silica gel column chromatography, and Gypensapogenin III 2.4.4 g is obtained through recrystallization.

Preparation example two

50Kg of gynostemma pentaphylla medicinal material is added with 150L of water for reflux extraction for 1.5 hours, reduced pressure concentration is carried out until 5L is reached, 0.5% hydrochloric acid aqueous solution is added for 1L, reflux is carried out for 0.5 hour, filtration and collection are carried out, sediment is subjected to silica gel column chromatography, elution is carried out by n-hexane-ethyl acetate (20:1), and Gypensapogenin III 2.1.1 g is obtained through recrystallization.

Preparation example III

50Kg of gynostemma pentaphylla medicinal material is added with 50L of water for reflux extraction for 2.5 hours, reduced pressure concentration is carried out until 5L is reached, 1L of 1.5% hydrochloric acid aqueous solution is added, reflux is carried out for 2 hours, filtration and collection are carried out, sediment is chromatographed by a silica gel column, eluting is carried out by n-hexane-ethyl acetate (15:1), and Gypensapogenin III 2.0.0 g is obtained through recrystallization.

Preparation example IV

50Kg of gynostemma pentaphylla medicinal material is added with 200L of water for reflux extraction for 1 hour, reduced pressure concentration is carried out to 5L, 2% hydrochloric acid aqueous solution is added for 1L, reflux is carried out for 2 hours, filtration is carried out, sediment is collected, silica gel column chromatography is carried out, elution is carried out by n-hexane-ethyl acetate (25:1), and Gypensapogenin III 2.0.0 g is obtained through recrystallization.

Preparation example five

50Kg of gynostemma pentaphylla medicinal material is added with 250L of water for reflux extraction for 0.5 hour, reduced pressure concentration is carried out until the concentration reaches 5L, 2.5% hydrochloric acid aqueous solution is added for 1L, reflux is carried out for 1.5 hours, filtration is carried out, sediment is collected, chromatography is carried out by a silica gel column, elution is carried out by n-hexane-ethyl acetate (25:1), and Gypensapogenin III 2.1.1 g is obtained through recrystallization.

Gypensapogenin III hydrogen, carbon and mass spectral data:

¹ H-NMR(C5D5N,400M)δ _H ：5.03,4.91,4.39,3.61,3.50,3.34,3.01,2.69,2.51,2.44,2.35,2.30；

¹³ C-NMR(C5D5N,100M)δ _C ：211.4,178.6,79.2,70.3,57.5,57.2,52.5,51.0,47.1,45.4,44.0,43.7,41.9,40.8,40.7,38.6,36.9,32.5,31.7,31.2,29.9,29.5,26.9,26.8,22.8,19.9,17.7,17.6,17.3,17.0；

ESI-MS：491.3(C ₃₀ H ₅₀ O ₅ +H)。

evaluation of pharmacological Activity

1 Experimental materials

1.1 Experimental drugs

Positive drug: salbutamol sulfate, lot number: 1305111 Jiangsu Pont epleren pharmaceutical Co., ltd;

test drug Gypensapogenin III was prepared as in the preparation example described above.

1.2 laboratory animals

1.2.1 sources, species, lines, eligibility: guinea pigs, of plain grade, purchased from the university of in-line medical science laboratory animal center, animal production license number: SCXK (qin) 2019-0001.

1.2.2 body weight: 150-180g

1.2.3 gender: the male and female parts are respectively half

1.2.4 number of animals per group: 8 pieces/group

1.2.5 raising: animal feeding at 25+ -3deg.C with relative humidity of 60-75%, day and night light and shade alternation time of 12 hr, free feeding and drinking water, and adaptive feeding for one week.

1.3 major reagents

Acetylcholine chloride: national drug group chemical reagent Co., ltd., lot number: w20101111;

histamine phosphate: national drug group chemical reagent Co., ltd., lot number: f20081222;

sodium carboxymethyl cellulose: national drug group chemical reagent Co., ltd., lot number: 30036328 purity grade: a CP;

vitamin C: nanjing Baijingyu pharmaceutical Limited liability company, lot number: 130903.

1.4 Main instruments

YLS-8A multifunctional cough and asthma inducing instrument: jinan Yiyan technology development Limited company;

sartorius analytical balance: beijing Sidoris balance Co., ltd;

KQ-250E type ultrasonic cleaner: kunshan, ultrosophy, inc.

2 Experimental methods

2.1 grouping of animals

The 40 guinea pigs were randomly divided into 5 groups of 8 each:

(1) model group: the solvent was administered by gavage as a 0.5% CMC-Na solution.

(2) Test drug Gypensapogenin III high dose group: the solution was administered by gavage at 0.9g/kg/d Gypensapogenin III.

(3) Test drug Gypensapogenin III high dose group: the solution was administered by gavage at 0.3g/kg/d Gypensapogenin III.

(4) Test drug Gypensapogenin III high dose group: the solution was administered by gavage at 0.1g/kg/d Gypensapogenin III.

(5) Salbutamol sulfate group as positive drug: 3mg/kg/d albuterol sulfate solution was administered by gavage.

Each of the above groups was continuously administered for 5 days, and the post-administration asthma-inducing latency was measured 1 hour after the last administration.

2.2 preparation of medicaments and reagents

(1) Solvent 0.5% cmc-Na solution: taking distilled water 500ml from a beaker, weighing 2.5g of sodium carboxymethylcellulose, sprinkling on the water surface, standing for about one day to obtain clear 0.5% CMC-Na solution, and preserving at 4 ℃ for later use.

(2) Test agent Gypensapogenin III high dose group drug solution: accurately weighing 0.9. 0.9g Gypensapogenin III powder, placing into a small beaker, adding a proper amount of 0.5% CMC-Na solution, placing the beaker into an ultrasonic cleaner to make the beaker fully suspended by ultrasonic, transferring the liquid medicine into a volumetric flask, fixing the volume to 20ml by using the 0.5% CMC-Na solution, shaking uniformly to obtain 45mg/ml high-concentration liquid medicine, and preserving at low temperature for later use.

(3) Dose group liquor in test drug Gypensapogenin III: sucking 5ml of the prepared high-concentration liquid medicine, adding 10ml of 0.5% CMC-Na solution for dilution, shaking uniformly to obtain 15mg/ml of medium-concentration liquid medicine, and preserving at low temperature for later use.

(4) Test agent Gypensapogenin III low dose group drug solution: sucking 5ml of the prepared medium-concentration liquid medicine, adding 10ml of 0.5% CMC-Na solution for dilution, shaking uniformly to obtain 5mg/ml of low-concentration liquid medicine, and preserving at low temperature for later use.

(5) Salbutamol sulfate liquor as positive medicine: taking a albuterol sulfate tablet (6 mg), fully grinding in a mortar, transferring into a small beaker, adding a proper amount of 0.5% CMC-Na solution, placing the beaker into an ultrasonic cleaner to fully ultrasonically suspend the albuterol sulfate tablet, transferring the liquid medicine into a volumetric flask, fixing the volume to 20ml by using the 0.5% CMC-Na solution, and shaking uniformly to obtain the liquid medicine with the concentration of 0.3mg/ml, and preparing the albuterol sulfate tablet for use.

(6) Asthma-relieving agent: weighing 1g of acetylcholine chloride, placing into a beaker, adding a proper amount of distilled water to dissolve the acetylcholine chloride, transferring the solution into a volumetric flask to constant volume to 50ml, and obtaining 2% acetylcholine chloride solution; weighing 0.05g of histamine phosphate, placing in a beaker, adding a proper amount of distilled water to dissolve the histamine phosphate, transferring the solution into a volumetric flask to a constant volume of 50ml to obtain a 0.1% histamine phosphate solution; and mixing the two solutions in equal volume to obtain equal volume mixed solution of 2% of acetylcholine chloride and 0.1% of histamine phosphate.

2.3 administration of drugs

The negative control group is administrated with solvent by stomach irrigation, and the other groups are administrated with corresponding drugs by stomach irrigation respectively, wherein the administration volume is 1ml/100g, and the administration is carried out for five days.

2.4 differential measurement of asthma-relieving latency

Animal screening was performed the day prior to the official experiment: a plurality of guinea pigs are taken and put into an animal box of a YLS-8A multifunctional cough and asthma inducing instrument one by one, the asthma inducing agent is a volume mixed solution of 2% of acetylcholine chloride, 0.1% of histamine phosphate and the like, and the asthma inducing time is set to be 15S. The pre-dose asthma latency (time from spraying to asthma onset, dyspnea until tics fall) of each guinea pig was recorded and was not selected for > 150S.

The asthma-inducing latency of each group of animals was measured 1h after the last administration as in the above method, and it was the asthma-inducing latency after the administration of each guinea pig. Observing 360S, the asthma latency of the non-falling person is calculated as 360S. The post-administration asthma-inducing latency was subtracted from the pre-administration asthma-inducing latency to give a difference.

3 statistical processing method

Mean ± standard deviation for each set of dataAnd (3) representing. Single-factor analysis of variance (one-way ANOVA) was performed on differences in pre-and post-administration incubation periods of guinea pigs in each group using Graphpad Prism 5 software, and the difference in pre-and post-administration incubation periods of each group was compared with the difference in negative control group for statistical significance, while the difference in pre-and post-administration incubation periods of each group was compared using t-test, p<0.05 indicates that the difference is statistically significant.

4 experimental results

The Gypensapogenin III high dose group was able to significantly prolong guinea pig asthma latency (P < 0.001) compared to the negative control group, and post-dosing priming latency was significantly different from pre-dosing (P < 0.001); the positive drug salbutamol sulphate significantly prolonged guinea pig asthma latency (P < 0.001) compared to the negative control group, and the post-dosing asthma latency of this group was significantly different (P < 0.01) compared to the pre-dosing.

Table 1 effect of Gypensapogenin III drug on guinea pig acetylcholine-histamine asthma-inducing latency (n=8,

p <0.05, < P <0.01, < P <0.001, compared to the negative control group;

in comparison to the pre-dose latency of each group, ^# p<0.05, ^## P<0.01, ^### P<0.001。

the foregoing is merely exemplary embodiments of the present invention, and specific structures and features that are well known in the art are not described in detail herein. It should be noted that modifications and improvements can be made by those skilled in the art without departing from the scope of the invention, which is also to be considered as the scope of the invention, and which does not affect the effect of the invention and the utility of the patent. The protection scope of the present application shall be subject to the content of the claims, and the description of the specific embodiments and the like in the specification can be used for explaining the content of the claims.

The foregoing has outlined rather broadly the more detailed description of embodiments of the present application, wherein specific examples have been provided herein to illustrate the principles and embodiments of the present application, and wherein the above examples are provided to assist in the understanding of the methods and concepts of the present application. Meanwhile, based on the ideas of the present application, those skilled in the art can make changes or modifications on the specific embodiments and application scope of the present application, which belong to the scope of the protection of the present application. In view of the foregoing, this description should not be construed as limiting the application.

Claims

1. A triterpene compound with the following structural formula or pharmaceutically acceptable salt thereof,

2. the triterpene compound according to claim 1, wherein the triterpene compound is obtained by hydrolyzing an aqueous extract of gynostemma pentaphylla by an acid hydrolysis method.

3. A pharmaceutical composition comprising a triterpene compound according to claim 1 or 2 and a pharmaceutically acceptable adjuvant.

4. A pharmaceutical composition according to claim 3, wherein the adjuvant is selected from one or more of the following: preservatives, lubricants, diluents, suspending agents, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, flavoring agents, coloring agents, buffers, disintegrants, thickening agents, masking agents and antioxidants.

5. The pharmaceutical composition of claim 3, wherein the pharmaceutical composition further comprises one or more of the following drugs: antiinfective, antitussive, antiasthmatic, expectorant and antihistamine.

6. The pharmaceutical composition of claim 3, wherein the pharmaceutical composition further comprises an antiasthmatic agent.

7. The pharmaceutical composition of claim 6, wherein the anti-asthmatic is selected from one or more of the following: bronchodilators, anti-inflammatory antiasthmatics and antiallergic antiasthmatics.

8. The pharmaceutical composition of claim 6, wherein the anti-asthmatic is one or more of the following: salbutamol sulphate, aminophylline, diprophylline, ipratropium bromide, oxitropium bromide, sodium tryptophanate, beclomethasone dipropionate, epinephrine, isoprenaline, ephedrine, albuterol and amicin.

9. A process for the preparation of the triterpene compound according to claim 1 or 2, wherein the process comprises the steps of:

(2) Adding 0.5 to 3 percent of acid solution with a certain volume ratio into the water extract, extracting for 0.5 to 3 hours by ultrasonic or reflux, filtering, and washing with water to obtain neutral precipitate; and

(3) The precipitate is subjected to silica gel column chromatography to obtain the triterpene compound shown in the claim 1.

10. The method according to claim 9, wherein the amount of water used for the extraction in the step (1) is 1 to 5 times by mass of the dry whole herb of gynostemma pentaphylla, respectively.

11. The method of claim 9, wherein the acid in step (2) is selected from one or more of hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, acetic acid, propionic acid, malonic acid, and butyric acid.

12. The method of claim 9, wherein the column chromatography in step (3) is performed with silica gel having a particle size of 100 to 400 mesh.

13. The method of claim 9, wherein the eluting mobile phase of the column chromatography is a mixed solvent with a volume ratio of n-hexane-ethyl acetate of 25:1 to 15:1.

14. The method of claim 9, wherein the volume ratio of the acid solution to the water extract is 1:3 to 1:10.

15. The method of claim 9, wherein the acid in step (2) is hydrochloric acid.

16. Use of a triterpene compound according to claim 1 or 2 or a pharmaceutical composition according to any one of claims 3 to 8 for the manufacture of a medicament for the treatment of respiratory diseases.

17. The use according to claim 16, wherein the respiratory disease is selected from one or more of the following: asthma, tracheitis, bronchitis, chronic obstructive pulmonary disease, pulmonary heart disease and tuberculosis.

18. The use of claim 17, wherein the asthma is selected from one or more of the following: allergic asthma, drug-induced asthma, senile asthma, cough variant asthma, chronic asthma, athletic asthma and childhood asthma.

19. The use according to claim 18, wherein the medicament in the pharmaceutical asthma is acetylcholine chloride and/or histamine phosphate.

20. The use according to claim 18, wherein the medicament in the medicament-induced asthma is a volume mixture of 2% acetylcholine chloride and 0.1% histamine phosphate.