Dammarane type triterpenes, preparation method thereof and application of dammarane type triterpenes in promoting ulcer healing

Abstract

The invention discloses a dammarane type triterpenoid compound or a pharmaceutically acceptable salt thereof. The dammarane type triterpenoid compound has a structural formula as shown in a formula I which is described in the specification. The triterpenoids which are extracted and separated from a gynostemma pentaphyllum medicinal material and have a new chemical structure have an obvious effect of promoting ulcer healing, the application range of gynostemma pentaphyllum can be expanded by developing the triterpenoids into medicines for treating diseases and symptoms related to ulcer, and the utilization value of gynostemma pentaphyllum plant resources is improved. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and particularly to dammarane-type triterpenes, their preparation methods, and their use in promoting ulcer healing. Background Art

[0002] Perianal inflammation refers to a skin disease in which the anus shows itching, pain, increased secretions, resulting in skin erosion, ulcers, skin papules, and lichenoid changes. It often occurs in young and middle-aged people, characterized by nodular ulcers and even the formation of fistulas. This disease is caused by the occlusion of the apocrine glands and secondary bacterial infection.

[0003] Perianal inflammation is mostly caused by Staphylococcus aureus, and sometimes can be caused by Streptococcus, Escherichia coli, and Proteus infections. This disease can also be caused by excessive sweating, poor hygiene, friction, scratching, and trauma. In the initial stage of perianal inflammation, single or several subcutaneous nodules the size of peas will appear, with local hemispherical bulges, which can increase to the size of a walnut, red, swollen, and painful, with mild systemic symptoms. After several weeks or months, the nodules soften, suppurate deeply, and form ulcers after rupture. There are also cases that last for a long time and show a chronic process, or form extensive fistulas.

[0004] Common drugs for treating ulcers caused by perianal inflammation include erythromycin, tetracyclines, etc. They can cause gastrointestinal symptoms such as diarrhea, nausea, vomiting, upper and middle abdominal pain, and loss of appetite. After taking the medicine, liver toxicity such as fatigue, jaundice, and abnormal liver function may occasionally occur, and allergic reactions such as drug fever, rash, and eosinophilia may occasionally occur after taking the medicine.

[0005] Gynostemma pentaphyllum Makino., also known as Gynostemma pentaphyllum and Gospel Herb, is a plant of the genus Gynostemma in the family Cucurbitaceae. Among them, gypenosides are the main active ingredients in Gynostemma pentaphyllum. Pharmacological experiments and clinical applications have shown that it has the effects of reducing blood lipids, anti-tumor, protecting the liver, and enhancing the body's immune function. It is mainly used to treat the proliferation of cancer cells such as liver cancer, lung cancer, uterine cancer, and skin cancer. In addition, it has the effects of nourishing the body, sedation, hypnosis, anti-tension, promoting appetite, reducing cholesterol, and reducing transaminase in the human body.

[0006] The inventors of the present invention have found through research that a class of triterpenoid compounds with a completely new chemical skeleton structure in Gynostemma pentaphyllum has a good effect on promoting ulcer healing and may have broad prospects in the preparation of drugs for preventing and / or treating diseases and disorders related to ulcers. The triterpenoid compounds in Gynostemma pentaphyllum itself have a wide range of pharmacological activities, are derived from natural plants, are safe and reliable, and there are currently no relevant reports on the use of the triterpenoid compounds involved in the present invention for treating and / or preventing diseases and disorders related to ulcers. Summary of the Invention

[0007] The main object of the present invention is to provide a drug for treating diseases and disorders related to ulcers, which has definite curative effects, few side effects, and is natural and safe, so as to solve the problems of common drugs in the prior art, such as easy occurrence of gastrointestinal discomfort, liver and kidney function damage, and easy occurrence of allergic reactions.

[0008] To achieve the above object, according to one aspect of the present invention, there is provided a dammarane-type triterpenoid compound or a pharmaceutically acceptable salt thereof, and the dammarane-type triterpenoid compound has a structural formula shown in Formula I:

[0009]

[0010] Further, the dammarane-type triterpenoid compound is obtained by hydrolyzing the ethanol extract of Gynostemma pentaphyllum with an acid hydrolysis method.

[0011] According to another aspect of the present invention, there is provided a method for preparing the dammarane-type triterpenoid compound, and the method comprises the following steps:

[0012] (1) Weigh an appropriate amount of Gynostemma pentaphyllum, add ethanol for extraction and then concentrate to obtain the ethanol extract of Gynostemma pentaphyllum;

[0013] (2) Add an acid solution with a certain volume ratio to the ethanol extract of Gynostemma pentaphyllum, reflux for a period of time, filter, and collect the precipitate; and

[0014] (3) Subject the precipitate to silica gel column chromatography, elute with a certain mobile phase, and recrystallize the eluate to obtain the dammarane-type triterpenoid compound.

[0015] Further, in step (1), the volume percentage concentration of the ethanol is 40% - 60%, for example, about 50%.

[0016] Further, in step (1), the volume / mass (L / Kg) ratio of the ethanol to the Gynostemma pentaphyllum is 1 - 5, for example, about 2.

[0017] Further, in step (1), the extraction method is reflux extraction, ultrasonic extraction, and / or shaking extraction, for example, reflux extraction.

[0018] Further, in step (1), the extraction time is 0.5 - 3 h, for example, about 2 h.

[0019] Further, in step (1), the concentration is vacuum concentration.

[0020] Further, in step (1), the volume / volume ratio of the ethanol extract of Gynostemma pentaphyllum to the ethanol is 0.05 - 0.2, for example, about 0.1.

[0021] Furthermore, in step (2), the acid is selected from one or more of hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, acetic acid, propionic acid, malonic acid and butyric acid, such as hydrochloric acid.

[0022] Furthermore, in step (2), the volume percentage concentration of the acid is 0.5% to 3%, for example, about 1%.

[0023] Furthermore, in step (2), the volume / volume ratio of the acid to the ethanol extract of Gynostemma pentaphyllum is 0.1 to 1, for example, about 0.2.

[0024] Furthermore, in step (2), the reflux time is 0.5 to 3 hours, for example, about 1 hour.

[0025] Furthermore, in step (3), the column chromatography uses silica gel with a particle size of 100 to 400 mesh.

[0026] Furthermore, in step (3), the elution mobile phase of the column chromatography is a mixed solvent of cyclohexane-ethyl acetate in a volume ratio of (40-60): (0.1-2).

[0027] Furthermore, in step (3), the elution mobile phase of the column chromatography is a mixed solvent of cyclohexane and ethyl acetate in a volume ratio of about 50: about 1.

[0028] According to another aspect of the present invention, a pharmaceutical composition comprising the dammarane-type triterpenoid compound is provided.

[0029] Furthermore, the pharmaceutical composition comprises one or more drugs and / or extracts for promoting ulcer healing.

[0030] Furthermore, the extract is a traditional Chinese medicine extract.

[0031] Furthermore, the Chinese medicine extract is Diyu Huaijiao Pills and / or Qiangli Zhigenduan.

[0032] Furthermore, the drug is nitroglycerin, nifedipine, diltiazem, zinc oxide, erythromycin and / or levofloxacin.

[0033] Furthermore, the pharmaceutical composition comprises one or more pharmaceutically acceptable excipients.

[0034] Furthermore, the auxiliary material is selected from one or more of the following: diluent, wetting agent, binder, disintegrant, inclusion agent, flavoring agent, sustained-release agent, retention agent, lubricant, dispersant, plasticizer, opacifier and antioxidant.

[0035] Furthermore, the dosage form of the pharmaceutical composition is ointment, powder, tablet, pill, capsule, film, lozenge, granule, injection or oral solution.

[0036] Furthermore, the paste is an ointment or a plaster.

[0037] According to another aspect of the present invention, there is provided the use of the above dammarane-type triterpenoids or the above pharmaceutical composition in the preparation of a drug for promoting ulcer healing.

[0038] Furthermore, the ulcer is an oral ulcer and / or a digestive tract ulcer.

[0039] Furthermore, the digestive tract ulcer is a gastric ulcer, a duodenal ulcer and / or a perianal ulcer.

[0040] Furthermore, the digestive tract ulcer is a perianal ulcer.

[0041] Furthermore, the digestive tract ulcer is an ulcer caused by perianal inflammation.

[0042] Advantages of the present invention:

[0043] Gynostemma pentaphyllum is the dried whole herb of the plant Gynostemma pentaphyllum (Thunb.) Makino of the genus Gynostemma in the family Cucurbitaceae. It is a perennial climbing herb, mainly distributed in China, Japan, Korea and other countries. Modern pharmacological research shows that its main active ingredient is gypenoside (Gyp). We have found that a class of triterpenoids with a new chemical structure isolated from Gynostemma pentaphyllum has an obvious effect of promoting ulcer healing. Developing it into a drug for treating ulcer-related diseases and disorders can expand the application range of Gynostemma pentaphyllum and improve the utilization value of Gynostemma pentaphyllum plant resources. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following will briefly introduce the drawings required for the description of the embodiments. Obviously, the following drawings are only some embodiments of the present invention. For those of ordinary skill in the art, other drawings can also be obtained according to these drawings without exceeding the scope of the present invention.

[0045] Figure 1 It is a schematic hydrogen spectrum diagram of the compound of formula I of the present invention.

[0046] Figure 2 It is a schematic carbon spectrum diagram of the compound of formula I of the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0047] The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are some, but not all, of the embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0048] Unless otherwise indicated, all technical and scientific terms and abbreviations used herein have the meanings commonly understood by those of ordinary skill in the art in the field of the present invention or in the field where the term is applied. Although any methods, conditions, substances or materials similar to or equivalent to those disclosed herein may be used in the practice of the present invention, the preferred methods, conditions, substances or materials are described herein.

[0049] The present invention is expected to cover all alternatives, variations and equivalents, which may be included in the field of the existing invention as defined in the claims. Those skilled in the art will recognize many methods and substances similar or equivalent to those described herein, which can be applied to the practice of the present invention. The present invention is not limited to the description of methods and substances.

[0050] The present invention will apply the following definitions unless otherwise indicated. For the purposes of the present invention, chemical elements are defined according to the Periodic Table of the Elements, CAS version, and the Handbook of Chemistry and Physics, 75th Ed., 1994. Additionally, general principles of organic chemistry are found in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March’s Advanced Organic Chemistry", Michael B. Smith and Jerry March, John Wiley & Sons, New York: 2007, and all content incorporates the references.

[0051] The singular forms "a", "an" and "the" as used in the specification and the appended claims include plural referents unless the context clearly dictates otherwise.

[0052] In the present invention, the terms "comprising" and "including" are synonymous. As used herein, the terms "comprising", "including", "having", "containing" or any other variation thereof are intended to cover non-exclusive inclusion. For example, a composition, step, method, article or device containing the listed elements is not necessarily limited to those elements, but may include other elements not expressly listed or elements inherent to such composition, step, method, article or device.

[0053] As described in the background art section, common drugs currently used to treat ulcer-related diseases and disorders are prone to problems such as gastrointestinal discomfort, liver and kidney function damage, and allergic reactions. To solve the above problems, the present invention provides a dammarane-type triterpenoid compound or a pharmaceutically acceptable salt thereof, and the dammarane-type triterpenoid compound has a structural formula shown in Formula I:

[0054]

[0055] The term "salt" used in the present invention refers to a salt of a compound of the present invention that is pharmaceutically acceptable and has the desired pharmacological activity of the parent compound. Suitable salts include all known pharmaceutically acceptable salts, including those formed with organic acids 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.

[0056] In a preferred embodiment, the dammarane-type triterpenoid compound is obtained by hydrolyzing the ethanol extract of Gynostemma pentaphyllum herbs by an acid hydrolysis method.

[0057] According to another aspect of the present invention, there is provided a method for preparing the dammarane-type triterpenoid compound, and the method includes the following steps:

[0058] (1) Weigh an appropriate amount of Gynostemma pentaphyllum herbs, add ethanol for extraction, and then concentrate to obtain an ethanol extract of Gynostemma pentaphyllum herbs;

[0059] (2) Add an acid solution with a certain volume ratio to the ethanol extract of Gynostemma pentaphyllum herbs, reflux for a period of time, filter, and collect the precipitate; and

[0060] (3) Subject the precipitate to silica gel column chromatography, elute with a certain mobile phase, and recrystallize the eluate to obtain the dammarane-type triterpenoid compound.

[0061] In a preferred embodiment, the volume percentage concentration of the ethanol in step (1) is 40% to 60%, for example, about 50%.

[0062] In the present invention, when a concentration, ratio, time, particle size, or other value or parameter is expressed as a range, a preferred range, or a range defined by a series of upper preferred values and lower preferred values, it should be understood that all ranges formed by any pairing of any range upper limit or preferred value with any range lower limit or preferred value are specifically disclosed, regardless of whether the range is disclosed individually. For example, when the range "40% - 60%" is disclosed, the described range should be interpreted to include ranges such as "40% - 60%", "40% - 55%", "40% - 50%", "40% - 45%", "45% - 60%", "45% - 55%", "45% - 50%", "50% - 60%", "50% - 55%", "55% - 60%", etc. When a numerical range is described herein, unless otherwise stated, the range is intended to include its end values and all integers and fractions within the range, and all numerical values within the range can achieve the effects of the present invention.

[0063] In a preferred embodiment, in step (1), the ratio of the volume / mass (L / Kg) of the ethanol to the Gynostemma pentaphyllum medicinal material is 1 - 5, for example, about 2.

[0064] In a preferred embodiment, in step (1), the extraction method is reflux extraction, ultrasonic extraction, and / or shaking extraction, for example, reflux extraction.

[0065] In a preferred embodiment, in step (1), the extraction time is 0.5 - 3 h, for example, about 2 h.

[0066] In a preferred embodiment, in step (1), the concentration is concentration under reduced pressure.

[0067] In a preferred embodiment, in step (1), the ratio of the volume / volume of the Gynostemma pentaphyllum medicinal material ethanol extract to the ethanol is 0.05 - 0.2, for example, about 0.1.

[0068] In a preferred embodiment, in step (2), the acid is selected from one or more of hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, acetic acid, propionic acid, malonic acid, and butyric acid, for example, hydrochloric acid.

[0069] In a preferred embodiment, in step (2), the volume percentage concentration of the acid is 0.5% - 3%, for example, about 1%.

[0070] In a preferred embodiment, in step (2), the ratio of the volume / volume of the acid to the Gynostemma pentaphyllum medicinal material ethanol extract is 0.1 - 1, for example, about 0.2.

[0071] In a preferred embodiment, in step (2), the reflux time is 0.5 - 3 h, for example, about 1 h.

[0072] In a preferred embodiment, in step (3), the column chromatography uses silica gel with a particle size of 100 to 400 mesh.

[0073] In a preferred embodiment, in step (3), the elution mobile phase of the column chromatography is a mixed solvent of cyclohexane - ethyl acetate with a volume ratio of (40 - 60):(0.1 - 2).

[0074] In a preferred embodiment, in step (3), the elution mobile phase of the column chromatography is a mixed solvent of cyclohexane - ethyl acetate with a volume ratio of approximately 50: approximately 1.

[0075] In the present invention, "about" refers to a value within the range of ±5% of a specific value. For example, "about 50" includes ±5% of 50, or from 47.5 to 52.5; "about 2" includes ±5% of 2, or from 1.9 to 2.1; "about 0.1" includes ±5% of 0.1, or from 0.095 to 0.105; "about 1" includes ±5% of 1, or from 0.95 to 1.05; "about 0.2" includes ±5% of 0.2, or from 0.19 to 0.21; "about 50" includes ±5% of 50, or from 47.5 to 52.5.

[0076] According to another aspect of the present invention, there is provided a pharmaceutical composition comprising the dammarane - type triterpenoid compound.

[0077] In a preferred embodiment, the pharmaceutical composition comprises one or more drugs and / or extracts for promoting ulcer healing.

[0078] In a preferred embodiment, the extract is a traditional Chinese medicine extract.

[0079] In a preferred embodiment, the traditional Chinese medicine extract is Zhuyu Huaijiao Pills and / or Qiangli Zhigen Duan.

[0080] In a preferred embodiment, the drug is nitroglycerin, nifedipine, diltiazem, zinc oxide, erythromycin, and / or levofloxacin.

[0081] In a preferred embodiment, the pharmaceutical composition comprises one or more pharmaceutically acceptable excipients.

[0082] The above - mentioned dammarane - type triterpenoid compound of the present invention or its pharmaceutically acceptable salt and pharmaceutically acceptable excipients are used in an effective dose for producing a pharmaceutical composition. The dose of the active compound may vary with the route of administration, the age and weight of the patient, the nature and severity of the disease to be treated, and similar factors.

[0083] The pharmaceutical compositions of the compounds of the present invention can be administered in any of the following ways: orally, by inhalation spray, rectally, nasally, buccally, topically, parenterally, such as subcutaneously, intravenously, intramuscularly, intraperitoneally, intrathecally, intraventricularly, intracoronarily and intracranially by injection or infusion, or by means of an implanted reservoir. Among them, the preferred administration routes are oral, topical, intraperitoneal or intravenous administration.

[0084] In a preferred embodiment, the pharmaceutical composition of the present invention contains at least one pharmaceutically acceptable excipient in an amount of from 0.00001 to 50 wt.%, or from 0.0001 to 10 wt.%, or from 0.0001 to 5 wt.%, or from 0.005 to 1 wt.%, or from 0.1 to 20 wt.%, or from 0.5 to 15 wt.%, or from 1 to 5 wt.% based on the total weight of the pharmaceutical composition.

[0085] In the present invention, the term "pharmaceutically acceptable" here refers to a substance, such as a carrier or diluent, that does not abolish the biological activity or properties of the compound and is relatively non-toxic, i.e., when administered to an individual, it does not cause unwanted biological effects or interact in a harmful manner with any of the components it contains.

[0086] In the present invention, the term "pharmaceutically acceptable excipient" refers to carriers and / or excipients that are compatible (i.e., capable of eliciting the desired therapeutic effect without causing any undesirable local or systemic effects) with the subject and the active ingredient pharmacologically and / or physiologically, and which are well known in the art (see, for example, Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995).

[0087] In a preferred embodiment, the excipient is selected from one or more of the following: diluents, wetting agents, binders, disintegrants, complexing agents, flavoring agents, sustained release agents, retention aids, lubricants, dispersants, plasticizers, opacifying agents and antioxidants.

[0088] Those skilled in the art will know how to select specific chemical substances within the scope of the above excipients. For example, the diluent can be selected from one or more of the following: powdered sugar, starch, compressible starch, lactose, dextrin, mannitol, sorbitol, microcrystalline cellulose, calcium sulfate, and calcium carbonate. The wetting agent can be selected from one or more of the following: polyoxymethylene stearate, poloxamer, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene hydrogenated castor oil, polyoxyethylene alkyl ether, polysorbate such as polysorbate 80, cetyl alcohol, glycerol fatty acid ester (such as triacetin, glycerol monostearate and the like), polyoxyethylene fatty acid ester, polyethylene glycol fatty acid ester, sodium lauryl sulfate, sorbitan fatty acid ester, sucrose fatty acid ester, polyoxyethylene ether, benzalkonium chloride, polyoxyethylene castor oil, and sodium docusate. The binder can be selected from one or more of the following: polyvinylpyrrolidone, hydroxypropyl cellulose, polyethylene glycol, and methyl cellulose. The disintegrant can be selected from one or more of the following: carboxymethyl cellulose, calcium carboxymethyl cellulose salt, and sodium carboxymethyl cellulose. The flavoring agent can be selected from one or more of the following: sorbitol, glucose, mannose, sucrose, and lactose. The dispersant can be selected from one or more of the following: croscarmellose sodium, sodium starch glycolate, and pregelatinized corn starch. The plasticizer can be dibutyl sebacate and / or various citric acid esters. The sustained-release agent can be selected from one or more of the following: sodium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose, hydroxypropyl methyl cellulose, microcrystalline cellulose, hydroxyethyl cellulose, gum arabic, gelatin, and shellac. The antioxidant can be selected from one or more of the following: sodium bisulfite, sodium metabisulfite, sodium sulfite, and sodium thiosulfate. The lubricant can be selected from one or more of the following: calcium stearate, talc, magnesium stearate, stearic acid, and colloidal silica.

[0089] These excipients are preferably pharmaceutically inert, or may have a synergistic or enhancing effect to enhance the therapeutic activity of the pharmaceutical composition. And the above excipients are only listed, and the excipients actually used in the present invention are not limited to the above excipients, and can be adjusted according to the actual situation, and the effects of the present invention can be achieved.

[0090] In a preferred embodiment, the dosage form of the pharmaceutical composition is ointment, powder, tablet, dripping pill, capsule, film, lozenge, granule, injection, or oral liquid.

[0091] In a preferred embodiment, the ointment is an ointment or a plaster.

[0092] The above drugs, excipients, and dosage forms of the present invention are only illustrative, and are not limited to those listed above in actual application. As long as they can be used in combination with the pharmaceutical composition of the present invention, they are all within the protection scope of the present invention.

[0093] According to another aspect of the present invention, there is provided the use of the dammarane-type triterpenoid compound or the pharmaceutical composition in the preparation of a medicament for promoting ulcer healing.

[0094] According to another aspect of the present invention, there is provided the use of the dammarane-type triterpenoid compound or the pharmaceutical composition in the preparation of a medicament for preventing and / or treating ulcers.

[0095] In a preferred embodiment, the ulcer is an oral ulcer and / or a digestive ulcer.

[0096] In a preferred embodiment, the digestive ulcer is a gastric ulcer, a duodenal ulcer and / or a perianal ulcer.

[0097] In a preferred embodiment, the digestive ulcer is a perianal ulcer.

[0098] In a preferred embodiment, the digestive ulcer is an ulcer caused by perianal inflammation.

[0099] In the present invention, the term "treatment" also includes "prevention" unless otherwise specifically stated to the contrary. The terms "treatment(al)" and "treatment(ly)" should be understood accordingly.

[0100] In the present invention, the term "treatment" includes alleviating, inhibiting or improving the symptoms or conditions of a disease; inhibiting the occurrence of complications; improving or preventing potential metabolic syndromes; inhibiting the occurrence of a disease or symptoms, such as controlling the development of a disease or condition; alleviating a disease or symptoms; reducing a disease or symptoms; alleviating the complications caused by a disease or symptoms, or preventing or treating the signs caused by a disease or symptoms. As used herein, a pharmaceutical composition or a pharmaceutical preparation, after administration, can improve a disease, symptom or condition, especially the severity thereof, delay the onset, slow down the progression of the disease, or reduce the duration of the disease. Whether administered regularly or temporarily, continuously or intermittently, it can be attributed to or related to the administration.

[0101] The present invention also provides a composition or a pharmaceutical preparation as described above for preventing and / or treating ulcers in a subject, such as oral ulcers and / or digestive ulcers.

[0102] The present invention also provides a method for preventing and / or treating ulcers in a subject, such as oral ulcers and / or digestive ulcers, comprising administering to the subject an effective amount of the composition or the pharmaceutical preparation as described above.

[0103] In the present invention, the term "subject" is a mammal. The mammal can be a human, a non-human primate, a mouse, a rat, a dog, a cat, a horse, or a cow, but is not limited to these examples. Mammals other than humans can be advantageously used as subjects representing ulcer models. Preferably, the subject is a human.

[0104] An "effective amount" of the pharmaceutical composition or formulation used in the present invention can achieve the desired therapeutic and / or prophylactic effect. The amount effective for this use will depend on, for example, the pharmaceutical composition, the mode of administration, the stage and severity of the disease being treated, the individual body weight and general health condition, and the judgment of the prescribing physician. The dose can be administered once a week, or once every two days or every day, or even several times a day. The dose unit can be administered over a short period (e.g., several weeks to several months) or a longer period (several months to several years).

[0105] The present invention will be further illustrated below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions noted in the following embodiments are generally carried out under conventional conditions or the conditions recommended by the manufacturer.

[0106] Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those familiar to persons skilled in the art. In addition, any methods and materials similar or equivalent to those described can be applied to the methods of the present invention. The preferred methods and materials described herein are for illustrative purposes only.

[0107] The above-mentioned features mentioned in the present invention, or the features mentioned in the embodiments, can be combined arbitrarily. All the features disclosed in this patent specification can be used in combination with any composition form, and each feature disclosed in the specification can be replaced by any alternative feature that provides the same, equivalent or similar purpose. Therefore, unless otherwise specified, the disclosed features are only general examples of equivalent or similar features.

[0108] Examples

[0109] Preparation Examples

[0110] 25 Kg of Gynostemma pentaphyllum medicinal materials were reflux-extracted with 50 L of 50% ethanol for 2 hours, concentrated under reduced pressure to 5 L, 1 L of 1% hydrochloric acid aqueous solution was added, refluxed for 1 hour, filtered, the precipitate was collected, chromatographed on a silica gel column, eluted with cyclohexane-ethyl acetate (50:1), and recrystallized to obtain 1.3 g of Gypensapogenin VI.

[0111] The hydrogen spectrum of Gypensapogenin VI (as Figure 1 shown) and the carbon spectrum (as Figure 2 shown) data:

[0112]

[0113] 1 1H NMR (400 MHz, CDCl3) δ H : 6.82 (m, 1H, 22-H), 5.58 (d, J = 9.0 Hz, 1H, 23-H), 4.90 (ddt, J = 9.0, 2.9, 1.4 Hz, 1H, 24-H), 4.48 (dd, J = 10.8, 5.5 Hz, 1H, 3-H), 2.54 (td, J = 10.8, 6.7 Hz, 1H, 17-H), 2.05 (s, 3H, 3-OCH3), 1.80 (d, J = 1.4 Hz, 3H, 27-CH3), 1.78 (d, J = 1.4 Hz, 3H, 26-CH3), 0.99 (s, 3H, 18-CH3), 0.90 (s, 3H, 30-CH3), 0.87 (s, 3H, 18-CH3), 0.86 (s, 3H, 28-CH3), 0.85 (s, 3H, 29-CH3).

[0114] 13 13C NMR (100 MHz, CDCl3) δ C : 173.94 (21-C), 171.07 (3-OAc), 145.90 (22-C), 140.31 (25-C), 137.87 (20-C), 119.02 (24-C), 80.84 (3-C), 78.06 (23-C), 55.89 (5-C), 50.63 (9-C), 49.84 (14-C), 46.74 (13-C), 40.40 (8-C), 38.72 (1-C), 37.88 (4-C), 37.66 (17-C), 37.09 (10-C), 35.31 (7-C), 31.34 (15-C), 28.77 (28-C), 27.97 (12-C), 25.81 (26-C), 24.83 (16-C), 23.67 (2-C), 21.38 (3-OCH3), 21.25 (11-C), 18.56 (27-C), 18.12 (6-C), 16.52 (29-C), 16.31 (18-C), 15.63 (19-C), 15.62 (30-C).

[0115] Pharmacological activity evaluation

[0116] Effect of Gypensapogenin VI on the healing of acetic acid-induced perianal ulcers in rats

[0117] 1 Experimental purpose

[0118] Taking Diyu Huaijiao Pills and Strong Hemorrhoid-Radical Tablets as positive controls, the effect of intragastric administration of Gypensapogenin VI on the healing of acetic acid-induced perianal ulcers in rats was observed.

[0119] 2 Experimental Materials

[0120] 2.1 Test Drugs

[0121] Gypensapogenin VI was prepared according to the preparation method of the above-mentioned example.

[0122] 2.2 Control Drugs

[0123] Name: Diyu Huaijiao Pills; Strong Hemorrhoid-Radical Tablets

[0124] Source: Diyu Huaijiao Pills, Tongrentang Pharmaceutical Factory of Beijing Tongrentang Co., Ltd.; Strong Hemorrhoid-Radical Tablets, Eversun Pharmaceutical Factory, Hamburg, Germany

[0125] Batch Number: Diyu Huaijiao Pills: 6035079; Strong Hemorrhoid-Radical Tablets: 11675

[0126] Specification: Diyu Huaijiao Pills: 10 g per 100 pills; Strong Hemorrhoid-Radical Tablets: 265 mg per tablet core

[0127] Preparation Method: Weigh 20 g of Diyu Huaijiao Pills and suspend them with 100 ml of 0.5% CMC-Na. Take 12 tablets of Strong Hemorrhoid-Radical Tablets (the active ingredient content per tablet is 0.2375 g) and suspend them with 100 ml of 0.5% CMC-Na.

[0128] 2.3 Main Reagents

[0129] Acetic Acid: Nanjing Chemical Reagent Co., Ltd., Batch Number: 160510438.

[0130] 2.4 Instruments

[0131] Sartorius Analytical Balance: Beijing Dorris Balance Co., Ltd.; TLS-2000A Type Electronic Scale: Changshu Shuangjie Testing Instrument Factory.

[0132] 3 Animals

[0133] 3.1 Source, Species, Strain, Certificate of Conformity:

[0134] SD rats, SPF grade, purchased from the Experimental Animal Center of Zunyi Medical University, Animal Production License Number: SCXK(Qian)2019-0001.

[0135] 3.2 Body Weight: 170 ± 10 g.

[0136] 3.3 Gender: Male.

[0137] 3.4 Quantity: 60.

[0138] 3.5 Feeding: The animals were raised at 25 ± 3°C, relative humidity of 60 - 75%, with a 12-hour day-night light-dark cycle. They had free access to food and water and were used for experiments after one week of adaptive feeding.

[0139] 4 Experimental methods and procedures

[0140] 4.1 Animal grouping and dose setting

[0141] The grouping table of experimental animals is shown in Table 1.

[0142] Table 1 Grouping table of experimental animals

[0143]

[0144] 4.2 Modeling and experimental procedures

[0145] 0.1 ml of 99.8% acetic acid was applied to the perianal area of the rats to form ulcers. On the second day after modeling, the rats were randomly grouped according to the ulcer degree. They were given drugs by gavage for 10 consecutive days. The ulcer healing conditions were observed and scored on the 3rd, 5th, 7th, 9th, 11th, and 13th days after the initial drug administration.

[0146] 4.3 Measurement indicators

[0147] Scoring criteria: Obvious ulcer exudate was seen, scored 1 point; A little ulcer exudate was seen, scored 2 points; There was eschar and it was basically healed, scored 3 points; Completely healed, scored 4 points.

[0148] 5 Drug administration method

[0149] Drug administration by gavage, with a dosing volume of 0.5 ml / 100 g. The model group was given a 0.5% CMC-Na solution with the same volume as the positive control.

[0150] 6 Statistical treatment

[0151] All data were expressed as mean ± standard deviation In Excel, a t-test was used for significant between-group tests. P < 0.05 indicated a significant difference, and P < 0.01 indicated a very significant difference.

[0152] 7 Experimental results

[0153] The healing scores of the positive control group of the Diyu Huaijiao Pills and the high-dose group of the test drug were similar, and there were extremely significant differences compared with the model group on the 9th, 11th, and 13th days (P < 0.05); the medium-dose group was similar to the positive control group of Qiangli Zhigen Duan, and there were significant differences in the scores on the 11th and 13th days after administration compared with the model group (P < 0.05); there was no statistical significance in the healing of the low-dose group compared with the model group (P > 0.05), but it was also found that its healing was relatively better than that of the negative control. The results are shown in Tables 2 and 3.

[0154] Table 2 Effects of Gypensapogenin VI on the healing of acetic acid-induced perianal ulcers in rats on the 3rd, 5th, and 7th days ( n = 10)

[0155]

[0156] Table 3 Effects of Gypensapogenin VI on the healing of acetic acid-induced perianal ulcers in rats on the 9th, 11th, and 13th days ( n = 10)

[0157]

[0158]

[0159] Among them, compared with the model group, *P < 0.05, **P < 0.01.

[0160] 8 Experimental conclusions

[0161] The results showed that the perianal ulcers of rats in the high-dose group of Gypensapogenin VI healed significantly on the 9th day after the first administration, and those in the medium-dose group healed significantly on the 11th day after the first administration. The scores were significantly different from those of the negative control group. The low-dose group showed a trend of promoting ulcer healing.

[0162] The above has introduced the embodiments of the present invention in detail. Specific examples are used in this article to elaborate on the principles and implementation manners of the present invention. The descriptions of the above embodiments are only used to help understand the method and its core idea of the present invention. At the same time, any changes or deformations made by those skilled in the art based on the idea of the present invention, within the specific implementation manners and application scopes of the present invention, all belong to the protection scope of the present invention. In summary, the content of this specification should not be construed as a limitation to the present invention.

Claims

1. A dammarane-type triterpenoid compound or a pharmaceutically acceptable salt thereof, characterized in that, The dammarane - type triterpenoid compound has the structural formula shown in Formula I:

2. The dammarane triterpenoid compound according to claim 1, wherein, The dammarane - type triterpenoid compound is obtained by hydrolyzing the ethanol extract of Gynostemma pentaphyllum herbs through an acid hydrolysis method.

3. A method for preparing the dammarane-type triterpenoid compound according to claim 1 or 2, characterized in that, The method comprises the following steps: (1) Weigh an appropriate amount of Gynostemma pentaphyllum herbs, add ethanol for extraction, and then concentrate to obtain the ethanol extract of Gynostemma pentaphyllum herbs; (2) Add an acid solution with a certain volume ratio to the ethanol extract of Gynostemma pentaphyllum herbs, reflux for a period of time, filter, and collect the precipitate; and (3) Subject the precipitate to silica gel column chromatography, elute with a certain mobile phase, and recrystallize the eluate to obtain the dammarane - type triterpenoid compound.

4. The method according to claim 3, characterized in that, In step (1), the volume percentage concentration of the ethanol is 40% - 60%, for example, about 50%; Preferably, in step (1), the volume / mass (L / Kg) ratio of the ethanol to the Gynostemma pentaphyllum herbs is 1 - 5, for example, about 2; More preferably, in step (1), the extraction method is reflux extraction, ultrasonic extraction, and / or shaking extraction, for example, reflux extraction; Also preferably, in step (1), the extraction time is 0.5 - 3 h, for example, about 2 h; Especially preferably, in step (1), the concentration is carried out under reduced pressure; Specifically preferably, in step (1), the volume / volume ratio of the ethanol extract of Gynostemma pentaphyllum herbs to the ethanol is 0.05 - 0.2, for example, about 0.

1.

5. The method according to claim 3, characterized in that, In step (2), the acid is selected from one or more of hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, acetic acid, propionic acid, malonic acid, and butyric acid, for example, hydrochloric acid; Preferably, in step (2), the volume percentage concentration of the acid is 0.5% - 3%, for example, about 1%; More preferably, in step (2), the volume / volume ratio of the acid to the ethanol extract of Gynostemma pentaphyllum herbs is 0.1 - 1, for example, about 0.2; Also preferably, in step (2), the reflux time is 0.5 - 3 h, for example, about 1 h.

6. The method according to claim 3, wherein In step (3), the column chromatography uses silica gel with a particle size of 100 - 400 mesh; Preferably, in step (3), the elution mobile phase for the column chromatography is a mixed solvent of cyclohexane - ethyl acetate with a volume ratio of (40 - 60):(0.1 - 2); More preferably, in step (3), the elution mobile phase for the column chromatography is a mixed solvent of cyclohexane - ethyl acetate with a volume ratio of about 50:about 1.

7. A pharmaceutical composition comprising the dammarane - type triterpenoid compound according to claim 1 or 2; Preferably, the pharmaceutical composition comprises one or more drugs and / or extracts for promoting ulcer healing; More preferably, the extract is a traditional Chinese medicine extract; Also preferably, the traditional Chinese medicine extract is Zhuyu Huaijiao Pills and / or Qiangli Zhigen Duan; 8. The pharmaceutical composition according to claim 7, wherein, The drug is nitroglycerin, nifedipine, diltiazem, zinc oxide, erythromycin, and / or levofloxacin.

9. The pharmaceutical composition according to claim 7, wherein The pharmaceutical composition comprises one or more pharmaceutically acceptable excipients; Preferably, the excipients are selected from one or more of the following: diluents, wetting agents, binders, disintegrants, inclusion agents, flavoring agents, sustained - release agents, retention aids, lubricants, dispersants, plasticizers, opacifiers, and antioxidants; More preferably, the dosage form of the pharmaceutical composition is an ointment, powder, tablet, dripping pill, capsule, film, lozenge, granule, injection or oral liquid; Even more preferably, the ointment is an ointment or a plaster.

10. Use of the dammarane triterpenoid compound according to claim 1 or 2 or the pharmaceutical composition according to any one of claims 7 to 9 in the preparation of a drug for promoting ulcer healing; Preferably, the ulcer is an oral ulcer and / or a digestive ulcer; More preferably, the digestive ulcer is a gastric ulcer, a duodenal ulcer and / or a perianal ulcer; Even more preferably, the digestive ulcer is a perianal ulcer; Even more preferably, the digestive ulcer is an ulcer caused by perianal inflammation.

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