Dammarane type triterpenes, preparation method thereof and antipyretic application of dammarane type triterpenes

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 pentaphylla medicinal material and have a new chemical structure have an obvious antipyretic effect, the application range of gynostemma pentaphylla can be expanded by developing the triterpenoids into medicines for treating diseases and symptoms related to body temperature rise, and the utilization value of gynostemma pentaphylla plant resources is improved. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technologies, and particularly to dammarane-type triterpenes, a preparation method thereof, and their use for antipyretic purposes. Background Art

[0002] Fever is a symptom in which body temperature exceeds the normal range caused by infectious and non-infectious diseases, and is one of the most common clinical disease symptoms. Exceeding the normal range of body temperature: Clinically, it is also called fever or pyrexia, which refers to an increase in body temperature caused by any reason such as excessive heat production, reduced heat dissipation, body temperature regulation disorders, or pyrogens acting on the body temperature regulation center, causing the set point to shift upward and exceeding the normal range, which is called an increase in body temperature. Generally speaking, when the axillary temperature exceeds 37°C or the oral temperature exceeds 37.5°C, and the body temperature fluctuates by more than 1°C within 24 hours, it can be called hyperthermia.

[0003] Fever is a clinical manifestation of an inflammatory reaction, mostly due to the infection of exogenous pathogenic microorganisms, which activates neutrophils, eosinophils, and the mononuclear-phagocyte system in the blood, such as interleukin (IL-1), tumor necrosis factor (TNF-α), and interferon, etc., causing them to produce and release endogenous pyrogens. Under the stimulation of pyrogens, the positive regulatory mediators (prostaglandin PGE2, cyclic adenosine monophosphate cAMP, and NO) and negative regulatory mediators (arginine vasopressin AVP) of the body temperature center change, and through various physiological mechanisms, the body temperature regulation set point increases, heat production increases while heat dissipation decreases, thus causing fever, and the fever caused by exogenous pyrogens is a delayed process. Hyperthermia is likely to cause many discomforts such as benign febrile convulsions, epileptic seizures, and tachycardia, and can also exacerbate the inflammation of infected wounds. A significantly high fever above 40°C may lead to permanent brain damage.

[0004] Antipyretic analgesics represented by non-steroidal anti-inflammatory drugs (NSAIDs) are the most widely used and applied type of drugs, such as paracetamol, ibuprofen, and aspirin. These non-steroidal analgesic anti-inflammatory drugs (NSAIDs) have advantages such as definite curative effects and good tolerance. However, the adverse reactions caused by NSAIDs, such as gastrointestinal discomfort, abnormal hematopoietic system function, kidney damage, and central nervous system abnormalities, limit the further application of NSAIDs. Therefore, exploring antipyretic analgesics with definite curative effects and few side effects has become a new research direction for relevant research scholars. Natural drugs represented by traditional Chinese medicine have become a research hotspot for new antipyretic analgesics due to their multi-target comprehensive action mechanism and few side effects.

[0005] Gynostemma pentaphyllum Makino., also known as Gynostemma pentaphyllum, Gospel Herb, etc., 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 immune function of the body. 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, sedating, hypnotizing, 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 triterpenoids with a completely new chemical skeleton structure in Gynostemma pentaphyllum has good antipyretic effects and may have broad prospects in the preparation of drugs for preventing or / and treating diseases and disorders related to elevated body temperature. The triterpenoids 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 reports on the use of the triterpenoids involved in the present invention for treating or / and preventing diseases and disorders related to elevated body temperature. Summary of the Invention

[0007] The main object of the present invention is to provide a drug for treating diseases and disorders related to elevated body temperature with definite curative effect, few side effects, natural safety, so as to solve the problems of common drugs in the prior art such as easy occurrence of gastrointestinal discomfort, abnormal hematopoietic system function, kidney damage, and central nervous system abnormality.

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

[0009]

[0010] Furthermore, the dammarane-type triterpenoid is obtained by hydrolyzing the ethanol extract of Gynostemma pentaphyllum medicinal materials by an acid hydrolysis method.

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

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

[0013] (2) Add an acid solution with a certain volume ratio to the ethanol extract of Gynostemma pentaphyllum medicinal materials, 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.

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

[0016] Further, in step (1), the ratio of the volume / mass (L / Kg) of the ethanol to the Gynostemma pentaphyllum medicinal material is 1 to 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 to 3 h, for example, about 2 h.

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

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

[0021] Further, 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.

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

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

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

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

[0026] Further, in step (3), the elution mobile phase for the column chromatography is a mixed solvent of cyclohexane - ethyl acetate with a volume ratio of (10 - 30):(0.1 - 2).

[0027] Further, 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 20:about 1.

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

[0029] Further, the pharmaceutical composition comprises one or more drugs and / or extracts for antipyretic use.

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

[0031] Further, the drug is a non-steroidal anti-inflammatory drug, a hormonal drug, and / or a traditional Chinese patent medicine.

[0032] Further, the non-steroidal anti-inflammatory drug is selected from one or more of the following: aspirin, ibuprofen, paracetamol, lysine acetylsalicylate, and diclofenac sodium.

[0033] Further, the hormonal drug is selected from one or more of the following: dexamethasone, methylprednisolone, hydrocortisone, triamcinolone, and prednisolone.

[0034] Further, the traditional Chinese patent medicine is Shufeng Jiedu Capsule and / or Tanreqing Capsule.

[0035] Further, the pharmaceutical composition comprises one or more pharmaceutically acceptable excipients.

[0036] Further, the excipient is selected from one or more of the following: diluent, wetting agent, binder, disintegrant, clathrant, flavoring agent, sustained-release agent, retention aid, lubricant, dispersant, plasticizer, light-blocking agent, and antioxidant.

[0037] Further, the dosage form of the pharmaceutical composition is ointment, powder, tablet, dripping pill, capsule, film, lozenge, granule, injection, or oral liquid.

[0038] Further, the ointment is an ointment or a plaster.

[0039] According to another aspect of the present invention, there is provided a use of the above-mentioned dammarane-type triterpenoid compound or the pharmaceutical composition in the preparation of a drug for antipyretic.

[0040] Advantages of the present invention:

[0041] Gynostemma pentaphyllum is the dried whole herb of Gynostemma pentaphyllum (Thunb.) Makino of the genus Gynostemma in the family Cucurbitaceae, and it is a perennial climbing herb. Modern pharmacological research shows that its main active ingredient is gypenoside (Gyp). We found that a class of triterpenoid compounds with a new chemical structure isolated from Gynostemma pentaphyllum herbs has obvious antipyretic effects. Developing it into a drug for treating diseases and disorders related to elevated body temperature can expand the application range of Gynostemma pentaphyllum and improve the utilization value of Gynostemma pentaphyllum plant resources. Description of the Drawings

[0042] To more clearly illustrate the technical solutions in the embodiments of the present invention, the following will briefly introduce the accompanying drawings required for the description of the embodiments. Obviously, the accompanying drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, other accompanying drawings can also be obtained based on these drawings without exceeding the scope of protection required by the present invention.

[0043] Figure 1 Schematic diagram of the hydrogen spectrum of the compound of formula I of the present invention.

[0044] Figure 2 Schematic diagram of the carbon spectrum of the compound of formula I of the present invention. Detailed implementation manners

[0045] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative efforts fall within the scope of protection of the present invention.

[0046] Unless otherwise stated, all technical and scientific terms and abbreviations used herein have the meanings commonly understood by those of ordinary skill in the field of the present invention or the field to which the term is applied. Although any methods, conditions, substances or materials similar to or equivalent to those disclosed herein can be used in the implementation of the present invention, the preferred methods, conditions, substances or materials are described herein.

[0047] The present invention is expected to cover all options, variations and equivalents, which may be included in the existing field of invention as defined by 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.

[0048] 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, the CAS version, and the Handbook of Chemistry and Physics, 75th Ed., 1994. In addition, general principles of organic chemistry can be 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. Therefore, all content incorporates the references.

[0049] As used in the specification and the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise.

[0050] In the present invention, the term "comprising" is synonymous with "including". 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 apparatus that comprises the recited elements need not be limited to those elements, but may include other elements not expressly listed or elements inherent to such composition, step, method, article, or apparatus.

[0051] As described in the background art section, common drugs currently used for treating diseases and disorders associated with elevated body temperature are prone to problems such as gastrointestinal discomfort, abnormal hematopoietic system function, kidney damage, and central nervous system abnormalities. 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:

[0052]

[0053] The term "salt" as 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.

[0054] In a preferred embodiment, the dammarane-type triterpenoid compound is obtained by hydrolyzing an ethanol extract of Gynostemma pentaphyllum herbs by an acid hydrolysis method.

[0055] According to another aspect of the present invention, there is provided a method for preparing the above-mentioned dammarane-type triterpenoid compound, and the method comprises the following steps:

[0056] (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;

[0057] (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

[0058] (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.

[0059] In a preferred embodiment, the volume percentage concentration of the ethanol in step (1) is 40% to 60%, for example, about 50%.

[0060] 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% to 60%" is disclosed, the described range should be interpreted as including the ranges "40% to 60%", "40% to 55%", "40% to 50%", "40% to 45%", "45% to 60%", "45% to 55%", "45% to 50%", "50% to 60%", "50% to 55%", "55% to 60%", etc. When a numerical range is described herein, unless otherwise specified, 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.

[0061] In a preferred embodiment, the ratio of the volume / mass (L / Kg) of the ethanol to the Gynostemma pentaphyllum medicinal material in step (1) is 1 to 5, for example, about 2.

[0062] In a preferred embodiment, the extraction method in step (1) is reflux extraction, ultrasonic extraction, and / or shaking extraction, for example, reflux extraction.

[0063] In a preferred embodiment, the extraction time in step (1) is 0.5 to 3 h, for example, about 2 h.

[0064] In a preferred embodiment, the concentration in step (1) is vacuum concentration.

[0065] In a preferred embodiment, the ratio of the volume / volume of the Gynostemma pentaphyllum medicinal material ethanol extract to the ethanol in step (1) is 0.05 to 0.2, for example, about 0.1.

[0066] 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, for example, hydrochloric acid.

[0067] In a preferred embodiment, the volume percentage concentration of the acid in step (2) is 0.5% to 3%, for example, about 1%.

[0068] In a preferred embodiment, the ratio of the volume / volume of the acid to the Gynostemma pentaphyllum medicinal material ethanol extract in step (2) is 0.1 to 1, for example, about 0.2.

[0069] In a preferred embodiment, the reflux time in step (2) is 0.5 to 3 h, for example, about 1 h.

[0070] In a preferred embodiment, in step (3), the column chromatography is performed using silica gel with a particle size of 100 to 400 mesh.

[0071] In a preferred embodiment, in step (3), the elution mobile phase for the column chromatography is a mixed solvent of cyclohexane - ethyl acetate with a volume ratio of (10 - 30):(0.1 - 2).

[0072] In a preferred embodiment, 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 20:about 1.

[0073] 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 20" includes ±5% of 20, or from 19 to 21.

[0074] According to another aspect of the present invention, there is provided a pharmaceutical composition comprising the above-mentioned dammarane - type triterpenoids.

[0075] In a preferred embodiment, the pharmaceutical composition comprises one or more drugs and / or extracts for antipyretic use.

[0076] In a preferred embodiment, the extract is a traditional Chinese medicine extract.

[0077] In a preferred embodiment, the drug is a non - steroidal anti - inflammatory drug, a hormonal drug and / or a traditional Chinese patent medicine.

[0078] In a preferred embodiment, the non - steroidal anti - inflammatory drug is selected from one or more of the following: aspirin, ibuprofen, paracetamol, lysine acetylsalicylate and diclofenac sodium.

[0079] In a preferred embodiment, the hormonal drug is selected from one or more of the following: dexamethasone, methylprednisolone, hydrocortisone, triamcinolone and prednisolone.

[0080] In a preferred embodiment, the traditional Chinese patent medicine is Shufeng Jiedu Capsule and / or Tanreqing Capsule.

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

[0082] An effective dose of the dammarane triterpenoid compound of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient are used 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 composition of the compound of the present invention can be administered in any of the following ways: orally, by spray inhalation, rectally, nasally, buccally, topically, parenterally, such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intracardiac, intrasternal and intracranial injection or infusion, or by means of an implantable reservoir. Among them, oral, topical, intraperitoneal or intravenous administration routes are preferred.

[0084] In a preferred embodiment, the pharmaceutical composition of the present invention contains at least one pharmaceutically acceptable excipient in an amount of 0.00001 to 50 wt.%, or 0.0001 to 10 wt.%, or 0.0001 to 5 wt.%, or 0.005 to 1 wt.%, or 0.1 to 20 wt.%, or 0.5 to 15 wt.%, or 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 cause the loss of the biological activity or properties of the compound and is relatively non-toxic. For example, when a substance is administered to an individual, it does not cause unwanted biological effects or interact with any of its components in a harmful manner.

[0086] In the present invention, the term "pharmaceutically acceptable excipient" refers to carriers and / or excipients that are compatible with the subject and the active ingredient pharmacologically and / or physiologically (i.e., capable of eliciting the desired therapeutic effect without causing any undesirable local or systemic effects), 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, inclusion agents, flavoring agents, sustained-release agents, retention aids, lubricants, dispersants, plasticizers, light-blocking 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 dioctyl sulfosuccinate. 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. Moreover, the above excipients are only illustrative, 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 all can achieve the effects of the present invention.

[0090] In a preferred embodiment, the dosage form of the pharmaceutical composition is an 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 during actual application. As long as they can be used in combination with the pharmaceutical composition of the present invention, they are within the protection scope of the present invention.

[0093] According to another aspect of the present invention, there is provided the use of the above-mentioned dammarane-type triterpenoids or the above-mentioned pharmaceutical composition in the preparation of a medicament for antipyretic use.

[0094] According to another aspect of the present invention, there is provided the use of the above-mentioned dammarane-type triterpenoids or the above-mentioned pharmaceutical composition in the preparation of a medicament for preventing and / or treating elevated body temperature.

[0095] In the present invention, the term "treatment" also includes "prevention" unless there is a specific indication to the contrary. The terms "treatment(al)" and "treatment(ly)" should be understood accordingly.

[0096] 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; reducing the complications caused by a disease or symptoms, or preventing or treating the signs caused by a disease or symptoms. As used herein, after administration of a pharmaceutical composition or a pharmaceutical preparation, a certain disease, symptom or condition can be improved, especially the severity thereof can be improved, the onset can be delayed, the progression of the disease can be slowed down, or the duration of the disease can be reduced. Whether administered regularly or temporarily, continuously or intermittently, it can be attributed to or related to the administration situation.

[0097] The present invention also provides the above-mentioned composition or the above-mentioned pharmaceutical preparation for preventing and / or treating elevated body temperature in a subject.

[0098] The present invention also provides a method for preventing and / or treating elevated body temperature in a subject, comprising administering to the subject an effective amount of the above-mentioned composition or the above-mentioned pharmaceutical preparation.

[0099] In the present invention, the term "subject" is a mammal. Mammals can be humans, non-human primates, mice, rats, dogs, cats, horses or cows, but are not limited to these examples. Mammals other than humans can be advantageously used as subjects representing the elevated body temperature model. Preferably, the subject is a human.

[0100] The "effective amount" of the pharmaceutical composition or preparation 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 overall 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).

[0101] The present invention will be further described 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 according to conventional conditions or conditions recommended by the manufacturers.

[0102] Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to the described content can be applied to the method of the present invention. The preferred methods and materials described herein are for illustrative purposes only.

[0103] 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 can achieve the same, equivalent or similar purpose. Therefore, unless otherwise specified, the disclosed features are only general examples of equivalent or similar features.

[0104] Embodiment

[0105] Preparation Example

[0106] 25 Kg of Gynostemma pentaphyllum medicinal materials were refluxed 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, subjected to silica gel column chromatography, eluted with cyclohexane-ethyl acetate (20:1), and recrystallized to obtain 2.1 g of Gypensapogenin IV.

[0107] The 1H NMR (as Figure 1 shown) and 13C NMR (as Figure 2 shown) data of Gypensapogenin IV:

[0108]

[0109] 1 H NMR (400 MHz, CDCl3) δH: 5.05 (d, J = 1.6 Hz, 1H, 21-H), 4.76 (s, 1H, 21-H), 4.24 (t, J = 7.2 Hz, 1H, 25-H), 3.13 (dd, J = 11.2, 5.0 Hz, 1H, 3-H), 2.02 (dt, J = 11.3, 6.0 Hz, 2H), 1.92 (dt, J = 13.6, 8.9 Hz, 1H), 1.22 (s, 3H), 1.19 (s, 3H), 0.91 (s, 3H), 0.91 (s, 3H), 0.80 (s, 3H), 0.77 (s, 3H), 0.71 (s, 3H).

[0110] 1313C NMR (100 MHz, CDCl3) δC: 155.24 (C-22), 106.73 (C-21), 81.32 (C-12), 80.72 (C-25), 78.93 (C-3), 55.81 (C-5), 50.81 (C-14), 49.40 (C-4), 47.30 (C-13), 43.71 (C-17), 40.38 (C-8), 39.05 (C-1), 38.95 (C-26), 38.60 (C-15), 37.17 (C-10), 35.37 (C-22), 32.97 (C-11), 31.38 (C-9), 31.05 (C-24), 28.77 (C-7), 28.15 (C-2), 28.00 (C-28), 27.37 (C-19), 24.80 (C-16), 21.34 (C-6), 18.25 (C-23), 16.20 (C-18), 15.84 (C-30), 15.65 (C-27), 15.38 (C-29).

[0111] Pharmacological activity evaluation

[0112] Antipyretic effect of Gypensapogenin IV on lipopolysaccharide-induced pyretic rats

[0113] 1 Experimental materials

[0114] 1.1 Experimental drugs

[0115] 1.1.1 Positive control drug

[0116] Ibuprofen, specification: 200 mg / tablet, Shandong Xinhua Pharmaceutical Co., Ltd., batch number: 1503230.

[0117] 1.1.2 Test drug

[0118] Gypensapogenin IV was prepared according to the above preparation examples.

[0119] 1.2 Experimental animals

[0120] 1.2.1 Source, species, strain, certificate: SD rats, SPF grade, purchased from the Experimental Animal Center of Zunyi Medical University, animal production license number: SCXK(Qian)2019-0001.

[0121] 1.2.2 Body weight: 170 ± 10 g.

[0122] 1.2.3 Gender: male.

[0123] 1.2.4 Quantity: 60.

[0124] 1.2.5 Feeding: The animals were raised at 25 ± 3 °C, with a relative humidity of 60 - 75%, a 12-hour light-dark cycle, and were allowed free access to food and water. After one week of adaptive feeding, they were used for experiments.

[0125] 1.3 Main Reagents

[0126] Lipopolysaccharide: Sigma L2880 - 10MG, Lot Number: 025M4040V;

[0127] Sodium carboxymethyl cellulose: Sinopharm Chemical Reagent Co., Ltd., Batch Number: 30036328, Purity Grade: CP;

[0128] 0.9% Sodium chloride solution: Anhui Shuanghe Pharmaceutical Co., Ltd., Batch Number: 150814UT;

[0129] White petrolatum: Nanjing Chemical Reagent Co., Ltd., Batch Number: 09101660604;

[0130] Chloral hydrate: Sinopharm Chemical Reagent Co., Ltd., Batch Number: 20130314. Before use, it was dissolved in 0.9% sodium chloride solution to prepare a 3% clear solution of chloral hydrate.

[0131] 1.4 Main Instruments

[0132] Mortar;

[0133] TELLMO Electronic thermometer BT - A21, Dongguan Fudakang Industrial Co., Ltd., Batch Number: 1309;

[0134] Sartorius analytical balance: Beijing Sartorius Co., Ltd.;

[0135] KQ - 250E Ultrasonic cleaner: Kunshan Ultrasonic Instruments Co., Ltd.

[0136] 2 Experimental Methods

[0137] 2.1 Animal Screening

[0138] After one week of adaptive feeding of the rats, starting at 9:00 every morning, the rectal temperature of the rats was measured every 1 hour for 8 hours, and this was done continuously for three days. The average body temperature at the same time points over 3 days was used as the basal body temperature, and rats with a body temperature between 36.6 - 38.3 °C and a temperature difference of no more than 0.5 °C over three measurements were selected.

[0139] 2.2 Animal Grouping

[0140] Forty-eight male rats that met the experimental requirements were randomly grouped into 6 groups, with 8 rats in each group.

[0141] ① Negative control group: intragastrically administered with the solvent 0.5% CMC-Na solution.

[0142] ② Model group: intragastrically administered with the solvent 0.5% CMC-Na solution.

[0143] ③ High-dose group of the test drug Gypensapogenin IV: intragastrically administered with 52.5 mg / kg / d Gypensapogenin IV solution.

[0144] ④ Medium-dose group of the test drug Gypensapogenin IV: intragastrically administered with 26.25 mg / kg / d Gypensapogenin IV solution.

[0145] ⑤ Low-dose group of the test drug Gypensapogenin IV: intragastrically administered with 13.13 mg / kg / d Gypensapogenin IV solution.

[0146] ⑥ Positive drug ibuprofen group: intragastrically administered with 0.09 g / kg / d ibuprofen solution.

[0147] The rats in each of the above groups were continuously administered for 3 days. 30 minutes after the last administration, 100 μg / kg lipopolysaccharide (LPS) was given to establish the model, and the body temperature changes of the rats in each group were measured after model establishment.

[0148] 2.3 Experimental method

[0149] Forty-eight qualified rats were randomly divided into 6 groups of 8 rats each, namely the negative control group, the model group, the high-dose Gypensapogenin IV group at 52.5 mg / kg, the medium-dose Gypensapogenin IV group at 26.25 mg / kg, the low-dose Gypensapogenin IV group at 13.13 mg / kg, and the positive drug ibuprofen group at 90 mg / kg. The negative control group and the model group were intragastrically administered an equal volume of CMC-Na solution. Administration was by gavage at a volume of 1 ml / 100 g once a day for 3 consecutive days. The rats were fasted for 6 h before administration but allowed to drink water to reduce the interference of feces during temperature measurement. Before each temperature measurement, the probe of the electronic thermometer was coated with vaseline and inserted 2.5 cm into the rectum of the rats (marked and fixed with adhesive tape at 2.5 cm to ensure the same insertion depth each time), and the reading was recorded after it stabilized. Thirty minutes before the last administration, the body temperature was measured three times continuously, once every 10 min. The high body temperature values due to stress were excluded, and the average value was taken as the basal body temperature. Thirty minutes after administration, the negative control group was intraperitoneally injected with 10 ml / kg of sterile normal saline, and the other groups were intraperitoneally injected with 100 μg / kg of lipopolysaccharide (LPS) to establish the model. The body temperatures of the rats in each group were monitored and recorded at 0.5, 1, 2, 3, 4, 5, 6, 7, and 8 h after modeling. The change value of body temperature △T (measured body temperature - basal body temperature) was calculated by measuring the body temperature changes of the animals in each group, the average body temperature change curve was plotted, and one-way analysis of variance was performed to compare the antipyretic effects of each group.

[0150] 3 Statistical processing methods

[0151] The data of each group were expressed as mean ± standard deviation (±SD). One-way analysis of variance (one-way ANOVA) and Turkey method were used for comparison of the mean values between groups. P < 0.05 indicated that the difference was statistically significant.

[0152] 4 Experimental results

[0153] As shown in Tables 1 - 4, compared with the negative control group, there were no differences in the normal body temperatures of rats in each group before modeling (P > 0.05); except at the 1 - hour time point, the model group maintained a higher body temperature at 8 time points after modeling, showing significant differences compared with the negative control group (P < 0.001, P < 0.01), indicating successful modeling; compared with the model group, except at the 1 - hour time point, the positive drug ibuprofen group had extremely significant antipyretic effects at 8 time points after modeling (P < 0.001); from 3h to 6h after modeling, each dose group of Gypensapogenin IV had significant antipyretic effects compared with the model group (P < 0.01, P < 0.001); at 3h after modeling, the temperature - lowering amplitude of each dose group of Gypensapogenin IV was the most significant (P < 0.001, P < 0.01), and then gradually weakened; compared with the medium - and low - dose groups of Gypensapogenin IV, the high - dose group showed a significant antipyretic effect at 2h after modeling (P < 0.05), and the antipyretic effect of the high - dose group was the strongest, with the time effect lasting up to 7h after modeling. Although the medium - and low - dose groups had a certain temperature - lowering trend in numerical values at local time points, they did not show statistical significance, indicating a certain dose - correlation.

[0154] Table 1 Effects of Gypensapogenin IV on the body temperature change values (0.5h - 3h) of LPS - induced febrile rats (±SD, n = 8)

[0155]

[0156] Table 2 Effects of Gypensapogenin IV on the body temperature change values (4h - 8h) of LPS - induced febrile rats (±SD, n = 8)

[0157]

[0158] Table 3 Effects of Gypensapogenin IV on the measured body temperature values (0.5h - 3h) of LPS - induced febrile rats (±SD, n = 8)

[0159]

[0160] Table 4 Effects of Gypensapogenin IV on the measured body temperature values (4h - 8h) of LPS - induced febrile rats (±SD, n = 8)

[0161]

[0162]

[0163] 5 Experimental conclusions

[0164] Compared with the model group, Gypensapogenin IV can start to exert its curative effect about 2 hours after modeling. Its antipyretic activity is the best at about 3 hours, and then weakens. The maintenance time is relatively long, showing a certain dose correlation and having a significant antipyretic effect.

[0165] The embodiments of the present invention have been introduced in detail above. Specific examples are used in this article to elaborate on the principle and implementation manner of the present invention. The description of the above embodiments is 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 manner and application scope of the present invention, all fall within 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 a structural formula shown in Formula I:

2. The dammarane-type triterpenoid compound according to claim 1, wherein The dammarane-type triterpenoid compounds are obtained by hydrolyzing the ethanol extract of Gynostemma pentaphyllum medicinal material by acid hydrolysis.

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) Weighing an appropriate amount of Gynostemma pentaphyllum medicinal material, adding ethanol to extract, and concentrating to obtain a Gynostemma pentaphyllum medicinal material ethanol extract; (2) adding a certain volume ratio of an acid solution to the ethanol extract of Gynostemma pentaphyllum, refluxing for a period of time, filtering, and collecting a precipitate; and (3) The precipitate is subjected to silica gel column chromatography, eluted with a certain mobile phase, and the eluate is recrystallized to obtain the dammarane-type triterpenoid compound.

4. The method according to claim 3, wherein The volume percentage concentration of ethanol in step (1) is 40% to 60%, for example, about 50%; Preferably, in step (1), the ratio of the volume / mass (L / Kg) of the ethanol to the Gynostemma pentaphyllum medicinal material is 1 to 5, for example, about 2; More preferably, the extraction method in step (1) is reflux extraction, ultrasonic extraction and / or shaking extraction, such as reflux extraction; Preferably, the extraction time in step (1) is 0.5 to 3 hours, for example, about 2 hours; Particularly preferably, the concentration in step (1) is concentration under reduced pressure; Particularly preferably, in step (1), the volume / volume ratio of the Gynostemma pentaphyllum medicinal material ethanol extract to the ethanol is 0.05 to 0.2, such as about 0.

1.

5. The method according to claim 3, 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, such as hydrochloric acid; Preferably, the volume percentage concentration of the acid in step (2) is 0.5% to 3%, for example, about 1%; More preferably, in step (2), the volume / volume ratio of the acid to the ethanol extract of Gynostemma pentaphyllum is 0.1 to 1, such as about 0.2; Preferably, the reflux time in step (2) is 0.5 to 3 h, such as about 1 h.

6. The method according to claim 3, characterized in that, The column chromatography in step (3) uses silica gel with a particle size of 100 to 400 mesh; Preferably, the elution mobile phase of the column chromatography in step (3) is a mixed solvent of cyclohexane-ethyl acetate in a volume ratio of (10-30):(0.1-2); More preferably, the elution mobile phase of the column chromatography in step (3) is a mixed solvent of cyclohexane and ethyl acetate in a volume ratio of about 20: 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 relieving fever; More preferably, the extract is a traditional Chinese medicine extract.

8. The pharmaceutical composition according to claim 7, wherein The drug is a non-steroidal anti-inflammatory drug, a hormone drug and / or a Chinese patent medicine; Preferably, the nonsteroidal anti-inflammatory drug is selected from one or more of the following: aspirin, ibuprofen, acetaminophen, lysine, and diclofenac sodium; More preferably, the hormone drug is selected from one or more of the following: dexamethasone, methylprednisolone, hydrocortisone, triamcinolone and prednisolone; Also preferably, the Chinese patent medicine is Shufengjiedu Capsule and / or Tanreqing Capsule.

9. The pharmaceutical composition according to claim 7, wherein, The pharmaceutical composition comprises one or more pharmaceutically acceptable excipients; Preferably, the adjuvant is selected from one or more of the following: diluent, wetting agent, binder, disintegrant, inclusion agent, flavoring agent, sustained-release agent, retention aid, lubricant, dispersant, plasticizer, light-shielding agent, and antioxidant; More preferably, the dosage form of the pharmaceutical composition is ointment, powder, tablet, dripping pill, capsule, film, lozenge, granule, injection, or oral liquid; Still more preferably, the ointment is an ointment or a plaster.

10. Use of the dammarane-type 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 medicament for antipyretic use.

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