CN118286306A - Abnormal black bile maturation agent extract and preparation method and application thereof - Google Patents

Abnormal black bile maturation agent extract and preparation method and application thereof Download PDF

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Publication number
CN118286306A
CN118286306A CN202311110158.XA CN202311110158A CN118286306A CN 118286306 A CN118286306 A CN 118286306A CN 202311110158 A CN202311110158 A CN 202311110158A CN 118286306 A CN118286306 A CN 118286306A
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preparation
abnormal black
abnormal
gall
agent
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吴乐和
韩玉玲
秦利彬
韩鹏军
阿卜杜喀哈尔·图尔荪江
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Hetian Uygur Pharmaceutical Co ltd
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Hetian Uygur Pharmaceutical Co ltd
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Abstract

The invention provides an abnormal black bile maturation agent extract, a preparation method and application thereof, belonging to the technical field of biological medicine, wherein the preparation method comprises the following steps: mixing dry powder of the abnormal black bile maturation agent with petroleum ether for degreasing, filtering to obtain filter residues, mixing the filter residues with water, stirring for dissolving, filtering to obtain filtrate, adding 80% -90% ethanol solution into the filtrate, standing at 3-8 ℃ for 12-36 h, and precipitating to obtain precipitate. The invention adopts the raw materials of the vitamin abnormal black gall maturing agent, has wide sources, is cheap and easy to obtain, and has no toxic or side effect on human bodies. In the preparation method, the preparation method has simple process, and the abnormal black gall maturing agent is separated and extracted by adopting petroleum ether, water and ethanol solution, so that the active ingredients in the abnormal black gall maturing agent can be effectively extracted, and the abnormal black gall maturing agent extract prepared by the preparation method can be used for effectively treating diabetes.

Description

Abnormal black bile maturation agent extract and preparation method and application thereof
Technical Field
The invention relates to the technical field of biological medicines, in particular to an abnormal black bile maturation agent extract, a preparation method and application thereof.
Background
Diabetes is a metabolic disease characterized by chronic hyperglycemia. Type 2 diabetics account for more than 90% of diabetics. A series of chronic protein, carbohydrate and fat metabolic disorders caused by type 2 diabetes cause insulin resistance can gradually cause damage to tissues and organs such as kidney, eye, heart and limbs, and acute metabolic disorder, hypertonic hyperglycemia syndrome, ketoacidosis and other diseases can occur to patients with serious failure. The hypoglycemic drugs are important means for treating type 2 diabetes, and although various effective hypoglycemic drugs can be selected by clinicians at present, the hypoglycemic drugs still cannot meet huge diabetes patient groups, and the overall blood sugar standard reaching rate is reduced. The scholars at home and abroad search active substances capable of assisting in reducing blood sugar from natural products, and hope to bring more choices to diabetics.
The abnormal black gall maturing agent is a compound preparation formed by ten medicinal materials, can condition abnormal black gall type body fluid quality of a vitamin, restore the balance of four body fluid qualities of black gall, mucic fluid quality, gall quality, blood quality and the like, and has the effects of preventing and treating difficult and complicated diseases such as abnormal black gall tumors, asthma, anxiety, cardiovascular diseases, psoriasis, aging, depression, psychosis and the like. Therefore, the detection of the effective components extracted from the abnormal black bile maturation agent, the biological activity and the medicinal value thereof has great significance.
Disclosure of Invention
The invention aims to provide an abnormal black gall maturing agent extract, a preparation method and application thereof.
In order to achieve the above object, the present invention provides the following technical solutions:
The invention provides a preparation method of an abnormal black bile maturation agent extract, which comprises the following steps:
mixing dry powder of the abnormal black gall maturing agent with petroleum ether for degreasing, filtering to obtain filter residues, mixing the filter residues with water, stirring for dissolving, filtering to obtain filtrate, adding 80% -90% ethanol solution into the filtrate, standing at 3-8 ℃ for 12-36 h for precipitation to obtain a precipitate, and obtaining the abnormal black gall maturing agent extract.
Preferably, the mass volume ratio of the filter residue to the water is 1:8-12 g/mL, the stirring and dissolving temperature is 65-95 ℃, and the stirring and dissolving time is 10-40 min.
Preferably, the volume ratio of the filtrate to the 80% -90% ethanol solution is 1:10-14.
Preferably, the mass-volume ratio of the dry powder of the abnormal black bile maturation agent to the petroleum ether is 1:4-6 g/mL.
Preferably, the degreasing temperature is 55-65 ℃, and the degreasing time is 1-2 h.
The invention also provides an abnormal black bile maturation agent extract prepared by the preparation method.
The invention also provides a medicine for treating diabetes mellitus, an active ingredient of the medicine and pharmaceutically acceptable auxiliary materials, wherein the active ingredient comprises the abnormal black bile maturation agent extract.
Preferably, the auxiliary materials comprise one or more of diluents, wetting agents, binders, lubricants, absorbents and preservatives.
The invention also provides application of the abnormal black bile maturation agent extract in preparing a medicament for treating diabetes.
Preferably, the diabetes is type 2 diabetes.
Compared with the prior art, the invention has the beneficial effects that:
The invention provides an abnormal black gall maturing agent extract and a preparation method and application thereof. In the preparation method, the preparation method has simple process, and the abnormal black gall maturing agent is separated and extracted by adopting petroleum ether, water and ethanol solution, so that the active ingredients in the abnormal black gall maturing agent can be effectively extracted, and the abnormal black gall maturing agent extract prepared by the preparation method can be used for effectively treating diabetes.
Detailed Description
The invention provides a preparation method of an abnormal black bile maturation agent extract, which comprises the following steps:
mixing dry powder of the abnormal black gall maturing agent with petroleum ether for degreasing, filtering to obtain filter residues, mixing the filter residues with water, stirring for dissolving, filtering to obtain filtrate, adding 80% -90% ethanol solution into the filtrate, standing at 3-8 ℃ for 12-36 h for precipitation to obtain a precipitate, and obtaining the abnormal black gall maturing agent extract.
In the invention, dry powder of the abnormal black bile maturation agent is mixed with petroleum ether for degreasing, and filter residues are filtered. In the invention, the raw material composition of the abnormal black bile maturing agent and the preparation method thereof are disclosed in Chinese patent application number 02130082.8. In the invention, the abnormal black bile maturing agent consists of the following raw materials in parts by weight: 7 parts of lavender, 7 parts of fennel, 15 parts of ragweed, 15 parts of jujube, 7 parts of herba hedyotis, 7 parts of melissa, 10 parts of liquorice, 7 parts of iron wire fern, 7 parts of humifuse euphorbia herb and 15 parts of thorn sugar. The preparation method of the abnormal black bile maturation agent comprises the steps of cutting other medicinal materials except for the glucose in the abnormal black bile maturation agent, adding 12 times of water, heating and decocting for 20min, stopping heating, soaking, standing for 18h, heating and decocting for 2h with slow fire, filtering, adding the glucose into the filtrate while the filtrate is hot, stirring and dissolving, filtering, and diluting the filtrate to 5 times with distilled water to obtain the abnormal black bile maturation agent. And (3) drying the abnormal black gall maturing agent at the temperature of 40-50 ℃ to obtain dry powder of the abnormal black gall maturing agent.
In the invention, the mass volume ratio of the dry powder of the abnormal black bile maturation agent to the petroleum ether is preferably 1:4-6 g/mL, more preferably 1:4.5-5.5 g/mL, the degreasing temperature is preferably 55-65 ℃, more preferably 58-63 ℃, and the degreasing time is preferably 1-2 h. According to the invention, petroleum ether degreasing is carried out on the dry powder of the abnormal black gall maturing agent, so that the fat-soluble impurity components of the dry powder of the abnormal black gall maturing agent can be effectively removed. The filtering method of the present invention is not particularly limited, and any method known in the art may be used.
In the invention, filter residues are mixed with water, stirred and dissolved, and the filtrate is filtered. The mass volume ratio of the filter residue to the water is preferably 1:8-12 g/mL, and more preferably 1:9-11 g/mL. The temperature of the stirring dissolution is preferably 65 to 95 ℃, more preferably 70 to 90 ℃, and the time of the stirring dissolution is preferably 10 to 40min, more preferably 15 to 35min. The invention can extract the active ingredients in the abnormal black gall maturing agent through water extraction.
In the invention, 80-90% ethanol solution is added into the filtrate, and the mixture is stood for 12-36 hours at the temperature of 3-8 ℃ to precipitate and obtain the precipitate, thus obtaining the abnormal black gall maturing agent extract. The volume ratio of the filtrate to the 80% -90% ethanol solution is preferably 1:10-14. The solvent of the ethanol solution is preferably water, and the concentration percentage of the ethanol solution is volume percentage. The invention also comprises drying at 40-50 ℃ until no ethanol smell exists after the precipitation is obtained. The invention obviously improves the content of the active ingredients of the abnormal black gall maturing agent by means of water extraction and alcohol precipitation.
The invention also provides an abnormal black bile maturation agent extract prepared by the preparation method.
The invention also provides a medicine for treating diabetes mellitus, an active ingredient of the medicine and pharmaceutically acceptable auxiliary materials, wherein the active ingredient comprises the abnormal black bile maturation agent extract.
In the invention, the dosage form of the medicine can be powder, granules, tablets, capsules, dripping pills, powder, freeze-dried powder injection or solution. The medicaments of the present invention may be administered in unit dosage form such as orally, intramuscularly, subcutaneously, nasally, oromucosally, dermally, peritoneally or rectally, etc. The route of administration of the medicament of the present invention may also be by injection, including intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection, and the like. The auxiliary materials comprise one or more of diluents, wetting agents, binders, lubricants, absorbents and preservatives. The diluent and the absorbent can be one or more of starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, kaolin, microcrystalline cellulose, aluminum silicate and the like; the wetting agent and the binding agent can be one or more of water, glycerol, polyethylene glycol, ethanol, propanol, starch slurry, dextrin, syrup, honey, glucose solution, acacia slurry, gelatin slurry, sodium carboxymethyl cellulose, shellac, methyl cellulose, potassium phosphate, polyvinylpyrrolidone and the like; the lubricant is one or more of talcum powder, silicon dioxide, corn starch, stearate, boric acid, liquid paraffin, polyethylene glycol and the like; the preservative can be one or more of sodium benzoate, nipagin, sorbic acid and the like.
The dosage of the agent of the present invention to be administered depends on many factors such as the nature and severity of the disease to be prevented or treated, the sex, age, weight, character and individual response of the patient or animal, the route of administration, the number of times of administration, the purpose of treatment, and thus the therapeutic dosage of the present invention may vary widely. Generally, the dosages of the medicaments used in the present invention are well known to those skilled in the art. The amount of the actual drug contained in the pharmaceutical formulation of the present invention can be appropriately adjusted to achieve the therapeutically effective amount thereof, thereby achieving the preventive or therapeutic object of the present invention. The amount of the drug of the present invention used per day is 0.001 to 150mg/kg body weight, preferably 0.01 to 100mg/kg body weight, more preferably 0.01 to 60mg/kg body weight, most preferably 0.1 to 10mg/kg body weight.
The invention also provides application of the abnormal black bile maturation agent extract in preparing a medicament for treating diabetes.
In the present invention, the diabetes is type 2 diabetes.
In the present invention, all raw material components are commercially available products well known to those skilled in the art unless specified otherwise.
The technical solutions of the present invention will be clearly and completely described in the following in connection with the embodiments of the present invention. It will be apparent that the described embodiments are only some, but not all, embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
A preparation method of an abnormal black bile maturation agent extract comprises the following steps:
Mixing 100g of dry powder of an abnormal black bile maturation agent with 500mL of petroleum ether, heating and degreasing for 1.5h at 60 ℃, filtering to obtain filter residues, and mixing the filter residues with water according to a mass volume ratio of 1: mixing 10g/mL, stirring at 80deg.C for 30min, filtering to obtain filtrate, adding 85% ethanol solution into the filtrate, mixing uniformly, standing at 5deg.C for 24 hr for precipitation, and oven drying the precipitate at 45deg.C until no ethanol smell is present to obtain abnormal black matrix maturation agent extract.
Example 2
A preparation method of an abnormal black bile maturation agent extract comprises the following steps:
Mixing 100g of dry powder of an abnormal black bile maturation agent with 400mL of petroleum ether, heating and degreasing for 2h at 55 ℃, filtering to obtain filter residues, and mixing the filter residues with water according to a mass volume ratio of 1: mixing 12g/mL, stirring at 95 ℃ for dissolving for 10min, filtering to obtain filtrate, adding 80% ethanol solution into the filtrate, mixing uniformly, standing for 12h at 3 ℃ for precipitation, and drying the precipitate at 50 ℃ until no ethanol smell exists, thus obtaining the abnormal black bile maturation agent extract.
Example 3
A preparation method of an abnormal black bile maturation agent extract comprises the following steps:
mixing 100g of dry powder of an abnormal black bile maturation agent with 600mL of petroleum ether, heating and degreasing for 1h at 65 ℃, filtering to obtain filter residues, and mixing the filter residues with water according to a mass volume ratio of 1: mixing 8g/mL, stirring at 65deg.C for 40min, filtering to obtain filtrate, adding 90% ethanol solution into the filtrate, mixing uniformly, standing at 8deg.C for 36 hr for precipitation, and oven drying the precipitate at 40deg.C until no ethanol smell is present to obtain abnormal black matrix maturation agent extract.
Test example 1
Therapeutic Effect of abnormal Heiban maturation agent extract prepared in example 1 on type 2 diabetic mice
Preparing an experimental reagent:
The abnormal black gall maturing agent extract prepared in the example 1 is dissolved in distilled water, and 30mg/mL, 60mg/mL and 120mg/mL of abnormal black gall maturing agent extract are prepared respectively.
1% Streptozotocin (STZ) configuration:
Adding 10mLddH 2 O into 0.2g of citric acid to obtain a citric acid solution, adding 10mL of ddH 2 O into 0.294g of sodium citrate to obtain a sodium citrate solution, mixing the citric acid solution and the sodium citrate solution according to a ratio of 1:1, and regulating the pH value of the buffer solution to 4.4 to obtain the citric acid-sodium citrate buffer solution. STZ and citric acid-sodium citrate buffer were weighed 1g:100mL of the mixture was gently swirled and mixed to prepare a 1% STZ solution.
Grouping and dosing animals:
experimental animal
C57/BL6J male mice (18-20 g, 3-5 weeks old) purchased from Hunan Laike Jingda laboratory animal Co., ltd., animal feeding temperature is 24+ -1deg.C, humidity is 50+ -10%, light and darkness rules are alternately ensured, free feeding and drinking water are ensured, and the animals are fed adaptively for 1 week.
Randomly dividing 60C 57/BL6J male mice fed for 1 week into 6 groups, namely a normal control group (NC), a type 2 diabetes model group, a low, medium and high concentration group of abnormal black gall maturation agent extract and a metformin positive control group; except for the normal control group, the other 5 groups are fed by a high-fat model feed (Nantong terlafei feed technology Co., ltd.) and the mice are fasted overnight for 12 hours, and 50mg/kg precooled 1% STZ solution is injected into the abdominal cavity of the high-fat diet mice for 3 days; NC group mice are injected with buffer solution in the abdominal cavity for 3 continuous days, and after stabilizing for 1 week, the fasting blood glucose of the tip of the mouse tail is measured to be more than 11.1mmo/L, so that the model is successfully modeled, and the model 2 type diabetes mice are continuously fed with 60% high-fat model feed.
And (3) taking the model-successfully-molded type 2 diabetes mice according to a normal control group (NC), a type 2 diabetes model group and a low, medium and high concentration group of abnormal black gall maturation agent extract, wherein 6 groups of the abnormal black gall maturation agent extract are respectively subjected to gastric lavage by adopting an abnormal black gall maturation agent extract solution of 30mg/mL, 60mg/mL and 120mg/mL in sequence, the gastric lavage dosage is calculated according to 0.2mL of gastric lavage liquid of each 20g of mice, and the normal control group and the type 2 diabetes model group are respectively subjected to gastric lavage by adopting distilled water of equal volume, and 300mg/kg of the metformin positive control group are respectively subjected to daily administration for 60 days, wherein the metformin positive control group is subjected to daily administration for 1 time.
And (3) index detection:
Mice body weight level detection: body weight was measured on days 10, 20, 30, 40, 50 and 60, respectively, throughout the experiment.
Mice fasting blood glucose level detection: mice were tested for fasting blood glucose on days 10, 20, 30, 40, 50 and 60, respectively. Each group was fasted for 12h in advance and the glucometer rapidly measured the blood from the mouse tail tip.
Sugar tolerance test: mice were fasted for 12h in advance. 20% glucose solution (2 g/kg) was intraperitoneally injected in the 60 th sky abdomen state. Blood glucose values were measured at time points of 0.5h, 1h and 2h.
TABLE 1 blood glucose change trend (Unit: nmol/L) of diabetic mouse model
From the results in Table 1, it can be seen that model establishment was successful in type 2 diabetic mice.
TABLE 2 blood glucose level measurement results (unit: nmol/L) after 60 days in different groups of mice
The results in table 2 show that the abnormal hepatoma agent extract was significantly reduced in fasting blood glucose levels in the high, medium, and low concentration groups and the metformin positive control group, as compared to the type 2 diabetes model group, wherein the high concentration group of the abnormal hepatoma agent extract was at normal blood glucose levels and the concentration group of the abnormal hepatoma agent extract was at normal blood glucose levels.
TABLE 3 results of blood glucose level changes in glucose tolerance after day 60 in mice of different groups (unit: nmol/L)
The results in table 3 show that the blood glucose level of the abnormal black bilinear maturation agent extract was decreased in the low, medium and high concentration groups and the metformin positive control group with the lapse of time, and the difference between the low, medium and high concentration groups of the abnormal black bilinear maturation agent extract after 2 hours was significant as compared with the type 2 diabetes model group.
The abnormal black gall maturing agent extract prepared by the preparation method disclosed by the invention treats a type 2 diabetes mouse model mouse, and the result shows that the abnormal black gall maturing agent extract disclosed by the invention can obviously reduce the fasting blood glucose level of each group of mice and improve the glucose tolerance, so that the abnormal black gall maturing agent extract prepared by the preparation method disclosed by the invention can effectively treat type 2 diabetes.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.

Claims (10)

1. The preparation method of the abnormal black bile maturation agent extract is characterized by comprising the following steps:
mixing dry powder of the abnormal black gall maturing agent with petroleum ether for degreasing, filtering to obtain filter residues, mixing the filter residues with water, stirring for dissolving, filtering to obtain filtrate, adding 80% -90% ethanol solution into the filtrate, standing at 3-8 ℃ for 12-36 h for precipitation to obtain a precipitate, and obtaining the abnormal black gall maturing agent extract.
2. The preparation method according to claim 1, wherein the mass-to-volume ratio of the filter residue to the water is 1:8-12 g/mL, the stirring and dissolving temperature is 65-95 ℃, and the stirring and dissolving time is 10-40 min.
3. The preparation method according to claim 1, wherein the volume ratio of the filtrate to the 80% -90% ethanol solution is 1:10-14.
4. The preparation method of claim 1, wherein the mass-to-volume ratio of the dry powder of the abnormal black bile maturation agent to the petroleum ether is 1:4-6 g/mL.
5. The method according to claim 1, wherein the degreasing temperature is 55 to 65 ℃ and the degreasing time is 1 to 2 hours.
6. An abnormal black matrix maturation agent extract prepared by the method according to any one of claims 1 to 5.
7. A medicament for treating diabetes, characterized in that the active ingredient of the medicament comprises the abnormal black bile maturation agent extract according to claim 6 and pharmaceutically acceptable auxiliary materials.
8. The medicament according to claim 7, wherein the auxiliary materials comprise one or more of diluents, wetting agents, binders, lubricants, absorbents, preservatives.
9. Use of the abnormal savda munziq agent extract according to claim 6, in the manufacture of a medicament for the treatment of diabetes.
10. The use according to claim 9, wherein the diabetes is type 2 diabetes.
CN202311110158.XA 2023-08-29 2023-08-29 Abnormal black bile maturation agent extract and preparation method and application thereof Pending CN118286306A (en)

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Application Number Priority Date Filing Date Title
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