CN117338825A - Application of cordyceps sinensis digestive gland part in preparation of antitumor drugs - Google Patents
Application of cordyceps sinensis digestive gland part in preparation of antitumor drugs Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/062—Ascomycota
- A61K36/066—Clavicipitaceae
- A61K36/068—Cordyceps
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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Abstract
The invention discloses an application of cordyceps sinensis digestive gland part in preparing antitumor drugs. The activity experiment result shows that the anti-tumor activity of the cordyceps sinensis digestive gland part extract is obviously higher than that of the whole cordyceps sinensis digestive gland, the fruiting body and the worm body, and in the process of extracting the cordyceps sinensis digestive gland part extract, the anti-tumor activity of the cordyceps sinensis digestive gland part extract obtained by extracting is higher when the cordyceps sinensis digestive gland is soaked by adopting an alcohol-water mixture and alcohol as solvents compared with the cordyceps sinensis digestive gland is soaked by adopting water as solvents.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to application of cordyceps sinensis digestive gland parts in preparation of antitumor drugs.
Background
Tumors are malignant diseases with highest global mortality, the annual morbidity and mortality are continuously increasing, and the lives and the health of human beings are seriously endangered. Many of the traditional Chinese medicines in China have anti-tumor effects, such as barbed skullcap herb, coix seed, selaginella doederleinii, ginseng, astragalus mongholicus and the like. The plant antitumor drugs such as paclitaxel, triptolide and camptothecine are widely applied in clinical treatment. Cordyceps sinensis is reported to have antitumor activity as well. In recent years, with the increase of demands and the increasing of the price of cordyceps sinensis, the research on the therapeutic functions of different parts of cordyceps sinensis is widely interesting for better playing the medicinal effects of cordyceps sinensis.
Cordyceps sinensis is called Cordyceps sinensis for short, belongs to Cordyceps plant of Clavipitaceae of Hypocreaceae, is a compound of stroma of Cordyceps sinensis and host hepialus larva cadaver, and is a traditional tonic Chinese medicine in China. Along with the development of medical science at home and abroad and the gradual popularization of Chinese medicaments worldwide, the efficacy of the cordyceps sinensis is found to be wider and wider in recent years. Proved by researches, the method comprises the following steps: it has effects in tranquilizing mind, stopping bleeding, relieving convulsion, lowering blood pressure, improving myocardial ischemia, resisting platelet aggregation, resisting aging, and regulating immunity and tumor. The linear structures in the Cordyceps sinensis sections are called digestive glands, which are important tissues for nutrition absorption and metabolism of Cordyceps sinensis in host insects. The nutrient substances are absorbed from host insects through the expansion of mycelia and are converted into the growth energy of cordyceps sinensis, the digestive glands of the cordyceps sinensis are one of the unique marks, and the digestive glands have strong activity, but the medicinal function of the digestive glands of the cordyceps sinensis is still blank.
Disclosure of Invention
The invention aims to provide the medicinal application of the cordyceps sinensis digestive gland part.
The invention provides a medicinal application of a cordyceps sinensis digestive gland part, which is an application of the cordyceps sinensis digestive gland part in preparing an anti-tumor medicament.
The tumor can be specifically non-small cell lung cancer and cervical cancer.
The invention aims at providing a preparation method of cordyceps sinensis digestive gland part extract.
The preparation method of the cordyceps sinensis digestive gland part extract provided by the invention comprises the following steps:
1) Taking fresh cordyceps sinensis as a raw material, longitudinally cutting the cordyceps sinensis, and stripping digestive glands to obtain cordyceps sinensis digestive glands;
2) Pulverizing extracted Cordyceps sinensis digestive gland, soaking in water or alcohol-water or alcohol as extraction solvent, decocting or ultrasonic extracting for 2-3 times, mixing extractive solutions, filtering, collecting supernatant, ultrafiltering, and lyophilizing to obtain Cordyceps sinensis digestive gland extract.
In the step 2), the alcohol-water may be an aqueous solution of ethanol with a mass concentration of 50%;
the amount of water or alcohol-water or alcohol is 6-10 times of the mass of the digestive gland of the Cordyceps sinensis;
the soaking time can be 2-3 hours;
the temperature of the ultrasonic wave is controlled at 25 ℃;
the extraction time can be 1-3 h.
The invention also aims to provide the application of the cordyceps sinensis digestive gland part extract in preparing antitumor drugs.
The tumor can be specifically non-small cell lung cancer and cervical cancer.
The invention also provides an anti-tumor medicament which takes the cordyceps sinensis digestive gland part or the extract of the cordyceps sinensis digestive gland part as an active ingredient,
in the medicine, the concentration of Cordyceps sinensis digestive gland extract is 20-40 μg/ml.
According to actual needs, the medicine can also comprise pharmaceutically acceptable auxiliary materials. Making into various dosage forms such as decoction, granule, pill, capsule, tablet, powder, and oral liquid according to conventional preparation process.
The activity experimental result shows that the anti-tumor activity of the extract of the cordyceps sinensis digestive gland is obviously higher than that of the whole cordyceps sinensis digestive gland, the fruiting body and the worm body, and in the process of extracting the extract of the cordyceps sinensis digestive gland, the anti-tumor activity of the extract of the cordyceps sinensis digestive gland obtained by extracting the extract of the cordyceps sinensis digestive gland is higher by adopting an alcohol-water mixture and alcohol as solvents to soak the cordyceps sinensis digestive gland compared with the extract of the cordyceps sinensis digestive gland by adopting water as solvents.
Drawings
FIG. 1 is a graph showing MTT test results of antitumor drugs extracted from different parts of Cordyceps sinensis in example 1 of the present invention.
FIG. 2 is a graph showing MTT test results of extracts of digestive gland parts of Cordyceps sinensis obtained by soaking and decocting in water in example 2 of the present invention.
FIG. 3 is a graph showing the results of MTT test of extract of digestive gland of Cordyceps sinensis obtained by ultrasonic extraction in water immersion in example 3 of the present invention.
FIG. 4 is a graph showing the results of MTT test of extract of digestive gland of Cordyceps sinensis obtained by ultrasonic extraction in alcohol-water immersion in example 4 of the present invention.
FIG. 5 is a graph showing MTT test results of extracts of digestive gland parts of Cordyceps sinensis obtained by alcohol-immersed ultrasonic extraction in example 5 of the present invention.
Detailed Description
The following detailed description of the invention is provided in connection with the accompanying drawings that are presented to illustrate the invention and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the invention in any way.
The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are carried out according to techniques or conditions described in the literature in the field or according to the product specifications. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
EXAMPLE 1 comparison of the anti-tumor Activity of Cordyceps sinensis digestive glands and other fractions
1. Preparation of antitumor drugs for digestive glands and other parts of cordyceps sinensis
Cleaning 100g of fresh cordyceps sinensis, longitudinally cutting, peeling the digestive gland part, crushing, sieving with a No. 3 screen (50 meshes), adding 1000ml of deionized water, soaking for 3 hours, extracting by a decoction mode for 1 hour, extracting for 3 times, combining extracting solutions, filtering, collecting supernatant, ultrafiltering, and performing freeze-drying treatment to obtain the cordyceps sinensis digestive gland part extract. The tablet of the Cordyceps sinensis digestive gland antitumor drug with the specification of 0.2g is prepared by taking Cordyceps sinensis digestive gland extract as an active ingredient and distilled water, starch, microcrystalline cellulose, sodium carboxymethyl cellulose and magnesium stearate as auxiliary materials. The same method can obtain extracts of other parts (whole Cordyceps, fruiting body, and insect body) of Cordyceps, and making into tablet.
2. Evaluating the activity of antitumor drugs at digestive gland part and other parts of Cordyceps sinensis
MTT experiment: heLa cells were inoculated into 96-well plates, and Cordyceps sinensis digestive gland antitumor drugs, whole drugs, fruiting body drugs and insect body drugs (active ingredient concentration is 100. Mu.g/ml) were added respectively for culturing for 72 hours, then after removing the culture medium, 110. Mu.l of 10% MTT DMEM medium was added, and culturing was continued for 4 hours. The medium was then aspirated, 110. Mu.l of DMSO was added, and after mixing, the absorbance at 490nm was measured to calculate the cell viability. The MTT assay results are shown in FIG. 1. As can be seen from FIG. 1, the extract of the digestive gland of Cordyceps sinensis has the highest antitumor activity compared with the extracts of the whole plant, fruiting body and insect body.
Example 2 comparison of antitumor Effect of Cordyceps sinensis digestive gland extract obtained by Water decoction
1. Preparation of Cordyceps sinensis digestive gland antitumor drug
Cleaning 100g of fresh cordyceps sinensis, longitudinally cutting, peeling the digestive gland part, crushing, sieving with a No. 3 screen (50 meshes), adding 1000ml of deionized water, soaking for 3 hours, extracting by a decoction mode for 1 hour, extracting for 3 times, combining extracting solutions, filtering, collecting supernatant, ultrafiltering, and performing freeze-drying treatment to obtain the cordyceps sinensis digestive gland part extract. The tablet of the Cordyceps sinensis digestive gland antitumor drug with the specification of 0.2g is prepared by taking Cordyceps sinensis digestive gland extract as an active ingredient and distilled water, starch, microcrystalline cellulose, sodium carboxymethyl cellulose and magnesium stearate as auxiliary materials.
2. Evaluation of the Activity of Cordyceps sinensis digestive gland part antitumor drug
MTT experiment: cells A549 and HeLa cells were inoculated into 96-well plates, then Cordyceps sinensis digestive gland antitumor drugs (Cordyceps sinensis digestive gland extracts at concentrations of 0. Mu.g/ml, 5. Mu.g/ml, 10. Mu.g/ml, 20. Mu.g/ml and 40. Mu.g/ml, respectively) were added at different concentrations, the culture was continued for 72 hours, then after removing the medium, 110. Mu.l of 10% MTT DMEM medium was added, and the culture was continued for 4 hours. The medium was then aspirated, 110. Mu.l of DMSO was added, and after mixing, the absorbance at 490nm was measured to calculate the cell viability. The MTT assay results are shown in FIG. 2. As can be seen from fig. 2, the survival rate of a549 and HeLa cells decreases with the increase of the concentration of the extract of the digestive gland of the Cordyceps sinensis obtained by water decoction, so that the extract of the digestive gland of the Cordyceps sinensis obtained by water decoction has remarkable anti-tumor activity and is positively correlated with the concentration thereof.
Example 3 preparation of Cordyceps sinensis digestive gland antitumor drug by soaking in water and ultrasonic extraction
100g of fresh cordyceps sinensis is taken, cleaned, longitudinally cut, stripped from the digestive gland part, crushed, screened by a No. 3 screen (50 meshes), soaked in 1000ml of deionized water for 3 hours, extracted by adopting an ultrasonic mode, and the extraction temperature is controlled at 25 ℃ for 3 hours. Extracting for 3 times, mixing the extractive solutions, filtering, collecting supernatant, ultrafiltering, and lyophilizing to obtain Cordyceps extract. The tablet of the Cordyceps sinensis digestive gland antitumor drug with the specification of 0.2g is prepared by taking Cordyceps sinensis digestive gland extract as an active ingredient and distilled water, starch, microcrystalline cellulose, sodium carboxymethyl cellulose and magnesium stearate as auxiliary materials.
2. Evaluation of the Activity of Cordyceps sinensis digestive gland part antitumor drug
MTT experiment: heLa cells and A549 cells were inoculated into 96-well plates, then, cordyceps sinensis digestive gland antitumor drugs (Cordyceps sinensis digestive gland extracts at concentrations of 0. Mu.g/ml, 5. Mu.g/ml, 10. Mu.g/ml, 20. Mu.g/ml and 40. Mu.g/ml, respectively) were added at different concentrations, the culture was continued for 72 hours, then, after removing the culture medium, 110. Mu.l of DMEM medium at 10% MTT was added, and the culture was continued for 4 hours. The medium was then aspirated, 110. Mu.l of DMSO was added, and after mixing, the absorbance at 490nm was measured to calculate the cell viability. The MTT assay results are shown in FIG. 3. As shown in fig. 3, the survival rate of a549 and HeLa cells is reduced with the increase of the concentration of the extract of the digestive gland of the Cordyceps sinensis obtained by the water immersion ultrasonic extraction, so that the extract of the digestive gland of the Cordyceps sinensis obtained by the water immersion ultrasonic extraction has remarkable anti-tumor activity and is positively correlated with the concentration thereof.
Example 4 comparison of antitumor Effect of Cordyceps sinensis digested glandular extract obtained by ethanol Water immersion ultrasonic extraction
1. Preparation of Cordyceps sinensis digestive gland antitumor drug
100g of fresh cordyceps sinensis is taken, cleaned, longitudinally cut, stripped from the digestive gland part, crushed, screened by a No. 3 screen (50 meshes), soaked in 1000ml of 50% ethanol water with mass concentration for 3 hours, extracted by adopting an ultrasonic mode, and the extraction temperature is controlled at 25 ℃ and the extraction time is 3 hours. Extracting for 3 times, mixing the extractive solutions, filtering, collecting supernatant, ultrafiltering, and lyophilizing to obtain Cordyceps extract. The tablet of the Cordyceps sinensis digestive gland antitumor drug with the specification of 0.2g is prepared by taking Cordyceps sinensis digestive gland extract as an active ingredient and distilled water, starch, microcrystalline cellulose, sodium carboxymethyl cellulose and magnesium stearate as auxiliary materials.
2. Evaluation of the Activity of Cordyceps sinensis digestive gland part antitumor drug
MTT experiment: heLa cells and A549 cells were inoculated into 96-well plates, then, cordyceps sinensis digestive gland antitumor drugs (Cordyceps sinensis digestive gland extracts at concentrations of 0. Mu.g/ml, 5. Mu.g/ml, 10. Mu.g/ml, 20. Mu.g/ml and 40. Mu.g/ml, respectively) were added at different concentrations, the culture was continued for 72 hours, then, after removing the culture medium, 110. Mu.l of DMEM medium at 10% MTT was added, and the culture was continued for 4 hours. The medium was then aspirated, 110. Mu.l of DMSO was added, and after mixing, the absorbance at 490nm was measured to calculate the cell viability. The MTT assay results are shown in FIG. 4. As can be seen from fig. 4, the extract of the digestive gland part of Cordyceps sinensis obtained by ultrasonic extraction after soaking in 50% ethanol water has remarkable anti-tumor activity and has positive correlation with the concentration thereof.
Example 5 comparison of antitumor Effect of Cordyceps sinensis digested glandular extract obtained by ethanol soaking ultrasonic extraction
1. Preparation of Cordyceps sinensis digestive gland antitumor drug
100g of fresh cordyceps sinensis is taken, cleaned, longitudinally cut, stripped from the digestive gland part, crushed, screened by a No. 3 screen (50 meshes), soaked in 1000ml of 100% ethanol for 3 hours, and then extracted by adopting an ultrasonic mode, wherein the extraction temperature is controlled at 25 ℃ and the extraction time is 3 hours. Extracting for 3 times, mixing the extractive solutions, filtering, collecting supernatant, ultrafiltering, and lyophilizing to obtain Cordyceps extract. The tablet of the Cordyceps sinensis digestive gland antitumor drug with the specification of 0.2g is prepared by taking Cordyceps sinensis digestive gland extract as an active ingredient and distilled water, starch, microcrystalline cellulose, sodium carboxymethyl cellulose and magnesium stearate as auxiliary materials.
2. Evaluation of the Activity of Cordyceps sinensis digestive gland part antitumor drug
MTT experiment: heLa cells and A549 cells were inoculated into 96-well plates, then, cordyceps sinensis digestive gland antitumor drugs (Cordyceps sinensis digestive gland extracts at concentrations of 0. Mu.g/ml, 5. Mu.g/ml, 10. Mu.g/ml, 20. Mu.g/ml and 40. Mu.g/ml, respectively) were added at different concentrations, the culture was continued for 72 hours, then, after removing the culture medium, 110. Mu.l of DMEM medium at 10% MTT was added, and the culture was continued for 4 hours. The medium was then aspirated by washing, 110. Mu.l of DMSO was added, and after mixing, the absorbance at 490nm was measured to calculate the cell viability. The MTT assay results are shown in FIG. 5. As can be seen from fig. 5, the extract of the digestive gland part of Cordyceps sinensis obtained by ultrasonic extraction with ethanol has remarkable antitumor effect, and the extract of the digestive gland part of Cordyceps sinensis obtained by ultrasonic extraction with ethanol soaking has the best antitumor effect compared with the extract of the digestive gland part of Cordyceps sinensis obtained by ultrasonic extraction with water soaking and 50% ethanol soaking.
The present invention is described in detail above. It will be apparent to those skilled in the art that the present invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with respect to specific embodiments, it will be appreciated that the invention may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains.
Claims (7)
1. Application of Cordyceps sinensis digestive gland part in preparing antitumor drug is provided.
2. Application of Cordyceps sinensis digestive gland extract in preparing antitumor drug is provided.
3. Use according to claim 1 or 2, characterized in that: the tumor is non-small cell lung cancer and cervical cancer.
4. The use according to claim 2, characterized in that: the Cordyceps sinensis digestive gland part extract is prepared by the following steps:
1) Taking fresh cordyceps sinensis as a raw material, longitudinally cutting the cordyceps sinensis, and stripping digestive glands to obtain cordyceps sinensis digestive glands;
2) Pulverizing extracted Cordyceps sinensis digestive gland, soaking in water or alcohol-water or alcohol as extraction solvent, decocting or ultrasonic extracting for 2-3 times, mixing extractive solutions, filtering, collecting supernatant, ultrafiltering, and lyophilizing to obtain Cordyceps sinensis digestive gland extract.
5. The use according to claim 4, characterized in that: in the step 2), the water or alcohol-water or alcohol amount is 6-10 times of the mass of the digestive gland of the cordyceps sinensis;
the soaking time is 2-3 hours;
the temperature of the ultrasonic wave is controlled at 25 ℃;
the extraction time is 1-3 h.
6. An antitumor drug comprises Cordyceps sinensis digestive gland part or Cordyceps sinensis digestive gland part extract as active ingredient.
7. The antitumor drug according to claim 6, wherein the concentration of extract of digestive gland part of Cordyceps sinensis is 20-40 μg/ml.
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