CN115919908A - Ganoderma spore oil-containing composition for improving tumor inhibition rate and preparation method thereof - Google Patents
Ganoderma spore oil-containing composition for improving tumor inhibition rate and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a ganoderma lucidum spore oil composition for improving tumor inhibition rate and a preparation method thereof, wherein the composition comprises the following components in parts by weight: 1-15 parts of ganoderma lucidum spore oil and 1-15 parts of ganoderma lucidum alcohol extract; wherein the ganoderma lucidum alcohol extract is prepared by mixing ganoderma lucidum fruit bodies according to the weight ratio of 1:10 to 12 weight percent of the extract is added with 75 percent ethanol, heated and extracted for 2 to 3 times, each time lasts for 1.5 to 2 hours, the mixture is filtered, the filtrate is combined, concentrated to the specific gravity of between 1.15 and 1.25, and the mixture is sprayed and dried to obtain the product. The invention takes ganoderma lucidum spore oil and ganoderma lucidum alcohol extract as main raw materials to prepare a composition containing ganoderma lucidum spore oil with high tumor inhibition rate. The composition has effects of enhancing immunity, inhibiting tumor, and has far stronger effect than single prescription, and novel drug development prospect.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a composition containing ganoderma lucidum spore oil and capable of improving the tumor inhibition rate and a preparation method thereof.
Background
Ganoderma lucidum is a fruiting body of Ganoderma lucidum of Polyporaceae. The shape of the utility model is umbrella shape, pileus kidney shape, semicircle or near circle. Ganoderma lucidum is recorded in Ben Cao gang mu (compendium of materia Medica) compiled by Li Shi Zhen of Ming Dynasty and university physicians. The ganoderma alcohol extract is prepared by collecting mature fresh sporophore at proper time, drying, extracting with alcohol, vacuum concentrating, spray drying, etc. to obtain ganoderma extract powder, which is ganoderma essence prepared by high-concentration of ganoderma powder. The Ganoderma alcohol extract contains main effective components such as ganoderan and triterpenoid ganoderic acid more abundant than Ganoderma powder. Has sleep improving, antioxidant, anticancer, and immunity enhancing effects.
The Ganoderma spore is tiny egg-shaped germ cells ejected from Ganoderma during mature period, and has all genetic material and health promotion effects of Ganoderma. The medicinal value of the ganoderma lucidum spore is increasingly valued, and researches show that the ganoderma lucidum spore has the effects of enhancing the immunity of organisms, inhibiting tumors, protecting liver injury and protecting radiation. The Ganoderma spore oil is oily lipid extracted from wall-broken Ganoderma spore by supercritical carbon dioxide fluid extraction technology, and integrates multiple active ingredients of Ganoderma spore. The ganoderma spore oil is yellow transparent liquid, mainly contains saturated fatty acid and unsaturated fatty acid, contains a small amount of ganoderma acid, and has effects of protecting liver, removing toxic substance, relieving pain, enhancing immunity, and inhibiting tumor.
The existing ganoderma lucidum spore oil products or ganoderma lucidum related products have low tumor inhibition rate and high dosage.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides the ganoderma lucidum spore oil composition for improving the tumor inhibition rate.
Another object of the present invention is to provide a method for preparing the above ganoderma lucidum spore oil composition with improved tumor inhibition rate.
The purpose of the invention is realized by the following modes:
a ganoderma lucidum spore oil composition for improving tumor inhibition rate comprises the following components in parts by weight:
1-15 parts of ganoderma lucidum spore oil and 1-15 parts of ganoderma lucidum alcohol extract; wherein the alcohol extract of ganoderma lucidum is prepared by mixing ganoderma lucidum fruit bodies according to the proportion of 1:10-12 weight parts of the raw materials are added with 75% ethanol, heated and extracted for 2-3 times, each time lasts for 1.5-2 hours, the mixture is filtered, the filtrate is combined, concentrated to the specific gravity of 1.15-1.25, and spray-dried to obtain the traditional Chinese medicine.
Preferably, in the above step, the ganoderma lucidum fruit body is prepared according to the ratio of 1:10 to 75% by weight of ethanol.
Preferably, the Ganoderma spore oil is prepared by mixing wall-broken Ganoderma spore powder with water, making into strip-shaped granule, drying at low temperature until water content is not higher than 6%, and adopting CO 2 Supercritical extraction of fat-soluble components; wherein, CO 2 The supercritical extraction conditions are that the extraction pressure is 25-28 MPa, the extraction temperature is 25-30 ℃, and the extraction time is 1-2 hours.
Preferably, said CO is 2 The supercritical extraction adopts the extraction pressure of 25MPa, the extraction temperature of 30 ℃ and the extraction time of 2 hours.
Preferably, the total triterpene content in the sporoderm-broken ganoderma lucidum spore powder is not less than 2.5%.
Preferably, the water amount adopted when the strip-shaped particles are prepared is 1/5-1/4 of the weight of the wall-broken ganoderma lucidum spore powder.
In the ganoderma lucidum spore oil composition, 1 to 3 parts by weight of ganoderma lucidum spore oil and 1 to 3 parts by weight of ganoderma lucidum alcohol extract are preferred.
The preparation method of the ganoderma lucidum spore oil composition for improving the tumor inhibition rate comprises the following steps:
(1) Adding water into wall-broken Ganoderma spore powder, making into strip granule, drying at low temperature until water content is not higher than 6%, and adopting CO 2 Supercritical extraction of fat-soluble components; wherein, CO 2 The supercritical extraction conditions comprise that the extraction pressure is 25-28 MPa, the extraction temperature is 25-30 ℃, and the extraction time is 1-2 hours, so as to obtain the ganoderma lucidum spore oil;
(2) The lucid ganoderma sporocarp is prepared according to the following steps of 1:10-12, adding 75% ethanol, heating and extracting for 2-3 times, each for 1.5-2h, filtering, mixing filtrates, concentrating to specific gravity of 1.15-1.25, and spray drying to obtain the Ganoderma extract.
(3) Mixing 1-15 parts by weight of ganoderma lucidum spore oil and 1-15 parts by weight of ganoderma lucidum alcohol extract.
Preferably, in the step (1), the total triterpenoids in the sporoderm-broken ganoderma lucidum spore powder are not less than 2.5%.
Preferably, in the step (1), the water amount is 1/5-1/4 of the weight of the sporoderm-broken ganoderma lucidum spore powder.
Preferably, in the step (2), CO 2 The supercritical extraction conditions were: the extraction pressure is 25MPa, the extraction temperature is 30 ℃, and the extraction time is 2 hours.
Compared with the prior art, the invention has the beneficial effects that: the invention takes ganoderma lucidum spore oil and ganoderma lucidum alcohol extract as main raw materials, and adopts a specific preparation method to prepare the composition containing ganoderma lucidum spore oil with high tumor inhibition rate. The composition has effects of enhancing immunity, inhibiting tumor, and has far stronger effect than single prescription, and novel drug development prospect.
Drawings
FIG. 1 shows typical chromatogram of Ganoderma triterpene.
Fig. 2 is a typical chromatogram for ergosterol.
Detailed Description
The invention is further illustrated by the following specific examples.
Ganoderma spore powder (total triterpene content not less than 2.5%) is from Anhui Huangshan Yunle Ganoderma Limited, and total triterpene content is not less than 3.0%.
Example 1
And uniformly mixing 1 part by weight of ganoderma lucidum spore oil and 1 part by weight of ganoderma lucidum alcohol extract to obtain a mixture. The main marker components are Ganoderma triterpene and ergosterol, and the chromatogram is shown in figure 1 and figure 2 respectively.
The preparation method comprises the following steps:
(1) Collecting Ganoderma spore powder with wall-broken rate of above 95% (total triterpene content is not less than 2.5%), adding water 1/5 of wall-broken Ganoderma spore powder weight to obtain strip granule, drying at low temperature until water content is not more than 6%, and adopting CO 2 Supercritical extraction of fat-soluble components; wherein, CO 2 The supercritical extraction conditions are that the extraction pressure is 25MPa, the extraction temperature is 30 ℃, and the extraction time is 2 hours, so as to obtain the ganoderma lucidum spore oil.
(2) The lucid ganoderma sporocarp is prepared according to the following steps of 1:10 weight percent of the ganoderma lucidum extract is added with 75 percent of ethanol by mass concentration, the mixture is heated and extracted for 2 times, each time lasts for 2 hours, the filtering is carried out, the filter liquor is combined, the concentration is carried out until the specific gravity is 1.15 to 1.25, and the spray drying is carried out, thus obtaining the ganoderma lucidum alcohol extract.
(3) Mixing 1 part by weight of ganoderma spore oil and 1 part by weight of ganoderma alcohol extract.
(4) Adding pharmaceutically acceptable adjuvants, and making into soft capsule by conventional soft capsule preparation method.
Example 2
And (3) uniformly mixing 15 parts by weight of ganoderma lucidum spore oil and 1 part by weight of ganoderma lucidum alcohol extract to obtain a mixture.
The preparation method comprises the following steps:
(1) Collecting Ganoderma spore powder with wall-broken rate of above 95% (total triterpene content is not less than 2.5%), adding water 1/4 of wall-broken Ganoderma spore powder weight to obtain strip granule, drying at low temperature until water content is not more than 6%, and adopting CO 2 Supercritical extraction of fat-soluble components; wherein, CO 2 The supercritical extraction conditions are that the extraction pressure is 25MPa, the extraction temperature is 30 ℃, and the extraction time is 2 hours, so as to obtain the ganoderma lucidum spore oil.
(2) The lucid ganoderma sporocarp is prepared according to the following steps of 1:12 weight percent, adding 75 percent ethanol, heating and extracting for 2 times, each time for 2 hours, filtering, combining the filtrates, concentrating to the specific gravity of 1.15-1.25, and spray drying to obtain the ganoderma alcohol extract.
(3) Mixing 15 parts by weight of ganoderma spore oil and 1 part by weight of ganoderma alcohol extract.
Adding pharmaceutically acceptable adjuvants, and making into soft capsule by conventional soft capsule preparation method.
Example 3
And (3) uniformly mixing 3 parts by weight of ganoderma lucidum spore oil and 2 parts by weight of ganoderma lucidum alcohol extract to obtain a mixture.
The preparation method comprises the following steps:
(1) Collecting Ganoderma spore powder with wall-broken rate of above 95% (total triterpene content is not less than 2.5%), adding water 1/4 of the weight of the wall-broken Ganoderma spore powder, making into strip-shaped granule, drying at low temperature until water content is not more than 6%, and adopting CO 2 Supercritical extraction of fat-soluble components; wherein, CO 2 The supercritical extraction conditions are that the extraction pressure is 25MPa, the extraction temperature is 30 ℃, and the extraction time is 2 hours, so as to obtain the ganoderma lucidum spore oil.
(2) The lucid ganoderma sporocarp is prepared according to the following steps of 1:12 weight percent, adding 75 percent ethanol, heating and extracting for 3 times, each time for 2 hours, filtering, combining the filtrates, concentrating until the specific gravity is 1.15-1.25, and spray drying to obtain the ganoderma alcohol extract.
(3) Mixing 3 parts by weight of ganoderma spore oil and 2 parts by weight of ganoderma alcohol extract.
Adding pharmaceutically acceptable adjuvants into the mixture, and making into capsule according to conventional capsule preparation method.
The effects of the present invention are further illustrated by pharmacodynamic experiments as follows: study and description of the efficacy test of the composition of the present invention: the inventor carries out experimental study on the technical scheme provided by the invention to prove the technical effect of the invention, and the following experiment is used for further explaining the technical effect of the invention but not limiting the invention.
Test examples
The ganoderma spore oil compositions used in the following experimental examples were prepared according to the method of example 1.
1. The tested drugs are: the resulting ganoderma spore oil composition, ganoderma spore oil, and alcohol-extracted ganoderma extract (i.e., ganoderma alcohol extract) were prepared as in example 1.
Ganoderma lucidum spore oil: collecting Ganoderma spore powder with wall-broken rate of above 95% (total triterpene content is not less than 2.5%), adding water 1/4 of the weight of the wall-broken Ganoderma spore powder, making into strip-shaped granule, drying at low temperature until water content is not more than 6%, and adopting CO 2 Supercritical extraction of fat-soluble components; wherein, CO 2 The supercritical extraction conditions are that the extraction pressure is 25MPa, the extraction temperature is 30 ℃ and the extraction time is 2 hours.
Alcohol extraction of ganoderma lucidum: the lucid ganoderma sporocarp is prepared according to the following steps of 1:10, adding 75 percent ethanol by weight, heating and extracting for 2 times, each time for 2 hours, filtering, combining the filtrates, concentrating to the specific gravity of 1.15-1.25, and spray drying to obtain the ganoderma lucidum alcohol extract.
The preparation method comprises the following steps: a proper amount of ganoderma spore oil composition is measured, and then diluted into working solutions of 50mg/mL, 25mg/mL and 12.5mg/mL respectively by soybean oil (manufacturer: auxiliary material for Shandong Rui crude drug, co., ltd., batch No. DD20210701, specification: 500 mL), the administration volume is 0.4mL/20g, namely the administration dosage is 1000mg/kg,500mg/kg and 250mg/kg respectively.
A proper amount of ganoderma spore oil is measured, and then diluted into working solutions of 50mg/mL, 25mg/mL and 12.5mg/mL respectively by soybean oil (manufacturer: shandong Rui crude drug adjuvant Co., ltd., lot: DD20210701, specification: 500 mL), wherein the administration volumes are 0.4mL/20g, namely the administration doses are 1000mg/kg,500mg/kg and 250mg/kg respectively.
Accurately weighing 0.065g of sample ethanol-extracted Ganoderma extract, adding 10mL of 0.5% CMC-Na to prepare 6.5mg/mL solution; accurately weighing 0.0325g of sample ethanol-extracted Ganoderma extract, adding 0.5% CMC-Na10mL to prepare 3.25mg/mL solution; accurately weighing 0.0163g of sample ethanol-extracted Ganoderma extract, adding 0.5% CMC-Na10mL to prepare 1.63mg/mL solution; a solution with a concentration of 6.5g/ml,3.25g/ml,1.63g/m was obtained. The administration volume is 0.4mL/20g, i.e., the administration dose is 130mg/kg,65mg/kg,32.5mg/kg.
2. Control drug
Name: 5-FU (5-fluorouracil)
Providing a unit: tianjin Yao pharmaceutical Co Ltd
The preparation method comprises the following steps: one bottle of the injection is 0.25g/10ml, 1ml of physiological saline (manufacturer: jiangsu Huai' an Shuanghe pharmaceutical industry, LLC; batch No. 2103223C; specification: 500ml, 4.5 g) is added to 7.33ml of physiological saline and uniformly shaken to obtain a 3mg/ml solution, and the solution is stored at 4 ℃, the administration volume is 0.2ml/20g, namely the administration dose is 30mg/kg.
3. Test animal
Source, strain: ICR mice, provided by Hangzhou medical college (production) (laboratory animal production license: SCXK (Zhe) 2019-0002).
The week age: 6-8 weeks old
Sex: female with a female
4. Grouping of laboratory animals (see Table 1-Table 2)
TABLE 1
TABLE 2
Animal model | Medicine | Group of | Dosage form | Mode of administration | Number of animals |
H22 | -- | Blank control group | -- | -- | 10 |
H22 | 5-FU | Positive control group | 30mg/kg | i.v.1 times/2 |
10 |
H22 | Ganoderma spore oil composition | High dose group | 130mg/kg | i.g.1 times per 1 |
10 |
H22 | Ganoderma spore oil composition | Middle dose group | 65mg/kg | i.g.1 times per 1 |
10 |
H22 | Ganoderma spore oil composition | Low dose group | 32.5mg/kg | i.g.1 times per 1 |
10 |
H22 | Ganoderma spore oil | High dose group | 1000mg/kg | i.g.1 times per 1 |
10 |
H22 | Ganoderma spore oil | Middle dose group | 500mg/kg | i.g.1 times per 1 |
10 |
H22 | Ganoderma spore oil | Low dose group | 250mg/kg | i.g.1 times per 1 |
10 |
H22 | Alcohol extracted glossy ganoderma extract | High dose group | 1000mg/kg | i.g.1 times per 1 |
10 |
H22 | Alcohol extracted glossy ganoderma extract | Middle dose group | 500mg/kg | i.g.1 times per 1 |
10 |
H22 | Alcohol extracted glossy ganoderma extract | Low dose group | 250mg/kg | i.g.1 times per 1 |
10 |
5. The experimental method comprises the following steps:
ICR mice were inoculated with ascites (ascites was collected under aseptic conditions, diluted with physiological saline to 1. Each group was administered 24 hours after inoculation (d 1) for the first dose, and samples No. 1-5 were administered by gavage 1 time/1 day for a total of 9 doses. The administration volume was 0.4ml/20g. Tumor-bearing mice were sacrificed at day 10 (d 10) after inoculation and the tumor masses were separated and weighed and the data obtained were statistically processed (t-test).
6. Results of the experiment
The experimental results of mouse sarcoma S180 cell transplanted tumor model are shown in Table 3, the Ganoderma spore oil composition is administrated by intragastric administration at 1000mg/kg,500mg/kg and 250mg/kg respectively, the administration is 1 time per day, the administration period is 10 days, the tumor inhibition rate of mouse sarcoma S180 cell transplanted tumor is 59.15%,48.00% and 25.28% respectively; the ganoderma lucidum spore oil is respectively administrated by intragastric administration at the doses of 1000mg/kg,500mg/kg and 250mg/kg, the administration is carried out for 1 time every day, the administration period is 10 days, and the tumor inhibition rates on mouse sarcoma S180 cell transplanted tumors respectively reach 48.26%,35.24% and 21.35%; the ethanol-extracted ganoderma lucidum extract is administrated by gastric lavage at the doses of 130mg/kg,65mg/kg and 32.5mg/kg respectively, the administration is carried out for 1 time every day, the administration period is 10 days, and the tumor inhibition rates on mouse sarcoma S180 cell transplanted tumors respectively reach 53.27%,35.35% and 22.45%; the positive drug 5-FU is administrated by tail vein injection of 30mg/kg, 1 time every 2 days, the administration period is 10 days, and the tumor inhibition rate on mouse sarcoma S180 cell transplantation tumor reaches 71.36%. In the test process, no obvious influence of each dosage group of No. 1-5 samples on the body weight of the experimental animal is found, but the positive drug 5-FU has an extremely obvious inhibiting effect on the body weight of the experimental animal.
* P <0.05 x P <0.01 in comparison to model control group
The experimental results of mouse liver cancer H22 cell transplantation tumor models are shown in Table 4, the ganoderma lucidum spore oil composition is administrated by intragastric administration at the doses of 1000mg/kg,500mg/kg and 250mg/kg respectively, the administration is carried out 1 time every day, the administration period is 10 days, and the tumor inhibition rates on mouse liver cancer H22 cell transplantation tumors respectively reach 57.17%,42.41% and 25.78%; the ganoderma spore oil is administrated by intragastric administration at the doses of 1000mg/kg,500mg/kg and 250mg/kg respectively, the administration is carried out for 1 time every day, the administration period is 10 days, and the tumor inhibition rates on mouse liver cancer H22 cell transplanted tumors respectively reach 43.03%,29.52% and 19.61%; the ethanol-extracted ganoderma lucidum extract is administrated by intragastric administration at doses of 130mg/kg,65mg/kg and 32.5mg/kg respectively, 1 time is taken every day, the administration period is 10 days, and the tumor inhibition rates on mouse liver cancer H22 cell transplanted tumors respectively reach 50.52%,33.47% and 18.85%; the positive drug 5-FU is administrated by tail vein injection of 30mg/kg, 1 time is administrated every 2 days, the administration period is 10 days, and the tumor inhibition rate on mouse liver cancer H22 cell transplantation tumor reaches 73.39%. In the test process, no obvious influence of each dosage group of No. 1-5 samples on the body weight of the experimental animal is found, but the positive drug 5-FU has an extremely obvious inhibiting effect on the body weight of the experimental animal.
* P <0.05 x P <0.01 in comparison to model control group
As can be seen from the data in tables 3 and 4, compared with the single prescription, the pharmaceutical composition provided by the invention has significantly improved tumor inhibition rate on mouse liver cancer H22 cell transplanted tumor and tumor inhibition rate on mouse sarcoma S180 cell transplanted tumor. Has synergistic effect.
Claims (8)
1. A ganoderma lucidum spore oil composition for improving the tumor inhibition rate is characterized by comprising the following components in parts by weight:
1-15 parts of ganoderma lucidum spore oil and 1-15 parts of ganoderma lucidum alcohol extract;
wherein the alcohol extract of ganoderma lucidum is prepared by mixing ganoderma lucidum fruit bodies according to the proportion of 1:10-12 weight parts of the raw materials are added with 75% ethanol, heated and extracted for 2-3 times, each time lasts for 1.5-2 hours, the mixture is filtered, the filtrate is combined, concentrated to the specific gravity of 1.15-1.25, and spray-dried to obtain the traditional Chinese medicine.
2. The composition of claim 1, wherein the ganoderma spore oil is prepared by mixing wall-broken ganoderma spore powder with water, granulating, drying at low temperature to water content of not higher than 6%, and adding CO 2 Supercritical extraction of fat-soluble components; wherein CO 2 The supercritical extraction conditions are that the extraction pressure is 25-28 MPa, the extraction temperature is 25-30 ℃, and the extraction time is 1-2 hours.
3. The ganoderma lucidum spore oil composition with improved tumor inhibition rate according to claim 2, wherein the CO is 2 The supercritical extraction adopts the extraction pressure of 25MPa, the extraction temperature of 30 ℃ and the extraction time of 2 hours.
4. The ganoderma lucidum spore oil composition for improving tumor inhibition rate according to claim 2, wherein the total triterpene content in the wall-broken ganoderma lucidum spore powder is not less than 2.5%.
5. The ganoderma lucidum spore oil composition for improving the tumor inhibition rate according to claim 2, wherein the water amount is 1/5 to 1/4 of the weight of the wall-broken ganoderma lucidum spore powder.
6. The ganoderma lucidum spore oil composition for improving tumor inhibition rate according to claim 1, wherein the ganoderma lucidum sporocarp is prepared from ganoderma lucidum spore oil according to a weight ratio of 1:10 to 75% by weight of ethanol.
7. The ganoderma lucidum spore oil composition for improving the tumor inhibition rate according to claim 1, wherein the ganoderma lucidum spore oil accounts for 1 to 3 parts by weight, and the ganoderma lucidum alcohol extract accounts for 1 to 3 parts by weight.
8. A preparation method of a ganoderma lucidum spore oil composition with high tumor inhibition rate is characterized by comprising the following steps:
(1) Adding water into wall-broken Ganoderma spore powder, making into strip granule, drying at low temperature until water content is not higher than 6%, and adopting CO 2 Supercritical extraction of fat-soluble components; wherein, CO 2 The supercritical extraction conditions comprise that the extraction pressure is 25-28 MPa, the extraction temperature is 25-30 ℃, and the extraction time is 1-2 hours, so as to obtain the ganoderma lucidum spore oil;
(2) The lucid ganoderma sporocarp is prepared according to the following steps of 1:10-12, adding 75% ethanol, heating and extracting for 2-3 times, each time for 1.5-2h, filtering, combining filtrates, concentrating to specific gravity of 1.15-1.25, and spray drying to obtain Ganoderma alcohol extract;
(3) Mixing 1-15 parts by weight of ganoderma lucidum spore oil and 1-15 parts by weight of ganoderma lucidum alcohol extract.
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