Reishi spore oil, method for preparing Reishi spore oil, its use in the preparation of anti-cancer fatigue drugs, and drugs containing Reishi spore oil

Abstract

Ganoderma lucidum spore oil and its use in preparing anti-cancer-related fatigue medicine. The Ganoderma lucidum spore oil has a triglyceride content of more than 90% and no Ganoderma lucidum triterpenoid compounds are detected. The Ganoderma lucidum spore oil is obtained by supercritical carbon dioxide extraction, eluting with a mixture of petroleum ether and ethyl acetate through a silica gel column, concentrating under reduced pressure, and drying in vacuum. The Ganoderma lucidum spore oil is found to significantly improve fatigue caused by cancer and has good safety, and therefore can be applied to preparing anti-cancer-related fatigue medicine.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and specifically relates to the use of Ganoderma lucidum spore oil and its anti-cancer related fatigue drugs in the preparation.

Background Art

[0002] Cancer-related fatigue (CRF) is one of the most common symptoms in tumor patients and the most common side effect in tumor treatment. According to the National Comprehensive Cancer Network (NCCN) in the United States, cancer-related fatigue is defined as "a subjective feeling of pain and persistence that is not consistent with recent activities, related to cancer or cancer treatment, and that hinders daily living functions, including physical, emotional and / or cognitive fatigue or exhaustion". According to the International Classification of Diseases (ICD), the symptoms of cancer-related fatigue are described as non-specific weakness, debility, general asthenia, drowsiness, fatigue, etc. According to statistics, the symptoms of cancer-related fatigue are found in more than 75% of tumor patients. However, currently, there is no clear conclusion regarding the mechanism of cancer-related fatigue, and there are also no therapeutic drugs for cancer-related fatigue.

[0003] In ancient traditional Chinese medicine literature, there is no record of "cancer-related fatigue", and most are expressed as "deficiency-induced fatigue". In "Plain Questions · General Discussion on Deficiency and Excess", it is stated that "if essence and qi are depleted, then there is deficiency", which is considered to be the first definition of deficiency-induced fatigue. In "Synopsis of the Golden Chamber · Treatise on Deficiency-induced Fatigue", it is believed that due to overwork, both the viscera, meridians, yin and yang qi and blood are deficient, and all a series of symptoms of overwork are deficiency-induced fatigue. Through the continuous development of modern traditional Chinese medicine, it is generally considered that most cancer-related fatigue is caused by emotional disorders, invasion of cancer toxins, improper diet, etc., which damage qi, blood, and body fluids, damage the qi of the viscera, and cannot recover for a long time. Therefore, the treatment of cancer-related fatigue is mainly based on the tumor itself, dialectically treated according to the difference in the deficiency and excess of qi, blood, yin and yang of the viscera, with the main treatment principle of strengthening the healthy qi and eliminating pathogenic factors, assisting the healthy qi and removing pathogenic factors, and in addition to eliminating the causes of fatigue as much as possible, overall adjustment should also be emphasized.

[0004] Reishi mushrooms are considered a rare and precious ingredient in traditional Chinese medicine, sometimes called "immortal herb," ​​and are used to treat mental malaise, insomnia, palpitations, lung deficiency, cough asthma, fatigue, shortness of breath, and loss of appetite. Reishi spores are extremely small spores that burst from the cap of the reishi mushroom when it is fully mature. They are the reproductive cells of the reishi mushroom and contain all the gene-active components of the reishi. Modern pharmacological research has shown that reishi spores have antitumor, immunomodulatory, blood glucose-lowering, blood lipid-lowering, anti-inflammatory, antioxidant, and radical scavenging effects. Reishi spore oil is a lipid-soluble active substance obtained by disrupting the cell wall of reishi spores or by supercritical carbon dioxide extraction. Modern research has shown that it has antitumor, immune-enhancing, and liver-protective effects.

[0005] Conventional research generally considers the main active ingredients of Reishi mushrooms to be Reishi triterpenoid compounds, and studies on the efficacy of Reishi have focused on these triterpenoids. Therefore, conventional spore oils typically have a high proportion of Reishi triterpenoid compounds, reaching 10-30%. However, compositional studies and efficacy analyses of other components in spore oils, such as the triglyceride compounds which make up the largest proportion, have not yet been reported. Furthermore, conventional research on the fatigue-relieving effects of Reishi has focused on the recovery of physiological fatigue, with little reported on its effectiveness in treating pathological fatigue, particularly cancer-related fatigue. [Overview of the project] [Problems that the invention aims to solve]

[0006] The present invention aims to provide the use of Reishi spore oil in the preparation of anti-cancer-related fatigue drugs. Based on the above reasons and needs, we have found that Reishi spore oil has a novel efficacy against anti-cancer-related fatigue, and we provide a new use for Reishi spore oil as a drug, namely its use in the preparation of drugs for the treatment or prevention of cancer-related fatigue. [Means for solving the problem]

[0007] To solve the above technical problems, the present invention employs the following technical solution.

[0008] Reishi spore oil shows no detectable levels of Reishi triterpenoid compounds.

[0009] Preferably, the triglyceride content in the reishi spore oil is greater than 90%.

[0010] The Reishi spore oil according to the present invention has a triglyceride content greater than 90%, an ergosterol content of 0% to 0.5%, and no detectable Reishi triterpenoid compounds. Currently, most studies attribute the immune-enhancing and antitumor effects of Reishi spore oil to the abundance of Reishi triterpenoid compounds, and therefore, compositional studies of Reishi spore oil focus on the abundance of Reishi triterpenoid compounds. According to previous research, it is assumed that as the content of triterpenoid compounds decreases, the various effects of the spore oil also weaken or disappear.

[0011] However, the present invention has unexpectedly discovered through research that increasing the triglyceride content of Reishi spore oil to more than 90% has a superior anti-cancer-related fatigue effect, particularly against pathological fatigue, especially cancer-related fatigue. In particular, it provides a superior anti-cancer-related fatigue effect compared to spore oil containing Reishi triterpenoid compounds, significantly improving the alleviation of mental and physical fatigue caused by cancer-related fatigue to an extent previously unimaginable.

[0012] The method for preparing the Reishi spore oil includes the steps of: A) disrupting the cell walls of Reishi spores, granulating and drying them, and performing supercritical carbon dioxide extraction to obtain crude spore oil; B) taking an appropriate amount of silica gel, dissolving it in a mixed solution of petroleum ether and ethyl acetate, packing it into a column by a wet method to obtain an elution column; and C) introducing the crude spore oil into the elution column, eluting it with the mixed solution of petroleum ether and ethyl acetate at atmospheric pressure, collecting and combining the eluates, concentrating them under reduced pressure to remove the solvent, and then vacuum drying to obtain the Reishi spore oil.

[0013] Measurements revealed that this Reishi spore oil has a triglyceride content greater than 90%, an ergosterol content of 0% to 0.5%, and no triterpenoids detected. This process yields spore oil with very little triterpenoid compound content, while also concentrating triterpenoid compounds, and is expected to lead to the creation of other novel products with high triterpenoid content.

[0014] Preferably, the mass ratio of the crude spore oil to the silica gel is 1:(10~40).

[0015] Preferably, in the petroleum ether-ethyl acetate mixed solution, the volume ratio of petroleum ether to ethyl acetate is (8.5-9.5):(0.5-1.5).

[0016] Preferably, in the petroleum ether-ethyl acetate mixed solution, the volume ratio of petroleum ether to ethyl acetate is 9.2:0.8.

[0017] Preferably, the ratio of the column inner diameter to the packing height of the elution column is 1:(10~20).

[0018] The aforementioned reishi mushroom spore oil is used in the preparation of an anti-cancer-related fatigue drug.

[0019] The drug containing the aforementioned reishi mushroom spore oil further contains a pharmaceutically acceptable carrier.

[0020] Preferably, the drug is one of the following: tablets, capsules, lozenges, granules, powders, pills, powders, ointments, elixirs, suspensions, solutions, injections, suppositories, creams, sprays, intravenous infusions, or patches. [Effects of the Invention]

[0021] Compared to the prior art, the present invention has the following beneficial effects.

[0022] As a result of experiments and research on Ganoderma lucidum components other than triterpenoid compounds, it was found that spore oil with a triglyceride content greater than 90% and an ergosterol content of 0% to 0.5% has a significant anti-fatigue effect on tumor-bearing mice. Compared with spore oil containing Ganoderma lucidum triterpenoid compounds, the Ganoderma lucidum spore oil according to the present invention unexpectedly found that it significantly improves the alleviation of mental fatigue and physical fatigue caused by cancer-related fatigue, and is expected to significantly improve the quality of life of cancer patients. In addition, when the Ganoderma lucidum spore oil according to the present invention is combined with a chemotherapeutic agent group, no animal deaths or significant weight loss are shown, and it does not affect the food intake of tumor-bearing mice, indicating that the safety of Ganoderma lucidum spore oil for animals is good. The Ganoderma lucidum spore oil according to the present invention is applicable to the preparation of anti-cancer-related fatigue drugs. In addition, by concentrating the isolated triterpenoid compounds, the development of other new products with a high triterpenoid content is also expected.

Brief Description of the Drawings

[0023] [Figure 1] It is a graph showing the change over time of the average body weight of a human gastric cancer cell transplanted mouse model in Effect Example 4. [Figure 2] It is a graph showing the change over time of the average daily intake of a human gastric cancer cell transplanted mouse model in Effect Example 4.

Modes for Carrying Out the Invention

[0024] In order to explain the object, technical solution and advantages of the present invention in more detail, the present invention will be further described below with reference to the drawings and specific examples.

[0025] Example 1 Reishi spore oil was obtained by disrupting the cell walls of Reishi spores, granulating and drying them, performing supercritical carbon dioxide extraction, and adsorption separation by silica gel column chromatography. The separation process was as follows: An appropriate amount of silica gel was taken, dissolved in a solution of petroleum ether:ethyl acetate = 9.2:0.8 (volume ratio), and packed into a column by a wet method, with a column inner diameter to packing height ratio of 1:10. Spore oil was introduced so that the mass ratio of spore oil to silica gel was 1:28, and eluted at atmospheric pressure with a solution of petroleum ether:ethyl acetate = 9.2:0.8 (volume ratio). The eluates were collected and combined, concentrated under reduced pressure to remove the solvent, and then vacuum dried to obtain a spore oil sample.

[0026] This Reishi spore oil contains more than 90% triglycerides, 0% to 0.5% ergosterol, and no Reishi triterpenoid compounds were detected.

[0027] Example 2 Reishi mushroom spores were subjected to cell wall disruption, granulation, drying, supercritical carbon dioxide extraction, and adsorption separation by silica gel column chromatography to obtain Reishi spore oil. The separation process was as follows: An appropriate amount of silica gel was taken, dissolved in a solution of petroleum ether:ethyl acetate = 9.2:0.8 (volume ratio), and packed into a column by a wet method, with a column inner diameter to packing height ratio of 1:13. Spore oil was introduced so that the mass ratio of spore oil to silica gel was 1:28, and eluted at atmospheric pressure with a solution of petroleum ether:ethyl acetate = 9.2:0.8 (volume ratio). The eluates were collected and combined, concentrated under reduced pressure to remove the solvent, and then vacuum dried to obtain a spore oil sample.

[0028] This Reishi spore oil contains more than 90% triglycerides, 0% to 0.5% ergosterol, and no Reishi triterpenoid compounds were detected.

[0029] Example 3 Reishi mushroom spores were subjected to cell wall disruption, granulation, drying, supercritical carbon dioxide extraction, and adsorption separation by silica gel column chromatography to obtain Reishi spore oil. The separation process was as follows: An appropriate amount of silica gel was taken, dissolved in a solution of petroleum ether:ethyl acetate = 9.2:0.8 (volume ratio), and packed into a column by a wet method, with a column inner diameter to packing height ratio of 1:15. Spore oil was introduced so that the mass ratio of spore oil to silica gel was 1:28, and eluted at atmospheric pressure with a solution of petroleum ether:ethyl acetate = 9.2:0.8 (volume ratio). The eluates were collected and combined, concentrated under reduced pressure to remove the solvent, and then vacuum dried to obtain a spore oil sample.

[0030] This Reishi spore oil contains more than 90% triglycerides, 0% to 0.5% ergosterol, and no Reishi triterpenoid compounds were detected.

[0031] Example 4 Reishi mushroom spores were subjected to cell wall disruption, granulation, drying, supercritical carbon dioxide extraction, and adsorption separation by silica gel column chromatography to obtain Reishi spore oil. The separation process was as follows: An appropriate amount of silica gel was taken, dissolved in a solution of petroleum ether:ethyl acetate = 9.2:0.8 (volume ratio), and packed into a column by a wet method, with a column inner diameter to packing height ratio of 1:15. Spore oil was introduced so that the mass ratio of spore oil to silica gel was 1:30, and eluted at atmospheric pressure with a solution of petroleum ether:ethyl acetate = 9.2:0.8 (volume ratio). The eluates were collected and combined, concentrated under reduced pressure to remove the solvent, and then vacuum dried to obtain a spore oil sample.

[0032] This Reishi spore oil contains more than 90% triglycerides, 0% to 0.5% ergosterol, and no Reishi triterpenoid compounds were detected.

[0033] Example 5 Reishi mushroom spores were subjected to cell wall disruption, granulation, drying, supercritical carbon dioxide extraction, and adsorption separation by silica gel column chromatography to obtain Reishi spore oil. The separation process was as follows: An appropriate amount of silica gel was taken, dissolved in a solution of petroleum ether:ethyl acetate = 9.3:0.7 (volume ratio), and packed into a column by a wet method, with a column inner diameter to packing height ratio of 1:15. Spore oil was introduced so that the mass ratio of spore oil to silica gel was 1:35, and eluted at atmospheric pressure with a solution of petroleum ether:ethyl acetate = 9.3:0.7 (volume ratio). The eluates were collected and combined, concentrated under reduced pressure to remove the solvent, and then vacuum dried to obtain a spore oil sample.

[0034] This Reishi spore oil contains more than 90% triglycerides, 0% to 0.5% ergosterol, and no Reishi triterpenoid compounds were detected.

[0035] Example 6 Reishi mushroom spores were subjected to cell wall disruption, granulation, drying, supercritical carbon dioxide extraction, and adsorption separation by silica gel column chromatography to obtain Reishi spore oil. The separation process was as follows: An appropriate amount of silica gel was taken, dissolved in a solution of petroleum ether:ethyl acetate = 9.2:0.8 (volume ratio), and packed into a column by a wet method, with a column inner diameter to packing height ratio of 1:15. Spore oil was introduced so that the mass ratio of spore oil to silica gel was 1:33, and eluted at atmospheric pressure with a solution of petroleum ether:ethyl acetate = 9.2:0.8 (volume ratio). The eluates were collected and combined, concentrated under reduced pressure to remove the solvent, and then vacuum dried to obtain a spore oil sample.

[0036] This Reishi spore oil contains more than 90% triglycerides, 0% to 0.5% ergosterol, and no Reishi triterpenoid compounds were detected.

[0037] Example 7 Reishi spore oil was obtained by disrupting the cell walls of Reishi spores, granulating and drying them, performing supercritical carbon dioxide extraction, and adsorption separation by silica gel column chromatography. The separation process was as follows: An appropriate amount of silica gel was taken, dissolved in a solution of petroleum ether:ethyl acetate = 9.0:1.0 (volume ratio), and packed into a column by a wet method, with a column inner diameter to packing height ratio of 1:17. Spore oil was introduced so that the mass ratio of spore oil to silica gel was 1:35, and eluted at atmospheric pressure with a solution of petroleum ether:ethyl acetate = 9.0:1.0 (volume ratio). The eluates were collected and combined, concentrated under reduced pressure to remove the solvent, and then vacuum dried to obtain a spore oil sample.

[0038] This Reishi spore oil contains more than 90% triglycerides, 0% to 0.5% ergosterol, and no Reishi triterpenoid compounds were detected.

[0039] Example 8 Reishi mushroom spores were subjected to cell wall disruption, granulation, drying, supercritical carbon dioxide extraction, and adsorption separation by silica gel column chromatography to obtain Reishi spore oil. The separation process was as follows: An appropriate amount of silica gel was taken, dissolved in a solution of petroleum ether:ethyl acetate = 9.5:0.5 (volume ratio), and packed into a column by a wet method, with a column inner diameter to packing height ratio of 1:10. Spore oil was introduced so that the mass ratio of spore oil to silica gel was 1:10, and eluted at atmospheric pressure with a solution of petroleum ether:ethyl acetate = 9.5:0.5 (volume ratio). The eluates were collected and combined, concentrated under reduced pressure to remove the solvent, and then vacuum dried to obtain a spore oil sample.

[0040] This Reishi spore oil contains more than 90% triglycerides, 0% to 0.5% ergosterol, and no Reishi triterpenoid compounds were detected.

[0041] Example 9 Reishi mushroom spores were subjected to cell wall disruption, granulation, drying, supercritical carbon dioxide extraction, and adsorption separation by silica gel column chromatography to obtain Reishi spore oil. The separation process was as follows: An appropriate amount of silica gel was taken, dissolved in a solution of petroleum ether:ethyl acetate = 8.5:1.5 (volume ratio), and packed into a column by a wet method, with a column inner diameter to packing height ratio of 1:20. Spore oil was introduced so that the mass ratio of spore oil to silica gel was 1:40, and eluted at atmospheric pressure with a solution of petroleum ether:ethyl acetate = 8.5:1.5 (volume ratio). The eluates were collected and combined, concentrated under reduced pressure to remove the solvent, and then vacuum dried to obtain a spore oil sample.

[0042] This Reishi spore oil contains more than 90% triglycerides, 0% to 0.5% ergosterol, and no Reishi triterpenoid compounds were detected.

[0043] Example of effect 1 Anti-fatigue effect of Reishi spore oil in a mouse model of human lung cancer cells. 1. Laboratory animals We prepared a total of 32 SPF-grade nude mice, half male and half female, and raised them in the nude mouse room of the SPF animal experiment house under conditions of room temperature 16-26°C, relative humidity 40-70%, and a 12-hour light / 12-hour dark cycle, allowing them to feed freely.

[0044] 2. Experimental materials Test substance 1 (commercially available Reishi spore oil containing Reishi triterpenoid components) and test substance 2 (Reishi spore oil according to the present invention, with a triglyceride content greater than 90%, an ergosterol content of 0% to 0.5%, and no detectable Reishi triterpenoid compounds) were prepared with corn oil to the corresponding concentrations before administration and immediately used. Cell line: Human lung cancer cell line NCI-H460.

[0045] 3. Experimental Method (1) Creation of a nude mouse model with a tumor NCI-H460 cells were cultured to the required amount, digested and counted, and then inoculated at a rate of 0.15 ml / mice near the right upper limb of nude mice to create a model (the blank control group was not inoculated). The tumor size was 100 mm. 3 After reaching a certain stage, nude mice were randomly divided into four groups of eight mice each: a blank control group (Control), a tumor model group (Model), a test substance 1 group (Sample 1), and a test substance 2 group (Sample 2), with half being male and half female.

[0046] (2) Method of administration Route of administration: Test substance 1, test substance 2, and corn oil were administered orally. Weekly administration frequency: Test substance 1, test substance 2, and corn oil were administered once daily. Administration cycle: Test substance 1, test substance 2, and corn oil were administered consecutively for 4 weeks. Dosage: 10 mL / kg of test substance. Dosage: 0.1 mL / 10 g body weight, i.e., 10 mL / kg.

[0047] (3) Tail suspension test On day 28 after administration, a tail suspension test was performed on tumor-bearing mice. The mouse's tail (2 cm from the tip) was secured to a test device with tape, and the mouse was suspended. The mouse struggled to overcome the abnormal posture, and the state in which it completely stopped struggling and became still was defined as "immobile," and the immobile time of the mouse over a 6-minute period was recorded.

[0048] 4. Experimental Results Table 3 shows the results. On day 28 after administration, tumor-bearing mice in the tumor model group had a significantly higher mean immobility time than the blank control group, and there was a significant difference between the two groups (P<0.05). Tumor-bearing mice in the test substance 1 group and the test substance 2 group had a significantly lower mean immobility time than the tumor model group, and there were significant differences between both groups and the tumor model group in all cases (P<0.05). Tumor-bearing mice in the test substance 2 group had a significantly lower mean immobility time than the test substance 1 group, and there was a significant difference between the two groups (P<0.05).

[0049] [Table 3]

[0050] In the tail suspension test, mice are forced into an abnormal posture and struggle as much as possible, both mentally and physically, to overcome it. Therefore, if the mouse does not experience fatigue throughout the test, the theoretical immobility time is 0. Conversely, if the mouse experiences mental or physical fatigue while struggling, it stops struggling and rests, and this rest period is recorded. Consequently, the longer the average immobility time, the greater the mental and physical fatigue the mouse experiences during that period.

[0051] The results of the tail suspension test showed that administration of test substance 1 and test substance 2 significantly reduced the immobility time of tumor-bearing mice, and test substance 2 further significantly reduced the immobility time of tumor-bearing mice compared to test substance 1, ultimately reducing it to the level of tumor-free mice (blank control group). This indicates that the triterpenoid-free Reishi spore oil according to the present invention has a significant anti-fatigue effect on tumor-bearing mice compared to spore oil containing Reishi triterpenoid compounds, ultimately reaching the level of a cancer-free state, thus significantly improving the alleviation of mental and physical fatigue caused by cancer-related fatigue.

[0052] As a result, further drug combination efficacy and safety experiments were conducted using only the reishi spore oil that does not contain triterpenoids according to the present invention.

[0053] Example of effect 2 Anti-fatigue effect of Reishi spore oil in a mouse model of liver cancer cells transplanted. 1. Laboratory animals Forty female SPF-grade Kunming mice were prepared and raised in the mouse room of the SPF animal experiment house under conditions of room temperature 16-26°C, relative humidity 40-70%, and a 12-hour light / 12-hour dark cycle, with free access to food.

[0054] 2. Experimental materials The test substance (Reishi mushroom spore oil according to the present invention) was prepared with corn oil to the appropriate concentration before administration and used immediately. The chemotherapeutic agent (cyclophosphamide, batch number: 8D231A, 200 mg / vial, manufactured by Baxter Oncology GmbH) was prepared with physiological saline to the appropriate concentration before administration and used immediately. Cell line: Mouse liver cancer cell line H22.

[0055] 3. Experimental Method (1) Creation of a mouse model with tumors H 22 Cells were cultured to the required volume, digested and counted, and then inoculated at a rate of 0.1 ml / mouse near the right upper limb to create a model, with a tumor size of 100 mm. 3 After reaching a certain level, the mice were randomly divided into four groups of 10: a model control group (Control), a cyclophosphamide group (CTX), a ginger spore oil group (GLSO), and a cyclophosphamide + ginger spore oil group (CTX+GLSO).

[0056] (2) Method of administration Route of administration: Reishi mushroom spore oil was administered orally, followed by tail vein administration of cyclophosphamide. Weekly administration frequency: Administered once daily for 5 consecutive days, followed by a 2-day rest period. Administration cycle: Administered continuously for 3 weeks. Dosage: Reishi mushroom spore oil 10 mL / kg, cyclophosphamide 30 mg / kg. Dosage: 0.1 mL / 10 g body weight, i.e., 10 mL / kg.

[0057] (3) Limit swimming test On days 13 and 20 after administration, limit swimming tests were performed on tumor-bearing mice. The mice were placed in a water box at a depth of 30 cm or more and a water temperature of 25 ± 1°C and allowed to swim. The time from when the tumor-bearing mice began swimming until they became exhausted was recorded. Throughout the experiment, the limbs of each tumor-bearing mouse were kept moving. If a mouse floated on the surface without moving its limbs, the water near the surface was stirred with a wooden stick. Exhaustion was defined as the tumor-bearing mouse remaining submerged for 10 seconds or more and failing to surface on its own. If the average swimming time exceeded 360 seconds, the statistics were calculated using 360 seconds.

[0058] 4. Experimental Results Table 1 shows the results. On days 13 and 20 after administration, tumor-bearing mice administered only with Reishi spore oil had a significantly higher average swimming time than the model group, and there was a significant difference between the two groups (P<0.05). On days 13 and 20 after administration, tumor-bearing mice administered both cyclophosphamide and Reishi spore oil had a significantly higher average swimming time than the model group, and there was a significant difference between the two groups (P<0.05).

[0059] [Table 1]

[0060] As a result, Reishi spore oil significantly extended the critical swimming time of tumor-bearing mice. This indicates that the Reishi spore oil according to the present invention has a significant anti-fatigue effect on liver cancer-bearing mice.

[0061] Example of effect 3 Anti-fatigue effect of Reishi spore oil in a mouse model transplanted with human gastric cancer cells. 1. Laboratory animals Forty female SPF-grade nude mice were prepared and raised in the nude mouse room of the SPF animal experiment house under conditions of room temperature 16-26°C, relative humidity 40-70%, and a 12-hour light / 12-hour dark cycle, with free access to food.

[0062] 2. Experimental materials The test substance (Reishi mushroom spore oil according to the present invention) was prepared with corn oil to the appropriate concentration before administration and used immediately. The chemotherapeutic agent (oxaliplatin, batch number: 190911AM, 50 mg / vial, manufactured by Jiangsu Hengrui Pharmaceutical Co., Ltd.) was prepared with physiological saline to the appropriate concentration before administration and used immediately. Cell line: Human gastric cancer cell line MNK45.

[0063] 3. Experimental Method (1) Creation of a nude mouse model with a tumor MNK45 cells were cultured to the required amount, digested and counted, and then inoculated at a rate of 0.1 ml / mice near the right upper limb of nude mice to create a model, with a tumor size of 100 mm. 3 After reaching a certain stage, nude mice were randomly divided into four groups of 10 mice each: a model control group (Control), a chemotherapy agent group (Oxaliplatin), a chemotherapy agent + test substance group (Oxaliplatin + Sample), and a chemotherapy agent + corn oil group (Oxaliplatin + Oil).

[0064] (2) Method of administration Route of administration: The test substance and corn oil were administered orally, and oxaliplatin was administered via tail vein. Weekly administration frequency: The test substance and corn oil were administered once daily for 5 consecutive days, followed by a 2-day rest period. Oxaliplatin was administered twice a week, on days 1 and 4, respectively. Administration cycle: Chemotherapy agents were administered for 6 weeks, followed by administration of the test substance and corn oil for 6 weeks, and then continued until the end of the study. Dosage: 10 mL / kg of test substance, 2 mg / kg of oxaliplatin. Dosage: 0.1 mL / 10 g body weight, i.e., 10 mL / kg.

[0065] (3) Limit swimming test On days 37 and 49 after administration, limit swimming tests were performed on tumor-bearing mice. The mice were placed in a water box at a depth of 30 cm or more and a water temperature of 25 ± 1°C and allowed to swim. The time from when the tumor-bearing mice began swimming until they became exhausted was recorded. Throughout the experiment, the limbs of each tumor-bearing mouse were kept moving. If a mouse floated on the surface without moving its limbs, the surface was stirred with a wooden stick. Exhaustion was defined as the tumor-bearing mouse remaining submerged for 10 seconds or more and failing to surface on its own. If the average swimming time exceeded 360 seconds, the statistics were calculated using 360 seconds.

[0066] 4. Experimental Results Table 2 shows the results. On day 37 after administration, tumor-bearing mice that received a combination of the chemotherapy agents oxaliplatin and reishi mushroom spore oil had a significantly higher average swimming time than the model group, and there was a statistically significant difference between the two groups (P<0.05). One week after discontinuing the chemotherapy agents, i.e., on day 49 after administration, tumor-bearing mice had a significantly higher average swimming time than the model group, and there was a statistically significant difference (P<0.05).

[0067] [Table 2]

[0068] As a result, administration of the test substance significantly extended the critical swimming time of tumor-bearing mice. This indicates that the Reishi spore oil according to the present invention has a significant anti-fatigue effect on tumor-bearing mice.

[0069] Example of effect 4 Study on the in vivo safety of Reishi mushroom spore oil 1. Reishi spore oil does not affect the body weight of tumor-bearing mice. Figure 1 shows the results. No significant weight loss was observed in tumor-bearing mice after co-administration of Reishi spore oil and oxaliplatin.

[0070] 2. Reishi spore oil does not affect the food intake of tumor-bearing mice. Figure 2 shows the results. No significant decrease in food intake was observed in tumor-bearing mice after co-administration of Reishi spore oil and oxaliplatin.

[0071] Based on the above, the Reishi spore oil according to the present invention has a significant anti-fatigue effect on tumor-bearing mice. Furthermore, animals treated with the Reishi spore oil according to the present invention in combination with the group of chemotherapy agents did not show death or significant weight loss, and there was no effect on the food intake of tumor-bearing mice, demonstrating that the Reishi spore oil is safe for animals. The Reishi spore oil according to the present invention is applicable to the preparation of anti-cancer-related fatigue drugs.

[0072] The above disclosures are merely preferred embodiments of the present invention and do not limit the scope of the claims of the present invention. Equivalent substitutions to the claims of the present invention fall within the scope of the present invention.

Claims

1. A. Disrupting the cell walls of Reishi spores, granulating and drying them, and performing supercritical carbon dioxide extraction to obtain crude spore oil, B. Take an appropriate amount of silica gel, dissolve it in a mixed solution of petroleum ether and ethyl acetate, pack it into a column using a wet method, and obtain an elution column. The process includes the steps of: C, introducing the crude spore oil into the elution column, eluting it with the petroleum ether-ethyl acetate mixture at atmospheric pressure, collecting and combining the eluates, concentrating them under reduced pressure to remove the solvent, and then vacuum drying to obtain the reishi spore oil. The mass ratio of the crude spore oil to the silica gel is 1:10 to 40. In the aforementioned mixed solution of petroleum ether and ethyl acetate, the volume ratio of petroleum ether to ethyl acetate is 8.5 to 9.5:0.5 to 1.

5. The elution column is prepared by a preparation method in which the ratio of the column inner diameter to the packing height is 1:10 to 20. Reishi spore oil does not contain Reishi triterpenoid compounds. Reishi spore oil characterized by the following features.

2. The reishi spore oil according to claim 1, characterized in that the content of triglyceride-based components is greater than 90%.

3. A method for preparing Reishi spore oil, A. The steps involve disrupting the cell walls of Reishi spores, granulating and drying them, and then performing supercritical carbon dioxide extraction to obtain crude spore oil. B. Take an appropriate amount of silica gel, dissolve it in a mixed solution of petroleum ether and ethyl acetate, pack it into a column using a wet method, and obtain an elution column. The process includes the steps of: C, introducing the crude spore oil into the elution column, eluting it with the petroleum ether-ethyl acetate mixture at atmospheric pressure, collecting and combining the eluates, concentrating them under reduced pressure to remove the solvent, and then vacuum drying to obtain the reishi spore oil. The mass ratio of the crude spore oil to the silica gel is 1:10 to 40. In the aforementioned mixed solution of petroleum ether and ethyl acetate, the volume ratio of petroleum ether to ethyl acetate is 8.5 to 9.5:0.5 to 1.

5. The ratio of the column inner diameter to the packing height of the elution column is 1:10 to 20. A method for preparing Reishi mushroom spore oil, characterized by the following:

4. In the aforementioned mixed solution of petroleum ether and ethyl acetate, the volume ratio of petroleum ether to ethyl acetate is 9.2:0.

8. The method for preparing Reishi spore oil according to feature 3.

5. Use of Reishi spore oil according to claim 1 in the preparation of an anti-cancer-related fatigue agent.

6. A drug containing Reishi spore oil as described in claim 1, Further contains a pharmaceutically acceptable carrier. A drug characterized by the following features.

7. It is one of the following: tablets, capsules, lozenges, granules, powders, pills, powders, ointments, elixirs, suspensions, solutions, injections, suppositories, creams, sprays, intravenous drips, or patches. A drug containing Reishi spore oil as described in feature 1.

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