TWI484968B - Composites of phellinus linteus combine bleomycin with anti cancer and reduce pulmonary fibrosis effect - Google Patents

Description

Composite composition of mulberry yellow combined with anticancer drug for treating lung cancer and reducing pulmonary fibrosis

The invention relates to a composite composition of mulberry yellow combined with anticancer drug which has the advantages of adjuvant treatment of lung cancer and lowering pulmonary fibrosis, and is a composite composition combining Chinese and western medicine, which can be used for treating cancer and reducing pneumonia. , side effects of pulmonary fibrosis.

Cancer is now the leading cause of death in humans, and drug treatment in the face of cancer is also accompanied by side effects.

Bleomycin (BLM) is an approved chemotherapeutic drug on the market. It is usually used in combination with other anticancer drugs. It is commonly used in the treatment of testicular cancer, lymphoma and various squamous cell carcinomas such as lung cancer and head and neck cancer. , cervical cancer, penile cancer, skin cancer, female vulvar cancer, Bleomycin (BLM) will cause side effects of pulmonary fibrosis after long-term use, which is due to Bleomycin hydrolase (BH), BH is a hydrolase, and BLM The resistance of the drug has a great relationship, but the content of BH in the lungs or skin is quite small, which means that the two parts lack the resistance to BLM, thus making BLM cause lungs. Direct effects, in addition, BLM also affects the production of free radicals in the lungs, which also affect the normal cells of the lungs, causing apoptosis and destroying the stability of endothelial cells. Should cause pulmonary fibrosis.

In view of the above-mentioned shortcomings, the inventor believes that it has the urgent need to make corrections, and that it has been engaged in the design and manufacture of related products for many years, and has always adhered to the excellent design concept, and researched and created the above disadvantages. After continuous efforts, the present invention is a composite composition of the mulberry yellow combined with an anticancer drug which has the advantages of adjuvant treatment of lung cancer and reduction of pulmonary fibrosis, and improves the efficacy of the product by correcting the excellent product structure.

The main object of the present invention is to provide a composite composition which synergizes with the general anti-cancer effect of a general drug and on the other hand can alleviate the side effects.

In order to achieve the purpose of the foregoing disclosure, the present invention has a composite composition of mulberry yellow combined with an anticancer drug for assisting in the treatment of lung cancer and reducing pulmonary fibrosis, and the system is directed to the anticancer drug Bleomycin (BLM) in the market. The side effects such as pneumonia and pulmonary fibrosis are improved. The system combines mulberry extract and BLM drugs to use the Chinese herbal medicine in combination with Western medicine. On the one hand, it can cooperate with BLM drugs for anticancer effect. Yellow to alleviate the side effects of pneumonia and pulmonary fibrosis caused by BLM drugs.

1‧‧‧ mulberry extract

11‧‧‧ Solid fermentation of mulberry mycelium

12‧‧‧ alcohol dissolution extraction

13‧‧‧Drying

14‧‧‧Removing alcohol

2‧‧‧BLM drugs

3‧‧‧Composite composition

First figure: block diagram of the composite composition of the present invention

Second: Schematic diagram of the extraction process of the mulberry extract of the present invention

Figure 3: Data of the results of the evaluation of the antioxidant capacity of the solid-state fermented mulberry mycelium alcohol extract and the solid-state fermented mulberry mycelium water extract of the present invention (TEAC)

Figure 4: Solid-state fermented mulberry mycelium alcohol extract and solid fermented mulberry yellow Data of the results of the DPPH-capacity test for the mycelial water extract

Figure 5: Data of the results of cytotoxicity test of the composite composition of the present invention and other anticancer drugs Cisplatin, Etoposide and Doxorubicin

Figure 6: Data of the results of the cytotoxicity test of the anticancer drug Cisplatin combined with the composite composition of the present invention and the anticancer drug Etoposide

Figure 7: Sliced map of pulmonary fibrosis animal model

The present invention relates to a composite composition of mulberry yellow combined with an anticancer drug which has the advantages of adjuvant treatment of lung cancer and reduction of pulmonary fibrosis, [see the first figure]. The composite composition (3) mainly comprises mulberry yellow extract ( 1) and BLM drugs (2); wherein: mulberry extract (1) is an extract obtained by solid-state fermentation of mulberry mycelium for alcohol extraction; BLM drug (2) is Bleomycin (BLM), An antibiotic complex is an approved chemotherapy drug on the market; by the way, [see the first figure] the mulberry extract (1) and the BLM drug (2) are 5:1 (mulberry extract: BLM drug) The ratio of the mixture can be combined with the composite composition (3) of the invention.

The invention combines the composite composition of mulberry yellow combined with anticancer drug for assisting the treatment of lung cancer and reducing the pulmonary fibrosis effect, and the detailed process description is as follows (please refer to the first figure and the second figure together) Yellow extract (1) is a solid-state fermented mulberry mycelium alcohol extract. The extraction step is mainly from solid fermentation of mulberry mycelium (11), alcohol dissolution extraction (12), drying (13) Combined with the removal of alcohol (14), the first step is the solid fermentation of mulberry mycelium (11), which is Y GS (0.5% yeast extract, 1% glucose, 1% sucrose, 0.1% KH2PO4, 0.5% MgSO4.H2O, Agar 1.2%) is the basic liquid medium, and then the ampicana culture medium full of mulberry mycelium is integrated into 100ml. Disintegrate in sterile water, then add 5ml of broken bacteria liquid into 95ml liquid medium, place 30 °C constant temperature shaking incubator, take 1 shake flask every 3 days, pump air filtration, leave mycelium drying, scale Heavy, and fermentation, after scraping the mulberry strain on the acacia medium (plate), put it into the sterilized PDB (Potato Dextrose Broth) culture solution, using a conical flask with a baffle at a speed of 150 rpm, temperature 50 °C for 5 days of shake flask culture to activate the strain. This formula is 200g wheat as solid medium, with 0.8:1 water and wheat matrix ratio, and adding 1% calcium carbonate or calcium sulfate as buffer and regulating PH Value: 10 ml of the activated bacterial liquid was added to the solid medium of wheat or soybean for solid-state fermentation for 1 to 2 weeks, and the fermentation product was dried by hot air at 60 ° C for 8 hours, and the dried product was pulverized and the fermented mycelium product was collected. Drying to form the mycelium of the mulberry yellow, and then the hyphae The alcohol is dissolved and extracted (12), and the alcohol is dissolved and extracted (12). The mulberry mycelium is first taken out 5 g, and 40 ml of 75% alcohol is added for extraction for 48 hours, and then the supernatant is centrifuged, and the obtained solution is An aqueous extract of mulberry mycelium, followed by drying in an oven at 60 ° C (13) for 8 hours to remove alcohol (14) to complete the extract of mulberry (1), and the extract of mulberry (1) can be 5 The ratio of 1:1 (mulberry extract: BLM drug) can be mixed with the BLM drug (2) to complete the preparation of the composite composition (3), wherein the mulberry extract (1) is 200 mg/kg, and 40 mg/kg. The BLM drug (2) is combined into a composite composition (3) as the best.

The present invention has a composite composition of mulberry yellow combined with an anticancer drug for assisting in the treatment of lung cancer and reducing pulmonary fibrosis, and to ensure that the mulberry yellow extract (1) can effectively inhibit free radicals to inhibit pulmonary fibrosis, [Please refer to the second picture] used in this case Erythrocyte extract (1) as a test for assessing antioxidant capacity-TEAC (Trolox Equivalent Ant-ioxidant Capacity, TEAC), Trolox equivalent antioxidant capacity (TEAC) analysis: using Trolox as an antioxidant standard, various tests The product will be compared with Trolox, and this test is to look at the antioxidant capacity of the pure water extract of mulberry mycelium and the 75% alcohol extract of mulberry mycelium (the mulberry extract used in this case). Please refer to the third figure] The data shows that the pure water extract of Phellinus chinensis and the 75% alcohol extract of M. sinensis have the ability to inhibit free radicals, especially the free extract of 75% alcohol extract of Phellinus sinensis. The ability of the base has reached 100% at 50 mg/ml, that is, [see the fourth figure]. This is a DPPH-capacity test. Common antioxidants provide hydrogen to remove lipid peroxide free radicals. To achieve the inhibition of the oxidation chain reaction, DPPH is usually used to evaluate the hydrogen supply capacity of antioxidants in the study of antioxidants. Therefore, this experiment also uses the ability to remove DPPH to see the pure water extract of mulberry mycelium and mulberry yellow. Mycelium 75% alcohol extract (ie this The antioxidant capacity of the mulberry extract used in the case, the data shows that the pure water extract of mulberry mycelium and the 75% alcohol extract of mulberry mycelium have the ability to scavenge DPPH, especially the mycelium of mulberry The ability of 75% alcohol extract to remove DPPH has reached 50% inhibition at 500 mg/ml. Therefore, it is known that the 75% alcohol extract of M. sinensis is more effective.

The cytotoxicity test is carried out by administering a combination of the anticancer drugs Cisplatin, Etoposide and Doxorubicin, and the mulberry yellow extract compound of the present invention and the western medicine BLM drug to the mouse lung cancer cell LL2 and the human lung cancer cell A549 for 72 hours, respectively. Observing the cytotoxicity test, [see the fifth figure], it was found that Cisplatin has no inhibitory effect on mouse lung cancer cell LL2 and human lung cancer cell A549 compared with other anticancer drugs, but the complex of the present invention The composition has no therapeutic effect. By the above test, the following is to confirm that the composite composition of the present invention has an adjuvant treatment effect, and the anti-cancer drug Cisplatin which is less inhibitory than the combination of the present invention and Etoposide is added to the mouse. Lung cancer cell LL2 and human lung cancer cell A549 were observed for cytotoxicity test 72 hours later. [See Fig. 6] The results showed that when the anticancer drugs Cisplatine and Etoposide were given to cancer cells together with mulberry yellow, there was a significant therapeutic effect. Therefore, it can be seen that mulberry has the effect of adjuvant treatment.

Next, by trying several different concentrations and compounding other anticancer drugs (see Figure 7), the pulmonary fibrosis was found 7 days after the administration of BLM 40 mg/kg from the section, and it was only 14 days later. The inflamed part is reduced, the fibrotic parenchyma begins to be produced (such as the slice C, D, E, F circle), in addition, the BLM 40mg / kg plus solid fermented mulberry wine extract 200mg / kg, has been Effectively alleviate the fibrosis (such as the slice H, I, J circle).

The invention has the advantages that the compound composition of the mulberry yellow combined with the anticancer drug has the advantages of assisting the treatment of lung cancer and reducing the pulmonary fibrosis effect, and has the advantages that the use of the Chinese herbal medicine BLM drug is more remarkable than the western medicine. The effect can also reduce the occurrence of side effects of pulmonary fibrosis. The experimental composition proves that the composite composition does have the effect of adjuvant therapy. Therefore, the overall technology in the current cancer chemotherapy can provide mutual assistance between the drugs, which is quite equivalent. Helper.

In summary, it is understood that the present invention is novel, and the present invention is not found in any publication, and is in compliance with the provisions of Articles 21 and 22 of the Patent Law.

The above description is only a preferred embodiment of the present invention, and the scope of the present invention is not limited thereto. That is, the equivalent changes and modifications made by the invention in accordance with the scope of the invention are still within the scope of the invention.

1‧‧‧ mulberry extract

2‧‧‧BLM drugs

3‧‧‧Composite composition

Claims (1)

A composite composition of mulberry yellow combined with an anticancer drug for treating lung cancer and reducing pulmonary fibrosis, the composite composition mainly comprising mulberry yellow extract and BLM medicine; wherein: mulberry yellow extract is used It is a solid-state fermented mulberry mycelium alcohol extract. The extraction step of the extract is mainly composed of solid fermentation of mulberry mycelium, alcohol dissolution extraction, drying and alcohol removal, and the mulberry hyphae The body fermentation is a solid fermented mulberry mycelium, the mycelium is put into alcoholic extraction, and the alcoholic extraction is added to 75% alcohol for 48 hours, and then the supernatant is centrifuged, and the obtained solution is obtained. For the mycelium alcohol extract of mulberry mycelium, then put it in an oven at 60 ° C for 8 hours to remove alcohol to complete the extract of mulberry yellow; BLM drug is Bleomycin, which is an antibiotic complex; by the way, the mulberry yellow extract The compound and the BLM drug are mixed and combined in a ratio of 5:1 to form a composite composition.