Cordyceps cicadae mycelium active substances, preparation method, pharmaceutical composition and application thereof

Abstract

A method of preparing Cordyceps cicadae mycelium active substances for preventing and/or treating xerophthalmia is provided. The method comprises: (a)culturing a Cordyceps cicadae mycelium in a plate media at 15 to 35° C. for 5 to 14 days; (b)inoculating the mycelium of step(a) to a flask containing liquid media and culturing it at 15 to 35° C. with a pH of 2 to 8 for 3 days; (c)inoculating the mycelium of step(b) to a fermenter tank and culturing it at 15 to 35° C. with a pH of 2 to 8 for 3 days, so as to obtain a Cordyceps cicadae mycelium fermentation liquid; (d)freeze-drying and grating the fermentation liquid, so as to obtain a Cordyceps cicadae mycelium powder; (e)extracting the powder with solvent, so as to obtain a Cordyceps cicadae mycelium extract; and (f)drying the extract, so as to obtained the Cordyceps cicadae mycelium active substances.

What is claimed is: 
     
         1 . A method of preparing  Cordyceps cicadae  mycelium active substances for preventing and/or treating xerophthalmia, wherein the method comprises following steps:
 (a) culturing a  Cordyceps cicadae  mycelium in a plate media at 15 to 35° C. for 5 to 14 days;   (b) inoculating the  Cordyceps cicadae  mycelium of step(a) to a flask containing liquid media and culturing the mycelium at 15 to 35° C. with a pH of 2 to 8 for 3 days;   (c) inoculating the  Cordyceps cicadae  mycelium of step(b) to a fermenter tank and culturing the mycelium at 15 to 35° C. with a pH of 2 to 8 for 3 days, so as to obtain a  Cordyceps cicadae  mycelium fermentation liquid;   (d) freeze-drying and grating the  Cordyceps cicadae  mycelium fermentation liquid, so as to obtain a  Cordyceps cicadae  mycelium powder;   (e) extracting the  Cordyceps cicadae  mycelium powder with at least one solvent, so as to obtain at least one  Cordyceps cicadae  mycelium extract; and   (f) drying the  Cordyceps cicadae  mycelium extract, so as to obtained the  Cordyceps cicadae  mycelium active substances.   
     
     
         2 . The method of  claim 1 , wherein the culturing process of step(b) is shake flask cultivation, and the flask is shaking at a speed range between10 and 250 rpm. 
     
     
         3 . The method of  claim 1 , wherein in step(c), the gas feed in the fermenter tank comprises air, oxygen, carbon dioxide, helium or a combination thereof. 
     
     
         4 . The method of  claim 1 , wherein in step(c), the pressure of the fermenter tank is at 0.5 to 1.0 kg/cm 2 , and the gas flow rate of fermenter tank is 0.01 to 1.5 VVM. 
     
     
         5 . The method of  claim 1 , wherein the liquid media used in step(b) and step(c) are the same, and the liquid media comprises grains, beans, inorganic salts, carbohydrates, yeast extract, malt extract or a combination thereof. 
     
     
         6 . The method of  claim 1 , wherein the solvent of step(e) comprises water and alcohol. 
     
     
         7 . The method of  claim 1 , wherein the extracting process of step(e) uses two solvents, which comprise water and alcohol. 
     
     
         8 . The method of  claim 7 , wherein the alcohol is methanol or ethanol. 
     
     
         9 . The method of  claim 8 , wherein in step(f), water extracts and alcohol extracts from the  Cordyceps cicadae  mycelium powder are mixed to obtain the  Cordyceps cicadae  mycelium active substances. 
     
     
         10 . The method of  claim 8 , wherein the  Cordyceps cicadae  mycelium active substances comprise the same weight of water extracts and alcohol extracts from the  Cordyceps cicadae  mycelium powder. 
     
     
         11 . A method of preparing a  Cordyceps cicadae  mycelium active substance for preventing and/or treating xerophthalmia, wherein the method comprises following steps:
 (a) culturing a  Cordyceps cicadae  mycelium in a plate media at 15 to 35° C. for 5 to 14 days;   (b) inoculating the  Cordyceps cicadae  mycelium of step(a) to a flask containing liquid media and culturing the mycelium at 15 to 35° C. with a pH of 2 to 8 for 3 days; and   (c) inoculating the  Cordyceps cicadae  mycelium of step(b) to a fermenter tank and culturing the mycelium at 15 to 35° C. with a pH of 2 to 8 for 3 days, so as to obtain a  Cordyceps cicadae  mycelium fermentation liquid comprising the  Cordyceps cicadae  mycelium active substances.   
     
     
         12 . The method of  claim 11 , wherein the culturing method of step(b) is shake flask cultivation, and the flask is shaking at a speed range between 10 and 250 rpm. 
     
     
         13 . The method of  claim 11 , wherein in step(c), the gas feed in the fermenter tank comprises air, oxygen, carbon dioxide, helium or a combination thereof. 
     
     
         14 . The method of  claim 11 , wherein in step(c), the pressure of the fermenter tank is at 0.5 to 1.0 kg/cm 2 , and the gas flow rate of fermenter tank is 0.01 to 1.5 VVM. 
     
     
         15 . The method of  claim 11 , wherein the liquid media used in step(b) and step(c) are the same, and the liquid media comprises grains, beans, inorganic salts, carbohydrates, yeast extract, malt extract or a combination thereof. 
     
     
         16 . A  Cordyceps cicadae  mycelium active substances for preventing and/or treating xerophthalmia, which is prepared by the method of  claim 1 . 
     
     
         17 . A method for preventing and/or treating xerophthalmia, which comprises administrating to a subject an effective amount of the  Cordyceps cicadae  mycelium active substances of  claim 16 . 
     
     
         18 . A pharmaceutical composition for preventing and/or treating xerophthalmia, which comprises a therapeutically effective amount of the  Cordyceps cicadae  mycelium active substances of  claim 1  and a pharmaceutically acceptable carrier, excipient, diluents or adjuvant. 
     
     
         19 . A method for preventing and/or treating xerophthalmia, which comprises administrating to a subject an effective amount of the pharmaceutical composition of  claim 18 .