Extracts of Polygonum multiflorum Thunb., and preparation process and uses of the same

Abstract

Disclosed herein are processes for preparing extract products from the root of Polygonum multiflorum Thunb. and the extract products thus obtained, which are biologically active in promoting the proliferation, growth and/or differentiation of hematocytes and bone marrow cells in vitro and in vivo.

1 . An extract product from the root of  Polygonum multiflorum Thunb. , which is prepared from a process comprising the steps of: 
 (a) subjecting a suitable amount of a starting root material of  Polygonum multiflorum Thunb.  to a freezing treatment for a period of time;    (b) subjecting a frozen product obtained in step (a) to an extraction treatment with methanol;    (c) subjecting a resultant product from step (b) to a separating treatment to obtain a methanol solution free of extracted root debris of the starting root material of  Polygonum multiflorum Thunb.;  and    (d) removing methanol from the methanol solution obtained in step (c) to obtain a methanol-extracted product.    
     
     
         2 . An extract product from the root of  Polygonum multiflorum Thunb . according to  claim 1 , wherein the product has a reverse-phase HPLC elution profile as shown in  FIG. 1 .  
     
     
         3 . An extract product from the root of  Polygonum multiflorum Thunb . according to  claim 1 , wherein in step (a) of the process, the starting root material of  Polygonum multiflorum Thunb.  to be used therein is a processed form of  Polygonum multiflorum Thunb . root.  
     
     
         4 . A pharmaceutical composition comprising an extract product from the root of  Polygonum multiflorum Thunb . as defined in  claim 1 .  
     
     
         5 . A pharmaceutical composition for promoting the proliferation of cells selected from the group consisting of hepatocytes, bone marrow stem cells, bone marrow stromal cells, bone marrow osteoprogenitor cells, bone marrow hematopoietic progenitor cells, bone marrow hematogenic cells, leukocytes and erythrocytes, comprising a therapeutically effective amount of an extract product from the root of  Polygonum multiflorum Thunb.  as defined in  claim 1 .  
     
     
         6 . A pharmaceutical composition for treating a subject afflicted with a liver disease selected from liver dysfunction, liver fibrosis and liver cirrhosis, comprising a therapeutically effective amount of an extract product from the root of  Polygonum multiflorum Thunb.  as defined in  claim 1 .  
     
     
         7 . A pharmaceutical composition for treating a subject in need of proliferation of cells selected from the group consisting of hepatocytes, bone marrow stem cells, bone marrow stromal cells, bone marrow osteoprogenitor cells, osteoblasts, bone marrow hematopoietic progenitor cells, bone marrow hematogenic cells, leukocytes and erythrocytes, comprising a therapeutically effective amount of an extract product from the root of  Polygonum multiflorum Thunb . as defined in  claim 1 .  
     
     
         8 . A pharmaceutical composition for treating a disorder or disease associated with the deficiency of bone marrow stem cells, comprising a therapeutically effective amount of an extract product from the root of  Polygonum multiflorum Thunb . as defined in  claim 1 .  
     
     
         9 . The pharmaceutical composition of  claim 8 , wherein the disorder or disease is selected from the group consisting of aging, osteoporosis, cancer, anemia and leukopennia.  
     
     
         10 . A pharmaceutical composition for treating a subject in need of restoration of bone marrow cells, comprising a therapeutically effective amount of an extract product from the root of  Polygonum multiflorum Thunb.  as defined in  claim 1 .  
     
     
         11 . The pharmaceutical composition of  claim 10 , wherein the subject is one selected from a hemodialysis patient, a patient receiving bone marrow transplantation therapy, and a cancer patient receiving irradiation therapy or chemotherapy or both.  
     
     
         12 . A process for preparing an extract product from the root of  Polygonum multiflorum Thunb.  comprising the steps of: 
 (a) subjecting a suitable amount of a starting root material of  Polygonum multiflorum Thunb . to a freezing treatment for a period of time;    (b) subjecting a frozen product obtained in step (a) to an extraction treatment with methanol;    (c) subjecting a resultant product from step (b) to a separating treatment to obtain a methanol solution free of extracted root debris of the starting root material of  Polygonum multiflorum Thunb.;  and    (d) removing methanol from the methanol solution obtained in step (c) to obtain a methanol-extracted product.    
     
     
         13 . The process of  claim 12 , wherein the starting root material of  Polygonum multiflorum Thunb.  used in step (a) is a processed form of  Polygonum multiflorum Thunb . root.  
     
     
         14 . The process of  claim 12 , wherein the freezing treatment of step (a) is conducted at a temperature ranging from −20° C. to −70° C.  
     
     
         15 . The process of  claim 13 , wherein the freezing treatment of step (a) is conducted at a temperature of −70° C.  
     
     
         16 . The process of  claim 12 , wherein in step (b), the frozen product obtained in step (a) is crashed and immersed in methanol to allow extraction.  
     
     
         17 . The process of  claim 12 , wherein in step (c), the separating treatment is conducted by suction filtration.  
     
     
         18 . The process of  claim 12 , wherein in step (d), methanol is removed by evaporation in vacuo.  
     
     
         19 . The process of  claim 12 , wherein the methanol-extracted product obtained from step (d) is further lyophilized.  
     
     
         20 . The process of  claim 12 , wherein the methanol-extracted product obtained from step (d) is further subjected to a n-hexane extraction treatment comprising the steps of: 
 (i) admixing the methanol-extracted product obtained from step (d) with a suitable amount water;    (ii) partitioning a resultant mixture from step (i) with n-hexane to form a n-hexane layer and a water layer;    (iii) collecting the n-hexane layer formed in step (ii), and    (iv) removing n-hexane from the n-hexane layer collected in step (iii) to obtain a n-hexane -extracted product.    
     
     
         21 . The process of  claim 20 , wherein the water layer formed in step (ii) of the n-hexane extraction treatment is further subjected to an ethyl acetate extraction treatment comprising the steps of: 
 (i′) partitioning the water layer formed in step (ii) of the n-hexane extraction treatment with ethyl acetate to form an ethyl acetate layer and a water layer,    (ii′) collecting the ethyl acetate layer formed in step (i′); and    (iii′) removing ethyl acetate from the ethyl acetate layer collected in step (ii′) to obtain an ethyl acetate-extracted product.    
     
     
         22 . The process of  claim 21 , wherein the water layer formed in step (i′) of the ethyl acetate extraction treatment is further subjected to a n-butanol extraction treatment comprising the steps of: 
 (i″) partitioning the water layer formed in step (i′) of the ethyl acetate extraction treatment with n-butanol to form a n-butanol layer and a water layer;    (ii″) collecting the n-butanol layer formed in step (i″); and    (iii″) removing n-butanol from the n-butanol layer collected in step (ii″) to obtain a n-butanol-extracted product.    
     
     
         23 . A pharmaceutical composition, comprising an extract product from the root of  Polygonum multiflorum Thunb . prepared by a process according to any one of claims  12 - 22 .