Process for isolating physalins from plants and pharmaceutical compositions containing physalins

Abstract

The present invention relates to a process for obtaining Physalis angulata extracts, particularly to isolate ergostane-type steroidal derivatives, such as physalins, which are useful in the treatment of infections caused by protozoans. The physalins of the invention have also prophylactic and therapeutic properties for treating auto-immune diseases in mammals. The isolation process to obtain physalins comprises the steps of: (a) grinding roots, leaves or stem-bark of Physalis ssp; (b) extracting the material obtained in step (a) with solvents selected from the group consisting of water and alcohols, such as methanol, ethanol, 1-propanol, 2-propanol, isobutanol; (c) evaporating the extract obtained in step (b) and washing the syrup material with a suitable solvent; (d) rapid filtering of the extract of step (c) and fractionating the components of the extract according to a polarity gradient of organic solvents; and (e) separating the steroidal components contained in the fractions by chromatography.

1 ) A pharmaceutical composition comprising an effective amount of one or more physalins in combination with additives and a pharmaceutically acceptable carrier.  
     
     
         2 ) The pharmaceutical composition according to  claim 1  characterized by its use in the treatment of diseases caused by protozoans, preferably, in the treatment of Chagas Disease and Leishmaniasis.  
     
     
         3 ) The pharmaceutical composition according to  claim 1  characterized by its use as immunomodulator.  
     
     
         4 ) The pharmaceutical composition according to  claim 2  or  3  wherein physalin B or its mixtures with other physalins are used.  
     
     
         5 ) The pharmaceutical composition according to  claim 2  wherein physalin D or its mixtures with other physalins are used.  
     
     
         6 ) The pharmaceutical composition according to  claim 2  wherein physalin H or its mixtures with other physalins are used.  
     
     
         7 ) The pharmaceutical composition according to  claim 2  wherein physalin L or its mixtures with other physalins are used.  
     
     
         8 ) The pharmaceutical composition according to  claim 1  wherein the effective amount is from 10 to 50%.  
     
     
         9 ) A pharmaceutical composition comprising an effective amount of an alcoholic steroid extract from Physalis species in combination with a pharmaceutically acceptable carrier.  
     
     
         10 ) The pharmaceutical composition according to  claim 9  characterized by its use in the treatment of diseases caused by protozoans, preferably, in the treatment of Chagas Disease and Leishmaniasis.  
     
     
         11 ) The pharmaceutical composition according to  claim 9  wherein the alcoholic steroid extract is derived from  P. angulata.    
     
     
         12 ) A pharmaceutical composition comprising an effective amount of an aqueous steroid extract from Physalis species in combination with a pharmaceutically acceptable carrier.  
     
     
         13 ) The pharmaceutical composition according to  claim 12  characterized by its use in the treatment of diseases caused by protozoans, preferably, in the treatment of Chagas Disease and Leishmaniasis.  
     
     
         14 ) The pharmaceutical composition according to  claim 13  wherein the aqueous steroid extract is derived from  P. angulata.    
     
     
         15 ) An isolation process of physalin from plants belonging to the Solonaceae family comprising the steps of: (a) grinding roots, leaves or stem-bark of Physalis ssp; (b) extracting the material obtained in step (a) with solvents selected from the group consisting of water and alcohols, such as methanol, ethanol, 1-propanol, 2-propanol, isobutanol; (c) evaporating the extract obtained in step (b) and washing the syrup material with a suitable solvent; (d) rapid filtering of the extract of step (c) and fractionating the components of the extract according to a polarity gradient of organic solvents; and (e) separating the steroidal components contained in the fractions by chromatography.  
     
     
         16 ) The isolation process according to  claim 14  wherein the extraction is carried out at room temperature or by heating.  
     
     
         17 ) The isolation process according to  claim 15  wherein the heating extraction is conducted in a suitable extractor with ethanol for at least 24 hours.  
     
     
         18 ) The isolation process according to  claim 16  wherein the washing procedure of step (c) is accomplished with a non-polar organic solvent.  
     
     
         19 ) The isolation process according to  claim 17  wherein the non-polar organic solvent is chloroform.  
     
     
         20 ) The isolation process according to  claim 14  wherein the plant belonging to the family Solonacea is  P. angulata.    
     
     
         21 ) The isolation process according to  claim 19  wherein roots, leaves or stems from  P. angulata  are used to obtain the physalins.  
     
     
         22 ) The isolation process according to  claim 20  wherein roots from  P. angulata  are used to obtain the physalins.  
     
     
         23 ) The isolation process according to  claim 21  wherein the physalins are selected from the group consisting of physalin B, physalin D, physalin L and physalin H.  
     
     
         24 ) The isolation process according to  claim 14  wherein the rapid filtration of step (d) over silica gel H uses in a sequential fashion, hexane, dichloromethane, ethyl acetate and methanol as solvent extractor to provide the corresponding fractions.  
     
     
         25 ) The isolation process according to  claim 23  wherein the dichloromethane fraction is treated again in a chromatographic step using an ethyl acetate/hexane gradient.  
     
     
         26 ) The isolation process according to  claim 24  wherein physalin B is eluted.  
     
     
         27 ) The isolation process according to  claim 23  wherein the dichloromethane fraction is treated again in a chromatographic step using a chloroform/ethyl acetate gradient.  
     
     
         28 ) The isolation process according to  claim 26  wherein physalin D is eluted.