Method of Treating Neurological Conditions with Extract of Nerium Species or Thevetia Species

Abstract

A method of treating neurological condition in a subject by administration of an extract, or fraction or sub-fraction thereof, of Nerium species or Thevetia species is provided, wherein the extract, or fraction or sub-fraction thereof, excludes oleandrin and neriifolin. Alzheimer's disease, Huntington's disease or stroke are treated by administering a therapeutically effective amount of the extract, or a fraction or sub-fraction thereof, to a subject. The extract can be present in a pharmaceutical composition.

1 ) (canceled) 
     
     
         2 ) (canceled) 
     
     
         3 ) An extract, or a fraction or sub-fraction thereof of  Nerium  species or of  Thevetia  species, the extract, fraction or sub-fraction thereof comprising as major components at least one steroid and at least one triterpene extracted from the  Nerium  species or  Thevetia  species, and comprising less than 1% wt. of polysaccharide extracted from the  Nerium  species or  Thevetia  species, and excluding oleandrin and neriifolin. 
     
     
         4 ) A pharmaceutical composition comprising the extract, or fraction or sub-fraction thereof of  claim 3 . 
     
     
         5 ) (canceled) 
     
     
         6 ) (canceled) 
     
     
         7 ) (canceled) 
     
     
         8 ) (canceled) 
     
     
         9 ) (canceled) 
     
     
         10 ) (canceled) 
     
     
         11 ) (canceled) 
     
     
         12 ) (canceled) 
     
     
         13 ) (canceled) 
     
     
         14 ) (canceled) 
     
     
         15 ) The extract, or fraction or sub-fraction thereof of  claim 3  further comprising one or more therapeutically effective agents extracted from the  Nerium  species or  Thevetia  species. 
     
     
         16 ) The extract, or fraction or sub-fraction thereof, of  claim 3  further comprising one or more other therapeutically effective agents. 
     
     
         17 ) The extract, or fraction or sub-fraction thereof, of  claim 3 , wherein the extract is obtained by extraction of  Nerium  species or  Thevetia  species by hot water extraction, cold water extraction, supercritical fluid extraction, organic solvent extraction or a combination thereof. 
     
     
         18 ) The extract, or fraction or sub-fraction thereof, of  claim 3 , wherein the extract excludes a therapeutically effective amount of cardiac glycoside. 
     
     
         19 ) (canceled) 
     
     
         20 ) The extract, or fraction or sub-fraction thereof, of  claim 3 , wherein the fraction has been prepared by liquid chromatographic fractionation of the extract. 
     
     
         21 ) The extract, or fraction or sub-fraction thereof of  claim 3 , wherein the  Nerium  species is  Nerium oleander  and the  Thevetia  species is  Thevetia neriifolia.    
     
     
         22 ) The extract, or fraction or sub-fraction thereof of  claim 3 , wherein the sub-fraction has been prepared by liquid chromatographic fractionation of a fraction of the extract, and the sub-fraction excludes oleandrin and neriifolin. 
     
     
         23 ) (canceled) 
     
     
         24 ) A method for preparing a fraction of extract of  Nerium  species or  Thevetia  species comprising:
 extracting a mass comprising  Nerium  species or  Thevetia  species to form an extract thereof, the extract comprising one or more pharmacologically active components for the treatment of a neurological condition; and   fractionating the extract to form two or more fractions thereof, wherein at least one fraction comprises as major components at least one steroid and at least one triterpene extracted from the  Nerium  species or  Thevetia  species, comprises less than 1% wt. of polysaccharide extracted from the  Nerium  species or  Thevetia  species and excludes oleandrin and neriifolin.   
     
     
         25 ) The method of  claim 24 , wherein the extracting has been conducted by hot water extraction, cold water extraction, organic solvent extraction, supercritical fluid extraction or a combination thereof. 
     
     
         26 ) The method of  claim 24 , wherein the fractionating has been conducted by liquid chromatography. 
     
     
         27 ) The method of  claim 24 , wherein: a) the at least one fraction excludes cardiac glycoside; or b) the at least one fraction further comprises cardiac glycoside. 
     
     
         28 ) A method of fractionating an extract of  Nerium  species or  Thevetia  species in order to provide one or more therapeutically effective fractions thereof, the method comprising:
 a) providing an extract of  Nerium  species or  Thevetia  species, wherein the extract comprises at least one triterpene and at least one steroid and comprises less than 1% wt. of polysaccharide extracted from the  Nerium  species or  Thevetia  species;   b) fractionating the extract to provide two or more different fractions of the extract, a first extract fraction comprising one or more pharmacologically active agents, which is/are not a cardiac glycoside, and excluding oleandrin and neriifolin, and a second extract fraction comprising one or more cardiac glycosides and one or more pharmacologically active agents, which is/are not a cardiac glycoside.   
     
     
         29 ) The method of  claim 28 , wherein the first or second extract fraction is subjected to further fractionation to provide two or more different sub-fractions, wherein a first sub-fraction comprises one or more steroids and a second sub-fraction comprises one or more triterpenes. 
     
     
         30 ) The method of  claim 28 , wherein the fractionation is performed by liquid chromatography with a stationary phase and a mobile phase. 
     
     
         31 ) A composition comprising a sub-fraction of fraction of an extract obtained from  Nerium  species or  Thevetia  species, whereby the sub-fraction has been obtained by further fractionation of a fraction of the extract obtained from  Nerium  species or  Thevetia  species, and wherein the sub-fraction comprises as major components at least one steroid and at least one triterpene extracted from the  Nerium  species or  Thevetia  species, comprises less than 1% wt. of polysaccharide extracted from the  Nerium  species or  Thevetia  species and excludes oleandrin and neriifolin. 
     
     
         32 ) The composition of  claim 31 , wherein: a) the sub-fraction comprises one or more steroids, cardiac glycosides, the associated aglycones of cardiac glycosides, oleandrigenin, cardenolides, or triterpenoids, and one or more tritepenes; b) the sub-fraction comprises one or more triterpenes and excludes a steroid; and/or c) each sub-fraction independently excludes oleandrin and neriifolin. 
     
     
         33 ) The extract, or fraction or sub-fraction thereof, of  claim 3 , wherein the extract comprises at least two pharmacologically active agents obtained (extracted) from  Nerium  species or  Thevetia  species. 
     
     
         34 ) The invention according to  claim 33 , wherein the at least one steroid and at least one triterpene function additively or synergistically to contribute to the therapeutic efficacy of the extract when the extract is administered to a subject. 
     
     
         35 ) The method of  claim 28 , wherein the cardiac glycoside is selected from the group consisting of oleandrin, odoroside, neritaloside, ouabain, bufalin, digitoxin, cinobufatalin, cinobufagin, and resibufogenin. 
     
     
         36 ) A pharmaceutical composition comprising the composition of  claim 31 . 
     
     
         37 ) The composition of  claim 31 , wherein: a) the extract has been obtained from an  oleander  plant mass or  Neriifolia  plant mass; and b) the plant mass comprises  Nerium oleander, Thevetia neriifolia  or  Thevetia peruviana.    
     
     
         38 ) The composition of  claim 31 , wherein: a) the extract was prepared by supercritical fluid (SCF) extraction optionally in the presence of a modifier; b) the extract was prepared by hot water extraction; c) the extract was prepared by cold water extraction; d) the extract was prepared by organic solvent extraction; e) the extract was prepared by aqueous organic solvent extraction; or f) the extract was prepared by a combination of two or more of supercritical fluid extraction, hot water extraction, cold water extraction, or organic solvent extraction. 
     
     
         39 ) The extract, or fraction or sub-fraction thereof of  claim 3 , wherein the extract, or fraction thereof or sub-fraction thereof, of  Nerium  species or  Neriifolia  species, has a  1 HNMR spectrum as described any of  FIGS. 9B-9I . 
     
     
         40 ) The extract, or fraction or sub-fraction thereof of  claim 3 , wherein the extract, or fraction thereof or sub-fraction thereof, of  Nerium  species or  Neriifolia  species, exhibits therapeutic activity as described herein when administered to a subject. 
     
     
         41 ) The extract, or fraction or sub-fraction thereof of  claim 3 , wherein the extract, or fraction thereof or sub-fraction thereof, of  Nerium  species or  Neriifolia  species, has a HPLC chromatogram as described in  FIG. 8D  when chromatographic analysis is conducted as described herein. 
     
     
         42 ) The extract, or fraction or sub-fraction thereof of  claim 3  comprising at least one steroid, at least two triterpenes and less than 1% wt. of polysaccharide extracted from the  Nerium  species or  Thevetia  species, and excludes oleandrin and neriifolin. 
     
     
         43 ) The extract, or fraction or sub-fraction thereof of  claim 3  comprising at least two steroids, at least two triterpenes and less than 1% wt. of polysaccharide extracted from the  Nerium  species or  Thevetia  species, and excludes oleandrin and neriifolin. 
     
     
         44 ) The extract, or fraction or sub-fraction thereof of  claim 3 , wherein the extract, fraction thereof or sub-fraction thereof comprises betulin (urs-12-ene-3β,28-diol); 28-norurs-12-en-3β-ol; urs-12-en-3β-ol; 3β,3β-hydroxy-12-oleanen-28-oic acid; 3β,20α-dihydroxyurs-21-en-38-oic acid; 3β,27-dihydroxy-12-ursen-38-oic acid; 3β,13β-dihydroxyurs-11-en-28-oic acid; 3β, 12α-dihydroxyoleanan-28,13β-olide; 3β,27-dihydroxy-12-oleanan-28-oic acid; less than 1% wt. of polysaccharide extracted from the  Nerium  species or  Thevetia  species, and excludes oleandrin and neriifolin. 
     
     
         45 ) The extract, or fraction or sub-fraction thereof of  claim 3 , wherein the extract, fraction thereof or sub-fraction thereof comprises oleandrigenin, ursolic acid, betulinic acid, odoroside, neritaloside, oleanolic acid and one or more triterpenes and less than 0.5% by weight polysaccharide extracted from the  Nerium  species or  Thevetia  species, and excludes oleandrin and neriifolin.