Method of isolating biologically active chemical compounds from a chemical compound library

Abstract

A method of producing a chemical compound library comprises extracting at least one extract from at least one species of plant; processing at least one of the extract(s) to remove at least one type of chemical interference to produce a processed extract; chromatographically separating the processed extract into a plurality of chromatographic fractions, each containing an amount of chemical compounds; determining the amount of chemical compounds in at least one of the chromatographic fractions; and normalizing the chromatographic fractions in which the amounts were determined to produce normalized chromatographic fractions, each such fraction comprising from about 1 microgram to about 500 micrograms of each of from one to seven chemical compounds that were present in lower concentrations in the extract and that each have a log P of from about −1 to about 5 and a molecular weight less than about 1000 Daltons; thereby to produce a chemical compound library from at least one species of plant.

1 - 25 . (canceled) 
   
   
       26 . A method of isolating at least one of biologically active chemical compound from a chemical compound library, comprising isolating the at least one biologically active chemical compound from a chemical compound library produced by:
 (a) extracting at least one extract from at least one species of plant;   (b) processing at least one of the at least one extract from step (a) to remove at least one type of chemical interference to produce a processed extract;   (c) chromatographically separating the processed extract from step (b) into a plurality of chromatographic fractions, each containing an amount of chemical compounds;   (d) determining the amount of chemical compounds in at least one of the chromatographic fractions from step (c); and   (e) normalizing the chromatographic fractions in which the amounts were determined in step (d) to produce normalized chromatographic fractions, each such normalized chromatographic fraction comprising from about 1 microgram to about 500 micrograms of each of from about one to seven chemical compounds that were present in lower concentrations in the extract and that each have a log P of from about −1 to about 5 and a molecular weight less than about 1000 Daltons;   (f) thereby to produce the chemical compound library from at least one species of plant.   
   
   
       27 . A method as set forth in  claim 26  wherein the at least one extract in step (a) is extracted from a plurality of species of plant. 
   
   
       28 . A method as set forth in  claim 27  wherein the at least one extract in step (a) is extracted from the plurality of species of plant in an extraction with an organic solvent. 
   
   
       29 . A method as set forth in  claim 27  wherein the at least one extract in step (a) is extracted from the plurality of species of plant in a multiple step extraction comprising a first extraction step carried out with a first organic solvent and thereafter a second extraction step whereby insoluble material is extracted by use of a mixture of a second organic solvent, which may or may not be the same as the first organic solvent, and water. 
   
   
       30 . A method as set forth in  claim 29  wherein the first organic solvent is about 50% by weight ethanol and about 50% weight ethyl acetate and wherein the mixture of the second organic solvent and water is about 70% by weight methanol and about 30% by weight water. 
   
   
       31 . A method as set forth in  claim 26  wherein in step (b), the at least one of the at least one extract from step (a) is processed to remove a tetramer or greater of a polyphenolic compound. 
   
   
       32 . A method as set forth in  claim 26  wherein in step (b), the at least one of the at least one extract from step (a) is processed to remove a compound with a molecular weight of greater than 1000 daltons. 
   
   
       33 . A method as set forth in  claim 26  wherein the at least one extract is processed in step (b) by flash chromatography to remove the at least one type of chemical interference. 
   
   
       34 . A method as set forth in  claim 26  wherein the at least one extract is processed in step (b) by use of a size exclusion filter to remove the at least one type of chemical interference. 
   
   
       35 . A method as set forth in  claim 26  wherein the at least one extract is processed in step (b) by use of a polyamide column to remove the at least one type of chemical interference. 
   
   
       36 . A method as set forth in  claim 26  wherein in step (b), the at least one of the at least one extract from step (a) is processed to remove a compound with a with a log P of greater than about 5 or less than −1. 
   
   
       37 . A method as set forth in  claim 26  wherein the chemical compound library comprises at least about 100 compounds from one. 
   
   
       38 . A method as set forth in  claim 26  wherein the chemical compound library comprises at least two or more isomeric forms of a chemical compound. 
   
   
       39 . A method as set forth in  claim 26  wherein the chemical compound library is screened by testing the chromatographic fractions in parallel using at least one biological assay. 
   
   
       40 . A method as set forth in  claim 39  wherein the at least one biological assay detects efficacy in treatment of a disease or in control of biological pests. 
   
   
       41 . A method as set forth in  claim 26  wherein the concentration of each of the chemical compounds in the chromatographic fractions tested is at least one micromolar. 
   
   
       42 . A method as set forth in  claim 37  wherein the chemical compound library contains at least one triterpene compound, one lignan compound, one flavonoid compound and one alkaloid compound. 
   
   
       43 . A method as set forth in  claim 37  wherein the chemical compound library comprises at least about 200 compounds from one plant species. 
   
   
       44 . A method as set forth in  claim 26  wherein the chemical compound library comprises at least about 80 normalized chromatographic fractions. 
   
   
       45 . A method as set forth in  claim 44  wherein the chemical compound library comprises at least about 100 normalized chromatographic fractions. 
   
   
       46 . A method as set forth in  claim 45  wherein the chemical compound library comprises at least about 150 normalized chromatographic fractions. 
   
   
       47 . A method as set forth in  claim 26  wherein each of the compounds has a log P of from about −1 to about 5 and a molecular weight less than about 1000 Daltons. 
   
   
       48 . A method as set forth in  claim 26  wherein each of the at least one of the chemical compounds that has been isolated is purified greater than 85%. 
   
   
       49 . A method as set forth in  claim 26  wherein the at least one of biologically active chemical compound is isolated from the chemical compound library by screening the chemical compound library for biological activity of said chemical compounds to identify the at least one bioactive chemical compound; and isolating the at least one bioactive chemical compound. 
   
   
       50 . A method of isolating at least one chemical compounds from a plant comprising:
 producing a chemical compound library comprising the steps of:
 (a) extracting at least one extract from at least one species of plant; 
 (b) processing at least one of the at least one extract from step (a) to remove at least one type of chemical interference to produce a processed extract; 
 (c) chromatographically separating the processed extract from step (b) into a plurality of chromatographic fractions, each containing an amount of chemical compounds; 
 (d) determining the amount of chemical compounds in at least one of the chromatographic fractions from step (c); and 
 (e) normalizing the chromatographic fractions in which the amounts were determined in step (d) to produce normalized chromatographic fractions, each such normalized chromatographic fraction comprising from about 1 microgram to about 500 micrograms of each of from about one to seven chemical compounds that were present in lower concentrations in the extract and that each have a log P of from about −1 to about 5 and a molecular weight less than about 1000 Daltons; 
 (f) thereby to produce a chemical compound library from at least one species of plant; and 
   isolating at least one of the chemical compounds in the chemical compound library.   
   
   
       51 . A method as set forth in  claim 48  wherein each of the at least one of the chemical compounds that has been isolated is purified greater than 85%. 
   
   
       52 . A method of isolating at least one chemical compounds from a plant comprising
 producing a chemical compound library comprising the steps of:
 (a) extracting at least one extract from at least one species of plant; 
 (b) processing at least one of the at least one extract from step (a) to remove at least one type of chemical interference to produce a processed extract; 
 (c) chromatographically separating the processed extract from step (b) into a plurality of chromatographic fractions, each containing an amount of chemical compounds; 
 (d) determining the amount of chemical compounds in at least one of the chromatographic fractions from step (c); and 
 (e) normalizing the chromatographic fractions in which the amounts were determined in step (d) to produce normalized chromatographic fractions, each such normalized chromatographic fraction comprising from about 1 microgram to about 500 micrograms of each of from about one to seven chemical compounds that were present in lower concentrations in the extract and that each have a log P of from about −1 to about 5 and a molecular weight less than about 1000 Daltons; 
 (f) thereby to produce a chemical compound library from at least one species of plant; 
   screening the chemical compound library for biological activity of said chemical compounds to identify at least one bioactive chemical compound; and   isolating the at least one bioactive chemical compound.   
   
   
       53 . A method as set forth in  claim 52  wherein each of the at least one of the bioactive chemical compounds that has been isolated is purified greater than 85%.