Screening of chemical compounds purified from biological sources

Abstract

The invention provides a chemical compound library containing a collection of chromatographic fractions purified from biological sources suitable for high-throughput screening for biological activity, wherein: (a) the chromatographic fractions contain primarily from about 1 microgram to 100 micrograms of primarily one to five chemical compounds; (b) the compounds have log P of greater than-1 and less than 5 and molecular weights less than 3,000 Daltons; (c) the library can include chemical compounds produced from at least fifty different biological sources; and (d) the library can contain at least about 200 different chemical compounds from each different biological source.

What is claimed is:  
     
         1 . A chemical compound library comprising a plurality of chromatographic fractions from at least one biological source, 
 (a) said fractions being free of compounds that interfere with biological assays;    (b) the majority of said chromatographic fractions comprising a normalized quantity of from about 1 microgram to about 500 micrograms of seven or fewer chemical compounds; and    (c) the majority of said chemical compounds having log P of from about −1 to about 5 and a molecular weight less than about 6,000 Daltons.    
     
     
         2 . The library of  claim 1 , the library comprising at least about 80 chromatographic fractions from at least one biological source.  
     
     
         3 . The library of  claim 1 , the library comprising at least about 100 of said different chemical compounds from at least one biological source.  
     
     
         4 . The library of  claim 1 , the library comprising at least about 150 of said different chemical compounds from at least one biological source.  
     
     
         5 . The library of  claim 1 , each of said chromatographic fractions having associated analytical data comprising molecular ions and chromatographic elution conditions.  
     
     
         6 . The library of  claim 1 , each said chromatographic fraction having associated analytical data comprising molecular ions, chromatographic elution conditions, and NMR spectra.  
     
     
         7 . The library of  claim 1 , each said chromatographic fraction having associated analytical data comprising molecular ions, chromatographic elution conditions and MS/MS fragmentation patterns.  
     
     
         8 . The library of  claim 1 , each said chemical compound having a molecular weight of less than about 3,000 Daltons.  
     
     
         9 . The library of  claim 1 , each said chemical compound having a molecular weight of less than about 1,000 Daltons.  
     
     
         10 . The library of  claim 1 , each said chemical compound having a molecular weight of less than about 600 Daltons.  
     
     
         11 . The library of  claim 1 , said at least one biological source being a plant.  
     
     
         12 . The library of  claim 11 , said chromatographic fractions being from a plurality of biological sources.  
     
     
         13 . The library of  claim 12 , said plurality of biological sources comprising two or more species of organisms.  
     
     
         14 . The library of  claim 1 , each of said chromatographic fractions having associated data sufficient to identify the amounts of each of said chemical compounds in said chromatographic fraction.  
     
     
         15 . The library of  claim 1 , each of said chromatographic fractions being free of tetramers or greater of polyphenolics.  
     
     
         16 . The library of  claim 1 , the majority of said chromatographic fractions comprising less than about 100 micrograms of each of said chemical compounds.  
     
     
         17 . A subset of the library of  claim 1 , comprising chemical compounds having similar predetermined molecular ions.  
     
     
         18 . A subset of the library of  claim 1  comprising said chemical compounds having similar predetermined log Ps.  
     
     
         19 . A method of preparing a sublibrary of the library of  claim 1 , said fractions having associated data comprising at least one of molecular ion data, chromatographic elution data, NMR spectra MS/MS fragmentation patterns, comprising selecting fractions having at least one set of similar data.  
     
     
         20 . A method of preparing a diverse chemical compound library comprising providing the chemical library of  claim 1 , each of said chromatographic fractions having associated data comprising at least one of molecular ion data for said chemical compounds, chromatographic elution data for said fractions, NMR spectra for said fractions and MS/MS fragmentation patterns for said chemical compounds selecting fractions having at least one characteristic selected from 
 (a) similar molecular ions;    (b) similar chromatographic elution data;    (c) similar log Ps; and    (d) a plurality of biological sources.    
     
     
         21 . The library of  claim 1 , produced by a chromatographic process, wherein said process: 
 (a) produces groups of primarily one to five chemical compounds with related physical properties which elute from a reverse phase chromatography column during a flow rate of more than 0.48 and less than 0.80 column volumes per minute of a solvent system during which the solvent system was increasing in acetonitrile concentration of more than 0.008% per second and less than 0.035% per second; produces at least fifteen chromatographic fractions originating from the same preceding chromatographic fraction;    (b) produces at least fifteen chromatographic fractions originating from the same preceding chromatographic fraction;    (c) produces said chromatographic fractions from about four chemically and physically distinct said preceding chromatographic fractions of each different biological source; and    (d) employs at least one method on the said preceding chromatographic fractions which removes compounds with log Ps greater than six and less than negative 1, tetramers or greater of phenolics, and molecular weights greater than 3,000 Daltons.    
     
     
         22 . A method of identifying biologically active chemical compounds comprising screening the library of  claim 1  for biological activity of said chemical compounds.  
     
     
         23 . The method of  claim 22 , comprising testing said chromatographic fractions in parallel using at least one biological assay.  
     
     
         24 . The method of  claim 22 , said at least one biological assay detecting efficacy in the treatment of disease or in the control of biological pests.  
     
     
         25 . The method of  claim 22 , each of said chemical compounds in said chromatographic fractions being prepared at a concentration of at least one micromolar for biological testing.  
     
     
         26 . The method of  claim 22 , said screening comprising testing multiple biological mechanisms of a specific disease to identify the preferred biological mechanism for therapeutic intervention which is determined from the biological activity exhibited because of the design and structural modifications of said chemical compounds which have been evolutionarily selected to interact with the optimal biological mechanisms of biological systems.  
     
     
         27 . The library of  claim 1 , said majority of fractions comprising at least about 80% of said fractions.  
     
     
         28 . The library of  claim 1 , said majority of fractions comprising at least about 95% of said fractions.  
     
     
         29 . The library of  claim 1 , said majority of compounds comprising at least 80% of said compounds.  
     
     
         30 . The library of  claim 1 , said majority of compounds comprising at least about 95% of said compounds.  
     
     
         31 . The library of  claim 1 , at least some of said chromatographic fractions comprising a blank, said blank having no detectable chemical compounds in a normalizable quantity.  
     
     
         32 . The library of  claim 1 , comprising fractions from at least 5 different organisms.  
     
     
         33 . A chemical compound library comprising: a plurality of arrayed chromatographic fractions derived from one or more biological sources, 
 the majority of said fractions comprising seven or fewer individual nonproteinaceous chemical compounds, and each of said isolates being free of compounds that interfere with biological testing of said individual compounds; and    wherein each of said individual compounds is present in a normalized quantity and has a log P suitable for biological testing of said individual compound, said normalized quantity being sufficient to prepare a sample from said isolate having a concentration of each of said individual compounds suitable for biological testing.    
     
     
         34 . The library of  claim 1 , said chemical compounds comprising less than about 10% by weight of the extracted compounds from said biological source.  
     
     
         35 . The library of  claim 1 , said chemical compounds comprising less than about 20% by weight of the extracted compounds from said biological source.  
     
     
         36 . A method for preparing an array of compounds of normalized concentrations from a natural product comprising a mixture of compounds of various concentrations, the method comprising: 
 (a) normalizing concentrations of the compounds; and    (b) producing an array of samples of the compounds of normalized concentrations.    
     
     
         37 . A method as set forth in  claim 36  wherein the normalized compounds are drug-like compounds.  
     
     
         38 . A method as seth forth in  claim 37  wherein the mixture further comprises non-drug-like compounds and the method further comprises removing enough of the non-drug-like compounds from the mixture to render the mixture substantially free of the non-drug-like compounds.  
     
     
         39 . A method as set forth in  claim 38  wherein the non-drug-like compounds are interferences.  
     
     
         40 . A method as set forth in  claim 36  further comprising separating the compounds in the mixture into fractions and samples correspond to the fractions.  
     
     
         41 . A method as set forth in  claim 40  wherein each sample contains from one to about five of the compounds.  
     
     
         42 . A method as set forth in  claim 41  wherein the mixture is derived from at least one plant.  
     
     
         43 . A method as set forth in  claim 40  wherein the separation is carried out by liquid chromatography.  
     
     
         44 . A method for screening a natural product comprising a mixture of compounds of various concentrations, the method comprising: 
 (a) normalizing concentrations of the compounds;    (b) producing an array of samples of the compounds of normalized concentrations; and    (c) screening the array.    
     
     
         45 . A method as set forth in  claim 44  wherein the screening is carried out by a high-throughput screening technique.  
     
     
         46 . A method as set forth in  claim 45  wherein the screening is designed to identify bio-active compounds.  
     
     
         47 . An array of compounds prepared by the method of  claim 36.