Guggulphospholipid methods and compositions

Abstract

The present invention relates to the methods for preparing synthetic guggulphospholipids, their fatty acid analogues and other bioactive molecules. The present invention relates to E-guggulsterone and Z-guggulsterone or mixture of E- and Z-guggulsterones, and E-guggulsterol and Z-guggulsterol or mixture of E- and Z-guggulsterols synthetically modified to guggulphospholipids and analogues and salts thereof, fatty acid analogues of guggulsterols, guggulsulfate and salts thereof, guggulphosphate and salts thereof; and guggulsterols conjugated with drugs for use as prodrugs. Also the present invention provides a novel method for the preparation of E-guggulsterol and Z-guggulsterol or mixture of E- and Z-guggulsterols from a mixture of E- and Z-guggulsterones. The present invention further relates to guggulphospholipids and other bioactive molecules incorporated into complexes such as liposomes, complexes, emulsions, vesicles, micelles, and mixed micelles, which can include other active agents, such as hydrophobic or hydrophilic drugs for use, e.g., in treatment of human and animal diseases.

What is claimed is: 
     
         1 . A guggul-based lipid molecule having a general structure selected from the group consisting of structures III-XII. 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         wherein:
 i) in structures III and. IV, R 1  is a saturated or unsaturated acyl or alkyl groups having between 1 and 34 carbon atoms; 
 ii) in structures III and IV, R 1  and R 2  is a saturated alkyl or alkyloxy group optionally substituted with amino or substituted amino group. 
 ii) in structure IV, R 1  is a sugar; 
 iii) in structure V, R 1  and R 2  are the same or different and at least one of the R 1  or R 2  is a saturated or unsaturated acyl group or alkyl group having between 1 and 34 carbon atoms; 
 iv) in structures IV-X, X is hydrogen, methyl, ammonium, sodium, potassium, calcium, barium ion or any non-toxic ion; 
 v) in structure VI, X and Y are the same or different and are hydrogen, 1(i methyl, hydrogen, ammonium, sodium, potassium, calcium, barium ion or any non-toxic ion; 
 vi) in structure VIII, R 3 , R 4  and R 5  are the same or different and are hydrogen or methyl group; X is hydrogen or methyl group. 
 vii) in structure X, PEG (polyethylene glycol) is a long chain, linear or branched synthetic polymer composed of ethylene oxide units, HO(CH 2 CH 2 O) n CH 2 CH 2 OCH 3 , where n is between about 1 and about 1000; 
 viii) in structure XI, drug is an active agent; and 
 ix) in structure XII, R 6 , R 7 , and R 8  are the same or different, and are selected from the group consisting of hydrogen, methyl, alkyl, substituted alkyl, alkyoxy, substituted alkyloxy groups, and are optionally hydroxylated, aminated, or polyaminated such that the composition has an overall positive charge; 
 
         and wherein:
 x) structures III-XII are in the form selected from the group consisting o E-isomer, Z-isomer, or a mixture of E- and Z-isomers; 
 xi) structures III-XII are optically pure, or one or more of structures III-XII are mixtures of optical isomers; 
 xii) structure VIII is cationic when X is a methyl group; and 
 xiii) structure XII is cationic, 
 
       
     
     
         2 . A method of preparing a guggul derivative of a formula selected from the group consisting of formula III-XII, comprising reacting a guggulsterol having structure IIA 
       
         
           
           
               
               
           
         
         to form a composition comprising a guggul derivative of a formula selected from the group consisting of formula III-XII. 
       
     
     
         3 . A method of preparing a guggul derivative of a formula selected from the group consisting of formula III-XII, comprising reacting a guggulsterone having structure I 
       
         
           
           
               
               
           
         
         to form a composition comprising a guggul derivative of a formula selected from the group consisting of formula III-XII. 
       
     
     
         4 . The method of  claim 3 , wherein said guggulsterone I is an E-isomer or Z-isomer or a mixture of E and Z isomers. 
     
     
         5 . The method of  claim 2 , wherein said guggulsterol IIA is an E-isomer or Z-isomer, or a mixture of E- and Z-isomer. 
     
     
         6 . The method of  claim 2 , wherein said guggulsterol is optically pure, and wherein said guggulsterol is a mixture of optical isomers. 
     
     
         7 . A method of preparing guggulsterol IIA, comprising treating guggulsterone I in one of more steps to produce preparing guggulsterol IIA. 
     
     
         8 . A method of preparing a compound of formula IV, V, VI, VIII, IX or X of  claim 1 ,  2 , or  3 , wherein at least one step comprises the use of a phosphoramidite reagent or a phosphorylating agent. 
     
     
         9 . The method of  claim 8 , wherein said phosphoramidite reagent is selected from the group consisting of NN-diisopropylmethylphosphoramidic chloride, (benzyloxy)(N,N-diisopropylamino)chlorophosphine, benzyloxybis (diisopropylamino) phosphine, 2-cyanoethyl-N,N,N,N-tetraisopropylphosphoramidite, (2-cyanoethyl)(N,N-diisopropylamino)chlorophosphine, difluorenyl diisopropylphosphoramidite, methyl-N,N,N,N tetraisopropylphosphorodiamidite, dimethyl N,N-diisopropylphosphoramidite, dibenzyl diisopropylphosphoramidite, di-tert-butyl-N,N-diisopropylphosphoramidite, 2-(diphenylmethylsilyl)ethyl-N,N,N,N-tetraisopropylphosphoramidite, (N-trifluoroacetyl amino) butyl and (N-trifluoroacetylamino) pentyl-N,N,N,N-tetraisopropylphosphoramidites. 
     
     
         10 . The method of  claim 8 , wherein said phosphorylating agent is selected from the group consisting of 2-bromoethyldichlorophosphate, tri methylsilylethyl dichlorophosphate, methyl dichlorophosphate, 2-chloro-2-oxo-1,3,2-dioxaphospholane, 2-chlorophenyl dichlorophosphate, and phosphorus oxychloride. 
     
     
         11 . A method of producing a composition of  claim 1 , wherein compound XI is prepared by conjugating guggulsterol with a drug directly or through a linker. 
     
     
         12 . The method of claim wherein compound XI is prepared by conjugating guggulsterol through a linker, wherein the linker is an alkyl group optionally substituted with a functional group selected from the groups consisting of 
     
     
         13 . A method comprising complexing a guggul-based lipid molecule of  claim 1  into a. composition comprising a liposome or lipid complex. 
     
     
         14 . The method of  claim 13 , wherein said complex is selected from a group consisting of micelles, vesicles, and emulsions, wherein if said composition comprises a plurality of micelles, said micelles are in the form of monomeric, dimeric, polymeric or mixed micelles. 
     
     
         15 . A method of retaining drug in a liposome or complex comprising providing one or more guggul derivatives of  claim 1  or preparing one or more guggul derivatives by a method of  claims 2 - 11 , and complexing said one or more guggul derivatives and a drug in a composition comprising a liposome or complex. 
     
     
         16 . The methods of  claims 13 - 15 , wherein said composition further comprises at least one of a phospholipid, a pegylated phospholipid, polyethylene glycol (PEG), a fatty acid, a sterol, cholic acid, or α-tocopherol. 
     
     
         17 . The method of  claim 16 , wherein said composition comprises at least one phospholipid, wherein said phopholipid comprises a phosphatidylcholine, a phosphatidylethanolamine, a phosphatidylglycerol, a phosphatidylserine, a phosphatidylinositol, or a phosphatidic acid. 
     
     
         18 . The method of  claim 16 , wherein said composition comprises at least one polyethylene glycol (PEG), wherein said polyethylene glycol (PEG) has an average molecular weight of 200-20,000. 
     
     
         19 . The method of  claim 16 , wherein said composition comprises at least one fatty acid, wherein said fatty acids is selected from a group consisting of fatty acids having chain length of C 4 -C 34 . 
     
     
         20 . The method of  claim 19 , wherein said fatty acid is saturated or unsaturated, and wherein said fatty acid is in acidic form, or in salt form. 
     
     
         21 . The method of  claim 16 , wherein said composition comprises at least one pegylated phospholipid selected from the group consisting of a pegylated derivative of disteroylhosphatidylglycerol, dimyristoylhosphatidylglycerol, or dioleoylphosphtidylglycerol. 
     
     
         22 . The method of  claim 16 , wherein said cholic acid is sodium deoxycholate. 
     
     
         23 . The method of  claim 16 , wherein said composition further comprises a sterol selected from a group consisting of cholesterol, derivatives of cholesterol, cholesterol sulfate, cholesterol succinate, cholesterol hemisuccinate, cholesterol oleate, cortisol, corticosterone, hydrocortisone, cholesterol-PEG, coprostanol, cholestanol, cholestane, β-sitosterol, lanosterol, campesterol, lathosterol, stigmasterol, stigmastanol, calciferol, guggulsterols, derivatives of guggulsterols, and mixtures thereof. 
     
     
         24 . The method of  claims 15 , wherein said drug comprises a therapeutically active agent. 
     
     
         25 . The method of  claim 24 , wherein said therapeutically active agent comprises an anticancer drug, an antiviral drug, antibacterial drug an antifungal drug, anti-inflammation drug, cholesterol lowering drug, and wherein at least one of said therapeutically agent is to treat cardiovascular disease, to treat neoplasia, to improve memory lose, to treat skin infection or skin disease, to treat hyperglycemia, or to enhance cognitive function. 
     
     
         26 . A method wherein at least one cationic guggul derivative of formulas VIII and XII of  claim 1  is used to facilitate intercellular delivery of a polynucleotide. 
     
     
         27 . A method of transfecting a cell with a polynucleotide, comprising (a) preparing a composition comprising a cationic guggul derivative of  claim 1  and said polynucleotide, and (b) contacting said cell with said composition whereby said polynucleotide is taken up by the cell. 
     
     
         28 . The method of  claim 26  or  27 , wherein said polynucleotide is selected from a group consisting of an oligonucleotide comprising DNA, RNA or DNA and RNA, antisense oligonucleotides, small interfering RNA (siRNA), microRNA (miRNA), aptamers, and ribozymes, and wherein said polynucleotide is a single-stranded or double-stranded. 
     
     
         29 . The method of any of  claims 14 - 28 , wherein said composition comprises unilamellar vesicles, multilamellar vesicles, or mixtures thereof. 
     
     
         30 . The method of any of  claims 13 - 29  wherein said liposome or complex has an overall charge selected from a negative charge, a positive charge, or a neutral charge. 
     
     
         31 . The method of any of  claims 13 - 30 , wherein the liposome or complex has a mean diameter of about 20 micron to less than 1 micron. 
     
     
         32 . The method of any of  claims 13 - 30 , wherein the liposome or complex has a mean diameter of about 500 nm to less than 100 nm. 
     
     
         33 . The method of any of  claims 13 - 32 , wherein said composition is in lyophilized form. 
     
     
         34 . The method  claim 33 , wherein said composition in lyophilized form comprises a cryoprotectant, wherein said cryoprotectant comprises one or more sugars are selected from a group consisting of trehalose, maltose, lactose, sucrose, and dextran. 
     
     
         35 . The method of any of  claims 13 - 33 , wherein said liposome or complex is in a powder form or in a solution form, in a suspension form, in an emulsion form, and wherein said liposome or complex is in a gel form, in micelle form, or in a paste form. 
     
     
         36 . The method of any of  claims 13 - 35 , wherein said composition is a tablet or is tilled in a capsule. 
     
     
         37 . A method of treating a cell, comprising preparing composition according to any of the  claim 1 , or  13 - 36 , and exposing said cell to said composition, 
     
     
         38 . The method of  claim 37 , wherein said cell is in vivo in a subject. 
     
     
         39 . A method of treating a human or animal disease, comprising administering a therapeutically effective amount of the composition prepared according to a method of  claim 1 ,  13 - 36  and exposing the composition to a human or animal in need thereof such that the composition is delivered to the human or animal patient. 
     
     
         40 . A kit of transfecting cells, said kit comprising a cationic guggul derivative of  claim 1  and one or more elements selected from the group consisting of a polynucleotide, instructions for formulating the cationic guggul derivative and polynucleotide into a preparation, instructions for transfecting cells using the cationic guggul derivative, reagents for facilitating transfection, containers for storing the cationic guggul derivative, containers for storing the polynucleotide, containers for storing the reagents, containers for storing a preparation including the cationic guggul derivative and polynucleotide, or containers for preparing the preparation, and materials to facilitate transfection. 
     
     
         41 . A composition produced according to a method of any of  claim 2 - 25  or  27 - 36 .