Ex vivo methods for predicting and confirming in vivo metabolism of pharmaceutically active compounds

Abstract

Methods and compositions for the catalytic oxidation of pharmaceutically active compounds, and more particularly to ex vivo methods for predicting in vivo metabolism of pharmaceutically active compounds, including predicting in vivo interaction between two or more pharmaceutically active compounds.

1 .- 24 . (canceled) 
     
     
         25 . A method for preparing oxidative metabolites for a pharmaceutically active compound, the method comprising;
 contacting a solution comprising one or more pharmaceutically active compounds with (1) one or more catalysts immobilized on a substrate; and (2) a co-catalyst comprising a nitrogen- or sulfur-containing amino acid derivative in an aqueous solution for a period sufficient for catalytic oxidation of the one or more pharmaceutically active compounds by the catalyst;   separating the solution from the one or more catalysts immobilized on a substrate; and   identifying the oxidative metabolites for the one or more pharmaceutically active compounds,   wherein the one or more catalysts are selected from the group consisting of sterically hindered and electronically activated metallotetraphenylporphyrins, metallophthalocyanines and metallosalen complexes.   
     
     
         26 . The method of  claim 25 , wherein the one or more catalysts are immobilized to a polymeric resin solid support. 
     
     
         27 . The method of  claim 25 , wherein the one or more catalysts are encapsulated in a polystyrene matrix. 
     
     
         28 . The method of  claim 25 , wherein the one or more catalysts are immobilized on a substrate contained within the flow chamber of a sample processing device. 
     
     
         29 . The method of  claim 25 , wherein the oxidative metabolites are identified by NMR, CMR, MS, HRMS, IR, or UV spectroscopy. 
     
     
         30 . The method of  claim 25 , further comprising isolating the resulting oxidative products by HPLC, UPLC, Flash Chromatography or other preparative chromatographic techniques. 
     
     
         31 . The method of  claim 25 , wherein the catalyst is a sterically hindered and electronically activated metallotetraphenylporphyrin compound of formula 1: 
       
         
           
           
               
               
           
         
         wherein R 1  is selected from the group consisting of Cl, Br, CH 3 , SO 3   − , CN, [N(R′) 3 ] + , COOR′, —OCONR′ 2 , —OMOM, CON—R′, CONR′ 2 , CH═NR′, SO 2 NR′ 2 , SO 2 R, CF and NO 2 ; 
         R 2  is selected from the group consisting of H, Cl, Br, CH 3 , SO 3   − , CN, [N(R′) 3 ] + , COOR′, —OCONR′ 2 , —OMOM, CON—R′, CONR′ 2 , CH═NR′, SO 2 NR′ 2 , SO 2 R, CF and NO 2 ; 
         R 3  is selected from the group consisting of H, Cl, Br, CH 3 , SO 3   − , CN, [N(R′) 3 ] + , COOR′, —OCONR′ 2 , —OMOM, CON—R′, CONR′ 2 , CH═NR′, SO 2 NR′ 2 , SO 2 R, CF and NO 2 ; 
         R 4  is selected from the group consisting of H, Cl, Br, CH 3 , SO 3   − , CN, [N(R′) 3 ] + , COOR′, —OCONR′ 2 , —OMOM, CON—R′, CONR′ 2 , CH═NR′, SO 2 NR′ 2 , SO 2 R, CF and NO 2 ; 
         R′ is H or a C1-C6 alkyl; 
         M is a transition metal, such as Fe, Zn, Co, Ni, Cu, Mn, Rh, Mg, Ru, Pt, and Pd; and 
         optionally wherein one or more axial ligands X selected from the group halogens (F, Cl, Br), OH, OCl, CO, [N(R′) 3 ] + , substituted or unsubstituted nitrogen- or sulfur-containing amino acid derivatives selected from the group consisting of imidazole, alkyl-substituted imidazoles, mercapto-substituted imidazoles, trifluoromethyl-substituted imidazoles, pyridine, alkyl-substituted pyridines, mercapto-substituted pyridines, trifluoromethyl-substituted pyridines, pyrimidine, alkyl-substituted pyrimidines, mercapto-substituted pyrimidines, trifluoromethyl-substituted pyrimidines, isoquinoline, alkyl-substituted isoquinolines, mercapto-substituted isoquinolines, trifluoromethyl-substituted isoquinolines, acridine, alkyl-substituted acridines, mercapto-substituted acridine, trifluoromethyl-substituted acridine, quinoline, benzoquinolines, alkyl-substituted quinolines, mercapto-substituted quinolines, trifluoromethyl-substituted quinolines, benzylmercaptan and thiophenol and/or a counter ion is included to maintain charge neutrality. 
       
     
     
         32 . The catalyst of  claim 31 , wherein R 1  is Cl, R 2  is H, R 3  is H, R 4  is Cl, M is Fe and X is Cl. 
     
     
         33 . The catalyst of  claim 31 , wherein R 1  is Cl, R 2  is H, R 3  is H, R 4  is Br, M is Fe and X is Cl. 
     
     
         34 . The catalyst of  claim 31 , wherein R 1  is Cl, R 2  is H and one R 2  is SO 3 Na, R 3  is H, R 4  is Br, M is Fe and X is Cl. 
     
     
         35 . The catalyst of  claim 31 , wherein R 1  is Cl, R 2  is H, R 3  is H, R 4  is Cl or Br, M is Ru and X is Cl. 
     
     
         36 . The method of  claim 25 , wherein the catalyst is a metallophthalocyanine compound of formula 2: 
       
         
           
           
               
               
           
         
         wherein R 1  is selected from the group consisting of Cl, Br, CH 3 , SO 3   − , CN, [N(R′) 3 ] + , COOR′, —OCONR′ 2 , —OMOM, CON-R′, CONR′ 2 , CH═NR′, SO 2 NR′ 2 , SO 2 R, CF and NO 2 ; 
         R 2  is selected from the group consisting of H, Cl, Br, CH 3 , SO 3   − , CN, [N(R′) 3 ] + , COOR′, —OCONR′ 2 , —OMOM, CON—R′, CONR′ 2 , CH═NR′, SO 2 NR′ 2 , SO 2 R, CF and NO 2 ; 
         R 3  is selected from the group consisting of H, Cl, Br, CH 3 , SO 3   − , CN, [N(R′) 3 ] + , COOR′, —OCONR′ 2 , —OMOM, CON—R′, CONR′ 2 , CH═NR′, SO 2 NR′ 2 , SO 2 R, CF and NO 2 ; 
         R 4  is selected from the group consisting of H, Cl, Br, CH 3 , SO 3   − , CN, [N(R′) 3 ] + , COOR′, —OCONR′ 2 , —OMOM, CON—R′, CONR′ 2 , CH═NR′, SO 2 NR′ 2 , SO 2 R, CF and NO 2 ; 
         wherein R′ is H or a C1-C6 alkyl;
 M is a transition metal, such as Fe, Zn, Co, Ni, Cu, Mn, Rh, Mg, Ru, Pt, and Pd; 
 
         and optionally wherein one or more axial ligands L selected from the group consisting of halogens (F, Cl, Br), OH, OCl, CO, [N(R′) 3 ] + , and substituted or unsubstituted nitrogen- or sulfur-containing amino acid derivatives selected from the group consisting of imidazole, alkyl-substituted imidazoles, mercapto-substituted imidazoles, trifluoromethyl-substituted imidazoles, pyridine, alkyl-substituted pyridines, mercapto-substituted pyridines, trifluoromethyl-substituted pyridines, pyrimidine, alkyl-substituted pyrimidines, mercapto-substituted pyrimidines, trifluoromethyl-substituted pyrimidines, isoquinoline, alkyl-substituted isoquinolines, mercapto-substituted isoquinolines, trifluoromethyl-substituted isoquinolines, acridine, alkyl-substituted acridines, mercapto-substituted acridine, trifluoromethyl-substituted acridine, quinoline, benzoquinolines, alkyl-substituted quinolines, mercapto-substituted quinolines, trifluoromethyl-substituted quinolines, benzylmercaptan and thiophenol and/or a counter ion is included to maintain charge neutrality. 
       
     
     
         37 . The catalyst of  claim 36 , wherein R 1  and R 2  are Cl. 
     
     
         38 . The catalyst of  claim 36 , wherein R 1  and R 2  are H. 
     
     
         39 . The method of  claim 25 , wherein catalyst is a metallosalen complex compound of formula 3: 
       
         
           
           
               
               
           
         
         wherein R 1  is selected from the group consisting of Cl, Br, CH 3 , SO 3   − , CN, [N(R′) 3 ] + , COOR′, —OCONR′ 2 , —OMOM, CON—R′, CONR′ 2 , CH═NR′, SO 2 NR′ 2 , SO 2 R, CF and NO 2 , 
         R 2  is the same or different and is selected from the group consisting of H, Cl, Br, CH 3 , —C(CH 3 ) 3 , SO 3   − , CN, [N(R′) 3 ] + , COOR′, OCONR′ 2 , —OMOM, CON—R′, CONR′ 2 , CH═NR′, SO 2 NR′ 2 , SO 2 R, CF and NO 2 , 
         R 3  is selected from the group consisting of H, Cl, Br, CH 3 , SO 3   − , CN, [N(R′) 3 ] + , COOR′, OCONR′ 2 , —OMOM, CON—R′, CONR′ 2 , CH═NR′, SO 2 NR′ 2 , SO 2 R, CF and NO 2 ; 
         R 4  is selected from the group consisting of H, Cl, Br, CH 3 , SO 3   − , CN, [N(R′) 3 ] + , COOR′, OCONR′ 2 , —OMOM, CON—R′, CONR′ 2 , CH═NR′, SO 2 NR′ 2 , SO 2 R, CF and NO 2 ; 
         wherein R′ is H or a C1-C6 alkyl; 
         M is a transition metal, such as Fe, Zn, Co, Ni, Cu, Mn, Rh, Mg, Ru, Pt, and Pd, 
         and optionally wherein one or more axial ligands X selected from the group consisting of halogens (F, Cl, Br), OH, OCl, CO, [N(R′) 3 ] + , and substituted or unsubstituted nitrogen- or sulfur-containing amino acid derivatives selected from the group consisting of imidazole, alkyl-substituted imidazoles, mercapto-substituted imidazoles, trifluoromethyl-substituted imidazoles, pyridine, alkyl-substituted pyridines, mercapto-substituted pyridines, trifluoromethyl-substituted pyridines, pyrimidine, alkyl-substituted pyrimidines, mercapto-substituted pyrimidines, trifluoromethyl-substituted pyrimidines, isoquinoline, alkyl-substituted isoquinolines, mercapto-substituted isoquinolines, trifluoromethyl-substituted isoquinolines, acridine, alkyl-substituted acridines, mercapto-substituted acridine, trifluoromethyl-substituted acridine, quinoline, benzoquinolines, alkyl-substituted quinolines, mercapto-substituted quinolines, trifluoromethyl-substituted quinolines, benzylmercaptan and thiophenoland/or a counter ion is included to maintain charge neutrality. 
       
     
     
         40 . The method of  claim 25 , further comprising an oxidizing agent. 
     
     
         41 . The method of  claim 25 , wherein the catalyst is present at less than 5% wt/wt catalyst/organic substrate. 
     
     
         42 . The method of  claim 41 , wherein the catalyst is present at less than 1% wt/wt catalyst/organic substrate. 
     
     
         43 . The method of  claim 41 , wherein the catalyst is present at less than 0.5% wt/wt catalyst/organic substrate. 
     
     
         44 . The method of  claim 40 , wherein the oxidizing agent is selected from the group consisting of organic and inorganic peroxides, oxygen donor molecules, peracids, hypochlorites, ozone, potassium hydrogen persulfate, 2,6-dichloropyridine-N-oxide and molecular oxygen. 
     
     
         45 . The method of  claim 25 , wherein the one or more pharmaceutically active compounds are selected from the group consisting of acetylcholine receptor stimulants and antagonists; adrenoreceptor-activated compounds, adrenoreceptor-blocking compounds, antihypertensive agents, vasodilators, cardiac glycosides, diuretics, histamine, serotonin, antihistamines, antihypertensives, polypeptides, antibiotics, anti-infective agents, antimicrobials, anticonvulsants, antidiabetic agents, antiemetics, steroids, sedatives, antiepileptic compounds, anesthetics, skeletal muscle relaxants, antidepressants, antipsychotics, analgesics, lithium, anticoagulants, cholinesterase inhibitors, procoagulants, HMG-CoA reductase inhibitors (statins), nonsteroidal anti-inflammatory agents, antimitotic agents, protease inhibitors, thyroid and antithyroid compounds, hypnotics, fibrinolytic agents, recombinant proteins, peptides, adrenocorticosteroids, gonadal hormones and inhibitors, immunomodulators, immunosuppressives, erectile dysfunction therapeutics, penicillins, cephalosporins, chloramphenicol, tetracyclines, polymyxins, antimyobacterial compounds, sulfonamides, narcotics, trimethoprim, antifungal agents, antiviral agents, non-steroidal anti-inflammatory compounds, anticancer agents, vaccines, antiprotozoal compounds, antacids, antiarythmics, and antihelminthic compounds. 
     
     
         46 . (canceled) 
     
     
         47 . The method of  claim 25 , wherein at least two pharmaceutically active compounds are contacted simultaneously with the one or more catalysts immobilized on a substrate. 
     
     
         48 . The method of  claim 25 , wherein at least three pharmaceutically active compounds are contacted simultaneously with the one or more catalysts immobilized on a substrate. 
     
     
         49 .- 56 . (canceled)