GERLACH MATTHIAS (DE)
SCHUCH FALK (DE)
| WO2003057896A1 | 2003-07-17 |
| Claims 1. A process for preparation of an N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound according to formula I or a pharmaceutically acceptable salt thereof, wherein R is a dicarboxylic group: formula (I) formula (II); said process comprising reacting N-Fmoc-doxorubicin (formula II) with a bis-acyl donor compound in the presence of a lipase in liquid organic media to acylate said N- Fmoc-doxorubicin at its C14-OH group, under conditions effective to remove water from the reaction mixture. 2. The process of claim 1, wherein the organic media is (i) an organic solvent selected from a group (A) consisting of dichloromethane, chloroform, tetrachloromethane, benzene, toluene, diethyl ether, methyl-tert- butyl ether, diisopropyl ether, cyclopentyl methyl ether, acetone, methyl- ethylketone, methyl-isobutylketone, anisole, dimethylsulfoxide, N-methyl-2- pyrrolidone, 1,4-dioxane, pyridine, acetonitrile, N,N-dimethyl formamide, tetrahydrofuran, 2-methyltetrahydrofuran, ethyl acetate, n-pentane, n-hexane, n- heptane, 2,2,4-trimethylpentane, cyclopentane, cyclohexane, cycloheptane, tert- butanol, tert-amyl alcohol; (ii) a mixture comprising a first organic solvent from group (B) selected from methyl- tert-butyl ether, toluene, benzene, cyclopentyl methyl ether, acetonitrile, tetrahydrofuran, 2-methyltetrahydrofuran, methyl-ethylketone, ethyl acetate, 1,4- dioxane, n-hexane, n-heptane, n-pentane, or 2,2,4-trimethyl pentane; and a second organic solvent from group (C) selected from methyl-ethylketone, acetone, cyclopentyl methyl ether, anisole, tetrahydrofuran, 2- methyltetrahydrofuran, methyl-isobutylketone, N-methyl-2-pyrrolidone, tert-amyl alcohol, tert-butanol, cyclopentane or diethyl ether; a mixture comprising a first organic solvent from group (B), a second organic solvent from group (C) and a third organic solvent from group (B) or group (C); or a higher order mixture, comprising of solvents selected from groups (A), (B) or (C). 3. The process of claim 1, wherein the organic media is a solvent system selected from a) mixtures with MTBE: al MTBE/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a2 MTBE/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a3 MTBE/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a4 MTBE/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a5 MTBE/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a6 MTBE/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a7 MTBE/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a8 MTBE/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a9 MTBE/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 al 0 MTBE/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 al 1 MTBE/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 al2 MTBE/diethyl ether in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b) mixtures with toluene: bl Toluene/MEK in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b2 Toluene/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b3 Toluene/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b4 Toluene/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b5 Toluene/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b6 Toluene/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b7 Toluene/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b8 Toluene/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b9 Toluene/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b 10 Toluene/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4 : 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 bl 1 Toluene/cyclopentane in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 bl2 Toluene/diethyl ether in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c) mixtures with benzene: cl Benzene/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c2 Benzene/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c3 Benzene/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c4 Benzene/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c5 Benzene/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c6 Benzene/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c7 Benzene/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3,1:4,1:5, or 1:6 c8 Benzene/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c9 Benzene/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 cl 0 Benzene/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 cl 1 Benzene/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 cl2 Benzene/diethyl ether in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d) mixtures with CPME: dl CPME/MEK in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d2 CPME/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d3 CPME/anisole in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d4 CPME/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d5 CPME/Me-THF in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d6 CPME/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d7 CPME/NMP in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d8 CPME/tAmylOH in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d9 CPME/tBuOH in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 dlO CPME/cyclopentane in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 dl 1 CPME/diethyl ether in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 e) mixtures with MeCN: el MeCN/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 e2 MeCN/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 e3 MeCN/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 e4 MeCN/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 e5 MeCN/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 e6 MeCN/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 e7 MeCN/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 e8 MeCN/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 e9 MeCN/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 elO MeCN/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 ell MeCN/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 el2 MeCN/diethyl ether in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 f) mixtures with THF: fl THF/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 12 THF/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 13 THF/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 f4 THF/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 f5 THF/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 f6 THF/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 17 THF/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 f8 THF/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 f9 THF/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 fl 0 THF/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 fl 1 THF/diethyl ether in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 g) mixtures with Me-THF: gl Me-THF/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 g2 Me-THF/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 g3 Me-THF/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 g4 Me-THF/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 g5 Me-THF/THF in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 g6 Me-THF/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 g7 Me-THF/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 g8 Me-THF/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 g9 Me-THF/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 glO Me-THF/cyclopentane in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 gl 1 Me-THF/diethyl ether in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 h) mixtures with MEK: hi MEK/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 h2 MEK/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 h3 MEK/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 h4 MEK/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 h5 MEK/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 h6 MEK/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 h7 MEK/NMP in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 h8 MEK/tAmylOH in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 h9 MEK/tBuOH in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 hlO MEK/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 hi 1 MEK/diethyl ether in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 i) mixtures with EtOAc: 11 EtOAc/MEK in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 12 EtOAc/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 13 EtOAc/CPME in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 i4 EtOAc/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 i5 EtOAc/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 16 EtOAc/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 17 EtOAc/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 18 EtOAc/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 19 EtOAc/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 il 0 EtOAc/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 il 1 EtOAc/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 il 2 EtOAc/diethyl ether in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 j) mixtures with 1,4-dioxane: j 1 1 ,4-dioxane/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 j2 1 ,4-dioxane/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 j3 1 ,4-dioxane/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 j4 1 ,4-dioxane/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 j5 1 ,4-dioxane/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 j6 1 ,4-dioxane/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 j7 1 ,4-dioxane/MIBK in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 j8 1 ,4-dioxane/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 j9 1 ,4-dioxane/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 j 10 1 ,4-dioxane/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 j 11 1 ,4-dioxane/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1 :3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 j 12 1 ,4-dioxane/ diethyl ether in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, furthermore preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 k) mixtures with n-hexane (hex): kl Hex/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 k2 Hex/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 k3 Hex/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 k4 Hex/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 k5 Hex/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 k6 Hex/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 k7 Hex/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 k8 Hex/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 k9 Hex/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 klO Hex/tBuOH in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 kl 1 Hex/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 kl2 Hex/diethyl ether in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 1) mixtures with « -heptane (hept): 11 Hept/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 12 Hept/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 13 Hept/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 14 Hept/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 15 Hept/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 16 Hept/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 17 Hept/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 18 Hept/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 19 Hept/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 110 Hept/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 111 Hept/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 112 Hept/diethyl ether in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 m) mixtures with n-pentane (pent): ml Pent/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 m2 Pent/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 m3 Pent/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 m4 Pent/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 m5 Pent/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 m6 Pent/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 m7 Pent/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 m8 Pent/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 m9 Pent/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 ml 0 Pent/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 ml 1 Pent/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 ml 2 Pent/diethyl ether in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 n) mixtures with isooctane: nl Isooctane/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 n2 Isooctane/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 n3 Isooctane/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 n4 Isooctane/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 n5 Isooctane/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 n6 Isooctane/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 n7 Isooctane/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 n8 Isooctane/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 n9 Isooctane/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 nlO Isooctane/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 nil Isooctane/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 nl2 Isooctane/diethyl ether in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 process of claim 1, wherein the bis-acyl donor compound is a dicarboxylic acid selected from the group malonic acid, succinic acid, glutaric acid, adipic acid, pimelic acid, suberic acid, azelaic acid, sebacic acid, undecanedioic acid, dodecanedioic acid, 1,12-dodecanedicarboxylic acid, fumaric acid, maleic acid, glutaconic acid, traumatic acid, muconic acid, cis aconitic acid, phthalic acid, isophthalic acid, terephthalic acid, cyclohexan-l,4-dicarboxylic acid, 3-(2-carboxyethyldisulfanyl)propionic acid, diglycolic acid, thio diglycolic acid, 3-(2-carboxyethoxy)propionic acid, 3-[2-(2-carboxyethoxy)ethoxy]propionic acid, 3- {2-[2-(2-carboxyethoxy)ethoxy]ethoxy} propionic acid, 3-(2-{2-[2-(2- carboxy-ethoxy)ethoxy]ethoxy} ethoxy)propionic acid, 3-[2-(2- {2-[2-(2- carboxyethoxy)ethoxy]ethoxy} ethoxy)ethoxy]propionic acid, 3- {2-[2-(2- {2-[2-(2- carboxyethoxy)ethoxy] ethoxy}ethoxy)ethoxy]ethoxy}propionic acid and 3-(2- {2-[2-(2- {2-[2-(2-carboxyethoxy)ethoxy]ethoxy } ethoxy)ethoxy]ethoxy} ethoxy)- propionic acid; or is an anhydride selected from the group of succinic anhydride, glutaric anhydride, adipic anhydride, pimelic anhydride, suberic anhydride, maleic anhydride, cis aconitic anhydride, phthalic anhydride, diglycolic anhydride and thio diglycolic anhydride. 5. The process of claim 1, wherein the lipase (i) is a lipases from microbial sources selected from the group of Alicaligenes sp., Aspergillus, Aspergillus niger, Aspergillus oryzae, Alcaligenes sp., Candida antartica A, Candida antartica B, Candida cylindracea sp., Candida rugosa, Fusarium solanipisi, Mucor miehei, Mucor javanicus, Penicillium roqueforti, Pseudomonas cepacia, Pseudomonas fluorescens, Pseudomonas sp., Pseudomonas stutzeri, Rhizomucor miehei, Rhizopus arrhizus, Rhizopus delemar, Rhizopus niveus, Rhizopus oryzae, Thermomyces lanuginosus, Bacillus subtilis and Porcine pancreas; (ii) is a genetically modified lipase selected from the group of Alicaligenes sp., Aspergillus, Aspergillus niger, Aspergillus oryzae, Alcaligenes sp., Candida antartica A, Candida antartica B, Candida cylindracea sp., Candida rugosa, Fusarium solanipisi, Mucor miehei, Mucor javanicus, Penicillium roqueforti, Pseudomonas cepacia, Pseudomonas fluorescens, Pseudomonas sp., Pseudomonas stutzeri, Rhizomucor miehei, Rhizopus arrhizus, Rhizopus delemar, Rhizopus niveus, Rhizopus oryzae, Thermomyces lanuginosus, Bacillus subtilis and Porcine pancreas; wherein the lipase is immobilized to a solid support selected from the group of diatomaceous earth, silica, ceramic, polypropylene, polystyrene or acrylic resins or is immobilized by covalent cross-linking of enzyme aggregates. 6. The process of claim 1, wherein the lipase is added in 1 to 4.79 weight equivalents (wt. eq.), preferably in 2, 3, 3.5, 4, 4.5, 4.79 wt. eq. with respect to the N-Fmoc-doxorubicin starting material 7. The process of claim 1 , wherein the bis-acyl donor is added in 1 to 12 equivalents (wt. eq.), preferably 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 eq. with respect to the N-Fmoc- doxorubicin starting material. 8. The process of claim 1 , wherein the solvent volume is between 40 and 80 times, preferably 60 or 80 times as high as the weight of the N-Fmoc-doxorubicin starting material. 9. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 2 to 4.79 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising a first organic solvent selected from the group of methyl-fert-butyl ether, tetrahydrofuran, 1 ,4-dioxane or acetonitrile; and a second organic solvent selected from the group of methyl-ethylketone, acetone, CPME, anisole, 2-methyltetrahydrofuran, or methyl- isobutylketone; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the Fmoc-doxorubicin starting material; wherein the ratio between said first solvent and said second solvent is covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3: 1 to 1 :3, further more preferably from 1 : 1 to 1 :3, and most preferably the ratio is selected from the group consisting of 6:1, 5: 1, 4:1, 3:1, 2:1, 1 :1, 1 :2, 1 :3, 1 :4, 1 :5, or 1 :6; under conditions effective to remove water from the reaction mixture. 10. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 4.0 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising a first organic solvent selected from the group of methyl-fert-butyl ether, tetrahydrofuran, 1 ,4-dioxane or acetonitrile; and a second organic solvent selected from the group of methyl-ethylketone, acetone, CPME, anisole, 2-methyltetrahydrofuran, or methyl- isobutylketone; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the Fmoc-doxorubicin starting material; wherein the ratio between said first solvent and said second solvent is covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3: 1 to 1 :3, further more preferably from 1 : 1 to 1 :3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6; under conditions effective to remove water from the reaction mixture. 11. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 3.5 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising a first organic solvent selected from the group of methyl-fert-butyl ether, tetrahydrofuran, 1,4-dioxane or acetonitrile; and a second organic solvent selected from the group of methyl-ethylketone, acetone, CPME, anisole, 2-methyltetrahydrofuran, or methyl- isobutylketone; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the Fmoc-doxorubicin starting material; wherein the ratio between said first solvent and said second solvent is covered by the range from6:l to 1:6, preferably from4:l to 1:4, more preferably from3:l to 1:3, further more preferably from 1 : 1 to 1 :3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6; under conditions effective to remove water from the reaction mixture. 12. The process of claims 9-11, wherein the organic media is a solvent system selected from a) mixtures with MTBE: al MTBE/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a2 MTBE/acetone in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a3 MTBE/CPME in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a4 MTBE/anisole in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a5 MTBE/Me-THF in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a6 MTBE/MIBK in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b) mixtures with THF: bl THF/MEK in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b2 THF/acetone in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b3 THF/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b4 THF/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b5 THF/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b6 THF/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c) mixtures with 1,4-dioxane: cl 1 ,4-dioxane/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c2 1 ,4-dioxane/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c3 1 ,4-dioxane/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c4 1 ,4-dioxane/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c5 1 ,4-dioxane/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c6 1 ,4-dioxane/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d) mixtures with MeCN: dl MeCN/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d2 MeCN/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d3 MeCN/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d4 MeCN/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d5 MeCN/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d6 MeCN/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6. The process of claim 9, wherein the organic media is a solvent system selected from a) mixtures with MTBE: al MTBE/MEK in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a2 MTBE/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a3 MTBE/CPME in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a4 MTBE/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a5 MTBE/Me-THF in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a6 MTBE/MIBK in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b) mixtures with THF: bl THF/MEK in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b2 THF/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b3 THF/CPME in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b4 THF/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b5 THF/Me-THF in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b6 THF/MIBK in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c) mixtures with 1,4-dioxane: cl 1 ,4-dioxane/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c2 1 ,4-dioxane/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c3 1 ,4-dioxane/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c4 1 ,4-dioxane/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c5 1 ,4-dioxane/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c6 1 ,4-dioxane/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d) mixtures with MeCN: dl MeCN/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d2 MeCN/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d3 MeCN/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d4 MeCN/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d5 MeCN/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d6 MeCN/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6. 13) The process of claim 9, wherein the organic media is a solvent system selected from a) mixtures with MTBE: al MTBE/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a2 MTBE/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a3 MTBE/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a4 MTBE/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a5 MTBE/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 a6 MTBE/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b) mixtures with THF: bl THF/MEK in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b2 THF/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b3 THF/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b4 THF/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b5 THF/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b6 THF/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c) mixtures with 1,4-dioxane: cl 1 ,4-dioxane/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c2 1 ,4-dioxane/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c3 1 ,4-dioxane/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c4 1 ,4-dioxane/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 1 ,4-dioxane/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 1 ,4-dioxane/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d) mixtures with MeCN: dl MeCN/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d2 MeCN/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d3 MeCN/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d4 MeCN/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d5 MeCN/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d6 MeCN/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6: 1, 5: 1, 4:1, 3:1, 2: 1, 1 :1, 1 :2, 1 :3, 1 :4, 1 :5, or 1 :6. 14. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 4 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the Fmoc-doxorubicin starting material wherein the solvent is a mixture comprising methyl- fert-butyl ether and methyl- ethylketone in a ratio of 3 : 1 ; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is resulting in 76% of N-Fmoc-Doxorubicin-O-hemiglutarate after 20 h (as analyzed by HPLC). 15. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 3.5 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the Fmoc-doxorubicin starting material wherein the solvent is a mixture comprising methyl- fert-butyl ether and methyl- ethylketone in a ratio of 3 : 1 ; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is resulting in 73% of N-Fmoc-Doxorubicin-O-hemiglutarate after 20 h (as analyzed by HPLC). 16. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 4 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising methyl- fert-butyl ether and anisole in a ratio of 1 : 1 ; wherein the bis-acyl donor compound is glutaric acid, added in 5 weight equivalents with respect to the N-Fmoc-doxorubicin starting material, and glutaric anhydride, added in 5 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent volume is 80 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is resulting in 88% of N-Fmoc-Doxorubicin-O-hemiglutarate after 25h (as analyzed by HPLC). 17. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 4 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising methyl- fert-butyl ether and Me-THF in a ratio of 1 : 1 ; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is completed after 4 h to give an isolated yield of 91.1 % (as analyzed by HPLC). 18. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 3.5 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising methyl- tert-butyl ether and Me-THF in a ratio of 1 : 1 ; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is resulting in 83% of N-Fmoc-Doxorubicin-O-hemiglutarate after 20 h (as analyzed by HPLC). 19. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 4 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising methyl- tert-butyl ether and Me-THF in a ratio of 1 :4; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is resulting in 78% of N-Fmoc-Doxorubicin-O-hemiglutarate after 20 h (as analyzed by HPLC). 20. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 3.5 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising methyl- tert-butyl ether and Me-THF in a ratio of 1 :4; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is resulting in 63% of N-Fmoc-Doxorubicin-O-hemiglutarate after 20 h (as analyzed by HPLC). 21. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 4 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising methyl- fert-butyl ether and Me-THF in a ratio of 1 :6; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is resulting in 79% of N-Fmoc-Doxorubicin-O-hemiglutarate after 20 h (as analyzed by HPLC). 22. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 3.5 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising methyl- fert-butyl ether and Me-THF in a ratio of 1 :6; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is resulting in 79% of N-Fmoc-Doxorubicin-O-hemiglutarate after 20 h (as analyzed by HPLC). The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 4 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising methyl- fert-butyl ether and Me-THF in a ratio of 3:1; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is resulting in 87% of N-Fmoc-Doxorubicin-O-hemiglutarate after 20 h (as analyzed by HPLC). The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 3.5 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising methyl- fert-butyl ether and Me-THF in a ratio of 3:1; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is resulting in 89% of N-Fmoc-Doxorubicin-O-hemiglutarate after 20 h (as analyzed by HPLC). 25. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 4 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising methyl- fert-butyl ether and Me-THF in a ratio of 6:1; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is resulting in 88% of N-Fmoc-Doxorubicin-O-hemiglutarate after 20 h (as analyzed by HPLC). The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 3.5 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising methyl- fert-butyl ether and Me-THF in a ratio of 6:1; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is resulting in 88% of N-Fmoc-Doxorubicin-O-hemiglutarate after 20 h (as analyzed by HPLC). The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 3.5 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising 1,4-Dioxan and Me-THF in a ratio of 1 :5; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is resulting in 88% of N-Fmoc-Doxorubicin-O-hemiglutarate after 20 h (as analyzed by HPLC). 28. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 3.5 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising CPME and THF in a ratio of 1 :2; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is resulting in 84% of N-Fmoc-Doxorubicin-O-hemiglutarate after 20 h (as analyzed by HPLC). 29. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 3.5 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising CPME and THF in a ratio of 1 : 1 ; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is resulting in 85% of N-Fmoc-Doxorubicin-O-hemiglutarate after 20 h (as analyzed by HPLC). 30. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 3.5 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising CPME and THF in a ratio of 2:1; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is resulting in 88% of N-Fmoc-Doxorubicin-O-hemiglutarate after 20 h (as analyzed by HPLC). 31. The process of claim 1 , wherein said N-Fmoc-doxorubicin- 14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 3.5 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising CPME and THF in a ratio of 5 : 1 ; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is resulting in 90% of N-Fmoc-Doxorubicin-O-hemiglutarate after 20 h (as analyzed by HPLC). 32. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 4 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising methyl- fert-butyl ether and Me-THF in a ratio of from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 3.5 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising methyl- fert-butyl ether and Me-THF in a ratio of from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture. 34. The process of claim 1, wherein said N-Fmoc-doxorubicin- 14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 4 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the N-Fmoc-doxorubicin starting material wherein the solvent is a mixture comprising methyl- fert-butyl ether and Me-THF in a ratio offrom 6: l to 1 :6, preferably from 3: 1 to 1 :3, more preferably from 1 :1 to 1 :3, further more preferred from 1 : 1,5 to 1 :2,5 and most preferred 1 :2; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture. 35. The process of claim 1, wherein said N-Fmoc-doxorubicin- 14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 3.5 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the N-Fmoc-doxorubicin starting material wherein the solvent is a mixture comprising methyl- tert-butyl ether and Me-THF in a ratio of from 6: 1 to 1 :6, preferably from 3:1 to 1 :3, more preferably from 1 :1 to 1 :3, further more preferred from 1 : 1,5 to 1 :2,5 and most preferred 1 :2; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture. 36. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 4 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising methyl- tert-butyl ether and Me-THF in a ratio of 1 :2; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the Fmoc-doxorubicin starting material; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is completed after 4 h to give an isolated yield of 91% (as analyzed by HPLC). 37. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-14-O-hemiglutarate, wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 3.5 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising methyl- tert-butyl ether and Me-THF in a ratio of 1 :2; wherein the bis-acyl donor compound is glutaric acid, added in 10 equivalents with respect to the Fmoc-doxorubicin starting material; wherein the solvent volume is 60 times as high as the weight of N-Fmoc-doxorubicin; under conditions effective to remove water from the reaction mixture; wherein the reaction is completed after 6 h to give an isolated yield of 92% (as analyzed by HPLC). The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmoc-doxorubicin-O-hemisuccinate, N-Fmoc-doxorubicin- O-hemipimelate, N-Fmoc-doxorubicin-0-hemifumarate, N-Fmoc-doxorubicin-O-hemi- 3-(2-carboxyethoxy)propionate, N-Fmoc-doxorubicin-O-hemiadipate, N-Fmoc- doxorubicin-O-hemisuberate, N-Fmoc-doxorubicin-O-hemiazelate, N-Fmoc- doxorubicin-O-hemisebacate, N-Fmoc-doxorubicin-O-hemiundecanedioate, N-Fmoc- doxorubicin-O-hemidodecanedio ate, N-Fmo c-doxorubicin- O-hemimaleate, N-Fmoc- doxorubicin-O-hemiterephthalate, or N-Fmo c-doxorubicin- 0-hemi-(2- carboxyethyldisulfanyl)propionate; wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 4 weight equivalents with respect to the N-Fmoc-doxorubicin starting material; wherein the solvent is a mixture comprising a first organic solvent selected from the group of methyl-fert-butyl ether, CPME, tetrahydrofuran, 1 ,4-dioxane or acetonitrile; and a second organic solvent selected from the group of acetone, methyl-ethylketone, methyl- isobutylketone, anisole, or 2-methyltetrahydrofuran; wherein the ratio between said first solvent and said second solvent is covered by the range from 6:1 to 1 :6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1 :3, further more preferably from 1 :1 to 1 :3, and most preferably may be 1 :1, 2: 1, 3:1, 4: 1; 5:1, 6: 1, 1 :2, 1 :3, 1 :4, 1 :5, or 1 :6; wherein benzene or toluene or both may be added to remove water from the reaction mixture; wherein the bis-acyl donor compound is glutaric acid; wherein said glutaric acid is added in 10 equivalents with respect to the N-Fmoc- doxorubicin starting material; wherein the solvent volume is 60 times as high as the weight of N-Fmo c-doxorubicin; under conditions effective to remove water from the reaction mixture. 39. The process of claim 1, wherein said N-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester compound is N-Fmo c-doxorubicin- O-hemisuccinate, N-Fmoc-doxorubicin- O-hemipimelate, N-Fmoc-doxorubicin-O-hemifumarate, N-Fmo c-doxorubicin- O-hemi- 3 -(2-carboxyethoxy propionate, N-Fmoc-doxorubicin-O-hemiadipate, N-Fmoc- doxorubicin-O-hemisuberate, N-Fmoc-doxorubicin-O-hemiazelate, N-Fmoc- doxorubicin-O-hemisebacate, N-Fmoc-doxorubicin-O-hemiundecanedioate, N-Fmoc- doxorubicin-O-hemidodecanedio ate, N-Fmo c-doxorubicin- O-hemimaleate, N-Fmoc- doxorubicin-O-hemiterephthalate, or N-Fmo c-doxorubicin- 0-hemi-(2- carboxyethyldisulfanyl)propionate; wherein the lipase is Candida antartica B immobilized to a solid acrylic support, added in 3.5 weight equivalents with respect to the N-Fmo c-doxorubicin starting material; wherein the solvent is a mixture comprising a first organic solvent selected from the group of methyl-fert-butyl ether, CPME, tetrahydrofuran, 1,4-dioxane or acetonitrile; and a second organic solvent selected from the group of acetone, methyl-ethylketone, methyl- isobutylketone, anisole, or 2-methyltetrahydrofuran; wherein the ratio between said first solvent and said second solvent is covered by the range from 6:1 to 1 :6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1 :3, further more preferably from 1 :1 to 1 :3, and most preferably may be 1 :1, 2: 1, 3:1, 4: 1; 5: 1, 6:1, 1 :2, 1 :3, 1 :4, 1 :5, or 1 :6; wherein benzene or toluene or both may be added to remove water from the reaction mixture; wherein the bis-acyl donor compound is glutaric acid; wherein said glutaric acid is added in 10 equivalents with respect to the N-Fmoc- doxorubicin starting material; wherein the solvent volume is 60 times as high as the weight of N-Fmo c-doxorubicin; under conditions effective to remove water from the reaction mixture. |
Description
Technical field
The present invention describes a process for the use of enzymes for the selective synthesis ofN-Fmoc-doxorubicin-14-O-dicarboxylic acid mono ester derivatives using bis-acyl donor compounds such as dicarboxylic acids or anhydrides.
Prior Art
Starting from N-Fmoc-doxorubicin, the chemical acylation to formN-Fmoc-doxorubicin-14- O-hemiglutarate as intermediate in the synthesis of LHRH analogs containing doxorubicin is described b Nagy et al. (J. Proc. Natl. Acad. Sci. USA 1996, 93, 7269).
N-Fmoc-Doxorubicin acylated N-Fmoc-Doxorubicin
Figure 1
Addition of glutaric anhydride to N-Fmoc-doxorubicin (obtained from C3'-NH 2 - doxorubicin) in anhydrous DMF in the presence of DIPEA overnight yielded a crude product, which by HPLC analyses showed -75% of the desired hemiglutarate, 18% of unreacted N-Fmoc-doxorubicin and 7% other impurities, including 4% attributed to the 4'- OH-hemiglutarate. HPLC purification resulted in a yield of 64% hemiglutarate. The low solubility of N-Fmoc-doxorubicin hemiglutarate was identified as a problem and used as explanation, why the hemiglutarate was usually not isolated, but directly converted to the final product.
Corresponding data are disclosed in US5843903, filed by Schally, Nagy and Cai.
In an overall judgment, these results were experimentally confirmed as shown by reference experiment 1.
Reference experiment 1: Synthesis ofN-Fmoc-doxorubicin-O-hemiglutarate using glutaric anhydride catalyzed by DIPEA as described by Nagy et al. (J. Proc. Natl. Acad. Sci. USA 1996, 93, 7269)
N-Fmoc-doxorubicin (13.46 g), DIPEA (4.54 g) and glutaric anhydride (8.02g) were dissolved in DMF (128 mL). The reaction mixture was stirred for 7 h at room temperature. The progress of the reaction was monitored by HPLC. The reaction mixture was cooled to 15 °C and added into 662 mL of a aqueous acetic acid solution (w/w = 5 %). The product is filtered, washed with aqueous acetic acid solution (w/w = 5 %) (142 mL) and dried under vacuum to constant weight at 50 °C to give crude N-Fmoc-doxorubicin- 14-O-hemiglutarate. Yield: 15.2 g (95 %)
Purity: HPLC: 51.8 % (Impurity profile - see Appendix 1).
HPLC analysis of isolated, crude hemiglutarate showed 51.8% N-Fmoc-doxorubicin-14- O-hemiglutarate, 1.3% of unreacted N-Fmoc-doxorubicin and 11.1% of the 4'-0-, 14-0- dihemiglutarate.
While not surprising, an analogous experiment without base resulted in nearly no conversion. Reference experiment 2: Synthesis ofN-Fmoc-doxorubicin-O-hemiglutarate using glutaric anhydride without base
The same reaction conditions as used in Example 1 were applied for the acylation without DIPEA. The reaction was slow and 64 % N-Fmoc-doxorubicin remained unreacted after overnight stirring. Therefore no work-up was performed.
Yield: not isolated, Purity: n.a.
Selective acylation of the C14-position of doxorubicin derivatives using enzymes
The utility of enzymes achieving the selective modification of structurally complex substrates under mild conditions is also well documented (Wong et al, Enzymes in Synthetic Organic Chemistry, 1 st ed. Pergamon: Tarrytown, NY, Oxford, U.K., 1994).
Cotterill et al. (Organic Process Research & Development 2005, 9, 818) reported the use of lipases for the regioselective esterification of the C14-hydroxy group of N-trifluoroacetyl- doxorubicin with a series of (mono) carboxylic acids.
Furthermore, it is reported that,
• it has been necessary to protect the C3'-NH 2 group in order to regioselectively
acylate the C14-OH group, since the C3'-NH 2 moiety and the C4'-OH group of doxorubicin are known to be chemical more reactive than the C14-OH group
• the removal of water is essential in order to allow the reaction to complete
• the selectivity of acylation (C14-OH vs C4'-OH) is clearly dependent upon the reaction solvent
• the solubility of N-trifluoroacetyl-doxorubicin in the reaction solvents is a key
prerequisite for production of gram quantities
• the best conditions to conduct the reaction were found to be at 58°C in 8h in a 3 : 1 MTBE/2-butanone mixture to give the desired N-trifluoroacetyl-doxorubicin valerate in 94% yield, including 4% byproducts.
These teachings are confirmed by reference experiments 3 and 4.
Reference experiment 3 shows, that the lipase is essential for the process of adding glutaric acid to N-Fmoc-doxorubicin, as otherwise N-Fmoc-doxorubicin remains unreacted. Reference experiment 4 shows, that protection of the C3'-NH 2 group is essential as otherwise the yield of the desired doxorubicin hemiglutarate is only 14% (as found by HPLC analysis), while starting material doxorubicin remains to 7%. The main product (47%) was not structurally identified.
Reference experiment 3: Synthesis of N-Fmoc-doxorubicin-O-hemiglutarate using glutaric acid without lipase
N-Fmoc-doxorubicin (50 mg) was suspended in MTBE/MEK 3 : 1 (v/v) (10 mL) and to this was added glutaric acid (82 mg). The reaction flask was fitted with a dropping funnel and condenser assembly in which the funnel was filled with 3 A molecular sieves (ca. 10 g).
The reaction mixture was stirred vigorously and heated for 25 h at reflux temperature. The progress of the reaction was monitored by HPLC. No product could be detected.
Yield: not isolated
Purity: product not detectable (HPLC - see Appendix 2)
Reference experiment 4: Synthesis of doxorubicin-O-hemiglutarate using glutaric acid as bis-acyl donor under lipase conditions
Doxorubicin-O-hemiglutarate
Doxorubicin (50 mg) was suspended in MTBE/MEK 3 : 1 (v/v) (10 mL) and to this was added lipase acrylic resin from Candida Antarctica B (233 mg, 10500 units/g resin) and glutaric acid (109 mg). The reaction flask was fitted with a dropping funnel and condenser assembly in which the dropping funnel (but not the pressure equalizing tube) was filled with 3 A molecular sieves (ca. 10 g). The reaction mixture was stirred vigorously and heated for 25 h at reflux temperature. The progress of the reaction was monitored by HPLC. Only less than 15 % of product (doxorubicin-O-hemiglutarate) beside other reaction products could be detected via HPLC. Therefore no work-up was performed.
Yield: not isolated
Purity: 14.1 % (HPLC- see Appendix 3)
Michels, Rich, Meckler and Coterrill filed a patent application (WO03/057896) disclosing a process for preparation of doxorubicin derivatives, which may carry N-alkylated amino sugar or a non-basic sugar moieties, and which are acylated via non-chemically modified lipases on the C14-OH group. A broad range of solvents, non-activated and activated acyl donors as well as lipases are described to be applicable within the claimed process.
Process examples either start from N-trifluoroacetyl-doxorubicin or from N-alloc- doxorubicin and use two lipases (PS30 and Chirazym L2, C3) to acylate to the C14- butyrate, -valerate and a PEP derivative, respectively.
To overcome the low solubility of anthracycline derivatives containing a free amine, conversion to N-alloc doxorubicin is proposed, which is disclosed as "highly soluble in less polar organic solvents suitable for synthesis".
Cotterill et al. (Biotech and Bioeng. 2008, 101, 435) reported a three step synthesis on unprotected doxorubicin- 14-O-esters using lipases starting from doxorubicin HC1 salt. The low solubility of unprotected doxorubicin is described as the key problem to be solved for efficient synthesis of doxorubicin- 14-O-esters.
The alloc group was identified as a modifying agent for the synthesis of doxorubicin C14 esters due to the improved solubility of N-alloc doxorubicin in a variety of process solvents and the facile removal of the alloc group to afford the desired products.
With regard to regioselectivity, the introduction of small protecting groups, such as Boc or alloc, onto the doxorubicin-3'-NH 2 group did not provide sufficient steric hindrance to prevent acylation of the C4'-OH. The introduction of larger protection groups (e.g. N- Fmoc) is described to partially influence the acylation of the adjacent C4'-OH, but due to the poor solubility of N-Fmoc-doxorubicin it is stated to be an unpractical approach.
Since the chosen alloc group did not provide for specific C14 esterification, selectivity was reported to be achieved by carefully selecting the binary solvent system. While one solvent was chosen with respect to maximizing solubilization of N-alloc doxorubicin, the other solvent was selected in order to maintain catalytic activity and doxorubicin-C14 selectivity.
Summarizing the prior art, it was known, that for a C14-OH esterification of doxorubicin, its C3'-NH 2 group needs to be modified, as otherwise no clean esterification occurs. Said
C3'- NH 2 group should be modified preferable by a bulky protection group that can shield the C4'-OH group to prevent its undesired esterification. The N-trifluoroacetyl- and the N-alloc groups were used to modify the C3'-NH 2 group. However, the N-Fmoc protecting group was clearly stated as unsuited due to the poor solubility of N-Fmoc-doxorubicin.
Summary of the Invention
It has surprisingly been found, that N-Fmoc-doxorubicin can indeed be used as starting material in enzyme catalyzed acylation reactions with very good yield and high selectivity for C14-OH esterification, despite its low solubility. The present inventions therefore provides a solution to a problem clearly identified in the prior art for esterification of N-Fmoc- doxorubicin, being specifically expressed by the statement "the solubility of N-Fmoc- doxorubicin was ... not synthetically useful" (see Cotterill et al. Biotech and Bioeng. 2008, p 438, left column, 1 st paragraph).
The use of the N-Fmoc protecting group to modify the doxorubicin C3'-NH 2 group is advantageous compared to other commonly used nitrogen modifying groups, because this bulky group shields the doxorubicin C4'-OH group and thus provides due to enhanced selectivity for better yields in shorter reaction times, combined with a better impurity profile.
The lipase catalyzed esterification of N-alloc-doxorubicin or N-trifluoroacetyl-doxorubicin with glutaric acid yielded 51% and 70% of the corresponding products, respectively (see reference experiments 5 and 6). Under the same conditions, N-Fmoc-doxorubicin was converted to N-Fmoc-doxorubicin hemiglutarate in 92% yield (see example la).
Further optimization resulted in yields up to 95%, while reducing the reaction time to 4h (see example Id). This was achieved by organic binary solvent systems, wherein one component provided the possibility to remove water from the reaction vessel in order to drive the reaction to completion, while the other component ensured lipase activity and a sufficient solubility of the starting materials. The need for a solvent to direct regioselective esterification was overcome due to the directing effects of the bulky Fmoc group. Description of the invention
The present invention relates to a process for preparation of an N-Fmoc-doxorubicin-14-O- dicarboxylic acid mono ester compound according to formula (I) or a pharmaceutically acceptable salt thereof, wherein in formula (I) R is a dicarboxylic group:
formula (II);
said process comprising reacting N-Fmoc-doxorubicin (formula II) with a bis-acyl donor compound in the presence of a lipase in liquid organic media to acylate said N-Fmoc- doxorubicin with high selectivity at its C14-OH group, under conditions effective to remove water from the reaction mixture.
The reaction of the present invention is carried out in liquid organic media. Possible liquid organic media include but are not limited to organic solvents of a group (A) selected from dichloromethane, chloroform, tetrachloromethane, benzene, toluene, diethyl ether, methyl- tert-butyl ether ("MTBE"), diisopropyl ether, cyclopentyl methyl ether ("CPME"), acetone, methyl-ethylketone ("MEK"), methyl-isobutylketone ("MIBK"), anisole
(PhOMe), dimethylsulfoxide ("DMSO"), N-methyl-2-pyrrolidone ("NMP"), 1,4-dioxane, pyridine, acetonitrile ("MeCN"), N,N-dimethyl formamide ("DMF"), tetrahydrofuran ("THF"), 2-methyltetrahydrofuran ("Me-THF"), ethyl acetate ("EtOAc"), n-pentane ("pent"), n-hexane ("hex"), n-heptane ("hept"), 2,2,4-trimethylpentane ("isooctane"), cyclopentane, cyclohexane, cycloheptane, tert-butanol ("tBuOH"), tert-amyl alcohol ("tAmyl-OH"). Liquid organic media include mixtures of two organic solvents, wherein one solvent is chosen to facilitate azeotropic distillation, i.e. the removal of water from the reaction vessel by azeotropic distillation in order to drive the reaction to completion. The other solvent is chosen to ensure lipase activity and a sufficient solubility of the starting materials.
Solvents facilitating azeotropic distillation include, but are not limited to solvents from a group (B) selected from methyl-tert-butyl ether, toluene, benzene, cyclopentyl methyl ether, acetonitrile, tetrahydrofuran, 2-methyltetrahydrofuran, methyl-ethylketone, ethyl acetate, 1,4-dioxane, n-hexane, n-heptane, n-pentane, or 2,2,4-trimethyl pentane.
The use of other solvents being also able to facilitate azeotropic distillation is considered analogous to the mentioned solvents and is therefore included into the process of the present application.
Solvents to ensure lipase activity and a sufficient solubility of the starting materials include, but are not limited to solvents from a group (C) selected from methyl-ethylketone, acetone, cyclopentyl methyl ether, anisole, tetrahydrofuran, 2-methyltetrahydrofuran, methyl- isobutylketone, N-methyl-2-pyrrolidone, tert-amyl alcohol, tert-butanol, cyclopentane or diethyl ether.
The use of other solvents being also able to ensure lipase activity and a sufficient solubility of the starting materials is considered analogous to the mentioned solvents and is therefore included into the process of the present application.
The ratio between said first solvent and said second solvent is covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3: 1 to 1 :3, further more preferably from 1 :1 to 1 :3, and most preferably the ratio is selected from the group consisting of 6: 1, 5: 1, 4: 1, 3: 1, 2: 1, 1 : 1, 1 :2, 1 :3, 1 :4, 1 :5, or 1 :6.
Mixtures of two organic solvents include:
a) mixtures with MTBE:
al MTBE/MEK in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1 :3, further more preferably from 1 :1 to 1 :3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1 :1, 1 :2, 1 :3, 1 :4, 1 :5, or 1 :6
a2 MTBE/acetone in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1 :1 to 1 :3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
a3 MTBE/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2,
1:3, 1:4, 1:5, or 1:6
a4 MTBE/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
a5 MTBE/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
a6 MTBE/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
a7 MTBE/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
a8 MTBE/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
a9 MTBE/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
al 0 MTBE/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 al 1 MTBE/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from
4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
al2 MTBE/diethyl ether in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b) mixtures with toluene:
bl Toluene/MEK in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
b2 Toluene/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
b3 Toluene/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
b4 Toluene/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
b5 Toluene/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
b6 Toluene/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 b7 Toluene/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
b8 Toluene/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
b9 Toluene/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
b 10 Toluene/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4 : 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
bl 1 Toluene/cyclopentane in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
bl2 Toluene/diethyl ether in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, furthermore preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c) mixtures with benzene:
cl Benzene/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
c2 Benzene/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 c3 Benzene/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
c4 Benzene/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
c5 Benzene/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
c6 Benzene/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2,
1:3, 1:4, 1:5, or 1:6
c7 Benzene/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
c8 Benzene/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
c9 Benzene/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
cl 0 Benzene/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
cl 1 Benzene/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
Benzene/diethyl ether in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 d) mixtures with CPME:
dl CPME/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
d2 CPME/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
d3 CPME/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
d4 CPME/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
d5 CPME/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
d6 CPME/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
d7 CPME/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
d8 CPME/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
d9 CPME/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
dlO CPME/cyclopentane in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from
4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
dl 1 CPME/diethyl ether in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 e) mixtures with MeCN:
el MeCN/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
e2 MeCN/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
e3 MeCN/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
e4 MeCN/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
e5 MeCN/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
e6 MeCN/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
e7 MeCN/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
e8 MeCN/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
e9 MeCN/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
e 10 MeCN/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4 : 1 to 1 :4, more preferably from 3:1 to 1 :3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2,
1:3, 1:4, 1:5, or 1:6
ell MeCN/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from
4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
el2 MeCN/diethyl ether in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 f) mixtures with THF:
fl THF/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
12 THF/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
f3 THF/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
f4 THF/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
f5 THF/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2,
1:3, 1:4, 1:5, or 1:6
f6 THF/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
17 THF/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
f8 THF/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
f9 THF/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
fl 0 THF/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
fl 1 THF/diethyl ether in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 g) mixtures with Me-THF:
gl Me-THF/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
g2 Me-THF/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
g3 Me-THF/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
g4 Me-THF/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
g5 Me-THF/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
g6 Me-THF/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
g7 Me-THF/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
g8 Me-THF/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
g9 Me-THF/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
glO Me-THF/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
gl 1 Me-THF/diethyl ether in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 h) mixtures with MEK:
hi MEK/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
h2 MEK/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
h3 MEK/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
h4 MEK/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
h5 MEK/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
h6 MEK/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
h7 MEK/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
h8 MEK/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
h9 MEK/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2,
1:3, 1:4, 1:5, or 1:6
hlO MEK/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
hi 1 MEK/diethyl ether in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 i) mixtures with EtOAc:
11 EtOAc/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
12 EtOAc/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
i3 EtOAc/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
14 EtOAc/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
15 EtOAc/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
16 EtOAc/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
17 EtOAc/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
i8 EtOAc/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
i9 EtOAc/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
il 0 EtOAc/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
il 1 EtOAc/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from
4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
il 2 EtOAc/diethyl ether in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 j) mixtures with 1,4-dioxane:
j 1 1 ,4-dioxane/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
j2 1 ,4-dioxane/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
j3 1 ,4-dioxane/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
j4 1 ,4-dioxane/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
j5 1 ,4-dioxane/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
j6 1 ,4-dioxane/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from
4:1 to 1:4, more preferably from 3:1 to 1:3, furthermore preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
j7 1 ,4-dioxane/MIBK in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
j8 1 ,4-dioxane/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
j9 1 ,4-dioxane/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
j 10 1 ,4-dioxane/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
j 11 1 ,4-dioxane/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
jl2 1,4-dioxane/diethyl ether in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, furthermore preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 k) mixtures with n-hexane (hex):
kl Hex/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
k2 Hex/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
k3 Hex/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
k4 Hex/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
k5 Hex/THF in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
k6 Hex/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
k7 Hex/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
k8 Hex/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
k9 Hex/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 klO Hex/tBuOH in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, furthermore preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
kl 1 Hex/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
kl2 Hex/diethyl ether in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
1) mixtures with « -heptane (hept):
11 Hept/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
12 Hept/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
13 Hept/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
14 Hept/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
15 Hept/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 16 Hept/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
17 Hept/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
18 Hept/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
19 Hept/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
110 Hept/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
111 Hept/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
112 Hept/diethyl ether in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 m) mixtures with n-pentane (pent):
ml Pent/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 m2 Pent/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
m3 Pent/CPME in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
m4 Pent/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
m5 Pent/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
m6 Pent/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
ml Pent/MIBK in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
m8 Pent/NMP in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
m9 Pent/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
ml 0 Pent/tBuOH in a ratio (v/v) covered by the range from 6: 1 to 1 :6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
ml 1 Pent/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
ml 2 Pent/ diethyl ether in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4 : 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6 n) mixtures with isooctane:
nl Isooctane/MEK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
n2 Isooctane/acetone in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
n3 Isooctane/CPME in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
n4 Isooctane/anisole in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
n5 Isooctane/THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
n6 Isooctane/Me-THF in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
n7 Isooctane/MIBK in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2,
1:3, 1:4, 1:5, or 1:6
n8 Isooctane/NMP in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4 : 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
n9 Isooctane/tAmylOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
nlO Isooctane/tBuOH in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4:1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
nl 1 Isooctane/cyclopentane in a ratio (v/v) covered by the range from 6:1 to 1:6, preferably from 4: 1 to 1 :4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, or 1:6
nl2 Isooctane/diethyl ether in a ratio (v/v) covered by the range from 6:1 to 1 :6, preferably from 4:1 to 1:4, more preferably from 3:1 to 1:3, further more preferably from 1:1 to 1:3, and most preferably the ratio is selected from the group consisting of 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2,
1:3, 1:4, 1:5, or 1:6
The addition of further solvents to form a ternary, or higher order mixtures, wherein said solvent or solvents is
• selected from group (B)
• also suited to facilitate azeotropic distillation
• selected from group (C)
• also suited to ensure lipase activity and a sufficient solubility of the starting materials, or
• a combination of these options is considered analogously to the mentioned solvent or solvents and is therefore included into the process of the present application.
For example, the use of solvent systems like MTBE/MEK/toluene, MTBE/MEK/Me-THF, MTBE/anisole/acetone or MTBE/Me-THF/benzene is considered analogous and is therefore included into the process of the present application.
The use of ratios in between the mentioned ratios, e.g. ratios like [1.2 : 1], [3.6 : 1.2] or [1.1 : 2.2] is considered analogous and is therefore included into the process of the present application. Examples 2a, Id and 2b show the small impact of the used starting material concentrations (i.e. the ratio between weight of starting material to solvent) in the process of the invention with regard to the yield.
The process of the invention is carried out at a reaction temperature of 0 to 150 °C, preferably at 25 to 130 °C, more preferably at 40 to 110 °C, even more preferably at 50 to 100 °C and most preferably at 60 to 80 °C.
The process of the present invention is carried out under conditions effective to remove water from the reaction mixture in order to allow the reaction to proceed to completion. These conditions include but are not limited to adding dehydrating agents (e.g., molecular sieves, calcium carbonate, magnesium sulfate or sodium sulfate) directly to the reaction mixture, microwave heating, and azeotropic distillation using organic media as described above. The latter is used to conduct the reaction in refluxing organic media using a flask/reactor fitted with a distillation apparatus. The distilled solvent is then wasted and simultaneously replaced by fresh solvent, while maintaining the original volume and ratio of solvents in the vessel. The flask/reactor with the refluxing organic media can also be fitted with a Dean-Stark and condenser assembly or a higher-scale technical equivalent, where after separation of water, the organic media is looped back into the flask/reactor. Since azeotropic distillation is somewhat less effective in smaller scale, the solvent reservoir of the Dean-Stark apparatus may be filled with molecular sieves to finally absorb water transported out of the reaction vessel. It is also possible to use a dropping funnel and condenser assembly in which the dropping funnel (but not the pressure equalizing tube) is filled with a dehydrating agent, e.g. molecular sieves.
Other methods known to those skilled in the art to remove water may also be used. While the removal of water from the reaction vessel is critical for the invention, the method to separate and finally absorb the water is not important as long as the water removal is efficient.
Dicarboxylic acid suitable as bis-acyl donor compounds of the present invention can be but are not limited to free dicarboxy groups (free acids), dicarbonates and anhydrides.
Examples of bis-acyl donors include dicarboxylic acids such as malonic acid, succinic acid, glutaric acid, adipic acid, pimelic acid, suberic acid, azelaic acid, sebacic acid, undecanedioic acid, dodecanedioic acid, 1,12-dodecanedicarboxylic acid, fumaric acid, maleic acid, glutaconic acid, traumatic acid, muconic acid, cis aconitic acid, phthalic acid, isophthalic acid, terephthalic acid, cyclohexan-l,4-dicarboxylic acid, 3-(2- carboxyethyldisulfanyl)propionic acid, diglycolic acid, thio diglycolic acid, 3-(2- carboxyethoxy)propionic acid, 3-[2-(2-carboxyethoxy)ethoxy]propionic acid, 3-{2-[2-(2- carboxyethoxy)ethoxy]ethoxy} propionic acid, 3 -(2- {2-[2-(2-carboxy- ethoxy)ethoxy]ethoxy}ethoxy)propionic acid, 3-[2-(2-{2-[2-(2- carboxyethoxy)ethoxy]ethoxy} ethoxy)ethoxy]propionic acid, 3- {2-[2-(2- {2-[2-(2- carboxyethoxy)ethoxy] ethoxy}ethoxy)ethoxy]ethoxy}propionic acid and 3-(2-{2-[2-(2- {2- [2-(2-carboxyethoxy)ethoxy ] ethoxy } ethoxy)ethoxy] ethoxy } ethoxy)propionic acid.
Examples of bis-acyl donors include anhydrides such as succinic anhydride, glutaric anhydride, adipic anhydride, pimelic anhydride, suberic anhydride, maleic anhydride, cis aconitic anhydride, phthalic anhydride, diglycolic anhydride and thio diglycolic
anhydride.
Equivalents of bis-acyl donor with regard to the starting material can vary from
substoichiometric ratios over stoichiometric ratios to use of an excess of bis-acyl donor.
As demonstrated by examples 2n-p, the amount of weight equivalents of added glutaric acid with regard to N-Fmoc-doxorubicin is an important factor in order to arrive at an optimal yield. While varying the glutaric acid weight equivalents from 2.5 to 12 eq., the yield of N-Fmoc-doxorubicin hemiglutarate increased.
Glutaric acid weight equivalents in between the mentioned values, i.e. weight equivalents like e.g. 3.2 wt. eq. or e.g. 7.1 wt. eq. are incorporated in the invention analogous to the mentioned values.
Lipases of the present invention can either be native (wild-type) lipases or genetically- modified lipases engineered using standard methods of molecular biology (Berglund, "Controlling Lipase Enantioselectivity for Organic Synthesis," Biomol. Eng., 18(1): 13-22 (2001); Brocca et al, "Novel Lipases Having Altered Substrate Specificity, Methods for Their Preparation, and Their Use in Biocatalytic Applications," Eur. Pat. Appl. pp.33 (2001); Svendsen, "Lipase Protein Engineering," Biochim. Biophys. Acta, 1543(2): 223- 238 (2000); Wong et al, "Lipase Engineering: A Window into Structure-Function Relationships," Methods in Enzymology, 284: 171-84 (1997), which are hereby incorporated in their entirety).
Lipases that can be used for the selective C14-OH acylation of N-Fmoc-doxorubicin include lipases from microbial sources such as Alicaligenes sp., Aspergillus, Aspergillus niger, Aspergillus oryzae, Alcaligenes sp., Candida antartica, Candida cylindracea sp., Candida rugosa, Fusarium solani pisi, Mucor miehei, Mucor javanicus, Penicillium roqueforti, Pseudomonas cepacia, Pseudomonas fluorescens, Pseudomonas sp., Pseudomonas stutzeri, Rhizomucor miehei, Rhizopus arrhizus, Rhizopus delemar, Rhizopus niveus, Rhizopus oryzae, Thermomyces lanuginosus, Bacillus subtilis and Porcine pancreas.
The nonaqueous biocatalysis of the present invention can be carried out where the lipase is immobilized to a solid support. Examples of solid support include diatomaceous earth, silica, ceramic, polypropylene, polystyrene or acrylic resins. The lipase can be immobilized by covalent cross-linking of enzyme aggregates (CLEAs).
Other methods known to those skilled in the art to immobilize a lipase can also be used. The type of solid support or the way the lipase is immobilized to a solid support is not critical for the invention as long as the lipase retains its natural function on the solid support.
Weight equivalents of lipase with regard to the starting material can vary from
substoichiometric ratios over stoichiometric ratios to use of an excess of lipase.
Weight equivalents in the context of an immobilized lipase mean the use of a lipase with an activity of 10.000 units/g resin. In the case of a different activity the weight equivalents are corrected accordingly. As demonstrated by examples 2h-m, the amount of weight equivalents of added Candida Antarctica B with regard to N-Fmoc-doxorubicin is an important factor in order to arrive at an optimal yield. Varying the Candida Antarctica B weight equivalents from 0.5 to 4.79 eq. with regard to the N-Fmoc-doxorubicin starting material, increased the yield of N-Fmoc-doxorubicin hemiglutarate while the reaction time decreased.
Weight equivalents in between the mentioned values, e.g. weight equivalents like e.g. 1.2 wt. eq. or 3.0 wt.eq. or e.g. 3.5 wt. eq. are incorporated in the invention as analogous to the mentioned Candida Antarctica B weight equivalents values.
As also demonstrated by example 2au, the lipase chemistry is applicable in kg scale.
Examples
In order to compare the process of the invention with conditions known from the prior art, reference experiments 5 and 6 are described below.
Reference experiment 5: Synthesis of N-alloc-doxorubicin-O-hemiglutarate using glutaric acid as bis-acyl donor under lipase conditions
N-alloc-doxorubicin-O-hemiglutarate
N-alloc-doxorubicin (50 mg) was suspended in MTBE/MEK 3 : 1 (v/v) (10 mL) and to this was added lipase acrylic resin from Candida Antarctica B (233 mg, 5 weight equivalents) and glutaric acid (109 mg). The reaction flask was fitted with a dropping funnel and condenser assembly in which the dropping funnel was filled with 3 A molecular sieves (ca. 10 g). The reaction mixture was stirred vigorously and heated for 25 h at reflux
temperature. The progress of the reaction was monitored by HPLC. No work-up was performed.
Yield: not isolated
Purity: 50.9 % (HPLC- see Appendix 4)
Reference experiment 6: Synthesis ofN-trifluoroacetyl-doxorubicin-O-hemiglutarate using glutaric acid as bis-acyl donor under lipase conditions
N-trifluoroacetyl-doxorubicin-O-hemiglutarate The same reaction conditions as used in reference experiment 5 were applied for the acylation except using N-trifluoroacetyl-doxorubicin (Cotterill et al. Organic Process Research and Development 2005, 9, 818) (50 mg) instead of doxorubicin. No work-up was performed. Yield: not isolated
Purity: 70.2 % (HPLC- see Appendix 5)
In a process typical for the invention, the N-Fmoc-doxorubicin derivative (Formula II) is dissolved in suitable organic media, containing 2.5-12 weight equivalents of the desired dicarboxylic acid. To this mixture 1-12 weight equivalents resin bound lipase per g N-Fmoc- doxorubicin are added. The reaction flask can be incubated at 25-130 °C, or, alternatively, if an accumulation of the reaction product water inhibits the progress of the reaction, fitted with a condenser assembly and/or an assembly to remove water and stirred at the reflux temperature of the solvent, from 2 hours up to 2 days. Reaction progress is followed by high performance liquid chromatography ("HPLC") or any other suitable analytical method. Upon depletion of the substrate the reaction mixture is filtered to remove the insoluble enzyme powder. The enzyme is washed with an appropriate solvent and the combined solvent is washed with an appropriate aqueous medium, dried and evaporated under reduced pressure. The product is purified using a suitable method, most routinely recrystallization, precipitation, silica gel chromatography, liquid chromatography on other solid adsorbents, or preparative high performance liquid chromatography. Inventive Examples
The following examples are provided to illustrate embodiments of the present invention.
Example la: Synthesis of N-Fmoc-doxorubicin-O-hemiglutarate using glutaric acid as bis- acyl donor under lipase conditions in MTBE/MEK
N-Fmoc-doxorubicin- 14-O-hemiglutarate
N-Fmoc-doxorubicin (1.00 g) was suspended in MTBE/MEK 3 : 1 (v/v) (80 mL) and to this was added lipase acrylic resin from Candida Antarctica B (4.00 g, 4 wt. eq., all weight equivalents with regard to the N-Fmoc-doxorubicin starting material) and glutaric acid (1.73 g, 10 eq.). The reaction flask was fitted with a dropping funnel and condenser assembly in which the dropping funnel (but not the pressure equalizing tube) was filled with 3 A molecular sieves (ca. 60 g). The reaction mixture was stirred vigorously and heated for 25 h at reflux temperature. The progress of the reaction was monitored by HPLC. The enzyme was removed by filtration and washed with hot MTBE (4 x 10 mL). The filtrate was washed with water (5 x 40 mL) and evaporated to dryness under reduced pressure at a temperature of < 50 °C. The residue was redissolved in 5 mL of MEK and added drop wise into 60 mL of a vigorously stirred aqueous acetic acid solution (w/w = 5 %). After stirring for one hour, the product is filtered through a membrane filter (0.2 μιη), washed with water and dried under vacuum to constant weight at a temperature of < 50 °C to give crude N-Fmoc-doxorubicin- 14-O-hemiglutarate.
Yield: 1.06 g red solid (92 %)
Purity: 88.2 % (HPLC - see Appendix 6) Example lb: Synthesis ofN-Fmoc-doxorubicin-O-hemiglutarate using glutaric anhydride and glutaric acid as bis-acyl donor under lipase conditions in MTBE/PhOMe
N-Fmoc-doxorubicin (300 mg) was suspended in MTBE/PhOMe 1 : 1 (v/v) (24 mL) and to this was added lipase acrylic resin from Candida Antarctica B (1200 mg, 4 wt. eq., all weight equivalents with regard to the N-Fmoc-doxorubicin starting material), glutaric anhydride (223 mg, 5 eq.) and glutaric acid (258 mg, 5eq.). The reaction mixture was stirred vigorously and heated for 25 h at approx. 80 °C. The progress of the reaction was monitored by HPLC. Following the removal of the enzyme by filtration the reaction was worked-up analogously to Example la to give crude
N-Fmoc-doxorubicin- 14-O-hemiglutarate.
Yield: 301 mg red solid (88 %)
Purity: 87.6 % (HPLC - see Appendix 7)
Example lc: Synthesis of N-Fmoc-doxorubicin-O-hemiglutarate using glutaric acid as bis- acyl donor under lipase conditions in Me-THF
N-Fmoc-doxorubicin (300 mg) was suspended in Me-THF (24 mL) and to this was added lipase acrylic resin from Candida Antarctica B (1200 mg, 4 wt. eq., all weight equivalents with regard to the N-Fmoc-doxorubicin starting material) and glutaric acid (518 mg, 10 eq.).
The reaction flask was fitted with a column and condenser assembly in which the column was filled with 3 A molecular sieves (ca. 20 g). The reaction mixture was stirred vigorously and heated for 4 h at approx. 100 °C. The progress of the reaction was monitored by HPLC. Following the removal of the enzyme by filtration the reaction was worked-up analogously to Example la to give crude
N-Fmoc-doxorubicin- 14-O-hemiglutarate.
Yield: 254 mg red solid (74 %)
Purity: 74.2 % (HPLC - see Appendix 8) Example Id: Synthesis of N-Fmoc-doxorubicin-O-hemiglutarate using glutaric acid as bis- acyl donor under lipase conditions in MTBE/Me-THF
N-Fmoc-doxorubicin (300 mg) was suspended in MTBE/Me-THF 1 :2 (v/v) (18 mL) and to this was added lipase acrylic resin from Candida Antarctica B (1200 mg, 4 wt. eq., all weight equivalents with regard to the N-Fmoc-doxorubicin starting material) and glutaric acid (518 mg, 10 eq.). The reaction flask was fitted with a column and condenser assembly in which the column was filled with 3 A molecular sieves (ca. 20 g). The reaction mixture was stirred vigorously and heated for 4 h at approx. 90 °C. The progress of the reaction was monitored by HPLC. Following the removal of the enzyme by filtration the reaction was worked-up analogously to Example la to give crude
N-Fmoc-doxorubicin- 14-O-hemiglutarate.
Yield: 327 mg red solid (95 %)
Purity: 92.1 % (HPLC - see Appendix 9)
Example le: Synthesis under continuous distillation N-Fmoc-doxorubicin (3 g) was suspended in MTBE/Me-THF 1 :2 (v/v) (180 mL) and to this was added lipase acrylic resin from Candida Antarctica B (12 g, 4 wt. eq., all weight equivalents with regard to the N-Fmoc-doxorubicin starting material) and glutaric acid (5.2 g, 10 eq.). The reaction was performed in a reaction flask which was fitted with a Dean-Stark condenser assembly. The reaction mixture was stirred vigorously and heated for 10 h at approx. 90 °C, while the distillate was collected and removed from the side arm of the Dean-Stark apparatus. Fresh solvent MTBE/Me-THF 1 :2 (v/v) was added to replace the removed distillate. The progress of the reaction was monitored by HPLC.
Yield: not isolated
Purity: 75.7 % (HPLC - see Appendix 10) Examples lf-lk: Synthesis ofN-Fmoc-doxorubicin-O-hemiglutarate using different liqid organic media under lipase conditions
Examples lf-lk were performed analogously to Example la applying different liquid organic media. The results are summarized in the following table:
Examples 2a-b: Synthesis ofN-Fmoc-doxorubicin-O-hemiglutarate using different N-Fmoc- doxorubicin concentrations
Examples 2a-b were performed analogously to Example Id applying different starting material concentrations, i.e. varying the ratio between solvent volume to weight ofN-Fmoc- doxorubicin.
Examples 2c-g: Synthesis of N-Fmoc-doxorubicin-O-hemiglutarate using different water removing conditions
N-Fmoc-doxorubicin (300 mg) was suspended in MTBE/Me-THF 1 :2 (v/v) (18 mL) and to this was added lipase acrylic resin from Candida Antarctica B (1200 mg, 4 wt. eq., all weight equivalents with regard to the N-Fmoc-doxorubicin starting material), glutaric acid (518 mg, 10 eq. with regard to the N-Fmoc-doxorubicin starting material). The reaction was performed in a reaction flask which was fitted with a column and condenser assembly in which the column was filled with 3 A molecular sieves or a Dean-Stark condenser assembly or in a sealed tube. The reaction mixture was stirred vigorously and heated at approx. 90 °C. The progress of the reaction was monitored by HPLC. The results are summarized in the following table:
It is shown by these examples, that effective removal of water is an important factor in order to arrive at an optimal yield.
Examples 2h-m: Synthesis of N-Fmoc-doxorubicin-O-hemiglutarate using different amounts of lipase Candida Antarctica B
N-Fmoc-doxorubicin (300 mg) was suspended in MTBE/Me-THF 1 :2 (v/v) (18 mL) and to this was added lipase acrylic resin from Candida Antarctica B (0.5, 1.0, 2.0 or 4.0 wt. eq., all weight equivalents with regard to the N-Fmoc-doxorubicin starting material), glutaric acid (518 mg, 10 eq.). The reaction flask was fitted with a column and condenser assembly in which the column was filled with 3 A molecular sieves. The reaction mixture was stirred vigorously and heated at approx. 90 °C. The progress of the reaction was monitored by HPLC. The results are summarized in the following table:
It is shown by these examples, that the amount of weight equivalents of added lipase with regard to N-Fmoc-doxorubicin is an important factor in order to arrive at an optimal yield.
Examples 2n-p: Synthesis ofN-Fmoc-doxorubicin-O-hemiglutarate using different amounts of glutaric acid
N-Fmoc-doxorubicin (300 mg) was suspended in MTBE/Me-THF 1 :2 (v/v) (18 mL) and to this was added lipase acrylic resin from Candida Antarctica B (1200 mg, 4 wt. eq., all weight equivalents with regard to the N-Fmoc-doxorubicin starting material), glutaric acid (2.5, 5.0 or 10.0 eq.). The reaction flask was fitted with a column and condenser assembly in which the column was filled with 3 A molecular sieves. The reaction mixture was stirred vigorously and heated at approx. 90 °C. The progress of the reaction was monitored by HPLC. The results are summarized in the following table:
It is shown by these examples, that the amount of weight equivalents of added glutaric acid with regard to N-Fmoc-doxorubicin is an important factor in order to arrive at an optimal yield.
Examples 2q-r: Synthesis of N-Fmoc-doxorubicin-O-hemiglutarate using different polymorph forms of N-Fmoc-doxorubicin
Amorphous (XRPD pattern see Appendix 19) or crystalline (XRPD pattern see Appendix 20) N-Fmoc-doxorubicin (300 mg) was suspended in MTBE/Me-THF 1 :2 (v/v) (18 mL) and to this was added lipase acrylic resin from Candida Antarctica B (1200 mg, 4 wt. eq., all weight equivalents with regard to the N-Fmoc-doxorubicin starting material), glutaric acid (518 mg, 10 eq.). The reaction flask was fitted with a column and condenser assembly in which the column was filled with 3 A molecular sieves. The reaction mixture was stirred vigorously and heated at approx. 90 °C. The progress of the reaction was monitored by HPLC. The results are summarized in the following table:
It is shown by these examples, that the solid state of the N-Fmoc-doxorubicin is not critical to the process of the invention.
Examples 2s-2au: Synthesis of N-Fmoc-doxorubicin-O-hemiglutarate using different amounts of lipase Candida Antarctica B and different liquid organic media
N-Fmoc-doxorubicin (50 mg) was suspended in different organic media (3 mL) and to these were added lipase acrylic resin from Candida Antarctica B (3.0, 3.5, 4.0 or 4.5 wt. eq., all weight equivalents with regard to the N-Fmoc-doxorubicin starting material), glutaric acid (86 mg, 10 eq.). The reaction flask was fitted with a column and condenser assembly in which the column was filled with 3 A molecular sieves. The reaction mixture was stirred vigorously and refluxed. The progress of the reaction was monitored by
HPLC. The results are summarized in the following table:
It is shown by these examples, that the amount of weight equivalents of added lipase with regard to N-Fmoc-doxorubicin and the solvent mixture are important factors in order to obtain at an optimal yield. Examples 2au: Synthesis of N-Fmoc-doxorubicin-O-hemiglutarate in kg scale using lipase Candida Antarctica B
N-Fmoc-doxorubicin (1.07 kg), glutaric acid (10 eq.) and Me-THF (40 vol. eq., all volume and weight equivalents with regard to the N-Fmoc-doxorubicin starting material) are charged into a reactor connected to a column containing molecular sieves 3 A (40 wt.eq.). To this was added lipase acrylic resin from Candida Antarctica B (3.5 wt. eq.) suspended in MTBE (20 vol. eq). The resulting mixture was heated at reflux until completion of the reaction (monitored by HPLC; HPLC after 13h: 91.3%- see appendix 21). The enzyme was removed by filtration and then the reaction mixture was concentrated. After aqueous washes the organic layer was further concentrated and the product isolated by
precipitation in n-heptane to give N-Fmoc-doxorubicin-14-O-hemiglutarate.
Yield: 1.17 kg red solid (82 %)
Purity (isolated product): 88.3%
Further inventive examples
The examples below can be synthesized analogously to Example Id.
APPENDIX
Appendix 1: Impurity profile of reference experiment 1
RT %Area
6.62 4.2
11.04 4.6
12.16 1.6
15.79
1.3
(/V-Fmoc-Dox)
18.37
(/V-Fmoc-Dox- 51.8
hemiglutarate)
22.28 11.1
(Di-Hemi(A))
26.01 0.9
31.09 1.4
Appendix 2 (relating to Fig. 2): HPLC of reference experiment 3
AEterna Zentaris GmbH, Frankfurt
Method Name: C:\LCE_Daten\FS_Analyt\Method\20B-20min-100B-30min+5.met Data: C:\LCE_Daten\FS_Analyt\Daten\AEZS-108\mikrosynthese\FS2003_0 03.dat Acquired: 24.06.2014 11:30:01
Operator: AB
Column: LEO, LiChrospher 100, RP18, 250-4, 12 μηι, FluB 1 ml / min
Mobile Phase: ACN, Water, TFA
Sample: FS2003
Data description: t(RK)=17h
Results: 254 nm, 4 nm
Retention Time (min) Area Area Percent
21,01 139631981 98,90 Fmoc-Dox
21,53 816318 0,58
27,67 184082 0,13
28,05 559354 0,40
Totals 141191735 100,00
Appendix 3 (relating to Fig. 3): HPLC of reference experiment 4
AEterna Zentaris GmbH, Frankfurt
Method Name: C:\LCE_Daten\FS_Analyt\Method\20B-20min-100B-30min+5.met Data: C:\LCE_Daten\FS_Analyt\Daten\AEZS- 108\mikrosynthese\FS 1982_003.dat Acquired: 16.05.2014 16:08:03
Operator: AB
Column: LEO, LiChrospher 100, RP18, 250-4, 12 μηι, FluB 1 ml / min
Mobile Phase: ACN, Water, TFA
Sample: FS1982
Data description: {Data Description}
Results: 254 nm, 4 nm
Retention Time (min) Area Area Percent
5,86 53208 0,44
10,62 794237 6,64
10,85 35805 0,30
11,51 24315 0,20
12,05 852582 7,13 Dox
12,89 33901 0,28
13,37 24468 0,20
14,34 1688599 14,12 Dox-Hemiglutarate
15,11 598976 5,01
15,97 208749 1,75
16,67 1038509 8,68
17,21 197554 1,65
18,07 342505 2,86
20,37 42888 0,36
20,97 102799 0,86
21,67 91792 0,77
24,45 5693020 47,61
27,08 133654 1,12
Totals 11957561 100,00 Appendix 4 (relating to Fig. 4): HPLC of reference experiment 5
Data File C:\CHEM32\1\DATA\050614\005-0501.D
Sample Name: FS1990
Alloc-Dox-Hemiglutarat S155442 gem 6.6.2014
Injection Date : 06.06.2014 00:46:32 Seq. Line 5 Sample Name : FS1990 Location Vial 5 Acq. Operator : sh Inj 1 Acq. Instrument : Snoopy Inj Volume 5 μΐ Acq. Method : C:\CHEM32\l\METHODS\FMOC-DOX3.M
Last changed : 05.06.2014 17:09:36 by sh
Analysis Method C:\CHEM32M\METHODS\FMOC-DOX3.M
Last changed : 06.06.2014 09:54:20 by sh
(modified after loading)
Area Percent Report
Sorted By Signal
Calib Data Modified 06.06.2014 09:54:21
Multiplier 1.0000
Dilution 1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=226,4 Ref=off
Peak RetTime [min] Pype Width Area Area % Name
[min] [mAU*s]
1 0,753 BV 0,0447 1,97633 0,0470 ?
2 0,837 VB 0,0485 150,96184 3,5865 ?
3 1,106 BV 0,0878 1,51280 0,0359 ?
4 1,201 VV 0,0859 7,72252 0,0346 ?
5 1,507 VV 0,0859 7,72252 0,1835 ?
6 1,630 VV 0,0626 87,58825 2,0809 ?
7 1,875 VB 0,1196 3,57689 0,0850 ?
8 2,727 VV 0,0682 2,58242 0,0614 ?
9 3,466 BV 0,0663 281,90942 6,6975 ?
10 3,597 VV 0,0637 65,48560 1,5558 ?
11 3,799 VB 0,0696 2,36651 0,0562 ?
12 4,153 BB 0,0488 6,74245 0,1602 ?
13 4,586 BB 0,1339 1,23046 0,0292 ?
14 7,804 BV 0,1220 2,11469 0,0502 ?
15 9,704 BB 0,1218 1,15021 0,0273 ?
16 10,200 BB 0,1594 4,86588 0,1156 ?
17 11,857 BB 0,1450 1,81092 0,0430 ?
18 12,370 BV 0,1157 1,37531 0,0327 ?
19 12,775 VV 0,1162 3,61454 0,0859 ?
20 13,835 BB 0,1494 4,30754 0,1023 ? 21 14,543 BV 0,1465 297,70215 7,0727 ?
22 14,995 VB 0,1507 190,80873 4,5331 ?
23 17,006 BV 0,1524 377,45557 8,9674 ?
24 17,615 VB 0,2006 9,36009 0,2224 ?
25 18,876 BB 0,1502 1,64563 0,0391 ?
26 19,510 BV 0,1514 16,58601 0,3940 Alloc-Dox
27 19,875 VV 0,1551 4,78868 0,1138 ?
28 20,172 VV 0,1317 22,35423 0,5311 ?
29 20,710 VV 0,1874 24,22222 0,5755 ?
30 21,209 VV 0,1998 23,07651 0,5458 ?
Peak RetTime Pype Width Area Area % Name
[min] [min] [mAU*s]
31 21,770 VV 0,2521 8,79346 0,2089 ?
32 22,208 VB 0,1337 4,68051 0,1112 ?
33 22,817 BV 0,1169 1,18396 0,0281 ?
34 23,062 VB 0,1373 2,78298 0,0661 ?
35 23,545 BV 0,1110 1,83656 0,0436 ?
36 24,553 VB 0,1415 2142,18237 50,8928 Alloc-Dox-
Hemiglutarate
37 25,226 BB 0,1142 16,22691 0,3855 ?
38 25,751 BV 0,1920 22,16626 0,5266 ?
39 26,149 VB 0,1110 1,55222 0,0369 ?
40 26,685 BV 0,1511 15,32774 0,3641 ?
41 27,412 VV 0,1459 136,74002 3,2486 ?
42 27,802 VV 0,1754 11,61036 0,2758 ?
43 27,990 VV 0,1003 5,80815 0,1380 ?
44 28,158 VV 0,1303 18,84417 0,4477 ?
45 28,313 VB 0,1032 8,69689 0,2066 ?
46 29,390 BB 0,1895 1,98135 0,0471 ?
47 30,194 BB 0,1933 2,56765 0,0610 ?
48 30,765 BV 0,1366 4,03893 0,0960 ?
49 31,024 VV 0,1262 18,18313 0,4320 ?
50 31,261 VV 0,1294 48,29337 1,1473 ?
51 31,540 VV 0,1394 2,97163 0,0706 ?
52 32,048 VV 0,1502 4,93075 0,1171 ?
53 32,462 VB 0,2846 6,48672 0,1541 ?
54 33,476 BV 0,1712 2,18098 0,0518 ?
55 33,957 VV 0,2232 4,61230 0,1096 ?
56 34,349 VV 0,1248 3,24586 0,0771 ?
57 34,678 VB 0,1981 16,27567 0,3867 ?
58 35,447 BBA 0,1946 8,56773 0,2035 ?
59 37,673 VB 0,1700 13,83938 0,3288 ?
60 38,496 BB 0,1687 1,75126 0,0416 ?
61 41,369 BB 0,2294 68,49178 1,6272 ?
Totals: 4209,20193 Appendix 5 (relating to Fig. 5): HPLC of reference experiment 6
Data File C:\CHEM32\1\DATA\050614\006-0601.D
Sample Name: FS1991
Trifluoroacetyl-Dox-Hemiglutarat S 155443 6.6.2014 gem
Injection Date : 06.06.2014 02: 17:39 Seq. Line 6 Sample Name : FS1991 Location Vial 6 Acq. Operator : sh Inj 1 Acq. Instrument : Snoopy Inj Volume 5 μΐ Acq. Method : C:\CHEM32\l\METHODS\FMOC-DOX3.M
Last changed : 05.06.2014 17:09:36 by sh
Analysis Method C:\CHEM32M\METHODS\FMOC-DOX3.M
Last changed : 06.06.2014 09:46:42 by sh
(modified after loading)
Area Percent Report
Sorted By Signal
Calib Data Modified 06.06.2014 09:46:42
Multiplier 1.0000
Dilution 1.0000
Use Multiplier & Dilution Factor with ISTDs
Signal 1: DAD1 A, Sig=226,4 Ref=off
Peak RetTime Pype Width Area Area % Name
[min] [min] [mAU*s]
1 0,754 BV 0,0456 1,77998 0,0314 ?
2 0,839 VB 0,0501 148,12938 2,6157 ?
3 1,109 BB 0,1250 2,97684 0,0526 ?
4 1,353 BV 0,0849 2,45058 0,0433 ?
5 1,508 VV 0,0733 4,79460 0,00847 ?
6 1,614 VV 0,0687 126,67136 2,2368 ?
7 1,862 VB 0,1434 4,05598 0,0716 ?
8 2,510 BV 0,1169 1,52618 0,0269 ?
9 2,727 VV 0,0765 3,23060 0,0570 ?
10 3,464 VV 0,0590 277,31018 4,8968 ?
11 3,594 VV 0,0606 66,49511 1,1742 ?
12 3,784 VV 0,0620 1,2850 0,0227 ?
13 4,093 VB 0,0450 4,93195 0,0871 ?
14 7,798 BB 0,1164 2,42427 0,0428 ?
15 10,209 BB 0,2047 4,14380 0,0202 ?
16 11,858 BB 0,1474 1,81232 0,0320 ?
17 12,374 BB 0,1424 1,60666 0,0284 ?
18 12,780 BV 0,1253 4,28284 0,0756 ?
19 12,988 VV 0,1316 1,61949 0,0286 ?
20 13,319 VB 0,1772 7,40499 0,1308 ?
21 13,841 BB 0,1772 7,40499 0,1308 ?
22 14,548 BV 0,1492 302,99411 5,3504 ? 23 15,000 VB 0,1531 198,38055 3,5031 ?
24 17,066 BB 0,2274 27,45908 0,4849 ?
25 19,765 BB 0,1494 21,41344 0,3781 Trifluoroacetyl-Dox
26 21,223 BB 0,1782 19,42617 0,3430 ?
27 21,980 BB 0,1769 3,39589 0,0600 ?
28 22,792 BB 0,1125 1,78286 0,0315 ?
29 23,422 BB 0,1309 1,73974 0,0307 ?
30 23,949 BV 0,1252 2,14982 0,0380 ?
31 24,194 VV 0,1295 3,93702 0,0695 ?
32 25,007 VV 0,1540 3977,46851 70,2355 Trifluoroacetyl-Dox- Hemiglutarate
33 25,505 VV 0,1160 1,63344 0,0288 ?
34 25,793 VV 0,1092 53,76739 0,9494 ?
35 26,125 VV 0,1831 7,99116 0,1411 ?
36 26,712 VV 0,1503 2,07425 0,0366 ?
37 27,118 VV 0,1913 2,01177 0,0355 ?
38 27,840 VV 0,1431 58,37741 1,0308 ?
39 28,330 VV 0,1272 9,79523 0,1730 ?
40 28,678 VB 0,1395 21,45978 0,3789 ?
41 30,222 BB 0,1928 2,85024 0,0503 ?
42 31,736 BB 0,1355 46,18381 0,8155 ?
43 32,349 BV 0,1382 1,40643 0,0248 ?
44 32,597 VV 0,1310 1,90871 0,0337 ?
45 32,930 VB 0,1796 5,94618 0,1050 ?
46 33,553 BV 0,1239 1,12035 0,0198 ?
47 33,980 VB 0,1533 2,45584 0,0434 ?
48 34,816 VB 0,1531 14,16499 0,2501 ?
49 37,879 VV 0,1956 10,01494 0,1768 ?
50 38,251 VB 0,1874 17,91219 0,3163 ?
51 38,796 BB 0,1855 3,16269 0,0558 ?
52 40,571 BV 0,1794 2,43357 0,0430 ?
53 42,193 BV 0,2514 137,89462 2,4350 ?
54 42,767 VB 0,2811 25,40734 0,4487 ?
Totals: 5663,04418
Appendix 6 (relating to Fig. 6): HPLC of Example la
Legend: Ml = N-Fmoc-Dox, M2 = N-Fmoc-Dox-hemiglutarate Appendix 7 (relating to Fig. 7): HPLC of Example lb
Legend: Ml = N-Fmoc-Dox, M2 = N-Fmoc-Dox-hemiglutarate Appendix 8 (relating to Fig. 8): HPLC of Example lc
Legend: Ml = N-Fmoc-Dox, M2 = N-Fmoc-Dox-hemiglutarate Appendix 9 (relating to Fig. 9): HPLC of Example Id
Legend: Ml = N-Fmoc-Dox, M2 = N-Fmoc-Dox-hemiglutarate Appendix 10 (relating to Fig. 10): HPLC of Example le
Legend: Ml = N-Fmoc-Dox, M2 = N-Fmoc-Dox-hemiglutarate Appendix 11 (relating to Fig. 11): HPLC of Example If
Legend: Ml = N-Fmoc-Dox, M2 = N-Fmoc-Dox-hemiglutarate Appendix 12 (relating to Fig. 12): HPLC of Example lg
Legend: Ml = N-Fmoc-Dox, M2 = N-Fmoc-Dox-hemiglutarate Appendix 13 (relating to Fig. 13): HPLC of Example lh
Legend: Ml = N-Fmoc-Dox, M2 = N-Fmoc-Dox-hemiglutarate
Appendix 14 (relating to Fig. 14): HPLC of Example li
Legend: Ml = N-Fmoc-Dox, M2 = N-Fmoc-Dox-hemiglutarate Appendix 15 (relating to Fig. 15): HPLC of Example lj
Legend: Ml = N-Fmoc-Dox, M2 = N-Fmoc-Dox-hemiglutarate Appendix 16 (relating to Fig. 16): HPLC of Example lk
Legend: Ml = N-Fmoc-Dox, M2 = N-Fmoc-Dox-hemiglutarate Appendix 17 (relating to Fig. 17): HPLC of Example 2a
Legend: Ml = N-Fmoc-Dox, M2 = N-Fmoc-Dox-hemiglutarate Appendix 18 (relating to Fig. 18): HPLC of Example 2b
Legend: Ml = N-Fmoc-Dox, M2 = N-Fmoc-Dox-hemiglutarate Appendix 19 (relating to Fig. 19):
XRPD pattern of amorphous N-Fmoc-doxorubicin (example 2o)
Appendix 20 (relating to Fig. 20)
XRPD pattern of crystalline N-Fmoc-doxorubicin (example 2p)
Appendix 21 (relating to Fig. 21): HPLC of Example 2au
Legend: Ml = N-Fmoc-Dox, M2 = N-Fmoc-Dox-hemiglutarate