Method of purifying macrolides

Abstract

Provided is a method of purifying a macrolide, especially tacrolimus, that includes loading macrolide onto a bed of sorption resin and eluting with a suitable eluent such as a combination of water and tetrahydrofuran.

1 . A method of separating a macrolide from impurities therein comprising the steps of: 
 a) providing a loading charge of a macrolide having an initial level of impurities,    b) loading the loading charge to a bed of sorption resin,    c) eluting the bed of loaded sorption resin with an eluent comprising water and an organic solvent selected from tetrahydrofuran and acetonitrile, to obtain an effluent, and    d) collecting at least one fraction of effluent comprising the macrolide.    
   
   
       2 . The method of  claim 1 , wherein the loading charge of a macrolide further contains a loading portion of a sorption resin.  
   
   
       3 . The method of  claim 2 , wherein the loading charge is deposited on the loading portion from its solution in an organic solvent in a step that includes combining the solution with an anti-solvent.  
   
   
       4 . The process of  claim 3 , wherein the organic solvent is selected from the group consisting of tetrahydrofuran, acetone, acetonitrile, methanol, ethanol, n-butanol, n-propanol, iso-propanol, esters and dipolar aprotic solvents.  
   
   
       5 . The process of  claim 4 , wherein the organic solvent is selected from the group consisting of tetrahydrofuran, acetone and acetonitrile.  
   
   
       6 . The process of  claim 3 , wherein the anti-solvent is selected from the group consisting of water, straight or branched alkanes, or cycloalkanes.  
   
   
       7 . The process of  claim 6 , wherein the anti-solvent is water.  
   
   
       8 . The process of  claim 3 , wherein the ratio of combined solution to combined anti-solvent is 40% or less.  
   
   
       9 . The method of  claim 1  further comprising the step of isolating the macrolide from the at least one fraction, wherein the macrolide has a final level of impurities that is lower than the initial level of impurities.  
   
   
       10 . The method of  claim 9  wherein the isolating comprises the step of concentrating the at least one fraction at reduced pressure and a temperature of about 70° C. or below.  
   
   
       11 . The method of  claim 10  wherein the temperature is about 60° C. or below.  
   
   
       12 . The method of  claim 10  wherein the pressure is about 760 mm Hg.  
   
   
       13 . The method of  claim 10  further comprising the step of, prior to concentrating, combining the at least one fraction with an inorganic acid.  
   
   
       14 . The method of  claim 13  wherein the inorganic acid is phosphoric acid.  
   
   
       15 . The method of  claim 13  wherein the amount of the acid is 1 to about 10 mL per liter of eluent.  
   
   
       16 . The method of  claim 9  wherein the isolating comprises the step of combining an antisolvent with the at least one fraction of eluent.  
   
   
       17 . The method of  claim 16  wherein, prior to the combining, the at least one fraction of effluent is concentrated at reduced pressure.  
   
   
       18 . The method of  claim 1  wherein the sorption resin is a macroreticular resin.  
   
   
       19 . The method of  claim 18  wherein the sorption resin is a macroreticular nonionic synthetic polymer.  
   
   
       20 . The method of  claim 1  wherein the bed of sorption resin is confined in a column.  
   
   
       21 . The method of  claim 1  wherein the volume of effluent collected in at least one fraction comprises about 60% to about 90%, by weight, of the macrolide initially present in the loading charge.  
   
   
       22 . The method of  claim 1  wherein the eluent flow rate is less than about 25 cm/h.  
   
   
       23 . The method of  claim 22  wherein the eluent flow rate is less than about 15 cm/h.  
   
   
       24 . The method of  claim 1  wherein the macrolide is selected from the group consisting of acsomycin, sirolimus, everolimus, and pimecrolimus.  
   
   
       25 . The method of  claim 1  wherein the eluent comprises a mixture of tetrahydrofuran and water having about 20 vol-% to about 50 vol-% tetrahydrofuran.  
   
   
       26 . The method of  claim 25  wherein the eluent has about 31 vol-% to about 40 vol-% tetrahydrofuran.  
   
   
       27 . The method of  claim 26  wherein the eluent has about 33 vol-% to about 35 vol-% tetrahydrofuran.  
   
   
       28 . The method of  claim 1  wherein the eluent comprises a mixture of acetonitrile and water having about 30 vol-% to about 70 vol-% acetonitrile.  
   
   
       29 . The method of  claim 28  wherein the eluent has about 40 vol-% to about 65 vol-% acetonitrile.  
   
   
       30 . The method of  claim 28  wherein the eluent includes up to about 0.003 parts of an inorganic acid per 1 part eluent, by volume.  
   
   
       31 . The method of  claim 30 , wherein the inorganic acid is phosphoric acid.  
   
   
       32 . The method of  claim 24  wherein the macrolide is ascomycin.  
   
   
       33 . The method of  claim 24  wherein the macrolide is sirolimus.  
   
   
       34 . The method of  claim 24  wherein the macrolide is everolimus.  
   
   
       35 . The method of  claim 24  wherein the macrolide is pimecrolimus.  
   
   
       36 . The method of  claim 2  wherein the loading charge is loaded onto the loading portion of sorption resin in a recirculating system.  
   
   
       37 . The method of  claim 1  wherein at least one additional bed of sorption resin is connected to the bed of sorption resin of step b.  
   
   
       38 . The method of  claim 37  wherein after additional series of eluent fractions the bed of resin of step b is disconnected.  
   
   
       39 . A method of separating ascomycin from the impurity tacrolimus therein comprising the steps of: 
 a) providing a loading charge of a ascomycin having an initial level of the impurity tacrolimus on a loading portion of a sorption resin that is a macroreticular resin,    b) juxtaposing the loading portion bearing the loading charge of ascomycin to a bed of sorption resin,    c) eluting the loading portion and bed juxtaposed thereto with an eluent comprising water and an organic solvent selected from tetrahydrofuran and acetonitrile to obtain an effluent,    d) collecting at least one fraction of effluent, and    e) isolating ascomycin having a final level of the impurity tacrolimus from the at least one fraction, wherein the final level of the impurity tacrolimus is lower than the initial level of the impurity.    
   
   
       40 . The method of  claim 39  wherein the eluent comprises a mixture of tetrahydrofuran and water having about 20 vol-% to about 50 vol-% tetrahydrofuran.  
   
   
       41 . The method of  claim 40  wherein the eluent comprises a mixture of tetrahydrofuran and water having about 31 vol-% to about 40 vol-% tetrahydrofuran.  
   
   
       42 . The method of  claim 41  wherein the mixture has about 33 vol-% to about 35 vol-% tetrahydrofuran.  
   
   
       43 . The method of  claim 39  wherein the eluent comprises a mixture of acetonitrile and water having about 30 vol-% to about 70 vol-% acetonitrile.  
   
   
       44 . The method of  claim 43  wherein the eluent has about 40 vol-% to about 65 vol-% acetonitrile.  
   
   
       45 . The method of  claim 39  wherein the eluent further comprises up to about 0.003 parts of an inorganic acid per part of eluent, by volume.  
   
   
       46 . The method of  claim 45 , wherein the inorganic acid is phosphoric acid.