Microparticle Formulations for Delivery to the Lower and Central Respiratory Tract and Methods of Manufacture

Abstract

Microparticle formulations of a sialidase fusion protein are produced by contacting an aqueous solution of a protein or other active agent with an organic solvent, a counterion and a scavenging agent, and chilling the solution. The microparticles are useful for preparing stable, uniform pharmaceuticals of predetermined defined dimensions.

1 .- 78 . (canceled) 
     
     
         79 . A method for preparing a pharmaceutical composition comprising DAS181, the method comprising:
 a) providing microparticles comprising DAS181 having the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2, wherein the microparticles are about 60-70% wt/wt DAS181, 7-15% wt/wt histidine, 7-11% wt/wt trehalose, 4-8% wt/wt magnesium sulfate, and 8-12% wt/wt water; and   b) combining the microparticles with a pharmaceutically acceptable aqueous solution.   
     
     
         80 . The method of  claim 79  wherein the microparticles are 62% to 68% (wt/wt) DAS181. 
     
     
         81 . The method of  claim 79  wherein the microparticles have a Mass Median Aerodynamic Diameter (MMAD) of 1-8 microns, 2-8 microns or 3-8 microns. 
     
     
         82 . The method of  claim 79  wherein the microparticles have a Geometric Standard Deviation (GSD) of 1.5-1.7, 1.3-1.6, or 1.4-1.6. 
     
     
         83 . The method of  claim 79  wherein the weight percent of microparticles having a fine particle fractionation (FPF) below 5 microns is less than 10%. 
     
     
         84 . A method for preparing a pharmaceutical composition comprising DAS181, the method comprising:
 a) providing microparticles comprising DAS181 having the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2, wherein the microparticles are about 69-74% wt/wt DAS181, 9-17% wt/wt histidine, 8-12% wt/wt trehalose, and 4-8% wt/wt magnesium sulfate; and   b) combining the microparticles with a pharmaceutically acceptable aqueous solution.   
     
     
         85 . The method of  claim 84  wherein the microparticles have a GSD of 1.5-1.7, 1.3-1.6, or 1.4-1.6. 
     
     
         86 . The method of  claim 84  wherein the weight percent of microparticles having a FPF below 5 microns is less than 10%. 
     
     
         87 . A method for preparing a pharmaceutical composition comprising DAS181, the method comprising:
 a) providing microparticles comprising DAS181 having the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2, wherein the microparticles are about 80-90% wt/wt DAS181, 1.5-3.5% wt/wt sodium sulfate, and 8-12% water; and   b) combining the microparticles with a pharmaceutically acceptable aqueous solution.   
     
     
         88 . The method of  claim 87  wherein the microparticles have a GSD of 1.2-1.8, 1.3-1.7 or 1.4-1.6 
     
     
         89 . The method of  claim 87  wherein the weight percent of microparticles having a fine particle fractionation (FPF) below 5 microns is less than 10%. 
     
     
         90 . The method of  claim 79  wherein the microparticles comprise 3-6% wt/wt histidine free base and 5-9% wt/wt histidine hydrochloride. 
     
     
         91 . The method of  claim 84  wherein the microparticles comprise 3-8% wt/wt histidine free base and 5-9% wt/wt histidine hydrochloride. 
     
     
         92 . A method for preparing a pharmaceutical composition comprising DAS181, the method comprising:
 a) providing microparticles comprising DAS181 having the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2, wherein the microparticles are about 60-70% wt/wt DAS181, 8-15% wt/wt histidine, 7-11% wt/wt trehalose, 4-8% wt/wt magnesium sulfate, and 8-12% water; and   b) combining the microparticles with a pharmaceutically acceptable aqueous solution.   
     
     
         93 . A method for preparing a pharmaceutical composition comprising DAS181, the method comprising:
 (a) providing microparticles comprising DAS181 having the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2, wherein the microparticles are about 69-74% wt/wt DAS181, 7-17% wt/wt histidine, 8-12% wt/wt trehalose, and 4-8% wt/wt magnesium sulfate; and   b) combining the microparticles with a pharmaceutically acceptable aqueous solution.