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 . A method of making a composition comprising microparticles comprising DAS181, the method comprising:
 a) providing a feedstock composition comprising DAS181, a counterion and an organic solvent; and   b) cooling the composition to below 25° C., whereby a composition comprising microparticles comprising DAS181 is formed.   
     
     
         2 . The method of  claim 1  wherein the DAS181 comprises a polypeptide comprising the amino acid sequence of SEQ ID NO:1. 
     
     
         3 . The method of  claim 1  wherein the DAS181 comprises a polypeptide comprising the amino acid sequence of SEQ ID NO:2. 
     
     
         4 .- 13 . (canceled) 
     
     
         14 . The method of  claim 1  wherein the step of providing a feedstock composition containing DAS181, a counterion and an organic solvent at a temperature at or above about 25° C. comprises providing a solution containing DAS181 and a counterion in an aqueous solvent and adding an organic solvent to the solution containing DAS181 and a counterion in an aqueous solvent. 
     
     
         15 . The method of  claim 1 , further comprising isolating the microparticles from the composition comprising microparticles, whereby a dry powder formulation of microparticles is formed. 
     
     
         16 . (canceled) 
     
     
         17 . The method of  claim 1 , wherein the feedstock composition comprises two or more counterions. 
     
     
         18 .- 36 . (canceled) 
     
     
         37 . The method of any of the forgoing claims, further comprising adding a scavenging agent to the mixture comprising the active agent, the counterion and the organic solvent or to the solution comprising the active agent and the counterion. 
     
     
         38 . The method of  claim 37 , wherein the scavenging agent is selected from among an amine, an antioxidant, a sugar, a chelator or a combination of two or more thereof. 
     
     
         39 . The method of  claim 38 , wherein the scavenging agent is histidine. 
     
     
         40 . The method of  claim 37 , wherein the scavenging agent further comprises a sugar. 
     
     
         41 . The method of  claim 40  wherein the sugar is trehalose. 
     
     
         42 .- 43 . (canceled) 
     
     
         44 . A composition comprising microparticles, wherein the microparticles are about 60-75% wt/wt DAS181, 3-6% wt/wt histidine, 4-8% wt/wt histidine hydrochloride, 7-11% wt/wt trehalose, 4-8% wt/wt magnesium sulfate, and 8-12% water. 
     
     
         45 .- 51 . (canceled) 
     
     
         52 . A composition comprising microparticles, wherein the microparticles are about 69-74% wt/wt DAS181, 4-8% wt/wt histidine, 5-9% wt/wt histidine hydrochloride, 8-12% wt/wt trehalose, and 4-8% wt/wt magnesium sulfate. 
     
     
         53 .- 55 . (canceled) 
     
     
         56 . The composition of  claim 52  wherein the microparticles have an MMAD of 1-8 microns, 4-8 microns, 2-8 microns or 3-8 microns. 
     
     
         57 .- 59 . (canceled) 
     
     
         60 . A composition comprising microparticles, wherein the microparticles are about 80-90% wt/wt DAS181, 1.5-3.5% wt/wt sodium sulfate, and 8-12% water. 
     
     
         61 .- 63 . (canceled) 
     
     
         64 .- 66 . (canceled) 
     
     
         67 . A capsule containing the composition of claim  64 . 
     
     
         68 . A composition comprising microparticles, wherein the microparticles are about 94-98% (wt/wt) DAS181 and 1.5-3.5% wt/wt sodium sulfate. 
     
     
         69 .- 78 . (canceled)