Insoluble compositions for controlling blood glucose

Abstract

The present invention relates to insoluble compositions comprising a protein selected from the group consisting of insulin, insulin analogs, and proinsulins; a derivatized protein selected from the group consisting of derivatized insulin, derivatized insulin analog, and derivatized proinsulin; a complexing compound; a hexamer-stabilizing compound; and a divalent metal cation. Formulations of the insoluble composition are suitable for both parenteral and non-parenteral delivery for treating hyperglycemia and diabetes. Microcrystal forms of the insoluble precipitate are pharmaceutically analogous to the neutral protamine Hagedorn (NPH) insulin crystal form. Surprisingly, it has been discovered that suspension formulations of such insoluble compositions possess unique and controllable dissolution properties that provide therapeutically advantageous glucodynamics compared with insulin NPH formulations.

I claim:  
     
         1 . An insoluble composition, comprising: 
 a) a protein selected from the group consisting of insulin, insulin analog, and proinsulin;    b) a derivatized protein selected from the group consisting of derivatized insulin, derivatized insulin analog, and derivatized proinsulin;    c) a complexing compound;    d) a hexamer-stabilizing compound; and    e) a divalent metal cation.    
     
     
         2 . The composition of  claim 1  which is an amorphous precipitate.  
     
     
         3 . The composition of  claim 1  which is a microcrystal.  
     
     
         4 . The composition of  claim 3 , wherein the microcrystal has rod-like morphology.  
     
     
         5 . The composition of  claim 3 , wherein the microcrystal has irregular morphology.  
     
     
         6 . The composition of  claim 1 , wherein the derivatized protein is selected from the group consisting of fatty acid-acylated insulin, fatty acid-acylated insulin analogs, and fatty acid acylated proinsulins.  
     
     
         7 . The composition of  claim 6 , wherein the completing compound is protamine, the hexamer-stabilizing compound is a phenolic preservative, and the divalent metal cation is zinc.  
     
     
         8 . The composition of  claim 1 , wherein the derivatized protein is selected from the group consisting of fatty acid-acylated insulin and fatty acid-acylated insulin analogs.  
     
     
         9 . The composition of  claim 8 , wherein the complexing compound is protamine, the hexamer-stabilizing compound is a phenolic preservative, and the divalent metal cation is zinc.  
     
     
         10 . The composition of  claim 1 , wherein the derivatized protein selected from the group consisting of acylated insulin, acylated insulin analogs, and acylated proinsulins.  
     
     
         11 . The composition of  claim 1 , wherein the derivatized protein is selected from the group consisting of acylated insulin and acylated insulin analogs.  
     
     
         12 . The composition of  claim 11 , wherein the derivatized protein is mono-acylated at its Lys-Nε-amino group.  
     
     
         13 . The composition of  claim 12 , wherein the complexing compound is protamine, which is present at about 0.15 mg to about 0.5 mg per 3.5 mg of total protein.  
     
     
         14 . The composition of  claim 13 , wherein the divalent metal cation is zinc, which is present at about 0.3 mole to about 0.7 mole per mole of total protein.  
     
     
         15 . The composition of  claim 14 , wherein the hexamer-stabilizing compound is a phenolic preservative selected from the group consisting of phenol, m-cresol, o-cresol, p-cresol, chlorocresol, methylparaben, and mixtures thereof, and is present at a ratio of at least 3 moles of phenolic preservative to 6 moles of total protein.  
     
     
         16 . The composition of  claim 15 , wherein the protein is selected from the group consisting of insulin and insulin analogs.  
     
     
         17 . The composition of  claim 16 , wherein the protein is insulin.  
     
     
         18 . The composition of  claim 17 , wherein the derivatized protein is insulin that is mono-acylated at the LysB29-Nε-amino group.  
     
     
         19 . The composition of  claim 18 , wherein the derivatized protein is insulin that is acylated with a straight-chain, saturated fatty acid.  
     
     
         20 . The composition of  claim 19 , wherein the straight-chain, saturated fatty acid is selected from the group consisting of n-hexanoic acid, n-heptanoic acid, n-octanoic acid, n-nonanoic acid, and n-decanoic acid.  
     
     
         21 . The composition of  claim 20 , wherein the mole ratio between the protein and the derivatized protein is from about 1.9 to about 9:1.  
     
     
         22 . The composition of  claim 21 , wherein the straight-chain, saturated fatty acid is selected from the group consisting of n-hexanoic acid, n-octanoic acid, and n-decanoic acid.  
     
     
         23 . The composition of  claim 22 , wherein the straight-chain, saturated fatty acid is selected from the group consisting of n-octanoic acid and n-decanoic acid  
     
     
         24 . The composition of  claim 23 , wherein the straight-chain, saturated fatty acid is n-octanoic acid.  
     
     
         25 . The composition of  claim 24 , wherein the mole ratio between the protein and the derivatized protein is from about 1:3 to about 3:1.  
     
     
         26 . The composition of  claim 22 , wherein the mole ratio between the protein and the derivatized protein is from about 1:9 to about 1:1.  
     
     
         27 . The composition of  claim 16 , wherein the protein is an insulin analog.  
     
     
         28 . The composition of  claim 27 , wherein the protein is LysB28, ProB29-human insulin analog.  
     
     
         29 . The composition of  claim 27 , wherein the protein is AspB28-human insulin analog.  
     
     
         30 . The composition of  claim 1 , wherein the protein is insulin or an insulin analog, and the derivatized protein is an acylated protein selected from the group consisting of acylated insulin and acylated insulin analogs.  
     
     
         31 . The composition of  claim 1 , wherein the derivatized protein is insulin that is mono-acylated at its LysB29-Nε amino group with a straight-chain, saturated fatty acid.  
     
     
         32 . The composition of  claim 31 , wherein the straight-chain, saturated fatty acid is selected from the group consisting of n-hexanoic acid, n-heptanoic acid, n-octanoic acid, n-nonanoic acid, and n-decanoic acid.  
     
     
         33 . The composition of  claim 32 , wherein the derivatized protein is selected from the group consisting of B29-Nε-hexanoyl-human insulin, B29-Nε-octanoyl-human insulin, and B29-Nε-decanoyl-human insulin.  
     
     
         34 . The composition of  claim 31 , wherein the straight-chain, saturated fatty acid selected from the group consisting of n-dodecanoic acid, n-tetradecanoic acid, and n-hexadecanoic acid.  
     
     
         35 . The composition of  claim 1 , wherein the derivatized protein is di-acylated at the Lys-Nε-amino group and is also acylated at one N-terminal Nα-amino group, and wherein the fatty acid is selected from the group consisting of n-hexanoic acid, n-heptanoic acid, n-octanoic acid, n-nonanoic acid, and n-decanoic acid.  
     
     
         36 . The composition of  claim 1 , wherein the derivatized protein is acylated with a branched-chain, saturated fatty acid.  
     
     
         37 . The composition of  claim 36 , wherein the derivatized protein is acylated with a branched-chain, saturated fatty acid having from three to ten carbon atoms in its longest branch.  
     
     
         38 . The composition of  claim 1 , wherein the derivatized protein is an insulin analog that is mono-acylated at its Lys-Nε-amino group with a straight-chain, saturated fatty acid.  
     
     
         39 . The composition of  claim 38 , wherein the fatty acid is selected from the group consisting of n-hexanoic acid, n-heptanoic acid, n-octanoic acid, n-nonanoic acid, and n-decanoic acid.  
     
     
         40 . The composition of  claim 38 , wherein the derivatized protein is an insulin analog that is mono-acylated at the Nε-amino group with a fatty acid selected from the group consisting of n-dodecanoic acid, n-tetradecanoic acid, and n-hexadecanoic acid.  
     
     
         41 . The composition of  claim 38 , wherein the derivatized protein is selected from the group consisting of fatty acid-acylated animal insulins, fatty acid-acylated monomeric insulin analogs, fatty acid-acylated deletion analogs, and fatty acid-acylated pI-shifted insulin analogs.  
     
     
         42 . The composition of  claim 41 , wherein the derivatized protein is fatty acid-acylated des(B30)-human insulin analog, fatty acid-acylated LysB28, ProB29-human insulin analog, or fatty acid-acylated AspB28-human insulin analog.  
     
     
         43 . The composition of  claim 42 , wherein the derivatized protein is fatty acid-acylated des(B30)-human insulin analog.  
     
     
         44 . The composition of  claim 43 , wherein the derivatized protein is B29-Nε-myristoyl-des(B30)-human insulin analog.  
     
     
         45 . The composition of  claim 42 , wherein the derivatized protein is fatty acid-acylated LysB28, ProB29-human insulin analog.  
     
     
         46 . The composition of  claim 45 , wherein the derivatized protein is B28-Nε-myristoyl-LysB28, ProB29-human insulin analog.  
     
     
         47 . The composition of  claim 42 , wherein the derivatized protein is fatty acid-acylated AspB28-human insulin analog.  
     
     
         48 . The composition of  claim 1 , wherein the mole ratio between the protein and the derivatized protein is from about 1:9 to about 9:1.  
     
     
         49 . The composition of  claim 48 , wherein the ratio is from about 1:3 to about 3:1.  
     
     
         50 . The composition of  claim 48 , wherein the ratio is from about 1:9 to about 1:1.  
     
     
         51 . The composition of  claim 1 , wherein the protein is insulin.  
     
     
         52 . The composition of  claim 1 , wherein the protein is an insulin analog.  
     
     
         53 . The composition of  claim 52 , wherein the protein is a monomeric insulin analog.  
     
     
         54 . A suspension formulation, comprising an insoluble phase and a solution phase, wherein the insoluble phase comprises the insoluble composition of  claim 1 , and the solution phase comprises an aqueous solvent.  
     
     
         55 . The suspension formulation of  claim 54 , wherein the solution phase further comprises a phenolic preservative at a concentration of about 0.5 mg per mL to about 6 mg per mL of solution, a pharmaceutically acceptable buffer, and an isotonicity agent.  
     
     
         56 . The suspension formulation of  claim 54 , wherein the solution phase further comprises insulin, an insulin analog, a derivatized insulin, or a derivatized insulin analog.  
     
     
         57 . The suspension formulation of  claim 56 , wherein the solution phase comprises insulin.  
     
     
         58 . The suspension formulation of  claim 56 , wherein the solution phase comprises derivatized insulin.  
     
     
         59 . The suspension formulation of  claim 56 , wherein the solution phase comprises an insulin analog.  
     
     
         60 . The suspension formulation of  claim 56 , wherein the insulin analog is a monomeric insulin analog.  
     
     
         61 . The suspension formulation of  claim 56 , wherein the insulin analog is LysB28, ProB29-human insulin analog.  
     
     
         62 . The suspension formulation of  claim 56 , wherein the insulin analog is AspB28-human insulin analog.  
     
     
         63 . The suspension formulation of  claim 54 , wherein the solution phase further comprises a protein selected from insulin and insulin analogs and a derivatized protein selected from derivatized insulin and derivatized insulin analogs.  
     
     
         64 . The suspension formulation of  claim 63  wherein the protein in the solution phase is the same protein that is in the insoluble phase, and wherein the derivatized protein in the solution phase is the same derivatized protein that is in the insoluble phase.  
     
     
         65 . The suspension formulation of  claim 54 , wherein the insoluble phase consists essentially of an amorphous precipitate.  
     
     
         66 . The suspension formulation of  claim 54 , wherein the insoluble phase consists essentially of a microcrystal.  
     
     
         67 . The suspension formulation of  claim 54 , wherein the insoluble phase consists of a mixture of amorphous precipitate and microcrystal.  
     
     
         68 . The suspension formulation of  claim 54 , wherein the solution phase further comprises zinc and protamine, wherein the ratio of zinc to total protein in the suspension formulation is from about 5 to about 7 mole of zinc atoms per mole of total protein, and the ratio of protamine to total protein in the suspension formulation is from about 0.25 mg to about 0.5 mg per mg of total protein.  
     
     
         69 . A method of treating diabetes comprising administering the composition of  claim 1  to a patient in need thereof in a quantity sufficient to regulate blood glucose levels in the patient.  
     
     
         70 . A method of treating hyperglycemia comprising administering the composition of  claim 54  to a patient in need thereof in a quantity sufficient to regulate blood glucose levels in the patient.  
     
     
         71 . A hybrid hexamer composition, comprising six monomers and zinc, wherein at least one monomer is selected from the group consisting of insulin, insulin analogs, and proinsulins, and at least one monomer is selected from the group consisting of derivatized insulin, derivatized insulin analogs, and derivatized proinsulins.  
     
     
         72 . A mixed hexamer composition, comprising zinc protein hexamers and zinc derivatized protein hexamers, wherein the zinc protein hexamers comprise zinc and a protein selected from the group consisting of insulin, insulin analogs, and proinsulins, and wherein the zinc derivatized protein hexamers comprise zinc and a derivatized protein selected from the group consisting of derivatized insulin, derivatized insulin analogs, and derivatized proinsulins.  
     
     
         73 . A process for preparing the insoluble composition of  claim 1  comprising: 
 a) dissolving a protein, a derivatized protein, a hexamer-stabilizing compound, and a divalent metal cation in an aqueous solvent having a pH that will permit the formation of hexamers, and  
 b) adding a complexing compound.  
 
     
     
         74 . A process for preparing the insoluble composition of  claim 1  comprising: 
 a) dissolving a protein, a derivatized protein, a hexamer-stabilizing compound, and a divalent metal cation in an aqueous solvent having a pH that will not permit the formation of hexamers, and  
 b) adjusting the pH to between about 6.8 and about 7.8; and  
 c) adding a complexing compound.  
 
     
     
         75 . A process for preparing the insoluble composition of  claim 1  comprising: 
 a) dissolving a protein, a hexamer-stabilizing compound, and a divalent metal cation in an aqueous solvent having a pH that will permit the formation of hexamers;  
 b) separately, dissolving a derivatized protein, a hexamer-stabilizing compound, and a divalent metal cation in an aqueous solvent having a pH that will permit the formation of hexamers;  
 c) thoroughly mixing together the solutions from steps a) and b); and  
 d) adding a complexing compound to the solution produced in step c).  
 
     
     
         76 . A process for preparing the insoluble composition of  claim 1  comprising: 
 a) dissolving a protein, a hexamer-stabilizing compound, a divalent metal cation, and a complexing compound in an aqueous solvent, wherein the resulting solution has a pH at which precipitation does not occur;  
 b) separately, dissolving a -derivatized protein, a hexamer-stabilizing compound, a divalent metal cation, and a complexing compound in an aqueous solvent, wherein the resulting solution has a pH at which precipitation does not occur;  
 c) thoroughly mixing together the solutions from steps a) and b); and  
 d) adjusting the pH of the solution of step c) to a value at which precipitation occurs.  
 
     
     
         77 . A process for preparing the insoluble composition of  claim 1  comprising: 
 a) dissolving a protein, a derivatized protein, a hexamer-stabilizing compound, a divalent metal cation, and a complexing compound in an aqueous solvent, wherein the resulting solution has a pH at which precipitation does not occur; and  
 b) adjusting the pH of the solution of step a) to a value at which precipitation occurs.  
 
     
     
         78 . A process for preparing the insoluble composition of  claim 1  comprising: 
 a) dissolving a protein, a derivatized protein, a hexamer-stabilizing compound, and a divalent metal cation, in an aqueous solvent, wherein the resulting solution has a pH at which precipitation will not occur when a complexing agent is added;  
 b) adding a complexing compound; and  
 c) adjusting the pH of the solution of step b) to a value at which precipitation occurs.  
 
     
     
         79 . A process for preparing the insoluble composition of  claim 1  comprising: 
 a) dissolving a protein, a hexamer-stabilizing compound, and a divalent metal cation in an aqueous solvent, wherein the resulting solution has a pH at which precipitation will not occur when a complexing compound is added;  
 b) separately, dissolving a derivatized protein, a hexamer-stabilizing compound, and a divalent metal cation in an aqueous solvent, wherein the resulting solution has a pH at which precipitation will not occur when a complexing compound is added;  
 c) thoroughly mixing together the solutions from steps a) and b);  
 d) adding complexing compound to the solution of step c); and  
 e) adjusting the pH of the solution of step d) to a value at which precipitation occurs.  
 
     
     
         80 . A process for preparing the insoluble composition of  claim 1  comprising: 
 a) dissolving a protein, a protein derivative, a hexamer-stabilizing compound, and a divalent metal cation in an aqueous solvent, wherein the resulting solution has a pH at which precipitation will not occur when a complexing compound is added;  
 b) adjusting the pH of the solution of step a) to a value at which precipitation will occur when a complexing compound is added; and  
 c) adding a complexing compound to the solution of step b).  
 
     
     
         81 . A process for preparing the insoluble composition of  claim 1  comprising: 
 a) dissolving a protein, a hexamer-stabilizing compound, and a divalent metal cation in an aqueous solvent, wherein the resulting solution has a pH at which precipitation will not occur when a complexing compound is added;  
 b) separately, dissolving a derivatized protein, a hexamer-stabilizing compound, and a divalent metal cation in an aqueous solvent, wherein the resulting solution has a pH at which precipitation will not occur when a complexing compound is added;  
 c) thoroughly mixing together the solutions from steps a) and b);  
 d) adjusting the pH of the solution of step c) to a value at which precipitation will occur when a complexing compound is added; and  
 e) adding a complexing compound to the solution of step d).  
 
     
     
         82 . A process for preparing hybrid hexamers, comprising dissolving a protein, a derivatized protein, a hexamer-stabilizing compound, and a divalent metal cation in an aqueous solvent having a pH that will permit the formation of hexamers.  
     
     
         83 . A process for preparing hybrid hexamers, comprising: 
 a) dissolving a protein, a derivatized protein, a hexamer-stabilizing compound, and a divalent metal cation in an aqueous solvent having a pH that will not permit the formation of hexamers, and    b) adjusting the pH to between about 6.8 and about 7.8.