Pegylated Insulin Lispro Compounds

Abstract

The present invention relates to the field of diabetes. More particularly, the invention relates to PEGylated insulin lispro compounds that are PEGylated with high molecular weight poly(ethylene glycol), are highly soluble at physiological pH, have an extended duration of action, and characterized by pharmacokinetic, pharmacodynamic, and/or activity peak-trough ratios of less than 2. The invention also relates to methods of providing such molecules, to pharmaceutical compositions containing them, and to their therapeutic uses.

1 - 33 . (canceled) 
     
     
         34 . A method of treating hyperglycemia or diabetes in a patient in need of such a treatment, comprising administering to the patient a therapeutically effective amount of a PEGylated insulin lispro compound of the formula: P-[(A)-(B)], or a pharmaceutically acceptable salt thereof, wherein:
 A is the A-chain of insulin lispro (SEQ ID NO: 1);   B is the B-chain of insulin lispro (SEQ ID NO: 3); and   P is a PEG having a molecular weight in the range from about 17.5 kDa to about 40 kDa and is attached via a urethane covalent bond to the epsilon-amino group of the lysine at position 28 of B, and wherein A and B contain a disulfide bond between the cysteine at position 7 of A (SEQ ID NO: 1) and the cysteine at position 7 of B (SEQ ID NO: 3), a disulfide bond between the cysteine at position 20 of A (SEQ ID NO: 1) and the cysteine at position 19 of B (SEQ ID NO: 3), and a disulfide bond between the cysteine at position 6 of A (SEQ ID NO: 1) and the cysteine at position 11 of A (SEQ ID NO: 1).   
     
     
         35 . The method of  claim 34 , wherein the patient is treated for diabetes mellitus. 
     
     
         36 . The method of  claim 34 , wherein the patient is treated for gestational diabetes. 
     
     
         37 . The method of  claim 34 , wherein the PEG has a molecular weight in the range from about 17.5 kDa to about 25 kDa. 
     
     
         38 . The method of  claim 34 , wherein the PEG has a molecular weight of about 20 kDa. 
     
     
         39 . A method of treating hyperglycemia or diabetes in a patient in need of such a treatment, comprising administering to the patient a therapeutically effective amount of a pharmaceutical composition, said pharmaceutical composition comprising a PEGylated insulin lispro compound, zinc, and a phenolic preservative, the PEGylated insulin lispro compound having the formula: P-[(A)-(B)], or a pharmaceutically acceptable salt thereof, wherein:
 A is the A-chain of insulin lispro (SEQ ID NO: 1);   B is the B-chain of insulin lispro (SEQ ID NO: 3); and   P is a PEG having a molecular weight in the range from about 17.5 kDa to about 40 kDa and is attached via a urethane covalent bond to the epsilon-amino group of the lysine at position 28 of B, and wherein A and B contain a disulfide bond between the cysteine at position 7 of A (SEQ ID NO: 1) and the cysteine at position 7 of B (SEQ ID NO: 3), a disulfide bond between the cysteine at position 20 of A (SEQ ID NO: 1) and the cysteine at position 19 of B (SEQ ID NO: 3), and a disulfide bond between the cysteine at position 6 of A (SEQ ID NO: 1) and the cysteine at position 11 of A (SEQ ID NO: 1).   
     
     
         40 . The method of  claim 39 , wherein the patient is treated for one of diabetes mellitus or gestational diabetes. 
     
     
         41 . The method of  claim 39 , wherein the PEG has a molecular weight in the range from about 17.5 kDa to about 25 kDa. 
     
     
         42 . The method of  claim 39 , wherein the pharmaceutical composition further comprises a TRIS buffer at a concentration in the range from about 10 mM to about 25 mM TRIS, wherein the pH of said pharmaceutical composition is from about pH 7.0 to about pH 8.0. 
     
     
         43 . The method of  claim 42 , wherein the pharmaceutical composition further comprises at least one isotonicity agent producing a solution with a tonicity between about 270 mOsm and about 330 mOsm. 
     
     
         44 . The method of  claim 39 , wherein the pharmaceutical composition further comprises a phosphate buffer at a concentration in the range from about 5 mM to about 10 mM, wherein the pH of said pharmaceutical composition is from about pH 7.0 to about pH 7.5. 
     
     
         45 . The method of  claim 39 , wherein the concentration of PEGylated insulin lispro compound in the pharmaceutical composition is from about 15 mg/mL to about 40 mg/mL. 
     
     
         46 . The method of  claim 39 , wherein the phenolic preservative is m-cresol, said pharmaceutical composition comprising about 30 mM m-cresol, wherein the pharmaceutical composition further comprises between about 2.5 mM and about 25 mM calcium and a surfactant. 
     
     
         47 . The method of  claim 39 , wherein the pharmaceutical composition comprises between about 0.3 and 5 moles of zinc per mole of PEGylated insulin lispro compound. 
     
     
         48 . The method of  claim 39 , wherein the pharmaceutical composition further comprises between about 100 mM and about 150 mM NaCl, between about 2.5 mM and about 25 mM calcium, and between about 0.5 mg/mL and about 5 mg/mL of poloxamer 188. 
     
     
         49 . The method of  claim 39 , wherein the pharmaceutical composition further comprises:
 a TRIS buffer at a concentration in the range from about 10 mM to about 25 mM TRIS, wherein the pH of said pharmaceutical composition is from about pH 7.0 to about pH 8.0;   at least one isotonicity agent producing a solution with a tonicity between about 270 mOsm and about 330 mOsm; and   between about 0.3 and 5 moles of zinc per mole of PEGylated insulin lispro compound, wherein the phenolic preservative is m-cresol, said pharmaceutical composition comprising about 30 mM m-cresol.   
     
     
         50 . A process of making a PEGylated insulin lispro compound of the formula:
 P-[(A)-(B)], or a pharmaceutically acceptable salt thereof, wherein:   A is the A-chain of insulin lispro (SEQ ID NO: 1);   B is the B-chain of insulin lispro (SEQ ID NO: 3); and   P is a PEG having a molecular weight in the range from about 17.5 kDa to about 40 kDa, and wherein A and B are properly cross-linked and P is attached via an urethane covalent bond to the epsilon-amino group of the lysine at position 28 of B which comprises reacting the epsilon-amino group of the lysine at position 28 of B with monomethoxypoly(ethylene glycol) p-nitrophenyl carbonate (mPEG-NPC) having a weight average molecular weight of between about 17.5 kDa and about 40 kDa in an aqueous solvent at a pH between about 8.5 and about 11.5 and at between about 25° C. and about 30° C. for a period of time between about 3 and about 6 hours.   
     
     
         51 . The process of  claim 50 , wherein the PEG:insulin lispro molar ratio is in the range between about 2.5 and about 5.0. 
     
     
         52 . The process of  claim 51 , wherein the weight average molecular weight of the mPEG-NPC is about 20 kDa. 
     
     
         53 . The process of  claim 52 , wherein the pH of the reaction is maintained at between about 10.5 and about 11.5. 
     
     
         54 . The process of  claim 53 , wherein the reaction is conducted at between about 25° C. and about 30° C. for about 3 hours.