Keratinocyte growth factor-2 formulations

Abstract

The invention is directed to liquid and lyophilized forms of Keratinocyte Growth Factor-2 (KGF-2) and derivatives thereof. This invention further relates to the formulation of KGF-2 for therapeutic use, for example, to promote or accelerate wound healing.

What is claimed is:  
     
         1 . A pharmaceutical composition, comprising: 
 (1) a KGF-2 polypeptide in a concentration range of about 0.02 to about 40 mg/ml (w/v);    (2) a buffer having a buffering capacity of between about 5.0 and about 8.0 at a concentration range of about 5 mM to about 50 mM; and    (3) a pharmaceutically acceptable diluent to bring the composition to a designated volume;    or a reaction product thereof.    
     
     
         2 . The pharmaceutical composition of  claim 1 , further comprising: 
 (1) a chelating agent at a concentration range of about 1 mM to about 10 mM; and    (2) NaCl at a concentration range of about 0.1 mM to about 150 mM.    
     
     
         3 . The pharmaceutical composition of  claim 1 , further comprising one of: 
 (1) about 0.5% to about 2% w/v glycerol,    (2) about 0.1% to about 1% w/v methionine, or    (3) about 0.1% to about 2% w/v monothioglycerol.    
     
     
         4 . The pharmaceutical composition of  claim 1 , wherein said KGF-polypeptide is present in a concentration range of about 0.05 to about 30 mg/ml (w/v).  
     
     
         5 . The pharmaceutical composition of  claim 4 , wherein said KGF-polypeptide is present in a concentration range of about 0.1 to about 20 mg/ml (w/v).  
     
     
         6 . The pharmaceutical composition of  claim 5 , wherein said KGF-polypeptide is present in a concentration range of about 0.2 to 4 mg/ml.  
     
     
         7 . The pharmaceutical composition of  claim 1 , wherein said KGF-2 polypeptide is KGF-2-Δ33.  
     
     
         8 . The pharmaceutical composition of  claim 1 , wherein said diluent is water.  
     
     
         9 . The pharmaceutical composition of  claim 2 , wherein said chelating agent is EDTA at a concentration of about 1 mM, and said NaCl is present at a concentration of about 125 mM.  
     
     
         10 . The pharmaceutical composition of  claim 1 , wherein said pH is from about pH 5.5 to about pH 6.5.  
     
     
         11 . The pharmaceutical composition of  claim 10 , wherein said pH is about pH 6.2.  
     
     
         12 . The pharmaceutical composition of  claim 1 , wherein said buffer is selected from the group consisting of phosphonic, acetic, aconitic, citric, glutaric, malic, succinic carbonic acid, and an alkali or alkaline earth salt thereof.  
     
     
         13 . The pharmaceutical composition of  claim 12 , wherein said buffer is a phosphate, acetate or citrate salt.  
     
     
         14 . The pharmaceutical composition of  claim 13 , wherein said buffer is a citrate salt.  
     
     
         15 . The pharmaceutical composition of  claim 1 , wherein said buffer is present in a concentration range of about 5 mM to about 30 mM.  
     
     
         16 . The pharmaceutical composition of  claim 15 , wherein said buffer is a citrate salt present in a concentration of from about 10 mM to about 20 mM.  
     
     
         17 . The pharmaceutical composition of  claim 1 , further comprising a stabilizing amount of one or more of (a) an antioxidant or (b) a thiol-compound.  
     
     
         18 . The pharmaceutical composition of  claim 1 , wherein said composition is maintained at a temperature at or below −20° C.  
     
     
         19 . The pharmaceutical composition of  claim 1 , comprising: 
 (1) 2 mg/ml KGF-2 Δ33 polypeptide (w/v);    (2) 20 mM sodium citrate;    (3) 125 mM NaCl;    (4) 1 mM EDTA; and    (5) water as diluent, or    a reaction product thereof.    
     
     
         20 . A pharmaceutical composition, comprising: 
 (1) a KGF-2 polypeptide in a concentration range of about 0.02 to about 40 mg/ml (w/v);    (2) a buffer having a buffering capacity of between about 5.0 and about 8.0 at a concentration range of about 5 mM to about 50 mM;    (3) a bulking agent; and    (4) a pharmaceutically acceptable diluent to bring the composition to a designated volume;    or a reaction product thereof.    
     
     
         21 . The pharmaceutical composition of  claim 20 , wherein said bulking agent is selected from the group consisting of sucrose, glycine, mannitol, trehalose, and mixtures thereof.  
     
     
         22 . The pharmaceutical composition of  claim 1 , further comprising: 
 (1) a chelating agent at a concentration range of about 1 mM to about 10 mM; and    (2) NaCl at a concentration range of about 0.1 mM to about 125 mM.    
     
     
         23 . The pharmaceutical composition of  claim 21 , wherein said bulking agent is sucrose or a mixture of sucrose and glycine.  
     
     
         24 . The pharmaceutical composition of  claim 21 , wherein said bulking agent is present in a concentration of about 2% to about 10% w/v.  
     
     
         25 . The pharmaceutical composition of  claim 21 , wherein said bulking agent is 5% mannitol, 7% sucrose, 8% trehalose, or 2% glycine+0.5% sucrose.  
     
     
         26 . The pharmaceutical composition of  claim 20 , wherein said pH is about pH 6.2.  
     
     
         27 . The pharmaceutical composition of  claim 20 , wherein said diluent is water.  
     
     
         28 . The pharmaceutical composition of  claim 20 , wherein said buffer is selected from the group consisting of phosphonic, acetic, aconitic, citric, glutaric, malic, succinic carbonic acid, and an alkali or alkaline earth salt thereof.  
     
     
         29 . The pharmaceutical composition of  claim 28 , wherein said buffer is a phosphate or citrate salt.  
     
     
         30 . The pharmaceutical composition of  claim 29 , wherein said buffer is a citrate salt.  
     
     
         31 . The pharmaceutical composition of  claim 27 , wherein over 90% of the water is removed by lyophilization.  
     
     
         32 . The pharmaceutical composition of  claim 31 , which is reconstituted in with an amount of sterile water effective to maintain isotonic conditions of 290 mOsm.  
     
     
         33 . The pharmaceutical composition of  claim 20 , wherein said KGF polypeptide is KGF-2-Δ33.  
     
     
         34 . The pharmaceutical composition of  claim 20 , wherein said buffer is added in a concentration from about 5 mM to about 50 mM  
     
     
         35 . The pharmaceutical composition of  claim 34 , wherein said buffer is citrate at a concentration of about 10 mM.  
     
     
         36 . The pharmaceutical composition of  claim 20 , further including a stabilizing amount of one or more of (a) an antioxidant, or (b) a thiol-compound.  
     
     
         37 . The pharmaceutical composition of  claim 31 , wherein said composition is reconstituted in sterile water containing a stabilizing amount of an antioxidant comprising: a) about 0.01% to about 2% w/v monothioglycerol, b) about 0.01% to about 2% w/v ascorbic acid, c) about 0.01% to about 2% w/v methionine or d) combinations thereof.  
     
     
         38 . A pharmaceutical composition, comprising: 
 (1) a KGF-2 polypeptide in a concentration range of about 0.02 to about 40 mg/ml (w/v);    (2) an effective amount of citric acid or a pharmaceutically acceptable salt thereof, at a concentration range of about 5 mM to about 20 mM;    (3) NaCl at a concentration range of about 0 mM to about 125 mM,    (4) EDTA at a concentration range of about 1 mM to about 10 mM and    (5) one or more of sucrose, mannitol, glycine or trehalose at a concentration range of about 2% w/v to about 15% w/v; and    (6) water.    
     
     
         39 . The pharmaceutical composition of  claim 38 , wherein said KGF-2 polypeptide is present at a concentration of about 2 mg/ml, about 4 mg/ml, or about 10 mg/ml.  
     
     
         40 . The pharmaceutical composition of  claim 38 , wherein over 90% of the water is removed by lyophilization.  
     
     
         41 . The pharmaceutical composition of  claim 1 , further comprising a thickening agent in an amount effective to raise the viscosity to about 50 to about 10,000 cps.  
     
     
         42 . The pharmaceutical composition of  claim 20 , further comprising a thickening agent in an amount effective to raise the viscosity to about 50 to about 10,000 cps.  
     
     
         43 . The pharmaceutical composition of  claim 41 , wherein said thickening agent is present in an amount effective to raise the viscosity to about 50 to about 1,000 cps.  
     
     
         44 . The pharmaceutical composition of  claim 43 , wherein said thickening agent in an amount effective to raise the viscosity to about 200 to about 300 cps.  
     
     
         45 . The pharmaceutical composition of  claim 41 , wherein said thickening agent is present in a concentration of 0 to 5% (w/w).  
     
     
         46 . The pharmaceutical composition of  claim 41 , wherein said thickening agent is a water soluble etherified cellulose or a high molecular weight polymer of acrylic acid cross-linked with allylsucrose or an allyl ether of pentaerythritol.  
     
     
         47 . The pharmaceutical composition of  claim 46 , wherein said etherified cellulose is an alkyl cellulose, hydroxyalkyl cellulose, carboxyalkyl cellulose or alkylhydroxyalkyl cellulose.  
     
     
         48 . The pharmaceutical composition of  claim 41 , wherein said etherified cellulose is methylcellulose, hydroxyethyl cellulose, hydroxy propyl cellulose, hydroxy propyl methylcellulose, or carboxymethyl cellulose.  
     
     
         49 . The pharmaceutical composition of  claim 47 , wherein said etherified cellulose derivative has a molecular weight of about 50,000 to about 700,000 and is present in a concentration of about 0 to about 20% by weight.  
     
     
         50 . The pharmaceutical composition of  claim 49 , wherein said etherified cellulose derivative has a molecular weight of about 80,000 to about 240,000 and is present in a concentration of about 2% to about 8% by weight.  
     
     
         51 . The pharmaceutical composition of  claim 42 , wherein said buffer is citrate in a concentration of about 10 mM to about 50 mM.  
     
     
         52 . The pharmaceutical composition of  claim 51 , wherein said buffer is citrate in a concentration of about 10 mM to about 20 mM citrate.  
     
     
         53 . The pharmaceutical composition of  claim 51 , wherein said bulking agent is sucrose in a concentration of about 0% to about 5% sucrose.  
     
     
         54 . The pharmaceutical composition of  claim 53 , wherein said thickening agent is added directly to a liquid formulation and thereafter lyophilized.  
     
     
         55 . The pharmaceutical composition of  claim 53 , wherein said thickening agent is added to a lyophilized formulation by reconstituting said formulation by adding a suitable diluent having a thickening agent dissolved therein.  
     
     
         56 . A thickened KGF-2 polypeptide solution composition formed by mixing: 
 (1) a topically effective amount of a KGF polypeptide;    (2) about 10 mM to about 500 mM sodium citrate buffer;    (3) about 0.1 to about 150 mM NaCl;    (4) 1 mM EDTA;    (5) about 0.1 to about 7% sucrose;    (6) about 0.75 to about 1.5% (w/w) carboxy methyl cellulose or about 0.5 to about 1.5% hydroxy propyl methyl cellulose or about 0.25 to about 0.75% hydroxy ethyl cellulose or about 0 to 1% carbomer or any combination thereof.    
     
     
         57 . The composition of  claim 1 , further comprising a gelling agent in an amount effective to raise the viscosity to about 0 to about 10,000 cps at room temperature.  
     
     
         58 . The composition of  claim 20 , further comprising a gelling agent in an amount effective to raise the viscosity to about 0 to about 10,000 cps at room temperature.  
     
     
         59 . The composition of  claim 57 , wherein said gel forming agent is a water-soluble polymer capable of forming a viscous aqueous solution, or non-water soluble, water-swellable polymer capable of forming a viscous solution.  
     
     
         60 . The composition of  claim 59 , wherein said gel forming agent is a high molecular weight polymer selected from the group consisting of vinyl polymer, polyoxyethylene-polyoxypropylene copolymer, polysaccharide, protein, poly(ethylene oxide), acrylamide polymer or a salt thereof.  
     
     
         61 . The composition of  claim 60 , wherein said gel forming agent is (1) a vinyl polymer selected from the group consisting of polyacrylic acid, polymethacrylic acid, polyvinyl pyrrolidone polyvinyl alcohol, and salts and esters thereof; or (2) a polysaccharide selected from the group consisting of a cellulose derivative, a glycosaminoglycan, agar, pectin, alginic acid, dextran, α-amylose, amylopectin, chitosan, and salts esters thereof.  
     
     
         62 . The composition of  claim 60 , wherein said gel forming agent is a glycosaminoglycan selected from the group consisting of hyaluronic acid, chondroitin, chondroitin-4-sulfate, heparan sulfate, heparin and salts and esters thereof.  
     
     
         63 . The composition of  claim 62 , wherein said glycosaminoglycan is present in combination with collagen, gelatin, or fibronectin.  
     
     
         64 . The composition of  claim 60 , wherein said gel forming agent is an acrylamide polymer selected from the group consisting of a polyacrylamide or a polymethacrylamide.  
     
     
         65 . The composition of  claim 60 , wherein said gel forming agent is a polyoxyethylene-polyoxypropylene block copolymer.  
     
     
         66 . The composition of  claim 65 , which comprises about 10 to about 60% by weight of a polyoxyethylene-polyoxypropylene block copolymer having an average molecular weight of about 500 to 50,000.  
     
     
         67 . The composition of  claim 66 , which comprises about 14 to about 18% by weight of a polyoxyethylene-polyoxypropylene block copolymer having a molecular weight in the range 1,000 to 15,000.  
     
     
         68 . The composition of  claim 67 , wherein said gel forming agent is a polyoxyethylene-polyoxypropylene block copolymer is Pluronic F108, Pluronic F127, or Poloxamer 407.  
     
     
         69 . The composition of  claim 1 , wherein said KGF-2 polypeptide is present in a concentration of about 0.01 mg/ml to about 10 mg/ml.  
     
     
         70 . The composition of  claim 57 , wherein said composition is formed by mixing: 
 (1) a KGF-2 polypeptide, in a final calculated concentration of 0.01 mg/ml to about 10 mg/ml;    (2) an effective amount of a buffering agent;    (3) about 10% to about 60% by weight of a polyoxyethylene-polyoxypropylene block copolymer having an average molecular weight of about 500 to 50,000; and    (4) a pharmaceutically acceptable diluent, preferably water.    
     
     
         71 . The composition of  claim 70 , wherein polyoxyethylene-polyoxypropylene block copolymer is present at a concentration of about 14% to about 18%.  
     
     
         72 . A KGF-2 gel formulation, comprising: 
 (1) a pharmaceutically active amount of KGF-2 polypeptide;    (2) about 10 mM to about 500 mM sodium citrate;    (3) about 0.1 mM to about 150 mM NaCl;    (4) about 1 mM EDTA;    (5) about 0.1% to about 7% sucrose;    (6) about 14% to about 18% Pluronic F127; and    (7)N about pH 6.2    
     
     
         73 . A KGF-2 gel formulation, comprising: 
 (1) a KGF-2 polypeptide at a concentration range of about 0.01 mg/ml to about 10 mg/ml (w/v),    (2) sodium citrate at a concentration range of about 5 mM to about 20 mM;    (3) about 10% to about 25% (w/v), of Pluronic 127 or Poloxamer 407; and    (4) water to volume.    
     
     
         74 . The gel formulation of  claim 73 , further comprising: 
 (1) EDTA at a concentration range of about 1 mM to about 10 mM.    (2) NaCl at a concentration range of about 0.1 mM to about 125 mM.    
     
     
         75 . The pharmaceutical composition of  claim 1 , wherein said KGF-2 polypeptide is a N-terminal deletion selected from the group consisting of Ala (63)—Ser (208) (KGF-2Δ28) and Ser (69)—Ser (208) (KGF-2Δ33).  
     
     
         76 . The pharmaceutical composition of  claim 1 , wherein said KGF-2 polypeptide is a N-terminal or C-terminal deletion mutant selected from the group consisting of Ala (39)—Ser (208); Pro (47)—Ser (208); Val (77)—Ser (208); Glu (93)—Ser (208); Glu (104)—Ser (208); Val (123)—Ser (208); Gly (138)—Ser (208); Met (1), Thr (36); and Cys (37)—Lys (153).