Liquid, Aqueous Pharmaceutical Compositions of Factor VII Polypeptides

Abstract

The present invention is directed to liquid, aqueous pharmaceutical compositions stabilised against chemical and/or physical degradation containing Factor VII polypeptides, and methods for preparing and using such compositions, as well as vials containing such compositions, and the use of such compositions in the treatment of a Factor VII-responsive syndrome. The main embodiment is represented by a liquid, aqueous pharmaceutical composition comprising at least 0.01 mg/mL of a Factor VII polypeptide (i); a buffering agent (ii) suitable for keeping pH in the range of from about 4.0 to about 9.0; and at least one stabilising agent (iii) comprising a —C(═N—Z 1 —R 1 )—NH—Z 2 —R 2 motif, e.g. benzamidine compounds and guanidine compounds such as arginine.

1 . A liquid, aqueous, and pharmaceutically acceptable composition comprising
 (i) at least 0.01 mg/mL of a Factor VII polypeptide;   (ii) a buffering agent suitable for keeping pH of the composition in the range of about 4 to about 9; and   (iii) a stabilizing composition comprising a benzamidine compound according to the formula —C 6 H 4 —C(═N—Z 1 —R 1 )—NH—Z 2 —R 2 , wherein   (a) C6H 4  denotes a substituted benzene ring, and   (b) Z 1  and Z 2  independently are selected from the group consisting of —O—, —S—, —NR H — and a single bond, where R H  is selected from the group consisting of hydrogen, C 1-4 -alkyl, aryl and arylmethyl, and R 1  and R 2  independently are selected from the group consisting of hydrogen, optionally substituted C 1-6 -alkyl, optionally substituted C 2-6 -alkenyl, optionally substituted aryl, optionally substituted heterocyclyl,
 —C═N—Z 1 —R 1  forms part of a heterocyclic ring, 
 —C—NH—Z 2 —R 2  forms part of a heterocyclic ring, or 
 —C(═N—Z 1 —R 1 )—NH—Z 2 —R 2  forms a hetercyclic ring wherein —Z 1 —R 1 —R 2 —Z 2 — is a biradical. 
   
     
     
         2 . The composition of  claim 1 , wherein the benzamidine compound comprises the motif >N—C 6 H 4 —C(═N—Z 1 —R 1 )—NH—Z 2 —R 2 , wherein C 6 H 4  denotes a substituted benzene ring. 
     
     
         3 . The composition of  claim 2 , wherein the molecular weight of the benzamidine compound is 1000 Da or less. 
     
     
         4 . The composition of  claim 3 , wherein the benzamidine is p-amino-benzamidine. 
     
     
         5 . The composition of  claim 1 , wherein the concentration of the benzamidine compound is at least 1 μM. 
     
     
         6 . The composition of  claim 5 , wherein the concentration of the benzamidine compound is at least 1 mM. 
     
     
         7 . The composition of  claim 4 , wherein the concentration of p-amino-benzamidine is at least 1 μM. 
     
     
         8 . The composition according to  claim 1 , wherein the benzamidine compound is S-2-[3-(4-Carbamimidoylphenyl)-ureido]-N-[1-(3-methoxyphenyl)-ethyl]-acetamide. 
     
     
         9 . The composition of  claim 8 , wherein the concentration of S-2-[3-(4-Carbamimidoylphenyl)-ureido]-N-[1-(3-methoxyphenyl)-ethyl]-acetamide is at least 1 μM. 
     
     
         10 . The composition of  claim 1 , wherein the Factor VII polypeptide is human Factor VIIa. 
     
     
         11 . The composition of  claim 1 , wherein the Factor VII polypeptide is a Factor VII sequence variant. 
     
     
         12 . The composition of  claim 1 , wherein the Factor VII polypeptide is present in a concentration of 0.01-20 mg/mL. 
     
     
         13 . The composition of  claim 1 , wherein the buffering agent (ii) comprises at least one component selected from the group consisting of acids and salts of MES, PIPES, ACES, BES, TES, HEPES, TRIS, histidine, imidazole, glycine, glycylglycine, glycinamide, phosphoric acid, acetic acid, lactic acid, glutaric acid, citric acid, tartaric acid, malic acid, maleic acid, and succinic acid. 
     
     
         14 . The composition of  claim 13 , wherein the concentration of the buffering agent (ii) is about 1-100 mM. 
     
     
         15 . The composition of  claim 1 , wherein the composition further comprises a non-ionic surfactant (iv). 
     
     
         16 . The composition of  claim 15 , wherein the non-ionic surfactant (iv) is at least one selected from the group consisting of polysorbates, poloxamers, polyoxyethylene alkyl ethers, ethylene/polypropylene block co-polymers and polyethyleneglycol (PEG). 
     
     
         17 . The composition of  claim 16 , wherein the concentration of the non-ionic surfactant (iv) is 0.005-2% by weight. 
     
     
         18 . The composition of  claim 1 , wherein the composition further comprises a tonicity modifying agent (v). 
     
     
         19 . The composition of  claim 15 , wherein the composition further comprises a tonicity modifying agent (v). 
     
     
         20 . The composition of  claim 19 , wherein the tonicity modifying agent (v) is at least one selected from the group consisting of neutral salts, amino acids, peptides of 2-5 amino acid residues, monosaccharides, disaccharides, polysaccharides, and sugar alcohols. 
     
     
         21 . The composition of  claim 20 , wherein at least one tonicity modifying agent (v) is a neutral salt selected from the group consisting of sodium salts, potassium salts, calcium salts, and magnesium salts. 
     
     
         22 . The composition of  claim 20 , wherein the tonicity modifying agent (v) is sodium chloride in combination with at least one selected from the group consisting of calcium chloride, calcium acetate, magnesium chloride and magnesium acetate. 
     
     
         23 . The composition of  claim 22 , wherein the tonicity modifying agent (v) is present in a concentration of at least 1 mM. 
     
     
         24 . The composition of  claim 19 , wherein at least one tonicity modifying agent (v) is an ionic strength modifying agent (v/a). 
     
     
         25 . The composition of  claim 24 , wherein the composition has an ionic strength of at least 50 mM. 
     
     
         26 . The composition according to  claim 1 , wherein the composition has an osmolality of 300±50 milliosmol/kg. 
     
     
         27 . The composition of  claim 1 , wherein the composition further comprises an antioxidant (vi). 
     
     
         28 . The composition of  claim 21 , wherein the composition further comprises an antioxidant (vi). 
     
     
         29 . The composition of  claim 28 , wherein the antioxidant (vi) is selected from L-methionine, D-methionine, methionine analogues, methionine-containing peptides, methionine-homologues, ascorbic acid, cysteine, homocysteine, gluthatione, cystine, and cysstathionine. 
     
     
         30 . The composition of  claim 29 , wherein the antioxidant (vi) is present in a concentration of 0.1-5.0 mg/mL. 
     
     
         31 . The composition of  claim 1 , wherein the composition further comprises a preservative (vii). 
     
     
         32 . The composition of  claim 28 , wherein the composition further comprises a preservative (vii). 
     
     
         33 . The composition of  claim 32 , wherein the preservative (vii) is selected from the group consisting of phenol, benzyl alcohol, orto-cresol, meta-cresol, para-cresol, methyl paraben, propyl paraben, benzalkonium chloride, and benzaethonium chloride. 
     
     
         34 . The composition of  claim 1 , wherein the composition comprises 0.1-20 mg/mL of a Factor VII polypeptide; the stabilizing agent comprises an effective amount of p-amino-benzamidine, S-2-[3-(4-Carbamimidoylphenyl)-ureido]-N-[1-(3-methoxyphenyl)-ethyl]-acetamide, or a mixture thereof, in a concentration of at least 5 μM; and the composition further comprises a non-ionic surfactant (iv) and a tonicity modifying agent (v) in a concentration of at least 5 mM. 
     
     
         35 . The composition of  claim 1 , wherein the composition is adapted for parenteral administration. 
     
     
         36 . The composition of  claim 1 , wherein the composition is adapted for subcutaneous, intramuscular, or intravenous injection. 
     
     
         37 . A method for treating a Factor VII-responsive syndrome comprising administering to a subject in need thereof an effective amount of the composition of  claim 1 . 
     
     
         38 . The method of  claim 37 , wherein the method comprises administering an effective amount of the composition of  claim 4  to the subject. 
     
     
         39 . The method of  claim 37 , wherein the method comprises administering an effective amount of the composition of  claim 8  to the subject. 
     
     
         40 . The method of  claim 37 , wherein the method comprises administering an effective amount of the composition of  claim 34  to the subject. 
     
     
         41 . An air-tight container containing the composition of  claim 1 , and optionally an inert gas. 
     
     
         42 . The container of  claim 41 , wherein the composition does not comprise an antioxidant.