Abstract:
The invention relates to formulations containing the peptide p277 or variants thereof which are characterized by having increased stability during storage and transport.

Description:
FIELD OF THE INVENTION  
       [0001]     The invention relates to formulations containing the peptide p277 or variants thereof which are characterized by having increased stability during storage and transport.  
       BACKGROUND OF THE INVENTION  
       [0002]     The p277 peptide and its variants consist of 24 amino acids and have the structure Val-Leu-Gly-Gly-Gly-X 1 -Ala-Leu-Leu-Arg-X 2 -Ile-Pro-Ala-Leu-Asp-Ser-Leu-X 3 -Pro-Ala-Asn-Glu-Asp (SEQ. I.D. No. 1), where X 1 , X 2  may be Cys or Val and X 3  may be Thr or Lys. Said peptide is described in EP 0 820 303. Further formulations of p277 and its variants are disclosed in EP 0 837 672.  
         [0003]     The p277 peptide and its variants are not stable at room temperature and must therefore be stored in the deep-freeze state below −10° C. The known formulations of p277 and its variants, which are the corresponding lyophilizates, are also not stable at room temperature and must therefore be stored and transported in the deep-freeze state. In order to achieve a stability of 12 months, said formulations must be permanently stored below −10° C.  
         [0004]     It was the object of the invention to find formulations of the p277 peptide and its variants, which are stable at relatively high temperatures for at least 6 months. It was particularly desirable to find those formulations which are stable at refrigerator temperatures of from +2° C. to +8° C. or even at room temperature (from +15° C. to +25° C.). Stable means that the total proportion of secondary products does not increase by more than 1% over a period of 6 months.  
         [0005]     It was intended, in particular, to find a stable formulation of this kind for p277 peptide (Val 6 -Val 11 -Thr 19 ) having the structure Val-Leu-Gly-Gly-Gly-Val-Ala-Leu-Leu-Arg-Val-Ile-Pro-Ala-Leu-Asp-Ser-Leu-Thr-Pro-Ala-Asn-Glu-Asp.  
       SUMMARY OF THE INVENTION  
       [0006]     The invention therefore relates to formulations which contain p277 or its variants having the structure Val-Leu-Gly-Gly-Gly-X 1 -Ala-Leu-Leu-Arg-X 2 -Ile-Pro-Ala-Leu-Asp-Ser-Leu-X 3 -Pro-Ala-Asn-Glu-Asp (SEQ. I.D. No. 1), where X 1 , X 2  may be Cys or Val and X 3  may be Thr or Lys, and a citrate buffer. 
     
    
     DETAILED DESCRIPTION  
       [0007]     Preference is given to formulations containing p277 (Val 6 -Val 11 -Thr 19 ) having the structure Val-Leu-Gly-Gly-Gly-Val-Ala-Leu-Leu-Arg-Val-Ile-Pro-Ala-Leu-Asp-Ser-Leu-Thr-Pro-Ala-Asn-Glu-Asp (SEQ. I.D. No. 2), and a citrate buffer.  
         [0008]     The invention furthermore relates to formulations containing p277 or its variants, an excipient from the group consisting of trehalose, maltose, lactose and mannitol, and a citrate buffer.  
         [0009]     Preference is given to formulations containing the excipient mannitol. Preference is furthermore given to a mannitol content of from 5 mg/ml to 200 mg/ml. Particular preference is given to a mannitol content of from 25 mg/ml to 50 mg/ml. Very particular preference is given to a mannitol content of 40 mg/ml.  
         [0010]     Further preference is given to formulations containing p277 and its variants, mannitol and a citrate buffer, wherein the pH may be from pH 3 to pH 6.  
         [0011]     Particular preference is given to formulations containing p277 and its variants, mannitol and a citrate buffer, wherein the pH may be from pH 4.5 to pH 6.  
         [0012]     A citrate buffer means an aqueous solution of citrates, preferably alkali metal citrates, particularly preferably sodium citrate or potassium citrate, and a strong acid, preferably hydrochloric acid (HCl).  
         [0013]     The formulations of the invention may be present in the form of a solution or a lyophilizate, preferably a lyophilizate.  
         [0014]     The examples listed below are intended to illustrate but not limit the invention.  
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   TABLE 1                                       Example   Comparative                1   2   3   4   5   6   Example 1                        p277 (Val 6 -Val 11 -Thr 19 )   1.3   mg   1.3   mg   1.3   mg   1.3   mg   1.3   mg   1.3   mg   1.3   mg            Trehalose   40.0   mg   —   —   —   —   —                    Maltose   —   40.0   mg   —   —   —   —                    Lactose   —   —   40.0   mg   —   —   —                    Mannitol   —   —   —   40.0   mg   40.0   mg   40.0   mg   40.0   mg            Citrate buffer pH 3.0*   —   —   —   ad 1.0   ml   —   —                    Citrate buffer pH 4.5*   ad 1.0   ml   ad 1.0   ml   ad 1.0   ml   —   ad 1.0   ml   —                    Citrate buffer pH 6.0*   —   —   —   —   —   ad 1.0   ml                    Water for injection purposes                                                   ad 1.0   ml                  
 
         [0015]    
       
         
               
             
               
               
               
               
             
               
               
               
             
               
               
               
               
             
           
               
                   
               
               
                   
               
               
                 * Citrate buffer composition: 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 NaOH 
                 8.000 
                 g 
               
               
                   
                 Citric acid * 1 H 2 O 
                 21.056 
                 g 
               
             
          
           
               
                   
                 HCl 
                 ad pH 3.0; pH 4.5; pH 6.0 
               
             
          
           
               
                   
                 Water for injection purposes 
                 ad 1.0 
                 l 
               
               
                   
                   
               
             
          
         
       
     
         [0016]     Stability of the formulations was determined after storage at temperatures of −20° C., +5° C. and +25° C. for 6 months. For this purpose, in each case 1 ml of said formulations was introduced into 2 ml clear glass vials and lyophilized therein. The vials were sealed with bromobutyl rubber stoppers and a flip-off flange cap.  
         [0000]     Storage took place in a temperature- and humidity-monitored/controlled area with protection from light.  
         [0017]     Table 2 depicts total contamination of the formulations in %.  
                                                                   TABLE 2                                       Time                T0   T6 months   T6 months   T6 months                        Temperature       −20° C.   +5° C.   +25° C.       Example 1   1.704   1.7968   1.7563   1.8570       Example 3   0.550   0.4808   0.4929   0.6714       Example 5   0.563   0.4776   0.4932   0.5667       Comparative Example 1   0.643   0.8872   1.9531   7.4321                  
 
         [0018]     The table indicates that the formulations of the invention are stable at −20° C., +5° C. and at +25° C. over a period of 6 months. At +5° C., total contaminations increase by no more than 0.0523% and at +25° C. by no more than 0.153%. In Comparative Example 1, total contaminations increase at +5° C. by 1.3101% and at +25° C. by 6.789%. The formulations of the invention are thus distinctly more stable than the formulation of the Comparative Example.