Abstract:
Compositions having excellent blood lipid peroxide lowering activity are provided. In particular, compositions for lowering lipid peroxides in the blood which contain pravastatin together with at least one substance selected from the group consisting of taurine, pantethine and inositol hexanicotinate. Use of these compositions enables the provision of excellent preventive or remedial agents capable of lowering the concentration of lipid peroxides in the blood which show effects of injuring vascular endothelial cells, accelerating platelet aggregation, forming foam cells, etc.

Description:
This is a Continuation-in-Part application of International Application No. PCT/JP01/09662 filed Nov. 5, 2001 which is incorporated herein by reference in its entirety. 
    
    
     BACKGROUND OF THE INVENTION 
     The present invention relates to blood lipid peroxide-lowering compositions consisting of pravastatin in combination with one or more substances selected from taurine, pantethine and inositol hexanicotinate. 
     An increase in blood lipid peroxide levels causes damage to endothelial cells, enhances platelet aggregation, and promotes foam cell forming, all of which contribute to arteriosclerosis. Thus lipid peroxide-lowering agents are considered to be useful agents. 
     Pravastatin reduces total cholesterol levels in the blood by inhibiting HMG-CoA reductase activity. However, it is not known that pravastatin reduces lipid peroxide levels in the blood. 
     Taurine and pantethine are known to reduce lipid peroxide levels in the blood (Reference: Sulfur Amino Acids, Vol. 7, No.1, 1984, p. 201–205; Geriatr. Med., Vol. 19, No. 3, p. 415–422). 
     However, it is not known that inositol hexanicotinate decreases lipid peroxide levels in the blood. 
     BRIEF DESCRIPTION OF THE INVENTION 
     The present invention relates to drug compositions containing pravastatin in combination with one or more substances selected from taurine, pantethine and inositol hexanicotinate, which reduce levels of lipid peroxides in the blood. 
     The present inventors investigated drug compositions that decrease lipid peroxide levels in the blood, and found that co-administration of pravastatin with taurine, pantethine or inositol hexanicotinate all decrease lipid peroxide levels in the blood and completed the present invention. 
     DETAILED DESCRIPTION OF THE INVENTION 
     Pravastatin (chemical name: (+)-(3R,5R)-3,5-dihydroxy-7-[(1S,2S,6S,8S,8aR)-6-hydroxy-2-methyl-8-[(S)-2-methylbutyryloxy]-1,2,6,7,8,8a-hexahydro-1-naphthyl]heptanoate) is a compound represented by the following chemical structure. It may exist in the form of a salt (particularly a sodium salt). The manufacturing methods of pravastatin have been disclosed in Japanese Patent Kokai Application No. SHO 57-2240 and so forth. Since pravastatin is commercially available, it is easily acquired.                         
 
     Blood lipid peroxides are lipid peroxides present in the blood, and involve hyperoxidated LDL (low-density lipoproteins) and so on. 
     The term “lowering” in the expression lipid peroxide-lowering agents indicates that the levels are decreased by clinically significant amounts following administration of the agents. 
     The weight percent of pravastatin contained in a tablet of the present invention of the lipid peroxide-lowering composition drug is 0.01 to 5%, preferably 0.05 to 3.0%. The weight percent of taurine in the tablet is typically 0.3 to 50%, preferably 1 to 25%. The weight percent of pantethine in the tablet is typically 1.3 to 50%, preferably 2.7 to 20%. The weight percent of inositol hexanicotinate in the tablet is typically 0.05 to 50%, preferably 0.5 to 25%. 
     The content of pravastatin contained in liquid and solution preparations of lipid peroxide-lowering composition according to the present invention is typically 1 to 100 mg/mL and preferably 3 to 7 mg/mL; that of taurine is typically 2.5–50 mg/mL, preferably 8 to 35 mg/mL. In addition, the content of pantethine in the liquid and solution preparation is typically 1 to 200 mg/mL, preferably 5 to 100 mg/mL, and that of inositol hexanicotinate typically 1 to 40 mg/mL, preferably 2 to 20 mg/mL. 
     Practical preparations of the drug compositions to reduce lipid peroxide levels in the blood are tablets, granules (involving powders), capsules, and liquids and solutions, etc., and they are manufactured following addition of the required additive agents or materials, if necessary, according to conventional methods described in The Pharmacopeia of Japan. 
     In the preparations described above, additive agents that are conventionally used can be employed based on the preparation. 
     For instance, in the case of tablets, lactose and crystalline cellulose are used as diluents, magnesium aluminometasilicate, etc., are used as stabilizing agents, hydroxypropylcellulose, etc., are used as binders, and magnesium stearate is used as a lubricant. 
     In granules and capsules, lactate and purified sucrose are used as diluents, magnesium aluminometasilicate is used as a stabilizing agent, corn starch, etc., are used as adsorbents, and hydroxypropylcellulose and polysorbate, etc., are used as binders. 
     In liquids and solutions, D-sorbitol solution and honey, etc., are used as sweeteners, dl-malic acid, etc., are used as flavoring agents, disodium dihydrogen ethylenediamine tetraacetate, etc., are used as stabilizing agents, ethanol is used as a co-solvent, and polyoxyethylene hydrogenated castor oil stearate 60, etc., are used as a solubilizer. 
     In the preparations described above, disintegrators such as crospovidone, etc.; adsorbents such as calcium silicate, etc.; coloring agents such as red ferric oxide, caramel, etc.; pH modifiers such as sodium benzoate, etc.; and a flavor may be used, if necessary. 
    
    
     EXAMPLES 
     The present invention is described in more detail by way of the following examples. However, the present invention is not limited to these examples. 
     Test Example 1 
     Tablets 
     (1) 
                                                       TABLE 1                               Inositol           Taurine   Pantethine   hexanicotinate           4 tablets   4 tablets   4 tablets           (680 mg)   (1,440 mg)   (1,400 mg)                                    Pravastatin sodium    20 mg    20 mg    20 mg       Taurine   500 mg   —   —       Pantethine   —   500 mg   —       Inositol hexanicotinate   —   —   500 mg       Crystalline cellulose   120 mg    12 mg    12 mg       Magnesium   144 mg   —   —       aluminometasilicate       Sucrose esters of fatty acids   —   140 mg   140 mg       Hydroxypropylcellulose    96 mg    48 mg    96 mg       Magnesium stearate    24 mg    24 mg    24 mg       Crospovidone   100 mg    48 mg   100 mg       Lactose   a   a   a               a: appropriate quantity            
(2) Manufacturing Methods
 
     The amount of each component described above is weighed and prepared according to the methods described in the “General Rules for Preparations of Tablets” in “The Pharmacopeia of Japan”. 
     (1) Composition 
                                                       TABLE 2                       Taurine   Pantethine   Inositol           4   4   hexanicotinate           packages   packages   4 packages           (4 g)   (5.2 g)   (5 g)                                    Pravastatin sodium     20 mg     20 mg     20 mg       Taurine     500 mg   —   —       Pantethine   —   1,000 mg   —       Inositol hexanicotinate   —   —   1,000 mg       Purified sucrose   1,400 mg   1,600 mg   1,400 mg       Stevia extracts   —     16 mg     16 mg       Corn starch   1,200 mg   1,200 mg   1,200 mg       Polysorbate 80     80 mg     48 mg     80 mg       Magnesium     144 mg   —     144 mg       aluminometasilicate       Magnesium stearate     24 mg     24 mg     24 mg       Lactose   a   a   a               a: appropriate quantity            
2) Manufacturing Methods
 
     The amount of each component described above is weighed and prepared according to the methods described in the “General Rules for Preparations of Granules” in “The Pharmacopeia of Japan”. 
     Test Example 3 
     Capsules 
     (1) Components 
                                                       TABLE 3                       Taurine   Pantethine   Inositol           4   8   hexanicotinate           capsules   capsules   8 capsules                                    Pravastatin sodium     20 mg     20 mg     20 mg       Taurine     500 mg   —   —       Pantethine   —     500 mg   —       Inositol hexanicotinate   —   —     500 mg       Corn starch     960 mg     960 mg     960 mg       Polysorbate 80     80 mg     48 mg     80 mg       Magnesium     144 mg   —     144 mg       aluminometasilicate       Magnesium stearate     24 mg     24 mg     24 mg       Lactose   a   a   a       Subtotal   1,520 mg   1,940 mg   2,000 mg       Capsule     320 mg     640 mg     640 mg       Total   1,840 mg   2580 mg   2,640 mg               a: appropriate quantity            
(2) Manufacturing Methods
 
     The amount of each component described above is weighed and prepared according to the methods described in the “General Rules for Preparations of Granules” in “The Pharmacopeia of Japan”, and hard capsules are prepared by filling the granules into capsules. 
     Test Example 4 
     Liquids and Solutioins 
     (1) Components 
                                                       TABLE 4                               Inositol           Taurine   Pantethine   Hexanicotinate           100 mL   100 mL   100 mL                                    Pravastatin sodium    20 mg    20 mg    20 mg       Taurine   500 mg   —   —       Pantethine   —   500 mg   —       Inositol hexanicotinate   —   —   500 mg       D-Sorbitol solution (70%)    4 g    6 g    4 g       Honey    7 g    8 g    7 g       dI-Malic acid   200 mg   —   200 mg       Disodium dihydrogen-    20 mg    20 mg    20 mg       ethylenediamine       tetraacetate       Ethanol    2 mL    2 mL    2 mL       Polyoxyethylene   100 mg   100 mg   100 mg       hydrogenated-       castor oil stearate 60       Sodium benzoate    60 mg    60 mg    60 mg       Flavor   b   b   b       Distilled water   a   a   a               a: appropriate quantity,       b: trace amount            
(2) Manufacturing Methods
 
     The amount of each component described above is weighed and prepared according to the methods described in the “General Rules for Preparations of Liquids and Solutions” in “The Pharmacopeia of Japan”. 
     Test Examples 
     (1) Test Compounds 
     Pravastatin with a purity of 99.4%, manufactured at Sankyo Co., Ltd. was used. 
     Taurine, pantethine and inositol hexanicotinate were purchased from Nacalai Tesque, Inc., Dai-ich Pharmaceutical Co., Ltd., and SHIRATORI PHARMACEUTICAL CO., LTD., respectively and were used. 
     (2) Test Animals 
     Beagle dogs aged 5 months were purchased from Covance Research Products Inc. and used after 1 month of quarantine and acclimatisation breeding. 
     (3) Form of the Preparation for Administration, Method of Preparation of the Formulation and Method of Stocking the Formulation 
     The required amounts of pravastatin or each component of the combination drug calculated from the body weight of each dog were weighed and filled in a gelatin capsule purchased from TORPAC Inc. Capsules filled with pravastatin were stocked in a refrigerator and those filled with combination drugs stocked at room temperature until use. 
     The combination drugs were filled in identical gelatin capsules. 
     (4) Route of Administration and Administration Period 
     Pravastatin or combination drug capsules were forcibly orally administered to each of the test animals once daily between 9:00 and 12:30. Animals were fasted for 2 or 3 hr prior to administration of the capsules. 
     The administration period was 11 successive days. 
     (5) Preparation of Test Samples and Procedures 
     Blood (10 mL) was collected from the superficial radial vein 3 or 4 days prior to administration (−3 or −4 days in the first week prior to capsule administration) and 4 and 8 days after administration of the capsule. Animals were fasted for approximately 18 hr prior to blood collection. Collected blood was placed in a test tube and left at room temperature for 0.5–1 hr and centrifuged (3,000 rpm, for 10 min). The obtained serum was used for assays of blood lipid peroxides according to Yagi&#39;s methods. 
     A fluorometer (Hitachi, Ltd., F3000) was used for assay of the lipid peroxides. 
     Results 
     Lipid peroxide levels in blood collected from dogs treated with pravastatin or one compound selected from taurine, pantethine and inositol hexanicotinate, as well as combinations of pravastatin together with one of the above drugs, were converted to their relative ratios against their averaged pre-dosing levels (100 as determined 2 and 1 weeks prior to drug administration). The averaged value in each group was obtained from 5 animals a group. 
     
       
         
               
             
               
               
               
             
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 5 
               
             
             
               
                   
               
               
                 (Effects of co-administration of pravastatin and taurine) 
               
             
          
           
               
                   
                   
                 Blood Lipid Peroxide Levels 
               
               
                   
                 Dose 
                 after administration 
               
             
          
           
               
                 Test Substance 
                 (mg/kg) 
                 4 days 
                 8 days 
               
               
                   
               
             
          
           
               
                 Pravastatin alone 
                 2 
                 110.8 
                 116.2 
               
               
                 Taurine alone 
                 1,000 
                 95.8 
                 93.8 
               
               
                 Pravastatin 
                 2 
                 89.9 
                 77.5 
               
               
                 + taurine 
                 1,000 
               
               
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 6 
               
             
             
               
                   
               
               
                 (Effects of co-administration of pravastatin and pantethine) 
               
             
          
           
               
                   
                   
                 Blood Lipid Peroxide Levels 
               
               
                   
                 Dose 
                 after administration 
               
             
          
           
               
                   
                 Test Substance 
                 (mg/kg) 
                 4 days 
                 8 days 
               
               
                   
                   
               
             
          
           
               
                   
                 Pravastatin alone 
                 2 
                 110.8 
                 116.2 
               
               
                   
                 Pantethine alone 
                 300 
                 82.5 
                 105.0 
               
               
                   
                 Pravastatin 
                 2 
                 83.6 
                 75.4 
               
               
                   
                 + pantethine 
                 300 
               
               
                   
                   
               
             
          
         
       
     
     
       
         
               
             
               
               
               
             
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 7 
               
             
             
               
                   
               
               
                 (Effects of co-administration of pravastatin and 
               
               
                 inositol hexanicotinate) 
               
             
          
           
               
                   
                   
                 Blood Lipid Peroxide Levels 
               
               
                   
                 Dose 
                 after administration 
               
             
          
           
               
                 Test Substance 
                 (mg/kg) 
                 4 days 
                 8 days 
               
               
                   
               
             
          
           
               
                 Pravastatin alone 
                 2 
                 110.8 
                 116.2 
               
               
                 Inositol hexanicotinate 
                 400 
                 98.5 
                 96.5 
               
               
                 alone 
               
               
                 Pravastatin 
                 2 
                 83.8 
                 81.3 
               
               
                 + inositol 
                 400 
               
               
                 hexanicotinate 
               
               
                   
               
             
          
         
       
     
     The composition of the present invention can be used for lowering blood lipid peroxide levels in a mammal and in particular a human. When administering to a human (or other mammal) in need of treatment, an effective amount of an agent which is pravastatin in combination with one or more of taurine, pantethine and inositol hexanicotinate, is used. 
     Although the dose of compounds used according to the invention may widely vary depending on the extent of diseases and age of patients, (e.g. human patients), the dose of one administration of pravastatin is normally within the range of from 0.01 mg/kg to 10 mg/kg, preferably from 0.1 mg/kg to 1 mg/kg, administered once or several times a day dependent on the extent of diseases. 
     The dose of one administration of taurine is normally within the range of from 1 mg/kg to 600 mg/kg, preferably from 10 mg/kg to 60 mg/kg, administered once or several times a day dependent on the extent of diseases. 
     The dose of one administration of pantethine is normally within the range of from 0.06 mg/kg to 120 mg/kg, preferably from 0.6 mg/kg to 12 mg/kg, administered once or several times a day dependent on the extent of diseases. 
     The dose of one administration of inositol hexanicotinate is normally within the range of from 0.16 mg/kg to 36 mg/kg, preferably from 1.6 mg/kg to 3.6 mg/kg, administered once or several times a day dependent on the extent of diseases. 
     The present invention, drug compositions of pravastatin in combination with a drug selected from taurine, pantethine and inositol hexanicotinate, exert excellent blood lipid peroxide-lowering effects.