JP4949661B2 - Pharmaceutical composition containing HMG-CoA reductase inhibitor and glutathione - Google Patents

Description

  The present invention relates to a pharmaceutical composition for treating cardiovascular diseases.

  Although it has been said that “humans are old with blood vessels” for a long time, in recent years, an increase in blood homocysteine (Hcy) level has attracted attention and has become widely known as an independent risk factor for arteriosclerosis.

  Hcy is an amino acid produced in the process of metabolism of methionine, which is one of the essential amino acids. It has been found that the amount of Hcy in blood increases due to genetic predisposition, vitamin cofactor deficiency, aging, sex, decreased renal function, diabetes, other diseases, drugs, smoking, etc. (see, for example, Non-Patent Document 1) ), This symptom is called hyperhomocysteinemia.

At present, as a treatment for hyperhomocysteinemia, folic acid administration for smoothly promoting Hcy metabolism is the first stage treatment, and vitamin B ₆ and vitamin B ₁₂ are administered as the second stage. (For example, refer nonpatent literature 1).

  The HMG-CoA reductase inhibitor (hereinafter referred to as statin) is a drug that specifically and antagonistically inhibits the pathway from HMG-CoA to mevalonic acid in the living body to lower the blood cholesterol level. There is no clear conclusion about the effect on blood Hcy levels.

  In addition, glutathione is known to have an antioxidant action and is also known as an antidote such as drug addiction, but its effect on blood Hcy levels is unknown.

  In addition to the above-mentioned Hcy, it has recently been found that “vascular aging” is deeply related to a decrease in vascular endothelial cell-derived nitric oxide (NO) production, that is, a decrease in endothelial NO synthase (eNOS) activity. It was.

  Vascular endothelium dysfunction is deeply related to the onset and progression of arteriosclerosis, and it is thought that the decrease in NO produced from eNOS is the major cause. It is said that it plays a role such as anti-arteriosclerosis action from the standpoints of inhibiting adhesion of neutrophils to endothelial cells, inhibiting migration and proliferation of vascular smooth muscle cells, inhibiting LDL oxidation, etc. (Refer to many other prior art).

  Animal studies have already confirmed that NO synthesis inhibition contributes directly to the onset and progression of arteriosclerosis (see, for example, Non-Patent Document 3). Even in humans, vascular endothelial cell-derived NO functions in many ways in terms of vascular protection, and is therefore considered to play an important role in the treatment of cardiovascular diseases.

  In addition, there is a report that specific statins have an effect of improving vascular endothelial cell function (for example, see Non-Patent Document 4).

  It is unclear whether glutathione has an effect of eNOS activation and / or blood concentration maintenance and increase of vascular endothelial cell-derived NO.

  Glutathione peroxidase (GPx) is an antioxidant enzyme important for the body that detoxifies hydrogen peroxide and peroxides in the presence of glutathione, and arteriosclerosis and cerebrovascular disease in which an increase in blood lipid peroxide is observed. It is useful for the prevention or treatment of diseases such as myocardial infarction (see, for example, Non-Patent Document 5).

  The presence or absence of the GPx activity increasing action of statin or glutathione is unknown.

So far, there is no known pharmaceutical composition containing a statin and glutathione, and there is no report on the combined use of a statin and glutathione.
Progress in Medicine, Vol. 19 No. 8, 1999 p. 49-53 Angiology, Vol. 38 No. 4 1998 p. 215-219 Vascular Medicine, Vol. 2 No. 2 2001 p. 189-194 Pharmacology and treatment, Vol. 29 No. 10 2001 p. 715-718 http: // www. hokudai. ac. jp / pharma / eisei / takahasi. htm

  As described above, (1) hyperhomocysteinemia, (2) decreased eNOS activity and / or decreased blood concentration of vascular endothelial cell-derived NO, and (3) improved decrease in GPx activity are particularly arterial. This is extremely important in the treatment or prevention of diseases related to “vascular aging” such as sclerosis. Furthermore, it would be even more effective and economical to find a means to improve these three etiologies simultaneously.

  Therefore, the present inventors simultaneously (1) decreased the amount of Hcy in the blood, (2) increased the activity of eNOS and / or increased the blood concentration of vascular endothelial cell-derived NO, and (3) increased the GPx activity simultaneously. We have conducted extensive research to find safe drugs to be realized.

  As a result, by using a combination of statin and glutathione, (1) a decrease in blood Hcy level, (2) an increase in eNOS activity and / or an increase in blood concentration of vascular endothelial cell-derived NO, and (3) It has been found that an increase in GPx activity can be achieved at the same time, and the present invention has been completed.

The present invention
(1) A pharmaceutical composition containing an HMG-CoA reductase inhibitor and glutathione,
(2) The pharmaceutical composition according to (1), wherein the HMG-CoA reductase inhibitor is one or more selected from the group consisting of pravastatin, lovastatin, simvastatin, fluvastatin, atorvastatin, pitavastatin, and rosuvastatin,
(3) The pharmaceutical composition according to (1), wherein the HMG-CoA reductase inhibitor is pravastatin,
(4) The pharmaceutical composition according to any one of (1) to (3), for reducing the amount of homocysteine in the blood,
(5) The pharmaceutical composition according to any one of (1) to (3), for maintaining the blood homocysteine level,
(6) The pharmaceutical composition according to any one of (1) to (3), for treating or preventing hyperhomocysteinemia,
(7) The pharmaceutical composition according to any one of (1) to (3), for treating or preventing a disease caused by hyperhomocysteinemia,
(8) Diseases resulting from hyperhomocysteinemia include arteriosclerosis, ischemic heart disease, myocardial infarction, thrombosis, peripheral vascular disorder, Buerger's disease, Raynaud's disease, cerebral infarction, cerebral circulation failure, cerebrovascular dementia, The pharmaceutical composition according to (7), which is one or more selected from the group consisting of Alzheimer's disease and Parkinson's disease,
(9) The pharmaceutical composition according to (7), wherein the disease caused by hyperhomocysteinemia is arteriosclerosis,
(10) The pharmaceutical composition according to any one of (1) to (3), which promotes nitric oxide synthesis of vascular endothelial cells,
(11) The pharmaceutical composition according to any one of (1) to (3), for increasing the blood concentration of vascular endothelial cell-derived nitric oxide,
(12) The pharmaceutical composition according to any one of (1) to (3), for maintaining the blood concentration of vascular endothelial cell-derived nitric oxide,
(13) The pharmaceutical composition according to any one of (1) to (3), for treating or preventing a disease caused by a decrease in blood concentration of vascular endothelial cell-derived nitric oxide,
(14) Diseases caused by a decrease in blood concentration of vascular endothelial cell-derived nitric oxide are high blood pressure, exertional hypertension, arteriosclerosis, ischemic heart disease, heart failure, myocardial infarction, thrombosis, peripheral vascular disorder, pulmonary hypertension (1), one or more selected from the group consisting of: Buerger disease, Raynaud's disease, cerebral ischemia, cerebral infarction, cerebral insufficiency, cerebrovascular dementia, impotence, pregnancy toxemia, diabetes A pharmaceutical composition of
(15) The pharmaceutical composition according to (13), wherein the disease caused by a decrease in blood concentration of vascular endothelial cell-derived nitric oxide is arteriosclerosis,
(16) The pharmaceutical composition according to any one of (1) to (3), for increasing the activity of glutathione peroxidase,
(17) The pharmaceutical composition according to any one of (1) to (3), for maintaining the activity of glutathione peroxidase,
(18) The pharmaceutical composition according to any one of (1) to (3), for treating or preventing a disease caused by a decrease in activity of glutathione peroxidase,
(19) The disease caused by the decreased activity of glutathione peroxidase is one or more selected from the group consisting of arteriosclerosis, ischemic heart disease, ischemic brain disease, and myocardial infarction. A pharmaceutical composition of
(20) The pharmaceutical composition according to (18), wherein (21) the disease caused by the decreased activity of glutathione peroxidase is arteriosclerosis, and (21) decreased blood homocysteine level, blood of vascular endothelial cell-derived nitric oxide The pharmaceutical composition according to any one of (1) to (3), which simultaneously achieves an increase in medium concentration and an improvement in activity of glutathione peroxidase.

Furthermore, the present invention provides
(22) The pharmaceutical composition according to any one of (1) to (21), which is a combination drug,
(23) The pharmaceutical composition according to any one of (1) to (21), which is a kit comprising a HMG-CoA reductase inhibitor and a drug containing glutathione,
(24) The method for producing a pharmaceutical composition according to any one of (1) to (3), wherein the HMG-CoA reductase inhibitor and glutathione are contained in the same pharmaceutical composition,
(25) Use of the pharmaceutical composition according to any one of (1) to (21) for producing a therapeutic agent for arteriosclerosis,
(26) A method for administering an HMG-CoA reductase inhibitor and glutathione simultaneously, sequentially or separately, and (27) a medicament described in any one of (1) to (3) selected from (1) to (3) to a mammal There is provided a method for preventing or treating arteriosclerosis, which comprises administering an effective amount of the composition.

  In the present invention, the “HMG-CoA reductase inhibitor” is a drug that specifically and antagonistically inhibits HMG (3-hydroxy-3-methylglutaryl) -CoA reductase, which is the rate-limiting enzyme of cholesterol biosynthesis. (It is called a statin as a generic name for this drug because the ending of the name is a statin), and it is used as a therapeutic agent for hyperlipidemia because it lowers blood cholesterol. Such statins include all natural substances derived from microorganisms, semisynthetic substances derived therefrom, and all synthetic compounds, and can be, for example, pravastatin, lovastatin, simvastatin, fluvastatin, atorvastatin, pitavastatin or rosuvastatin .

  The planar structural formulas of typical statins are shown below.

  Of these statins, pravastatin, simvastatin or atorvastatin is preferred, and pravastatin is more preferred.

  In the present invention, “glutathione” is a tripeptide consisting of three amino acids, glutamic acid, cysteine and glycine.

  In the present invention, “the amount of homocysteine in the blood” refers to the weight of homocysteine present in the blood.

  In the present invention, “maintaining the amount of homocysteine in the blood” means keeping the original state by suppressing an increase in the amount of homocysteine in the blood.

In the present invention, “hyperhomocysteinemia” means, for example, a lack of vitamin B ₆ , B ₁₂ or folic acid in the body, resulting in abnormal metabolism and an increase in blood homocysteine level. State.

  In the present invention, the “disease caused by hyperhomocysteinemia” is not particularly limited as long as it is a disease that develops due to an increase in blood Hcy level. Cardiovascular diseases such as heart disease, myocardial infarction, thrombosis, peripheral vascular disorder, Buerger's disease, Raynaud's disease and cerebrovascular disorders such as cerebral infarction, cerebral circulation failure, cerebrovascular dementia, nervous system such as Alzheimer's disease, Parkinson's disease Disease, etc., more preferably, arteriosclerosis.

  Factors that cause hyperhomocysteinemia include, for example, aging, smoking, nutritional disorders related to Hcy metabolism, drug properties, decreased renal function, chronic renal failure, diabetes, insulin resistance, malignant tumor, decreased thyroid function And pernicious anemia.

  In the present invention, “to promote nitric oxide synthesis in vascular endothelial cells” means to activate eNOS present in vascular endothelial cells and promote synthesis of NO.

  In the present invention, “maintaining the blood concentration of vascular endothelial cell-derived nitric oxide” refers to keeping the original state by suppressing the decrease of vascular endothelial cell-derived nitric oxide in blood.

  In the present invention, the “disease caused by a decrease in blood concentration of vascular endothelial cell-derived nitric oxide” means that the activity of eNOS is decreased and / or the blood concentration of vascular endothelial cell-derived NO is decreased. There are no particular limitations as long as the disease develops, but preferably, hypertension, exertional hypertension, arteriosclerosis, ischemic heart disease, heart failure, thrombosis, pulmonary hypertension, cerebral ischemia, cerebral infarction, cerebral circulation failure, There are senile dementia, impotence, pregnancy toxemia, diabetes and the like, more preferably arteriosclerosis.

  Examples of causes that cause a decrease in the blood concentration of vascular endothelial cell-derived NO include aging, menopause, hypertension, hyperlipidemia, smoking, insulin resistance, diabetes, oxidative stress, and drug properties. There is also a duplication of results and a vicious circle.

  In the present invention, “maintaining the activity of glutathione peroxidase” refers to maintaining the original state by suppressing a decrease in the activity of glutathione peroxidase.

  In the present invention, the “disease caused by a decrease in activity of glutathione peroxidase” is not particularly limited as long as it is a disease caused by an increase in active oxygen or an increase in lipid peroxide. There are sclerosis, cerebrovascular disease, ischemic heart disease, myocardial infarction, carcinogenesis, inflammation, aging, etc. More preferably, there is arteriosclerosis.

  In addition, although the factor causing the decrease in GPx activity has not been sufficiently elucidated, there are selenium deficiency, malnutrition, aging, and the like at this stage.

  In the present invention, it was conspicuous in the early stages of “disease caused by hyperhomocysteinemia”, “disease caused by decreased blood concentration of vascular endothelial cell-derived NO” or “disease caused by decreased activity of GPx”. Although there are few subjective symptoms and self-judgment is not easy, examples of the subjective symptoms include headache, migraine, dizziness, numbness or numbness, coldness of limbs, and stiff shoulders. Therefore, when these subjective symptoms occur, the above-mentioned disease can be treated at an early stage by taking the pharmaceutical composition containing the statin and glutathione of the present invention.

  When administering the pharmaceutical composition of the present invention, the respective components can be administered simultaneously, sequentially or separately.

  In the present invention, “simultaneously” administration includes administration at exactly the same time as pharmacologically acceptable as well as administration at the same time. The dosage form is not particularly limited as long as it can be administered at approximately the same time, but it is preferably a single composition.

  “Sequential or separate” administration in the present invention is not particularly limited as long as it is a dosage form that can be administered separately at different times. For example, one component is administered, and then, after a predetermined time, There are ways to administer other ingredients.

  In the present invention, “treat” means to cure or ameliorate a disease or symptom or to suppress a symptom.

  The pharmaceutical composition containing a statin and glutathione of the present invention comprises (1) a decrease in blood Hcy level, (2) an increase in eNOS activity and / or an increase in blood concentration of vascular endothelial cell-derived NO, and (3) It is useful for simultaneously increasing the GPx activity and treating or preventing diseases caused by each phenomenon.

  Statin, for example, pravastatin, lovastatin, simvastatin, fluvastatin, atorvastatin, pitavastatin or rosuvastatin contained in the composition of the present invention is disclosed in JP-A-57-2240 (USP4346227), JP-A-57-163374 (USP4231938), JP 56-122375 (USP 44444784), JP 60-500015 (USP 4739073), JP 3-58967 (USP 527939), JP 1-2279866 (US Pat. No. 5,854,259 and USP 5856336) or JP 51787881. According to the method described in (USP 5260440) etc., it can manufacture easily.

  Glutathione is listed in the Japanese Pharmacopoeia Standard for Drugs 2002.

  The composition of the present invention containing statin and glutathione contains statin and glutathione as essential components, and may optionally contain additives for formulation, and further comprises statin and glutathione. Other components may be contained within a range that does not adversely affect the combined action. The pharmaceutical composition preferably contains only statin and glutathione as active ingredients, and further contains additives for formulation.

  Specific dosage forms of the pharmaceutical composition of the present invention include, for example, tablets, fine granules (including powders), capsules, liquids (including syrups) and the like, and are suitable for each dosage form. An additive and a base material can be used as appropriate, and can be produced according to the usual method described in the Japanese Pharmacopoeia.

  In the above dosage forms, various commonly used additives can be used depending on the dosage form.

  For example, in the case of tablets, lactose, crystalline cellulose or the like as an excipient, magnesium aluminate or magnesium oxide as a stabilizer, hydroxypropyl cellulose or the like as a coating agent, magnesium stearate or the like as a lubricant Can be used.

  In the case of fine granules and capsules, lactose or purified sucrose is used as an excipient, magnesium aluminate or magnesium oxide as a stabilizer, corn starch or the like as an adsorbent, hydroxypropylcellulose or the like as a binder As can be used.

  In each of the above dosage forms, a disintegrant such as crospovidone; a surfactant such as polysorbate; an adsorbent such as calcium silicate; a colorant such as iron sesquioxide and caramel; a stabilizer such as sodium benzoate; A pH adjuster; a fragrance, etc. can also be added.

  The single dose of statin varies depending on the type of statin, indication, purpose of use and age, but is preferably 0.01 mg / kg to 10 mg / kg, more preferably 0.05 mg / kg to This is 5 mg / kg and is administered once to three times a day.

  When the pharmaceutical composition of the present invention is a solid preparation, the content of statin is usually 0.05 mg to 200 mg, and preferably 0.5 mg to 100 mg. The glutathione content is 1 to 100 parts by weight, preferably 3 to 30 parts by weight per 1 part by weight of the statin.

  The statin contained when the composition of the present invention is a liquid preparation is usually 0.001 mg / mL to 200 mg / mL, and preferably 0.01 mg / mL to 50 mg / mL. The glutathione content is 1 to 100 parts by weight, preferably 3 to 30 parts by weight per 1 part by weight of the statin.

  Hereinafter, the present invention will be described in more detail with reference to examples and test examples, but the scope of the present invention is not limited thereto.

(Example 1) Tablet (1) Components in a single dose (Table 1) Weight per tablet (mg)
Ingredients (1a) (1b) (1c)
――――――――――――――――――――――――――――――――――――
Pravastatin sodium or atorvastatin calcium 10 − −
Simvastatin-5
Pitavastatin -1
Glutathione 100 100 100
Magnesium oxide 90 70 50
Hydroxypropyl cellulose 60 60 60
Croscarmellose sodium 20 20 20
Magnesium stearate 15 15 15
Lactose appropriate amount appropriate amount appropriate amount ――――――――――――――――――――――――――――――――――――
(2) Manufacturing method Taking the said component and quantity, the tablet was manufactured according to the item of the Japanese Pharmacopoeia general rule "tablet".

(Example 2) Fine granules (1) Components in a single dose (Table 2) Weight in 2 packages (mg)
Ingredient (2a) (2b) (2c)
――――――――――――――――――――――――――――――――――――
Pravastatin sodium or atorvastatin calcium 20 − −
Simvastatin-10-
Pitavastatin --2
Glutathione 200 200 200
Magnesium oxide 100 80 60
Sucrose 950 950 950
Corn starch 920 920 920
Polysorbate 80 70 70 70
Magnesium stearate 20 20 20
Lactose appropriate amount appropriate amount appropriate amount ――――――――――――――――――――――――――――――――――――
(2) Production method The above ingredients and amounts were taken, and a fine granule was produced in accordance with the section of the Japanese Pharmacopoeia General Rules “Granule”.

(Example 3) Capsule (1) Components in a single dose (Table 3) Weight in 2 capsules (mg)
Ingredient (3a) (3b) (3c)
――――――――――――――――――――――――――――――――――――
Pravastatin sodium or atorvastatin calcium 10 − −
Simvastatin-5
Pitavastatin -1
Glutathione 100 100 100
Magnesium oxide 90 70 50
Corn starch 440 440 450
Polysorbate 80 60 60 60
Magnesium stearate 15 15 15
Lactose appropriate amount appropriate amount appropriate amount ――――――――――――――――――――――――――――――――――――
(2) Manufacturing method After taking the said component and quantity, and manufacturing a fine granule according to the term of a Japanese pharmaceutical formulation general rule "granule", the hard capsule was manufactured by filling it with a capsule.

(Example 4) Syrup (1) Components in a single dose (Table 4) Weight in 50 mL (mg)
Ingredients (4a) (4b) (4c)
――――――――――――――――――――――――――――――――――――
Pravastatin sodium or atorvastatin calcium 20 − −
Simvastatin-10-
Pitavastatin --2
Glutathione 200 200 200
Sodium benzoate 180 180 180
Sucrose 980 980 980
Concentrated glycerin 630 630 630
Polyvinyl alcohol 120 120 120
Purified water Remaining Remaining Remaining Remaining ――――――――――――――――――――――――――――――――――――
(2) Manufacturing method The syrup agent was manufactured by taking the said component and quantity, and manufacturing a syrup agent according to the term of a Japanese Pharmacopoeia general rule "syrup agent", and filling it into a brown glass bottle.
(Test example)
(Test Example 1) Evaluation test of blood GPx activity, Hcy amount and total NOx concentration (1) Test substance Pravastatin sodium was manufactured by Sankyo Co., Ltd., and glutathione manufactured by Kojin Co., Ltd. was used.
(2) Animals As test animals, Covance Research Products Inc. Beagle dogs were purchased at 5 months of age and used after approximately one month of quarantine and acclimatization.
(3) Dosage Form, Preparation Method and Preservation Method The required amount of the test substance calculated based on the body weight of each test animal was filled into a gelatin capsule (1/2 ounce) manufactured by TORPAC. After filling, the capsules were placed in cases separated by animal and stored refrigerated until administration.
(4) Administration route and administration period Capsules filled with the test substance were orally administered by gavage to test animals once a day between 9:00 and 12:30. The test animals were fasted for 2 to 3 hours before administration.

The administration period was 7 days.
(5) Preparation of test sample About 10 mL of blood was collected from the cephalic vein on the 14th and 7th days before the capsule administration (2nd week and 1st week before the administration start), and on the 4th and 8th days after the administration start. . The test animals were fasted for about 18 hours before blood collection.

The obtained blood was placed in a test tube, allowed to stand at room temperature for 30 minutes to 1 hour, and then serum obtained by centrifugation (about 1600 × g, 10 minutes) was used.
(6) Test method The blood GPx activity was determined by the NADPH method, and the total blood Hcy amount was determined by the HPLC method.

On the other hand, NO produced by eNOS is known to be quickly converted into nitrate ions (NO ₃ ⁻ ) and nitrite ions (NO ₂ ⁻ ), so the total blood nitrogen oxide (NOx) concentration is It was calculated as the sum of NO ₃ ⁻ and NO ₂ ⁻ determined using the HPLC method.
(7) Test results The results of the test values for the single agent and the combination drug for each dose of pravastatin sodium and glutathione were obtained by converting the average of the test values on the 14th day and the 7th day before administration to 100. .

  The obtained results are shown in Tables 5 to 7. Each value is an average value of 5 animals per group.

(Table 5)
Blood GPx activity (%)
Test substance (mg / Kg) 4 days after administration 8 days after administration ―――――――――――――――――――――――――――――――――
Pravastatin sodium (2) 87.2 88.6
Glutathione (20) 98.8 100.0
Pravastatin sodium (2)
+ Combination of glutathione (20) 104.1 107.2
―――――――――――――――――――――――――――――――――

  From the results of Table 5, an increase in GPx activity that was not predicted from the results of pravastatin and glutathione single agent was observed.

(Table 6)
Blood Hcy level (%)
Test substance (mg / Kg) 4 days after administration 8 days after administration ―――――――――――――――――――――――――――――――――
Pravastatin sodium (2) 105.6 105.2
Glutathione (20) 102.3 99.4
Pravastatin sodium (2)
+ Combination of glutathione (20) 97.9 88.2
―――――――――――――――――――――――――――――――――

  From the results of Table 6, an effect of decreasing the amount of Hcy in the blood, which could not be predicted from the results of each single agent of pravastatin and glutathione, was observed.

(Table 7)
Total blood NOx concentration (%)
Test substance (mg / Kg) 4 days after administration 8 days after administration ―――――――――――――――――――――――――――――――――
Pravastatin sodium (2) 97.2 103.7
Glutathione (20) 101.8 92.2
Pravastatin sodium (2)
+ Combination of glutathione (20) 123.6 133.8
―――――――――――――――――――――――――――――――――

  From the results in Table 7, an effect of increasing the total concentration of NOx in blood, which could not be predicted from the results of each single agent of pravastatin and glutathione, was observed.

  The pharmaceutical composition containing a statin and glutathione of the present invention comprises (1) a decrease in blood Hcy level, (2) an increase in eNOS activity and / or an increase in blood concentration of vascular endothelial cell-derived NO, and (3) It is useful for simultaneously increasing the GPx activity and treating or preventing diseases caused by each phenomenon.

Claims (1)

A pharmaceutical composition for increasing blood concentration of vascular endothelial cell-derived nitric oxide, comprising pravastatin and glutathione.