COMPLEX FORMULATION COMPRISING HMG-COA REDUCTASE INHIBITOR AND CLOPIDOGREL

Abstract

The present invention relates to a complex preparation comprising clopidogrel, an HMG-CoA reductase inhibitor, and a separation membrane containing a hydrophobic compound. More particularly, an objective of the present invention is to provide the complex preparation comprising clopidogrel and the HMG-CoA reductase inhibitor, wherein the complex preparation is intended for preventing or treating a cardiovascular disease, which has excellent storage stability by preventing a decrease in the stability of the HMG-CoA reductase inhibitor.

Claims

1. A complex preparation, comprising: a pharmaceutically acceptable acid salt of clopidogrel, optical isomers thereof, hydrates or solvates thereof, or mixtures thereof; an HMG-CoA reductase inhibitor, pharmaceutically acceptable salts thereof, optical isomers thereof, hydrates or solvates thereof, or mixtures thereof; and a separation membrane containing a hydrophobic compound.

2. The complex preparation according to claim 1, wherein said complex preparation comprises a first active component-containing layer including a first active component, a separation membrane layer containing a hydrophobic compound, and a second active component-containing layer including a second active component; wherein said first and second active components are different from each other; and wherein said first and second active components are one of i) a pharmaceutically acceptable acid salt of clopidogrel, optical isomers thereof, hydrates or solvates thereof, or mixtures thereof; or ii) an HMG-CoA reductase inhibitor, pharmaceutically acceptable salts thereof, optical isomers thereof, hydrates or solvates thereof, or mixtures thereof.

3. The complex preparation according to claim 2, wherein said complex preparation comprises: a) a first layer containing said first active component; b) a second layer, i.e., a separation membrane, formed on said first layer and containing a hydrophobic compound; and c) a third layer formed on said second layer and including said second active component.

4. The complex preparation according to claim 2 or 3, wherein said separation membrane prevents said first active component and second active component from being in contact with each other.

5. The complex preparation according to claim 1, wherein said complex preparation comprises a first particle including a first active component, a second particle including a second active component, and a separation membrane containing a hydrophobic compound; wherein the separation membrane containing the hydrophobic compound separates the first particle and the second particle from each other, and thus prevents said first active component and second active component from being in contact with each other; and wherein said first and second active components are one of i) a pharmaceutically acceptable acid salt of clopidogrel, optical isomers thereof, hydrates or solvates thereof, or mixtures thereof; or ii) an HMG-CoA reductase inhibitor, pharmaceutically acceptable salts thereof, optical isomers thereof, hydrates or solvates thereof, or mixtures thereof.

6. The complex preparation according to claim 5, wherein said separation membrane is formed on one or more of a first or second particle.

7. The complex preparation according to claim 5, wherein said complex preparation comprises: a) an inactive pellet including a pharmaceutically acceptable carrier; b) a first particle including a first active component formed on said pellet; c) a separation membrane containing a hydrophobic compound formed on said first particle; and d) a second particle physically separated from the first particle and including a second active component.

8. The complex preparation according to claim 7, wherein said second particle consists of an active component only.

9. The complex preparation according to claim 7, wherein said second particle comprises an active component and a pharmaceutically acceptable carrier.

10. The complex preparation according to claim 1, wherein said complex preparation comprises said first active component, said second active component, and a particle including a separation membrane containing a hydrophobic compound, which prevents said first active component and said second active component from being in contact with each other; and wherein said particle comprises: a first layer including said first active component; a second layer including said second active component; and a separation membrane present between said first layer and second layer.

11. The complex preparation according to claim 1 or 5, wherein said pharmaceutically acceptable salt of clopidogrel is one or more selected from the group consisting of hydrogensulfate, hydrochloride, napadisilate, besylate, bromate, taurocholate and acetate.

12. The complex preparation according to claim 1 or 5, wherein said HMG-CoA reductase inhibitor is one or more selected from the group consisting of rosuvastatin, atorvastatin, simvastatin, lovastatin, mevastatin, pravastatin, fluvastatin, cerivastatin, pitavastatin, bervastatin, dalvastatin, glenvastatin, salts thereof and isomers thereof.

13. The complex preparation according to claim 1 or 5, wherein said HMG-CoA reductase inhibitor is one or more selected from the group consisting of rosuvastatin, atorvastatin, pravastatin and fluvastatin, salts thereof and isomers thereof.

14. The complex preparation according to claim 1 or 5, wherein said clopidogrel, pharmaceutically acceptable salts thereof, hydrates or solvates thereof, optical isomers thereof, or mixtures thereof are included in an amount of 75 to 300 mg.

15. The complex preparation according to claim 1 or 5, wherein said separation membrane comprises a hydrophobic compound.

16. The complex preparation according to claim 1 or 5, wherein said separation membrane comprises a hydrophobic compound in an amount of 0.0 to 0.8 parts by weight per 1 part by weight of the separation membrane.

17. The complex preparation according to claim 16, wherein said hydrophobic compound is one or more selected from the group consisting of glyceryl palmitostearate, glyceryl stearate, glyceryl behenate, cetyl palmitate, lecithin, glyceryl monooleate, stearic acid, cetostearyl alcohol, cetyl alcohol, stearyl alcohol, carnauba wax, cera and microcrystalline wax.

18. The complex preparation according to claim 16, wherein said hydrophobic compound is one or more selected from the group consisting of glyceryl behenate, sodium stearyl fumarate, carnauba wax and lecithin.

19. The complex preparation according to claim 1 or 5, wherein said separation membrane comprises a hydrophilic compound in an amount of 0.2 to 0.99 parts by weight per 1 part by weight of the separation membrane.

20. The complex preparation according to claim 19, wherein said hydrophilic compound is one or more selected from the group consisting of hydroxypropyl methylcellulose, hydroxypropyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, methyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl methylcellulose acetate succinate, hydroxyethyl methylcellulose, guar gum, locust bean gum, tragacanth, carrageenan, acacia gum, arabic gum, gellan gum, xanthan gum, gelatin, casein, zein, polyvinyl alcohol, polyvinyl pyrrolidone, copolyvidone, polyvinyl acetal dietylamino acetate, poly(butylmethacrylate-(2-dimethylaminoethyl)methacrylate-methylmethacrylate)copolymer, poly(methacrylic acid-methylmethacrylate)copolymer, poly(methacrylic acid-ethylacrylate)copolymer, polyethylene glycol, polyethylene oxide and carbomer.

21. The complex preparation according to claim 1 or 5, wherein said separation membrane comprises a hydrophobic compound and a hydrophilic compound in an amount of 1:10 to 3:1 parts by weight.

22. The complex preparation according to claim 1 or 5, wherein said complex preparation further comprises one or more additives selected from the group consisting of a stabilizer, binder, disintegrant, lubricant, diluent, coating agent, pH-adjusting agent, dissolution aid and surfactant.

23. The complex preparation according to claim 22, wherein said stabilizer is an antioxidant, alkali metal salt, organic salt or mixtures thereof.

24. The complex preparation according to claim 23, wherein said stabilizer is calcium carbonate, sodium carbonate, sodium hydrogen carbonate, magnesium oxide, magnesium carbonate, sodium citrate, meglumine, triethanolamine, arginine, glycine, butylated hydroxytoluene (BHT), dibutyl hydroxytoluene (DHT), butylated hydroxylanisole (BHA), sodium sulfite, sodium pyrosulfite, sodium hydrogensulfite, propyl gallate, calcium phosphate or mixtures thereto.

25. The complex preparation according to claim 1 or 5, wherein said preparation is a plain tablet, coated tablet, multi-layered tablet, cored tablet, powder preparation, granule preparation, pellet preparation or capsule preparation.

26. The complex preparation according to claim 1 or 5, wherein said complex preparation is intended for preventing or treating a cardiovascular disease selected from the group consisting of angina, hypertension, arteriospasm, deep vein, cerebral infarction, cardiomegaly, congestive heart failure and myocardial infarction.

27. A method for preparing a complex preparation, wherein the method comprises steps of: (a) preparing a plain tablet including a first active component; (b) forming a separation membrane by coating a surface of said plain tablet with a composition including a hydrophobic compound; and (c) forming a membrane including a second active component on said separation membrane, wherein said first and second active components are different from each other; and wherein said first and second active components are one of i) clopidogrel, pharmaceutically acceptable salts thereof, hydrates or solvates thereof, optical isomers thereof, or mixtures thereof; or ii) an HMG-CoA reductase inhibitor, pharmaceutically acceptable salts thereof, hydrates or solvates thereof, optical isomers thereof, or mixtures thereof.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0093] FIG. 1 shows a content of lactone related substances of rosuvastatin as a result of the stability test on the inventive complex preparation (tablet) under an accelerated condition for four months.

[0094] FIG. 2 shows a content of total related substances of rosuvastatin as a result of the stability test on the inventive complex preparation (tablet) under the accelerated condition for four months.

[0095] FIG. 3 shows a content of lactone related substances of rosuvastatin as a result of the stability test on the inventive complex preparation (capsule preparation) under the accelerated condition for one month.

[0096] FIG. 4 shows a content of total related substances of rosuvastatin as a result of the stability test on the inventive complex preparation (capsule preparation) under the accelerated condition for one month.

[0097] FIG. 5 shows a comparative elution rate of rosuvastatin between the inventive complex preparation and Crestor Tab. 10 mg at pH 1.2.

[0098] FIG. 6 shows a comparative elution rate of clopidogrel between the inventive complex preparation and Plavix Tab. 75 mg at pH 1.2.

[0099] FIG. 7 shows a change of rosuvastatin concentration in blood with a beagle dog after an oral administration of the inventive complex preparation and Crestor Tab. 10 mg.

[0100] FIG. 8 shows a change of clopidogrel concentration in blood with a beagle dog after an oral administration of the inventive complex preparation and Plavix Tab. 75 mg.

MODE FOR INVENTION

[0101] The features and advantages of the present invention as well as methods for achieving them will be apparent with reference to exemplary embodiments described in detail hereinafter. However, the present invention is not limited to the exemplary embodiments disclosed hereinafter, but will be implemented in various different forms. Hereinafter, the following exemplary embodiments will be suggested for better understanding of the present invention and be provided only for the purpose of completely illustrating the scope of the present invention to those skilled in the art, and thus the present invention will be defined only by the scope of the claims thereto.

<Example 1> Preparation of a Clopidogrel Plain Tablet

[0102] In accordance with components and contents as shown in a following table 1, clopidogrel, L-HPC, lactose, copovidone, colloidal silicon dioxide, talc and magnesium stearate were mixed, compressed and prepared into a plain tablet. At that time, the plain tablet was made by means of a tablet press (ERWEKA APPARATEBAU).

TABLE-US-00001 TABLE 1 Component Amount Compartment Component (mg/Tab.) (1) Clopidogrel plain tablet Clopidogrel hydrogensulfate 97.9 L-HPC 25.0 Lactose 110.1 Copovidone 12.0 Colloidal silicon dioxide 2.5 Talc 4.0 Magnesium stearate 3.5 Solid content amount 255.0

<Examples 2 to 5> Tablet Including a Separation Membrane Containing a Hydrophobic Compound

[0103] In accordance with components and contents as shown in a following table 2, (2) a coating solution for a hydrophobic separation membrane containing a hydrophobic compound and (3) a coating solution containing rosuvastatin were respectively prepared, after which (1) a clopidogrel plain tablet of Example 1 was sequentially coated with (2) the coating solution for the hydrophobic separation membrane and (3) the coating solution containing rosuvastatin, such that a coating layer was prepared. At that time, coating was performed at a spray velocity of 20 g/min at an inlet temperature of 58-72 C. for 180 minutes by using a tablet coating machine (FREUND HCT-30).

TABLE-US-00002 TABLE 2 Component Amount (mg/Tab) Compartment Component Example 2 Example 3 Example 4 Example 5 (1) Clopidogrel Example 1 255.0 255.0 255.0 255.0 plain tablet (2) Coating solution Glyceryl behenate 10.0 for hydrophobic Sodium stearyl fumarate 10.0 separation Carnauba wax 10.0 membrane Lecithin 10.0 Copovidone 10.0 10.0 10.0 10.0 Titanium oxide 0.5 0.5 0.5 0.5 Talc 0.5 0.5 0.5 0.5 Purified water (volatile) 75.0 75.0 75.0 75.0 Ethanol (volatile) 300.0 300.0 300.0 300.0 (3) Rosuvastatin Rosuvastatin calcium 10.4 10.4 10.4 10.4 coating solution Copovidone 59.6 59.6 59.6 59.6 Talc 1.0 1.0 1.0 1.0 Purified water (volatile) 70.0 70.0 70.0 70.0 Ethanol (volatile) 200.0 200.0 200.0 200.0 Solid content amount 347.0 347.0 347.0 347.0

<Comparative Examples 1 to 3> Tablet without a Separation Membrane or Tablet Including a pH-Dependent Separation Membrane

[0104] In accordance with components and contents as shown in a following table 3, (4) a coating solution for a pH-dependent separation membrane and (3) a coating solution containing rosuvastatin were respectively prepared, after which (1) the clopidogrel plain tablet of Example 1 was sequentially coated with (4) the coating solution for the pH-dependent separation membrane and (3) the rosuvastatin coating solution. However, in case of a complex preparation without a separation membrane in Comparative Example 1, coating for the separation membrane was omitted and coating was performed with (3) the coating solution containing rosuvastatin. At that time, coating was performed at a spray velocity of 20 g/min at an inlet temperature of 58-72 C. for 180 minutes by using a tablet coating machine (FREUND HCT-30).

TABLE-US-00003 TABLE 3 Component Amount (mg/Tab.) Comparative Comparative Comparative Compartment Component Example 1 Example 2 Example 3 (1) Clopidogrel Example 1 255.0 255.0 255.0 plain tablet (4) Coating Eudragit EPO 15.0 solution for no Eudragit L100 17.0 separation Sodium lauryl sulfate 1.5 membrane and Triethylacetate 1.5 pH-dependent Stearic acid 2.0 separation Talc 7.5 7.5 membrane Purified water (volatile) 150.0 150.0 (3) Rosuvastatin Rosuvastatin calcium 10.4 10.4 10.4 coating solution Copovidone 59.6 59.6 59.6 Talc 1.0 1.0 1.0 Purified water (volatile) 70.0 70.0 70.0 Ethanol (volatile) 200.0 200.0 200.0 Solid content amount 321.0 347.0 347.0

<Comparative Examples 4 to 6> Tablet Including a Hydrophilic Separation Membrane

[0105] In accordance with components and contents as shown in a following table 4, (5) a coating solution for a hydrophilic separation membrane without containing a hydrophobic compound and (3) a coating solution containing rosuvastatin were respectively prepared, after which (1) the clopidogrel plain tablet of Example 1 was sequentially coated with (5) the coating solution for the hydrophilic separation membrane and (3) the coating solution containing rosuvastatin. At that time, coating was performed at a spray velocity of 20 g/min at an inlet temperature of 58-72 C. for 180 minutes by using a tablet coating machine (FREUND HCT-30).

TABLE-US-00004 TABLE 4 Component Amount (mg/Tab.) Comparative Comparative Comparative Compartment Component Example 4 Example 5 Example 6 (1) Clopidogrel Example 1 255.0 255.0 255.0 plain tablet (5) Coating layer Povidone 20.0 for hydrophilic Copovidone 20.0 separation PVA 20.0 membrane Titanium oxide 0.5 0.5 0.5 Talc 0.5 0.5 0.5 Purified water (volatile) 75.0 75.0 75.0 Ethanol (volatile) 300.0 300.0 300.0 (3) Rosuvastatin Rosuvastatin calcium 10.4 10.4 10.4 coating layer Copovidone 59.6 59.6 59.6 Talc 1.0 1.0 1.0 Purified water (volatile) 70.0 70.0 70.0 Ethanol (volatile) 200.0 200.0 200.0 Solid content amount 347.0 347.0 347.0

<Comparative Examples 7 and 8> Single Tablet

[0106] A commercially available rosuvastatin preparation (Crestor Tab. 10 mg, AstraZeneca, Comparative Example 7) and a clopidogrel preparation (Plavix Tab. 75 mg, Sanofi-Aventis, Comparative Example 8) were purchased and used.

<Example 6> Capsule Preparation Including a Hydrophobic Separation Membrane

[0107] In accordance with components and contents as shown in a following table 5, (3) a coating solution containing rosuvastatin and (2) a coating solution for a hydrophobic separation membrane containing a hydrophobic compound were respectively prepared, after which an inactive pellet (seed pellet) was sequentially coated with (3) the coating solution containing rosuvastatin and (2) the coating solution for the hydrophobic separation membrane, then mixed with clopidogrel hydrogensulfate powder, and then filled into a capsule. At that time, pellet coating was performed at a spray velocity of 20-50 mg/min at an inlet temperature of 35-65 C. for to hours by using a fluidized bed coating machine (FREUND GPC-G1) in a bottom mode.

<Comparative Example 9> Capsule Preparation without a Separation Membrane

[0108] In accordance with components and contents as shown in a following table 5, coating for the hydrophobic separation membrane was omitted and an inactive pellet (seed pellet) was coated with (3) a coating solution containing rosuvastatin, then mixed with clopidogrel hydrogensulfate powder, and then filled into a capsule. At that time, pellet coating was performed at a spray velocity of 20-50 mg/min at an inlet temperature of 35-65 C. for to hours by using a fluidized bed coating machine (FREUND GPC-G1) in a bottom mode.

TABLE-US-00005 TABLE 5 Component Amount (mg/Cap.) Compartment Component Example 6 Comparative Example 9 Inactive pellet Microcrystalline cellulose 150.0 150.0 pellet (3) Rosuvastatin Rosuvastatin calcium 10.4 10.4 coating solution Copovidone 20.6 20.6 Talc 4.0 4.0 Purified water (volatile) 80.0 80.0 Ethanol (volatile) 170.0 170.0 (2) Coating solution Opaglos 50.0 for hydrophobic (Lecithin = 6.1% w/w) (Lecithin = 3.05 mg/Tab.) separation Povidone K30 50.0 membrane Purified water (volatile) 900.0 Ethanol (volatile) 100.0 Clopidogrel Clopidogrel 97.9 97.9 hydrogensulfate Capsule Capsule no. 1 Capsule no. 1 Solid content amount (excluding the capsule) 382.9 282.9

<Experimental Example 1> Disintegration Test

[0109] In case of Examples 2 to 5 and Comparative Examples 1 to 6 above, a disintegration test was performed at pH 1.2 with a 1.sup.st test solution for disintegration test methods of the Korean Pharmacopoeia. The disintegration test method is as follows and results thereof are as shown in a following table 6.

[0110] [Test Method]

[0111] 1. Basis for disintegration test: Disintegration test of general test methods in the 10.sup.th revision of the Korean Pharmacopoeia

[0112] 2. Test method: Each of six samples was inserted into a glass tube of a testing machine, and the testing machine was put into operation for two hours with a test solution set at 372 C.

[0113] 3. Test solution: A 1.sup.st test solution (pH 1.2) for disintegration test methods of the Korean Pharmacopoeia

[0114] 4. Analysis method: A point of time, at which properties of all six samples completely disappear, was measured.

[0115] [Test Results]

TABLE-US-00006 TABLE 6 Disintegration time Example 2 15 min 17 sec Example 3 13 min 48 sec Example 4 25 min 12 sec Example 5 21 min 30 sec Comparative Example 1 10 min 26 sec Comparative Example 2 23 min 31 sec Comparative Example 3 Timeout Comparative Example 4 10 min 15 sec Comparative Example 5 10 min 21 sec Comparative Example 6 10 min 50 sec

[0116] As shown in the table 6 above, in case of Examples 2 to 5 and Comparative Examples 1, 2 and 4 to 6 according to the present invention, an disintegration time was 30 minutes or so at pH 1.2 similar to a gastrointestinal tract, and thus it was identified that a drug may be easily dissolved in the gastrointestinal tract.

[0117] On the other hand, in case of Comparative Example 3, disintegration was not done at pH 1.2 within two hours, and thus an elution of an active component was delayed. Eudragit L100, which was a separation membrane base of Comparative Example 3 above, may be dissolved at pH 6.0 or more, and thus it may be identified that Eudragit L100 is not desirable as a separation membrane of clopidogrel and the HMG-CoA inhibitor, which requires a release in the gastrointestinal tract.

<Experimental Examples 2> Stability Test

[0118] In case of Examples 2 to 4 and Comparative Examples 1 to 6, each of tablets was packed into an HDPE bottle along with silica gel, then stored under an accelerated condition, then taken out in zero (initial), two and four months later, and then evaluated by measuring an amount of rosuvastatine lactone related substances and total related substances, which were representative acid decomposition products of rosuvastatine out of the HMG-CoA reductase inhibitor. The stability test method is as follows and results thereof are as shown in a following table 7 and FIGS. 1 and 2.

[0119] [Test Method]

[0120] 1. Basis for stability test: Items of the accelerated test in the Ministry of Food and Drug Safety Notification No. 2014-59

[0121] 2. Test method: Each of 30 tablets was packed into a 100 mL HDPE bottle along with silica gel, then stored in a stability chamber under an accelerated condition (402 C., relative humidity of 755%), then opened in zero (initial), two and four months later, and then analyzed. At that time, the test was finished, if an amount of total related substances exceeds 5%.

[0122] 3. Analysis method: An amount of lactone related substances and total related substances, which were representative acid decomposition products, was analyzed according to a method for analyzing related substances of rosuvastatin.

[0123] [Test Results]

TABLE-US-00007 TABLE 7 Accelerated test Lactone related Total related Remark period substance substance (Separation membrane (month) (%) (%) form) Comparative 0 (initial) 0.050 0.304 No separation membrane Example 1 2 4.970 5.130 Comparative 0 (initial) 0.050 0.304 pH-dependent separation Example 2 2 7.723 7.923 membrane Comparative 0 (initial) 0.051 0.385 Example 3 2 6.180 6.378 Comparative 0 (initial) 0.051 0.307 Hydrophilic separation Example 4 2 5.841 6.115 membrane without Comparative 0 (initial) 0.055 0.366 containing a hydrophobic Example 5 2 3.007 3.661 compound 4 7.858 8.994 Comparative 0 (initial) 0.062 0.491 Example 6 2 9.139 10.168 Example 2 0 (initial) 0.050 0.377 Separation membrane 2 0.000 0.555 containing a hydrophobic 4 0.013 0.688 compound Example 3 0 (initial) 0.050 0.377 2 0.000 0.475 4 0.249 0.914 Example 4 0 (initial) 0.050 0.377 2 0.000 0.460 4 0.166 0.864

[0124] As shown in the table 7 above and FIG. 1, it was identified for Examples 2 to 4 of the present invention that a growth rate of lactone related substances was as very low as less than 1% for four months under the accelerated condition.

[0125] On contrary, it was identified for Comparative Examples 1 to 6 that the lactone related substances were greatly increased by 3% or more within two months under the accelerated condition.

[0126] In particular, in case of Comparative Examples 2 to 5, the lactone related substances were more greatly increased. That's because protons (H.sup.+) were accumulated in a pH-dependent separation membrane and a hydrophilic water-soluble separation membrane without containing a hydrophobic compound, and thus the lactone related substances with low stability in acid were greatly increased.

<Experimental Example 3> Stability Test on a Capsule Preparation

[0127] In case of Example 6 and Comparative Example 9, each of capsule preparations was individually packed with Alu-Alu blister, then stored under an accelerated condition, then taken out in zero (initial) and one month later, and then evaluated by measuring an amount of rosuvastatine lactone related substances and total related substances, which were representative acid decomposition products of rosuvastatine out of the HMG-CoA reductase inhibitor. The stability test method is as follows and results thereof are as shown in a following table 8 and FIGS. 3 and 4.

[0128] [Test Method]

[0129] 1. Basis for stability test: Items of the accelerated test in the Ministry of Food and Drug Safety Notification No. 2014-59

[0130] 2. Test method: Each of capsule preparations was packed with Alu-Alu blister, then stored in a stability chamber under an accelerated condition (402 C., relative humidity of 755%), then opened in zero (initial) and one month after, and then analyzed.

[0131] 3. Analysis method: An amount of lactone related substances and total related substances, which were representative acid decomposition products, was analyzed according to a method for analyzing related substances of rosuvastatin.

[0132] [Test Results]

TABLE-US-00008 TABLE 8 Lactone Total Remark Accelerated test related related (Separation period substance substance membrane (month) (%) (%) form) Example 6 0 (initial) 0.113 0.332 Hydrophobic 1 0.192 0.309 separation membrane Comparative 0 (initial) 0.145 0.368 No separation Example 9 1 0.512 0.710 membrane

[0133] As shown in the table 8 above and FIG. 3, it was identified for Example 6 of the present invention that an amount of increase in lactone related substances was as very low as less than 0.08% under the accelerated condition for one month.

[0134] On contrary, it was identified for Comparative Example 9 that an amount of increase in lactone related substances was 0.36% or more under the accelerated condition as early as one month, and such great increase was at least 4.5 times more than Example 6. Accordingly, it was also identified that an increase in lactone related substances might be inhibited even in a capsule preparation filled with a pellet coated with a hydrophobic separation membrane.

<Experimental Example 4> Elution Test

[0135] A comparative elution test was performed with each of four tablets of Example 2 and Comparative Examples 7 and 8 above. A method for the comparative elution test is as follows and results thereof are as shown in a following table 9 and FIGS. 5 and 6.

[0136] [Test Method]

[0137] 1. Basis for elution test: Elution test method of general test methods in the 10.sup.th revision of the Korean Pharmacopoeia

[0138] 2. Test method: Paddle method, 370.5 C., 50 revolutions/min, n=4

[0139] 3. Test solution: 900 ml of 1.sup.st test solution for disintegration test methods of the Korean Pharmacopoeia (pH 1.2)

[0140] 4. Analysis method: Method for analyzing an elution (content) of rosuvastatin and clopidogrel

[0141] [Test Results]

TABLE-US-00009 TABLE 9 0 5 10 15 30 45 60 90 120 min min min min min min min min min Rosuvastatin Example 2 Average 0.0 16.5 47.3 71.3 87.2 90.9 93.2 96.0 96.3 elution rate S.D. 0.0 5.5 9.5 7.3 4.9 4.0 4.8 3.5 3.1 (%) Comparative Average 0.0 49.9 77.5 90.1 97.2 98.7 98.5 99.3 99.1 Example 7 S.D. 0.0 2.3 3.0 3.1 2.7 2.2 1.8 1.3 1.3 Clopidogrel Example 2 Average 0.0 0.0 2.1 7.8 46.8 80.7 95.3 98.1 98.7 elution rate S.D. 0.0 0.0 0.5 2.3 8.4 7.6 5.1 2.0 1.1 (%) Comparative Average 0.0 22.4 46.4 67.9 86.1 91.6 93.9 94.5 95.4 Example 8 S.D. 0.0 6.2 8.5 9.9 8.9 6.1 5.4 5.2 2.9

[0142] As shown in the table 9 and FIG. 5, in case of both Example 2 and Comparative Example 7 of the present invention, a drug of rosuvastatin was eluted within 30 minutes and both preparations were eluted 80% or more within 30 minutes, which was shorter than about two hours, that is, a general drug retention time in the gastrointestinal tract. However, in case of Example 2 of the present invention, an elution rate of rosuvastatin was faster than Comparative Example 7 under the condition above, seemingly because a coated rosuvastatin of Example 2 may be released into eluate faster than rosuvastatin of the tablet of Comparative Example 7.

[0143] Meanwhile, as shown in the table 9 and FIG. 6, in case of both Example 2 and Comparative Example 8 of the present invention, clopidogrel was eluted 80% or more within 60 minutes, which was shorter than about two hours, that is, a general drug retention time in the gastrointestinal tract. However, in case of Example 2, an elution rate of clopidogrel was slower than Comparative Example 8 under the condition above, because clopidogrel is eluted after a complete dissolution of a rosuvastatin coating layer and the separation membrane due to structural properties of the preparation of Example 2.

<Experimental Example 5> Blood Concentration Test

[0144] With regard to Example 2 (test group) and Comparative Examples 7 and 8 (control group) above, each of preparations was orally administered into a beagle dog, and then a blood concentration test was performed on the test group and the control group according to the present invention.

[0145] Meanwhile, with regard to the test group and the control group, each group of four beagle dogs was subjected to a cross-matching test (22) twice at a drug-free interval of two weeks. A detailed experimental condition is as shown in a following table 10. Also, with regard to the control group and the test group, the results of the blood concentration test on rosuvastatin and clopidogrel are as shown in a table 11 and FIGS. 7 and 8.

TABLE-US-00010 TABLE 10 Title Oral PK test on a beagle dog for a hydrophobic separation membrane base and a single/combined preparation with rosuvastatin/clopidogrel Test design The present test was designed as follows. Control group: Two tablets of rosuvastatin 10 mg (Crestor Tab. 10 mg) + Two tablets of clopidogrel 75 mg (Plavix Tab. 75 mg) Test group: Two tablets of the hydrophobic separation membrane prepared in Example 2 Each of the control group and the test group was subjected to a cross- matching test twice at a drug-free interval of two weeks. Evaluation Evaluation of drug concentration in blood: method After administration of a test drug, a concentration of rosuvastatin and clopidogrel in blood was measured by means of LC/MS/MS, and a difference between groups was compared and evaluated with following items set as parameters. Area under the blood concentration-time curve up to a last measurable point of blood collection: AUC.sub.Iast Area under the blood concentration-time curve calculated by means of extrapolation from a last measurable point of blood collection to infinite time: AUC.sub.inf Max. blood concentration: C.sub.max Time to reach Max. blood concentration: T.sub.max Drug half-life: T.sub.1/2

[0146] [Test Results]

TABLE-US-00011 TABLE 11 Rosuvastatin Clopidogrel PK parameter Control group Test group Control group Test group C.sub.max (ng/ml) 138.563 24.745 121.908 47.555 249.14 90.056 223.618 107.886 T.sub.max (hr) 1.6 0.3 1.4 0.8 1.4 0.5 1.4 0.6 AUC.sub.last (ng/ml) 462.246 106.647 422.990 213.641 435.725 196.644 364.373 137.952 AUC.sub.inf (ng/ml) 552.449 138.648 513.212 244.548 448.219 200.370 381.545 142.895 t.sub.1/2 (hr) 2.3 0.6 2.5 0.5 1.2 0.5 1.6 0.8

[0147] According to the table 11 and FIGS. 7 and 8, in case of the test group (Example 2) and the control group (Comparative Examples 7 and 8) of the present invention, there was no significant difference in AUC, Cmax, Tmax and T.sub.1/2 values of rosuvastatin and clopidogrel.

[0148] In case of Experimental Example 3 above, there was a slight difference in elution rates of Example 2 and Comparative Examples 7 and 8. However, in case of the PK test on the beagle dog, there was no significant difference in AUC, Cmax, Tmax and T.sub.1/2 values. Thus, it might be identified that there is no great change in pharmacokinetic parameters of the drug through an in vivo experiment.

[0149] In the complex preparation containing an HMG-CoA reductase inhibitor and clopidogrel of the present invention, the preparation coated with a separation membrane containing a hydrophobic substance of the present invention may provide the complex preparation with excellent storage stability without any PK change of an active component in vivo, and thus may be utilized as a therapeutic agent for a cardiovascular disease with excellent stability.