TWI487524B - Novel therapeutic combinations of nicotinic acid and meldonium - Google Patents

Description

Novel therapeutic combination of nicotinic acid and rice koji

The present invention relates to a novel therapeutic combination of nicotinic acid and mitoxantrone and for preventing and/or treating a group comprising abnormal dyslipidemia, hyperlipemia, atherosclerosis, selected from the group consisting of angina pectoris and myocardial infarction Coronary heart disease, cerebrovascular accidents with temporary or permanent ischemic attack, and diseases such as stroke and peripheral arterial occlusive disease, methods for preventing platelet aggregation and thrombosis.

More specifically, the present invention relates to a novel therapeutic combination of niacin, nicotinic acid and rice koji, which synergistically enhances the therapeutic effect of nicotinic acid and improves some undesirable side effects of niacin. In particular, peripheral vasodilatation (flushing) and an increase in blood glucose concentration. The present invention also relates to a pharmaceutical composition comprising the pharmaceutical, and to the use thereof for the manufacture of a medicament for preventing and/or treating the aforementioned diseases.

Abbreviation used For the sake of simplicity, the following abbreviations are further used in this description:

ATP-adenosine triphosphate

C-cholesterol

GL-glucose

HDL-C-High Density Lipoprotein-Cholesterol

I/R-ischemia/reperfusion

LDL-C-Low Density Lipoprotein-Cholesterol

MD-米曲肼 (INN)

NA-nicotinic acid

NAMg-magnesium nicotinic acid salt

PI-pyridylamine (piracetam)

RPP-heart rate blood pressure product = mean blood pressure x heart rate x 1000 ^-1

SI-simvastatin

TG-triglyceride s

TR-tetrabutyraldehyde (Triton) WR1339 (Tyloxapol Tyloxapol)

VF-ventricular fibrillation

VT-ventricular tachycardia

NA is an important agent for the treatment of abnormal dyslipidemia. It is also the only agent available for all components that positively affect blood lipid analysis: it lowers the concentration of total cholesterol, TG and LDL-C in the blood, and changes lipids in various ways. The agent has the most significant activity to increase HDL-C (Pieper JA, American Journal of Nursing Management 2002; 8 (12 Supplement): page S308-14).

As early as 1955 (Altshul R, Hoffer A and Stephen JD, Journal of Biochemistry and Biophysics 1955; 54: 558-559) and 1959 (Parsons Jr WB and Flinn JH, American Medical Association internal scientific literature) 1959; 103: 783-790), NA has been reported for the treatment of abnormal dyslipidemia.

Because NA effectively increases HDL-C concentration (McKenney J, Internal Medicine, 2004; 164 (7): pp. 697-705. Carlson LA, Journal of Internal Medicine 2005; 258: pp. 94-114), currently NA binds to other lipid modifiers, most of which act on LDL-C concentrations in order to increase HDL-C concentration (Rosenson RS, American Medical Journal 2005; 118(10): pp. 1067-77).

NA is an effective lipid-modifying agent that prevents progression of atherosclerosis and reduces cardiovascular events (Savel'ev AA and Shershevskii MG, Surgical Medicine (Russian) 1996; 74: Page 48-52 Drexel H, European Heart Journal Supplement 2006; Issue 8, Supplement F: Page F23-F29. Brown BG and Zhao XQ, American Journal of Cardiology 2008; 101 (8A): Page 58B-62B. Increase the HDL-C concentration. NA reduces morbidity and mortality in patients with hyperlipidemia (Canner PL et al., Journal of the American College of Cardiology, 1986; 8: pp. 1245-55). In the case of hyperlipidemia, NA is the most effective agent for increasing HDL-C (Ellingworth DR et al., Internal Medicine 1994, 154: pp. 1586-95. Schectman G et al., American Heart) Journal of Diseases 1993; 71: 758-65). NA attenuates thrombosis, lowers blood viscosity and has cardioprotective effects that limit ischemia-reperfusion injury (Lamping KA et al., Journal of Pharmacology and Therapeutics, 1984; 231(3): 532-538 Trueblood NA et al., American Journal of Physiology - Cardiac and Circulatory System Physiology 2000; 279(2), pp. H764-H771. Rosenson RS, Atherosclerosis 2003; 171(1): Pages 87-96).

Because of the risk factors for stroke in low HDL-C (Wannamethee SG, Shaper AG, Ebrahim S, Stroke 2000; 31: page 1882. Sacco RL, Benson RT, Kargman DE, American Journal of Medicine 2001; 285: Page 2729-2735.Rizos E, Mikhailidis DP with vascular heart research 2001; 52 (2), pp 199-207.Sanossian N, Tarlov NE with, the current treatment options for cardiovascular medicine 2008; 10 (3) of, Pages 195-206), NA as an agent to improve HDL-C (Carlson LA, Cardiology, New 2006; 21(4): pp. 336-344), useful in preventing stroke and stroke sequelae (Koniukov) SG, Liberzon SP, Clinical Medicine (Mosk) 1975; 53(9): pp. 38-41). It has been recognized that HDL-C concentrations are reduced in acute ischemic strokes (Russman AN et al., Neuroscience Journal 2009; 279 (1-2): pp. 53-56). NA has been shown to improve functional recovery after stroke (Chen J et al., Neurobiology Yearbook 2007; 62(1): pp. 49-58). HDL-C itself has been proposed for the treatment of stroke and other ischemic symptoms (Kapur NK et al., Vascular Health and Risk Management 2008; 4(1): pp. 39-57. European Patent No. EP 1 425 031 ).

NA is available in 3 formulations (immediate release, extended release and long-acting effects). Immediate release of NA is associated with poor flushing and elevated blood glucose levels. Long-acting NA is associated with reduced flushing, but there is also a risk of hepatotoxicity. Prolonged release is associated with less flushing and low hepatotoxicity risk (Pieper JA, US Health Systems Pharmacy Journal 2003; 60 (13 Supplement 2): Page S9-14. McKenney J, Internal Medicine, 2004; 164 (7): pp. 697-705. Knopp RH, American Journal of Cardiology 2008; 86 (supplement); page 51L-56L). The use of sodium, potassium and magnesium salts of NA is also described.

A major disadvantage of NA is that large doses must be administered to substantially alter blood lipid levels. Almost 100% of subjects treated with NA experience uncomfortable flushing side effects that prevent treatment with NA. The release of prostaglandin D2 from the skin was identified as the direct cause of flushing induced by NA (Morrow JD et al., Journal of Dermatology Research 1992; 98: pp. 812-5). As a result of the activity of the flushing prostaglandins associated with NA, acetyl salicylic acid, which is a recognized inhibitor of prostaglandin synthesis, has been proposed to control flushing. In addition to acetaminophen, other NSAIDS are also effective (Oberwittler H and Baccara-Dinet M, International Journal of Clinical Practice 2006; 60(6): 707-715). However, NSAIDS itself is not completely free of side effects and can cause gastrointestinal irritation and ulceration.

Recently, a specific antagonist of prostaglandin D2 has been proposed (Parhofer KG, Vascular Health and Risk Management 2009; 5: 901-908). Receptor type 1 and laropiprant. Agents for reducing flushing induced by NA (Lai E et al., Journal of Clinical Medicines and Therapeutics, 2007; 81: 849-857. Davidson MH, American Journal of Cardiology 2008; 101 [Supplement]: Page 14B-19B). Although the addition of lauropirin will reduce the frequency of flushing, it does not completely eliminate this side effect. Laropilin does not alter the effects of niacin on lipids or other side effects of niacin. Therefore, the combination of nicotinic acid and lauropirin allows the high dose of niacin to be used, thus developing the full potential of this drug (Parhofer KG, Vascular Health and Risk Management 2009; 5: 901-908, Olsson AG, Pharmacological Therapeutic Expert Advice 2010; 11(10): 1715-1726).

Patients with type 2 diabetes often have altered lipid abnormalities characterized by an increase in TG concentration and a decrease in HDL-C concentration. Considering the pharmacological effects of NA on lipid metabolism, NA should counteract the abnormal changes in lipids in patients with type 2 diabetes. However, several reports indicate that NA increases insulin resistance (Garg A and Grundy SM, American Journal of Medicine 1990; 264: pp. 723-6. Kahn SE et al., Diabetes 1989; 38: page 562 -8) and increase the glucose concentration (Elam, MB et al., American Medical Journal 2000; 284 (10): 1263-1270). Therefore, a limited NA dose (<2 g/day) is recommended for diabetic patients (Shepher J, Betteridge J, and Van Gaal L, Current Medical Research and Opinions 2005; 21(5): pp. 665-682). Obviously, there is a need for novel agents that improve glycemic control in diabetic patients who use NA. Clinically, it has been found that the blood sugar lowering ability of mitoxantrone (Statsenko ME et al., Surgical Medicine (Russian) 2007; 85 (7): pp. 39-42), it is expected that this agent can be combined with NA as an additional clinical Advantage.

During coronary heart disease, platelets play a key role in the development of atherosclerosis and fatal thrombosis. Antiplatelet agents have become the first choice for the prevention and management of a variety of diseases associated with cardiovascular, cerebrovascular, and peripheral arterial systems (Meadows TA et al., Recycling Research 2007; 100(9): pp. 1261-75). NA is an effective lipid-modifying agent that prevents atherosclerosis and reduces cardiovascular events. NA has a variety of lipoprotein and anti-atherosclerotic effects, which improve endothelial function, reduce inflammation, increase plaque stability, and reduce thrombosis (Rosenson RS, atherosclerosis 2003; 171: 87 -96).

NA inhibits platelet aggregation (Lakin KM, Farmakol Toksikol, 1980; 43(5): pp. 581-5). NA affects platelet activity in vitro by gently inhibiting platelet aggregation and stimulates significant prostaglandin release, as well as most intact major platelet receptors. The efficacy of NA is unique, unlike other known antiplatelet agents, and offers potential opportunities for therapeutic combinations (Serebruany VL et al., Thrombosis and Hemostasis , 2010 (in press)).

NA almost completely prevents intravascular coagulation induced by thrombin and pituitrin, showing its effect of lysing thrombus (Baluda VP, Cardiology 1974; 14 (11): pp. 105-7 (Russian) ). Several authors have described the antithrombotic properties of NA (Shestakov VA, Probl Gematol Pereliv Krovi , 1977; 22(8): pp. 29-35. Chekalina SI, Sov Med 1982, 5: p. 105-8) . Niacin reduces the risk of blood clots (Chesney CM et al., American Heart Journal , 2000; 140: 631-36).

The MD system has certain drugs that have beneficial effects on the heart and blood vessels. Some desirable MD activities have been found in animal models of atherosclerosis (Veveris M, Smilsaraja B, Baltic Animal Science Laboratory 2000; 10, pp. 194-199. Veveris M et al., Baltic animals) Science Laboratory 2002; 12: pp. 116-122. Okunevich IV and Ryzhenkov VE, Patol Fiziol Eksp Ter 2002; (2): pp. 24-7), and clinically observed (Karpov RS et al. Human, Ter Arkh 1991; 63(4): pp. 90-3), it is therefore expected that this agent can be combined with NA as an additional clinical advantage.

It has also been noted that MD inhibits platelet aggregation (Tsirkin VI, Ros Kardiol Zh 2002; Phase 1: Pages 45-52). MD medical use for oral administration to rabbits and dogs for two weeks after experimental arterial thrombosis, showing the effect of thrombolytics (Logunova L et al., Experim Clin Pharmacoter 1991; 19: page 91-9 (Russian )). There is no known data on the preventive effect of MD in controlling or preventing thrombosis.

The current trend in the development of pharmaceutical compositions to treat diseases associated with lipid metabolism can improve the efficiency of clinical care (Black DM, Contemporary Therapy Research 2003; 5: pp. 39-32). Because fibrates, NA, and Studdin can each modulate serum lipids according to different mechanisms, combination therapy provides a particularly desirable benefit to others than monotherapy. As the development of novel agents for lowering LDL-C concentrations has progressed slowly, research has turned to developing better agents that increase HDL-C concentrations. Increasingly, a combination of NA, fibrate, Sterling, and bile acid sequestrants is used to treat lipid metabolism-related diseases because of the additive profiles of the products (Miller M, Mayo Hospital) set 2003; 78 (6) of: page 735-42.Backes JM, et al., vascular health and risk management 2005; 1 (4) of: page 317-331.Belsey L, et al., current Medical Research and Views 2008; 24(9), pp. 2703-9. Rosenson RS and Pitt B, Natural Cardiovascular Clinical Applications 2009; 6(2): pp. 98-100). Increasingly, a combination of NA and other agents such as acetylsalicylic acid and other NSAIDs as platelet inhibitors has been used (U.S. Patent No. 5,981,555). However, these compositions did not increase the activity of NA, i.e., no synergistic effects were reported. Therefore, any agent that enhances the therapeutic effect of NA in a lipid metabolism-related disease without increasing the adverse side effects of NA will have excellent clinical utility.

One object of the present invention is to provide a therapeutic combination for preventing and/or treating a disease associated with lipid metabolism, and a method for preventing and/or treating a lipid metabolism-related disease in a subject in need of such prophylactic treatment, by using NA binds to MD, which has a further beneficial effect of alleviating the adverse side effects of NA. The therapeutic combination of the present invention is expected to have synergistic effects in a method of treating and/or preventing a disease associated with lipid metabolism, including abnormal dyslipidemia, hyperlipidemia, atherosclerosis, selected from the group consisting of angina pectoris and myocardial infarction. Coronary heart disease, transient or permanent ischemic attack with cerebrovascular accidents and stroke and peripheral arterial occlusive disease, prevention of platelet aggregation and thrombosis.

According to the present invention, a therapeutic combination is defined as having a synergistic effect if it is therapeutically superior to NA or MD alone. Needless to say, the therapeutic combination as used herein means that the composition agent is administered simultaneously, sequentially or separately. Another object of the present invention is to provide a pharmaceutical composition comprising both NA and MD for the aforementioned purposes. Further objects of the present invention will become more apparent hereinafter, and other objects will be apparent to those skilled in the art.

Description of the invention

The present invention comprises a combination of NA and MD which provides the result of an effective synergistic combination for the treatment of diseases associated with lipid metabolism, preferably in the form of a single dosage unit. Alternatively, the two components can be administered separately, simultaneously or sequentially in any order. The exact form of administration of the active ingredient is not critical as long as the desired effect of the present invention can be obtained. The active ingredient may be in the form of a capsule, suspension, dispersion, elixir, syrup, or the like, which may be administered separately or in a single composition.

Now we have surprisingly found that NA and MD have synergistic effects on lipid metabolism-related diseases, and other beneficial effects. Unexpectedly, MD was the first agent to enhance the beneficial effects of NA, that is, to reduce the concentration of TG and LDL-C and increase the concentration of HDL-C, enhance the reverse polymerization of NA, and improve the defect of NA. Side effects, especially flushing and increased blood sugar levels.

The above composition is therefore expected to be a preferred agent for the treatment of abnormal dyslipidemia in diabetic patients. We have also surprisingly found that the compositions of the present invention improve the effects of experimental infarction and stroke.

Now we have surprisingly found that NA and MD have a synergistic effect on platelet aggregation. Unexpectedly, MD was the first agent to enhance the antiplatelet effect of NA.

The compositions of the invention may be in a form suitable for oral administration (for example as a tablet, capsule, aqueous suspension or dispersible powder or granule), for parenteral administration (for example as for intravenous, subcutaneous, or intramuscular injection) A sterile aqueous solution of the drug or as a suppository for rectal administration. Preferably, the therapeutic combination of the invention is in a form suitable for oral administration, for example as a tablet or capsule.

The therapeutic combination according to the invention also comprises a composition of separate pharmaceutical compositions of the active agent comprising a first NA composition or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient or carrier, And a second composition comprising MD or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient or carrier. The composition of this pharmaceutical composition provides for the simultaneous or sequential use of the therapeutic combinations of the invention. An advantage of this composition is that it allows the physician to adjust the proportion of active agent for individual patients. Therapeutic combinations of the invention are also contemplated to include pharmaceutically acceptable salts of NA and NA (sodium, potassium or magnesium) as well as sustained release or extended release dosage forms of MD and its salts.

The therapeutic combination of the present invention may also include other drugs having known activity against lipid metabolism-related diseases, namely, Sterling, particularly simvastatin.

The pharmaceutical compositions of the present invention can be obtained by conventional procedures using conventional pharmaceutically acceptable excipients or carriers and techniques.

The following examples are provided to illustrate the invention but not to limit the scope of the invention.

Instance

Test: The medicinal activity of the test substance was studied by standard methods used in the prior art. Animals were placed in appropriate cages in a climatic chamber at 22 ± 1 ° C and 60 ± 5% relative humidity for 12/12-hour light/night cycles, with free access to food and water. . All experiments were carried out in accordance with the provisions of the European Community Council Directive (86/609/EEC) on the care of laboratory animals on November 24, 1986. All efforts are made to minimize animal suffering and reduce animal use.

Substance: Cholesterol (Acros Organics), Miqu (Grindes Grindex), Animal Nutrition (R 70 Lactamin), Cream (commercially possible), Sodium Cholate (Akuros Fukuoka Organic), NA (Akuros Fukuoka organic), laropiprant (MK 0524, Cayman Chemicals). Other chemicals are commercially available.

Example 1 - Determination of anti-atherosclerotic activity

METHODS: A atherosclerotic model of C57BL/6J mice susceptible to atherosclerosis genes was used, as suggested by the literature (Smith JD and Breslow JL, Journal of Internal Medicine 1997; 242: page 99) -109). The control group received standard laboratory animal nutrition. Experimental atherosclerosis was induced by the addition of 1.25% cholesterol, 15% cream, and 0.5% sodium cholate to standard nutrients (Nishina P et al., Journal of Lipid Research 1993; 34: 1413-1422 ). The experimental group received the test substance in the form of the effective dose established by the lead experiment and the composition. Add the substance to be tested to drinking water. The preparation of the dose was corrected based on the amount of water actually consumed during the experiment. On average, mice drink about 12% of body weight of water a day. At 22 weeks, atherosclerotic changes were assessed in terms of morphology, biochemistry, and histology by standard methods and criteria (Paigen B et al., Atherosclerosis 1987; 68: page 231- 240.) Total C, HDL-C, LDL-C and TG in serum were determined using a commercially available test kit. The LDL-C/HDL-C ratio was accepted as a standard tool for assessing cardiovascular risk (Fernandez ML and Webb D, American University Nutrition Journal , 2008; 27(1): 1-5).

The atherosclerosis index (which reflects coronary atherosclerosis and is differentially identified from peripheral atherosclerosis) is calculated by the following formula: index = LDL-C/HDL-C.

The coefficient of atherosclerosis (reflecting coronary atherosclerosis) was calculated from the following equation: coefficient = total C/HDL-C.

Statistics: Data for 7 to 10 separate animals/quantity are expressed as mean ± standard deviation (Mean ± SEM). One-way ANOVA analysis and repeated comparisons (tower assays) were used to compare the differences between the experimental groups. P<0.05 is considered significant of.

Results: The first series of trials were arranged and the effects of different ratios and doses of the compositions of the compositions were determined. As shown in Table 1, after 22 weeks, C-controlled animals receiving lipid- and C-rich nutrients developed changes in atherosclerosis in the aorta, particularly the aortic arch. NA and MD, when used separately, show a tendency to reduce the damage area. Surprisingly, the combination of NA and MD results in a relatively high and statistically significant protective effect against atherosclerotic damage compared to individual substances, i.e., it has a synergistic effect.

As shown in Table 2 below, the combination of NA and MD showed a statistically significant effect on lowering the LDL-C and TG concentrations and increasing the HDL-C concentration. The composition dose NA50 + MD150, wherein the NA system is used at a dose which does not have a significant effect on the lipid concentration, and the summary effect is unexpectedly higher than that of each individual substance. This is particularly evident in the atherosclerosis index and total C/HDL-C ratio, and the synergistic effect of NA and MD is observable.

In another series of experiments, the anti-atherosclerotic activity of the compositions of the invention was evaluated in more detail and a known anti-lipemic agent SI was further added to the composition. NA and MD, when used separately, showed a tendency to reduce the damage area (Table 3). Surprisingly, the combination of NA+MD results in a relatively high and statistically significant protective effect against atherosclerotic damage compared to individual substances. The addition of the composition of the invention to SI further increases its protective effect against atherosclerotic damage.

In the same series of experiments, total C, HDL-C, LDL-C and TG in serum were also determined. As shown in Table 4 below, NA and MD, when used separately, only slightly improved the ratio of cholesterol content and the atherosclerosis index without any statistical significance. Surprisingly, the combination of NA and MD considerably improved the ratio of cholesterol levels and statistically significantly reduced the atherosclerosis index and total C/HDL-C ratio. This composition also prevents an increase in LDL-C and TG in the serum. Thus, the compositions of the present invention have a significantly higher protective effect on altered lipid metabolism relative to when used separately. The three recombinants with SI also maintain a significant protective effect.

In yet another series of experiments, NAMg and NA+SI compositions were included in the comparative evaluation (Table 5). Some have compared NAM and NA of cocks (Burstein J and Telkka A, Nordic Journal of Pathogenic Microbes, 1962; 56: 261-265). Based on the proportions used in clinical trials, the combination of NA (50 mg/kg) and MD (150 mg/kg) has the most significant beneficial effects on the aorta, better than NA and NAMg, and better than SI and NA. Composition (Pandian A et al., Vascular Health and Risk Management 2008; 4(5): pp. 1001-1009).

Similar to morphological data, biochemical tests confirmed that the NA+MD composition was significantly superior to NA or NAMg in normalizing lipid concentrations (Table 6).

NA50+SI2=NA 50 mg/kg+SI 2 mg/kg*P<0.05 vs C control group**P<0.005 vs C control group***P<0.0005 vs C control group ^$ P<0.05 vs NA ^@P <0.05 vs NAMg ^& P<0.05 vs MD

The composition of NA+MD is similar to the composition of NA+SI in reducing total C and LDL-C, but is substantially better in the effect on the beneficial effects of HDL-C and TG concentrations, and in arterial porridge. The hardening index and the total C/HDL-C ratio have more significant effects.

This series of tests confirmed that in the case of experimental atherosclerosis, the combination of NA and MD has significantly better beneficial effects than NA or NAMg and is also superior to the clinically used compositions of NA and SI.

Example 2 - Effects of NA and MD on the lipids of the rat hyperlipidemia model and the effects of the composition

Methods: Experimental chronic hyperlipidemia/hypercholesterolemia was induced with TR using the method described by Levine and Saltzman (Levine S and Saltzman A, Journal of Pharmacology and Toxicology 2007; 55: 224-226). Animals received 250 mg/kg TR solution via the tail vein three times a week for three weeks. The solution of the test substance for the experimental group or the water for the control group was orally introduced one hour before the injection of the TR solution or the blood sample.

Male Weiss rats weighing 220-240 grams were assigned to the following 8 groups (group, number of animals):

After 1, 2, and 3 weeks (from the next day after TR injection), blood for biochemical analysis was obtained by cardiac puncture under ether anesthesia. Serum was separated by centrifugation and total C, HDL-C, LDL-C and TG concentrations were analyzed in a commercially available kit.

Statistics: The data were mathematically processed using software Microsoft Excel and the results were expressed as mean ± mean standard deviation (SEM). ANOVA one-way analysis and Student's t-test were used to compare the average results of different groups. At P < 0.05, the difference in results was considered to be significant.

RESULTS: Repeated injection of TR developed significant and stable hypercholesterolemia and hyperlipidemia, characterized by a significant increase in total C, LDL-C, and TG concentrations compared to the control group. NA therapy, more significantly in the first week, limited the increase in total C, LDL-C, and TG, but significantly increased HDL-C concentrations only at 2 and 3 weeks. MD is slightly weaker in reducing total C and LDL-C concentrations and increasing HDL-C concentration, but does not prevent an increase in TG concentration. Surprisingly, the combination of NA+MD is more effective in reducing LDL-C and TG concentrations and further increasing HDL-C concentrations compared to individual components. Furthermore, the use of extended NA+MD (2 or 3 weeks in our experiments) demonstrates that, in contrast to the use of NA+SI, in terms of lowering LDL-C and TG concentrations and increasing HDL-C concentrations, More significant (see below). Therefore, a composition of NA + MD is expected to be useful in the prevention and/or treatment of hypercholesterolemia and hyperlipidemia.

The use of SI at a dose of 10 mg/kg significantly reduced the increase in total C and LDL-C induced by TR, but only slightly affected the concentrations of HDL-C and TG. The combination of SI and NA in a clinically acceptable ratio demonstrates significant protection against the increase in total C, LDL-C and TG in serum and increases HDL-C concentration. Surprisingly, the combination of NA, SI and MD, compared to the individual use of each component, still demonstrates better protection against TR-induced changes, and significantly lowers the concentration of TG and LDL-C compared to NA. +SI is better. Of particular importance for clinical practice is the fact that the combination of NA, SI and MD is substantially better than SI or NA in terms of increasing HDL-C concentration. Therefore, it is expected that a combination of NA+MD and NA+SI+MD is useful for clinically preventing and/or treating hypercholesterolemia and hyperlipidemia.

^# P<0.05 vs NA+SI

Example 3 - Determination of Cardiac Protection Characteristics

METHODS: Male Weiss rats were assigned to 6 groups (12 to 16 animals per group): 1) control group received oral 0.9% saline; 2) NA50 group received oral 50 mg/kg/day NA; MD50 group received 50 mg/kg/day MD; 4) NA50+MD50 group received 50 mg/kg/day NA plus 50 mg/kg MD; 5) MD150 group received 150 mg orally MD/kg/day MD; 6) NA50+MD150 group received 50 mg/kg/day NA plus 150 mg/kg/day MD.

Animals of the test group received an aqueous solution of the test substance via a gastric catheter at 48, 24 and 1 hour before the experiment. Animals in the control group received an equal amount of saline. Animals were anesthetized (intraperitoneal injection of pentobarbital sodium 60 mg/kg) and placed under mechanical ventilation to prepare for obstruction of the left coronary artery. Experimental arterial infarction was induced by occlusion of the coronal for 45 minutes and then reperfusion for 2 hours. The following data were recorded: VT, VF, number of dead animals, mean arterial pressure, RPP, functional status of the RPP myocardium, reflect the production of ATP in the myocardium, and widely used index for clinical and experimental data analysis (Broderick TL, Drug Research and Development 2008; 9(2): pp. 83-91). After the experiment, the areas of ischemia and necrosis were detected by triphenyltetramine-Evans blue perfusion staining. The left ventricle was incised and weighed. The criteria for calculating the morphology were: the proportion of the ischemic area of the left ventricle, the proportion of the necrotic area of the left ventricle, and the ratio of the necrotic area to the ischemic area (necrosis index).

Statistics: Results for each group are expressed as mean ± standard deviation (Mean ± SEM). Statistical analysis within the group was performed by Student's t test. Data from the recorded blood pressure and heart rate were calculated from animals surviving the ischemia-reperfusion experiment. The number of episodes of arrhythmia (VT and VF) and the number of deaths were calculated for all animals. One-way ANOVA analysis and repeated comparisons (tower assays) were used to compare the differences between the experimental groups. P < 0.05 was considered significant.

RESULTS: In the control group of 16 animals, cardiac ischemia followed by reperfusion caused severe heart rhythm disorders, with 7 deaths. The NA group was not significantly different from the control group in terms of heart rhythm disorders. The MD and NA+MD groups had less significant life-threatening heart rhythm disorders (VF and VT) relative to the control group, but the number of deaths was significantly lower in the NA50+MD50 group and the NA50+MD150 group (Table 8 below).

During reperfusion, mean arterial pressure and heart rate were similar in all experimental groups, but the decrease in RPP observed in the control group was significantly prevented only in the NA50+MD50 group and the NA50+MD150 group system ( Table 9). It is pointed out that the treatment of the composition of the present invention is carried out for a short period of time (3 days long), i.e., it has the protection against functional depletion caused by infarction.

This was also confirmed by the composition of the present invention (NA+MD of both dose combinations) statistically significantly reducing the percentage of necrotic areas to the left ventricle and to the ischemic area (Table 10), as confirmed.

Therefore, we have surprisingly found that the synergistic action of the combination of NA and MD drugs counteracts the high degree of protection against the onset of myocardial infarction. The NA+MD composition significantly maintained myocardial function and increased animal survival (Tables 8, 9), and compared with NA and MD alone, it significantly prevented myocardial necrosis due to ischemia and reperfusion (Table) 10).

Example 4 - Further testing of the anti-hypoxia and anti-ischemic effects of the brain to determine the composition of NA and MD in the experimental CNS ischemia, hypoxia and stroke models compared to individual The utility of the ingredient utility.

4.1. Mouse cerebral circulation hypoxia model

METHODS: Experimental circulatory hypoxia was induced by introducing MgCl ₂ (2% MgCl ₂ , dose 200 mg/kg) into the tail vein of male ICR mice within 3 seconds (Berga P et al., Drug Discovery 1986; 36 (9): Pages 1314-1320). Animals received the test substance in a single dose or an equal dose once daily for 7 days. The test substance is introduced through a gastric catheter. Animals were randomly divided into 7 groups (a group of 6 to 10 animals):

Control group receives water 0.01 ml / g

PI500 group (active control) accepts 500 mg/kg dose of pyridoxamine

NA50 group received NA at 50 mg/kg dose

MD50 group receives MD of 50 mg/kg dose

MD150 group accepts MD of 150 mg/kg dose

NA50+MD50 group accepts 50 mg/kg NA plus 50 mg/kg MD dose

NA50+MD150 group received 50 mg/kg NA plus 150 mg/kg MD dose

The last dose of the test substance was administered 1 hour before the start of the test. The period from the end of the injection of MgCl _{2 to} the end of the last respiratory action was recorded as the survival time.

Statistics: Results for each group are expressed as mean ± standard deviation (Mean ± SEM). Statistical analysis within the group was performed by Student's t test. One-way ANOVA analysis and repeated comparisons (tower assays) were used to compare the differences between the experimental groups. P < 0.05 was considered significant.

Results: The results are summarized in Table 11. When repeatedly introduced, clinically used PI and MD agents showed protective anti-hypoxia effects (Table 11). Unexpectedly, the composition of NA+MD has shown significant protective effects after a single application, especially at a dose of NA50 + MD150, where the effect is superior to individual substances. Repeated application of this composition has a more pronounced effect. The results obtained in this trial indicate that the composition of NA + MD is clinically useful for the treatment of hypoxia.

^* P <0.05 vs control group ^# P <0.005 vs control groups ^@ P <0.0005 vs Control Group ^$ P <0.05 vs MD same dose ^{& P <0.05 vs NA § P} <0.005 vs NA

4.2. Middle cerebral artery occlusion model

Methods: Male ICR mice weighing 21 to 25 grams were used. The cerebral artery (MCA) source was occluded by intracavitary filament technique according to the known method applied to mice by ZhangQ et al. ( Behavioural Brain Research 2006; 169: pp. 66-74) (Longa EZ et al. , Stroke 1989; 20: page 84-91). The scientific experimental program was processed using prophylactic (7 days a day for permanent MCA occlusion models) and therapeutic (starting treatment after temporary MCA occlusion).

Control group animals received only saline. This test is a good model for a true brain injury and stroke in a clinical setting, which often encounters occlusion of the middle cerebral artery. The experiment was further continued according to two scientific experiment plans. In the first plan, the occlusion was permanent. In the second plan, the occlusion of the intraluminal filaments was temporary and the filaments were removed after 2 hours and introduced into the reperfusion. The neurological status of the animals was assessed 24 hours after occlusion. On a one-level scale, animals were given 0 points for disease-free animals and 4 points for animals that were unable to spontaneously move (Longa EZ et al., Stroke 1989; 20: page 84-91). After assessing neurological deficits, the animals received an excess of sodium pentobarbital, which was isolated and sliced into 6 layers of approximately 1.5 mm each. The sections were stained with 2% triphenyltetrathene at 37 ° C for 30 minutes and photographed. A third section on the cerebral side of the optic nerve crossing layer was selected, which is best suited for the calculation of cerebral ischemic injury because it is entirely supplied by blood from the middle cerebral artery.

Statistics: Data for 7 to 9 separate animals are expressed as mean ± standard deviation (Mean ± SEM). One-way ANOVA analysis and repeated comparisons (tower assays) were used to compare the differences between the experimental groups. Student's t test was used to analyze the difference in neurological function scores between groups. P < 0.05 was considered significant.

RESULTS: After 24 hours of occlusion, all 8 control animals exhibited an average of 2.63 points of neurological disorder (Table 12 below).

In the sham operation group, only one animal showed a slight disorder. Repeated use of MD for a total of 7 days partially prevented the deterioration of the MCA occluded nerve state. Surprisingly, the NA+MD composition exhibited significant defense against neurological disorders, superior to the effect of NA (Table 12).

In the control group animals, occlusion of the middle cerebral artery resulted in ischemic injury, which accounted for 22.2% of the brain tissue in the optic nerve crossing layer (Table 13). Administration of the NA+MD composition for 7 days exhibited significant defense against brain tissue damage and was also superior to NA alone.

In the next experiment, the middle cerebral artery was temporarily occluded for 2 hours, followed by reperfusion, resulting in severe brain dysfunction in the control group of animals 24 hours after the test (Table 14 below). The test substance was introduced at 1, 3, and 6 hours of occlusion, and neither the NA25 (25 mg/kg x 3) group nor the MD75 (75 mg/kg x 3) group provided significant protection. Surprisingly, only the NA25+MD75 (25 mg/kg+75 mg/kg) group with the test combination was introduced 3 times after occlusion, and the brain function was substantially protected, significantly better than NA and MD alone. (Table 14 below).

Morphological analysis revealed that NA or MD alone did not have significant defense against brain tissue damage caused by temporary occlusion and reperfusion of MCA (Table 15 below). Surprisingly, the combination of NA+MD (25 mg/kg + 75 mg/kg), starting treatment after occlusion, demonstrated substantial defense against brain tissue damage, significantly better than NA and MD alone (Table 15).

Therefore, we have surprisingly found that the combination of NA plus MD, whether used before or after MCA occlusion, provides a significantly better functional and morphological impairment of brain tissue than the individual components. Defense. These results indicate that compositions of such drugs may be beneficial in the treatment and/or prevention of CNS hypoxic-ischemic conditions including stroke, also because of their inhibitory activity against platelet aggregation and thrombosis tests, as described below.

Antiplatelet aggregation and antithrombotic activity

Test: Platelet aggregation (in vitro); Rat thrombosis model (in vivo); Record changes in skin temperature in vivo.

Example 5 - Effect of MD and NA on platelet aggregation

METHODS: Whole blood was collected from a healthy donor B. (37 years old) who had never used ASA or any other antiplatelet agent. Multiplate (multiple platelet function analyzers, Dynabyte Medical, Germany) was used. Methods of establishment (Toth O et al., Journal of Thrombosis and Hemostasis, 2006; 96: pp. 781-788. Velik-Salchner C et al., Anesthesiology and Pain Relief Journal ; 107: Page 1798-1806) To study platelet aggregation. Blood samples were collected into plastic tubes covered with hirudin (Dynabyte Medical, Germany) for measurement between 30 minutes and 4 hours after collection. Measurements were made according to a modified scientific experiment by Dynabyte Medical. Isotonic sodium chloride solution (0.3 ml, or saline with the compound to be studied (to a final concentration of 10 ^-6 to 10 ^-4 mmol/ml)) was preheated to 37 ° C and pipetted to the test cells In addition, 0.3 ml of a whole blood sample with hirudin anticoagulation was added. After 5 minutes of incubation, stir at 37 ° C to add the appropriate agonist solution (obtained from Dynabyte Medical, Germany) to start the measurement:

1) Adenosine diphosphate (ADP)-ADP-test. ADP stimulates platelet activation by the ADP receptor (P2Y12 and others).

2) ADP HS test (combination of prostaglandin E ₁ and ADP). The addition of the endogenous inhibitor PGE ₁ made the ADPHS test more sensitive to clopidogrel and related drugs than the ADP test.

The aggregation curve was recorded for 6 minutes and analyzed using software from Dynabyte Medical. We calculated the following parameters for platelet aggregation:

1) Amax, the maximum value of platelet aggregation in aggregates in arbitrary units (AU);

2) AUC, the total area under the aggregation curve (AU*min). This is affected by the total height of the aggregation curve and its slope and is best suited to express overall platelet activity.

Statistics: Results are expressed as mean ± standard deviation (Mean ± SEM). To assess the significance of the differences, one-way ANOVA analysis was used. If the null hypothesis is ruled out, then the student Newman Muller check is used after the fact.

Results: The first series of tests were planned to determine the effect of different concentrations of the agent. As shown in Table 16, a wide range of concentrations of MD provided significant defense against platelet aggregation induced by ADP + PGE ₁ . Amax decreased from 100% of the control group to 55-58% of the MD 10 ^-5 group and the 10 ^-4 group. NA (in the 10 ^-4 and 10 ^-3 mM/ml groups) also reduced aggregation caused by ADP. The binding activity of the two substances provided a significantly more and significant decrease in platelet aggregation caused by ADP or ADP + PGE ₁ , which is shown in both AUC and Amax data.

^## P<0.0005 vs MD 10 ^{-4 $} P<0.005 vs NA 10 ^{-4 $$} P<0.0005 vs NA 10 ^-4

Example 6 - Effect of MD and NA on thrombosis

METHODS: We chose an experimental thrombosis model based on rat arterial thrombosis induced by ferric chloride (FeCl ₃ ) (Kurz K et al., Thrombosis Research 1990, 60: 269-280). Wang X and Xu L, Thrombosis Research 2005, 115: 95-100). Tissue damage initiated by the chemical oxidation of Tiezhongsuke causes injury sites that are susceptible to platelet adhesion and aggregation, followed by coagulation activation and fibrin deposition. Male Weiss rats weighing 350-420 grams were used in the experiment. Rats were randomly divided into experimental groups, each consisting of not less than 7 animals. Vehicle or test chemistry MD (25 mg/kg), NA (25 mg/kg) and composition MD+NA (25+25 mg/kg) were administered by oral route 2 hours prior to the onset of thrombosis.

Rats were anesthetized with an intraperitoneal injection of 50 mg/kg sodium pentobarbital and placed on a thermally controlled operating table to maintain a body temperature of 37 °C throughout the experiment. One of the carotid arteries was exposed by a neck incision, separated from the adhered tissue, the vagus nerve, and a first-class probe (electromagnetic blood flow meter MFV 1200, Nicon Kohden, Japan) was placed on the exposed part of the total carotid artery to record Blood flow. After a 15 minute stabilization period, thrombosis was induced by topical application (contact with the outer surface of the adventitia) two sheets (2 x 1 mm) of Whitman filter paper immersed in a 15% solution of ferric chloride (FeCl ₃ ). The time of carotid thrombosis is recorded as the time required to cause complete stop of blood flow, and is described here as time to block (TTO).

In addition, during the thrombosis experiment, the tail bleeding time of the rat was measured. The tail was transected 5 mm from the trailing end with a scalpel and the tail was immediately immersed in warm isotonic saline at 37 ° C until bleeding was noted. The point at which bleeding completely stopped and no more bleeding occurred in the next 30 seconds was defined as bleeding stop.

Statistics: Data was analyzed by software Microsoft Excel 2007. Data for 7 to 8 separate animals/quantities are expressed as mean ± standard deviation (Mean ± SEM). One-way ANOVA analysis and repeated comparisons (tower assays) were used to compare the differences between the experimental groups. P < 0.05 was considered significant.

RESULTS: In the control group, the mean time for vascular thrombosis caused by ferric chloride (FeCl ₃ ) and the resulting arterial flow arrest was 24.4 minutes (Table 17).

MD did not provide a significant increase in TTO at the 25 mg/kg dose. NA causes an increase in a small amount of TTO but is not obvious. NA has little effect on tail bleeding time. The combination of MD and NA unexpectedly caused a fairly significant increase in TTO (39%), with no significant increase in tail bleeding time.

Considering the positive benefits of a combination of MD and NA against platelet aggregation and prolongation of TTO in vivo, this composition can be used to reduce significant atherosclerosis, possible myocardial infarction and injury, and peripheral blood circulation. The risk of thrombosis in patients with disorders. The fact that the MD and NA compositions do not prolong the bleeding time indicates the potential use of this composition for patients with increased risk of bleeding before and during surgery.

Example 7 - Comparative study of MD/NA and LA/NA applied to reduce flushing

Nicotinic acid (niacin, NA) is effective in lowering serum cholesterol, LDL and triglycerides, and increasing HDL. However, a restrictive adverse effect on patients receiving immediate or sustained release of niacin is the apparent rapid development of skin warming and vasodilation, known as "flushing", which severely leads to discontinuation (Gupta EK and Ito MK) , Heart Disease 2002; 4: page 124-137). Laropiprant (MK-0524) has been proposed as one of the most active and prospective agents for reducing nicotinic acid flushing (Cheng K et al., Proceedings of the National Academy of Sciences, 2006; 103: Page 6682-6687). Our goal was to compare the effects of MD and LA on flushing (changes in skin temperature and blood flow) caused by NA in the experiment.

7.1.1. Determination of vasodilation of the skin

Model: Male Weiss rats were anesthetized with sodium pentobarbital (50 mg/kg, ip) and anesthetized at an additional dose per hour (10 mg/kg). The blood pressure of the left carotid artery was measured and the ECG was recorded in the standard II lead. The blood flow to the right ear artery was measured with a laser Doppler flowmeter (OXYFLOW 2000, USA). Blood flow, ECG and arterial pressure were recorded in an AD instrument PowerLab system and stored in a computer for further processing. After a 10-minute long baseline recording, the test substance was injected into the armor area by subcutaneous injection and recording continued for 30 minutes. The mean blood flow data for each animal was calculated taking into account the mean blood pressure and compared with the starting value and the control group. The results were calculated experimentally from 5 to 8 and expressed as a percentage as the largest blood flow to the baseline (Carballo-Jane E et al., Journal of Pharmacology and Toxicology 2007; 56(3): Page 308-316).

Statistics: Results for each group are expressed as mean ± standard deviation (Mean ± SEM). Statistical analysis between groups was performed by Student's t-test and chi-square test for unpaired data. One-way ANOVA analysis and repeated comparisons (tower assays) were used to compare the differences between the experimental groups. P < 0.05 was considered significant.

Results: As can be seen from Table 18 below, in this animal model, a dose of 15 mg/kg of NA resulted in a significant increase in blood flow to the auricular arteries. MD is similar to the control group, resulting in insignificant changes in blood flow. Together with MD, NA alone resulted in a delayed (slow increase) and a significantly statistically less significant increase in blood flow compared to NA alone (Table 18). MD can antagonize peripheral vasodilation caused by NA, and this potential can have clinically beneficial effects for attenuating the effects of niacin on the skin (flushing) and is therefore further studied in detail (described below).

Table 18 Effect of experimental substances on vasodilation of the skin; n = 5-8, mean ± standard deviation

7.1.2. Effects of MD and LA on vasodilation of skin induced by NA

Materials and methods: see section 7.1.

Experimental plan

Statistics: Results for each group are expressed as mean ± standard deviation (Mean ± SEM). Statistical analysis within the group was performed by Student's t test. One-way ANOVA analysis and repeated comparisons (tower assays) were used to compare the differences between the experimental groups. P < 0.05 was considered significant.

RESULTS: When MD was administered alone, similar to LA, only a significant change in blood flow was caused. When NA and MD were administered simultaneously (advance time = 0), the increase in blood flow became slower, and was less significant than the simultaneous administration of NA and LA (LA + NA [0], see Table 19). In terms of ear blood flow, there was no significant difference between MD+NA[0] (when MD was added with NA) and MD+NA[30] (pre-treatment before MD 30 mg/kg 30 minutes before NA) s difference. In our experiments, pre-LA treatment 30 minutes before NA (NA+LA [30]) significantly reduced the increase in NA-induced blood flow in rat ear vessels (Table 19).

MD can antagonize peripheral vasodilation caused by NA, and this potential can have clinically beneficial effects for attenuating the effects of NA on the skin (flushing). In our experiments, we were unable to establish the advantages of simultaneous administration of any of the NA and LA compositions compared to the simultaneous administration of NA and MD.

7.2. Assessment of changes in skin temperature induced by niacin

Materials and Methods: To record changes in skin temperature throughout the rat, a non-contact temperature recording method was used (Papaliodis D et al., British Journal of Pharmacology 2008; 153: 1382-1387). Temperature measurements were performed using a hand-held infrared thermometer (Model Proscan 510, TFA-Dostman). Animals were accustomed to being treated with the infrared probe three days prior to use. Under no anesthesia, three animals were read from the dorsal side of each ear before the animals were injected subcutaneously with NA (armor region) or solvent/test compound (tail region). The ear temperature was then measured every 5 minutes for a period of up to 60 minutes. Between measurements, animals are placed back into their cages. Six ear temperature measurements (three per ear) were averaged at each time point. On each day of the experiment, laropiprant (MK 0524, Cayman Chemicals Cayman Chemicals) was first dissolved in dimethylarsine (DMSO) followed by dilution with 0.9% sodium chloride (NaCl). . LA ratio NA and the composition is based on modified Tredaptive ^TM Summary of Product Characteristics of sustained-release tablets (Niacin / laropiprant (laropiprant)) 1000 mg / 20 mg.

Experimental plan

I test time and the effect of solvent on skin temperature

Study on the effect of II NA/LA and NA/MD compositions on skin temperature

Simultaneously introduce LA with LA such as LA+NA[0], or introduce LA such as LA+NA[30] 30 minutes before NA, import MD with NA+MD[0] simultaneously with NA, or 30 minutes before NA Introduce MD such as NA+MD [30]; also examine the effect of LA and MD alone on skin temperature.

Statistics: Data were analyzed by software Microsoft Excel and the results were expressed as mean +/- mean standard deviation. The average results of the different groups were compared according to ANOVA using single factor analysis and Student's t test. P < 0.05 was considered significant.

Results: From 10 AM to 2 PM, the recorded baseline mean ear temperature was 28.1-30.2 °C. A time response study of NA (15 mg/kg, subcutaneous injection) showed that at 10 minutes, the maximum temperature from the baseline was increased by 2.32 ± 0.37 ° C, and the maximum temperature was increased by 2.57 ± 0.43 compared with the solvent group (P < 0.005) (Figure 1). It was established that the effect of LA solvent on ear temperature was only substantially different from NA and MD solvents at the first 5 minutes after injection, so only one control group was used.

Subcutaneous injection of MD or LA did not cause significant changes in rat ear skin temperature (Table 21). Simultaneous administration of NA and MD (NA+MD[0] group; lead time = 0) caused a decrease in NA flushing, which was similar to simultaneous administration of NA and LA. The temperature increase caused by NA was reduced, correspondingly to 69% and 67% (Table 21). There was no change in temperature between MD+NA[0] (when MD was added with NA) and MD+NA[30] (pre-treatment before MD 45 mg/kg 30 minutes ago). In our experiments, only pre-LA treatment (when subcutaneously injected at a dose of 0.3 mg/kg, 30 minutes in advance) resulted in significant defense against skin temperature increase by NA (Table 21).

We have therefore established that the combination of NA and MD not only has unanticipated synergistic anti-platelet activity, but also reduces side effects - flushing.

Example 8 - Determination of the effect on blood glucose concentration

It is well known that even a single large oral dose of NA increases the blood GL concentration in animals (Thornton JH and Schultz LH, Journal of Dairy Science 1980; 63, pp. 262-268), and in the treatment of diabetic patients, NA needs to monitor sugar concentration (Goldberg RB and Jacobson TA, Mayo Hospital Proceedings 2008; 83(4): 470-8).

Methods: Male adult Weiss rats were used in the experiment. Animals were fasted one night before the experiment to stabilize the blood sugar concentration. The GL concentration in venous blood was measured before the experiment and 45 minutes after oral administration of the blank control group or the test substance. The GL concentration was determined in a commercially available kit ( Optium, Abbott Diabetes Care Ltd, USA ). A high dose of NA (300 mg/kg, orally introduced), which is known to cause a stable and long-term increase in blood GL concentration. Use the same dose of MD (300 mg/kg).

Animals were randomly divided into 4 groups (n=8):

1) Control group (accept 1% sodium chloride (NaCl) solution, dose 2 ml / kg)

2) NA group (accepting NA, dose 300 mg / kg)

3) MD group (accept MD, dose 300 mg / kg)

4) NA+MD group (accept 300 mg/kg NA and 300 mg/kg MD)

Statistics: Results for each group are expressed as mean ± standard deviation (Mean ± SEM). Statistical analysis within the group was performed by Student's t test. One-way ANOVA analysis and repeated comparisons (tower assays) were used to compare the differences between the experimental groups. P < 0.05 was considered significant.

Results: NA caused a statistically significant increase in GL concentration compared to the baseline and control groups (Table 21 below). MD caused a significant decrease in GL concentration, which is no different from the control group. The combination of NA and MD caused a notable, less significant increase in GL concentration (27.5% for the benchmark and 61.1% for NA). This positive benefit indicates that the combination of NA + MD drugs can cause less significant side effects in patients with unstable or disordered glycemic control.

Summary conclusion

Unexpectedly, it was found that the combination of NA and MD enhances the therapeutic effect of NA on diseases including abnormal dyslipidemia, hyperlipidemia, atherosclerosis, and a group selected from angina and myocardial infarction. In the case of coronary heart disease, transient or permanent ischemic attack with cerebrovascular accidents and stroke and peripheral arterial occlusive disease, the composition improves the heart and brain disorders under ischemia-anoxia. The composition also improves peripheral vasodilation caused by NA. Thus, this therapeutic combination, compared to NA, is expected to exhibit improved activity in the treatment of lipid metabolism-related disorders, such that the daily dose of NA can be reduced with less significant adverse side effects.

Mode for carrying out the invention

Herein, the treatment combines the terms to provide a component for simultaneous, sequential or separate administration of the composition. Accordingly, the present invention provides a therapeutic combination for simultaneous, sequential or separate use comprising NA and MD or a pharmaceutically acceptable salt thereof for preventing platelet aggregation and thrombosis. The therapeutic combination of the invention can be administered in the form of a pharmaceutical composition. According to this aspect of the invention, there is provided a pharmaceutical composition comprising NA or a pharmaceutically acceptable salt thereof and MD or a pharmaceutically acceptable salt thereof, in admixture with a pharmaceutically acceptable diluent or carrier.

The pharmaceutical composition according to the present invention also includes a separate composition comprising a first NA composition or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable diluent or carrier, and a second comprising MD or A pharmaceutically acceptable salt thereof and a pharmaceutically acceptable diluent or carrier. This composition provides for sequential or separate use. Since the treatment or prevention of diseases associated with lipid metabolism assumes long-term use of the drug, the mode of best practice of the present invention is to provide a form suitable for oral administration, such as a tablet or capsule. In a pharmaceutical composition, the amount of each active ingredient in the therapeutic combination will depend on the condition being treated. Those skilled in the art of treating lipid metabolism-related diseases can easily select an appropriate amount of each active ingredient and a suitable dosage formulation. Preferred ratios of active ingredients of NA and MD or their salts are from 3:1 to 1:3.

According to a further aspect of the invention there is provided a therapeutic combination as defined above or a pharmaceutical composition as defined above for the manufacture of a medicament for simultaneous, sequential or separate use to prevent platelet aggregation/thrombosis.

Figure 1 shows the effect of NA, solvent for NA (SolvNA) and solvent for LA (SolvLA) on skin temperature in rat ears.

Claims (9)

A niacin acid medical treatment composition comprising an effective amount of nicotinic acid or a pharmaceutically acceptable salt thereof, and an effective amount of mitrequinone or a pharmaceutically acceptable salt thereof, in combination with a pharmaceutically acceptable compound Forming agent or carrier. The pharmaceutical composition for niacin acid treatment according to claim 1, wherein the nicotinic acid or a pharmaceutically acceptable salt thereof is in the form of an immediate release, sustained release or extended release formulation. The niacin acid medical treatment composition of claim 1, wherein the rice koji or a pharmaceutically acceptable salt thereof is in the form of an immediate release, sustained release or extended release formulation. The pharmaceutical composition for niacin acid treatment according to claim 1, which comprises about 50-500 mg of nicotinic acid or a pharmaceutically acceptable salt thereof, and about 50-500 mg of rice koji or a medicinal thereof. Acceptable salts. The pharmaceutical composition for niacin acid treatment according to any one of claims 1 to 4, further comprising a stell. A pharmaceutical composition for niacin acid treatment according to claim 5, wherein the Schindler is simvastatin. A use of a niacin acid medical treatment composition according to any one of claims 1 to 4 for the preparation of a medicament for preventing and/or treating diseases: hyperlipemia, atherosclerosis, peripheral Arterial occlusive disease, myocardial infarction, angina with transient or permanent ischemic attack, cerebrovascular accident, stroke, and vascular embolism. A pharmaceutical composition for niacin acid therapeutic composition according to item 5 of the scope of the patent application for the preparation of a medicament for preventing and/or treating a group consisting of the following diseases Uses: abnormal dyslipidemia, hyperlipidemia and atherosclerosis. Use of a niacin acid medical treatment composition according to item 6 of the patent application for the preparation of a medicament for preventing and/or treating a group consisting of abnormal dyslipidemia, hyperlipemia and atherosclerosis .