WO2013176564A1

WO2013176564A1 - Terpenoid agent for the prophylaxis and treatment of atherosclerosis and the correction of metabolic syndrome

Info

Publication number: WO2013176564A1
Application number: PCT/RU2012/000420
Authority: WO
Inventors: Людмила Анатольевна ЛАЦЕРУС; Анатолий Федорович ПИНИГИН; Фания Иршатовна МАГАНОВА; Нина Максимовна ПИНИГИНА; Валерий Геннадьевич МАКАРОВ
Original assignee: Общество С Ограниченной Ответственностью "Инитиум-Фарм"
Priority date: 2012-05-25
Filing date: 2012-05-25
Publication date: 2013-11-28

Abstract

A terpenoid agent for the prophylaxis and/or treatment of atherosclerosis and the correction of metabolic syndrome is proposed, as well as the use of the pharmaceutical substance Abisil for the prophylaxis and/or treatment of atherosclerosis and the correction of metabolic syndrome, and the use of the pharmaceutical substance Abisil in preparing a terpenoid agent for the prophylaxis and/or treatment of atherosclerosis and the correction of metabolic syndrome. The proposed terpenoid agent provides an anti-atherosclerotic effect characterized by a decrease in total lipids, total cholesterol, triglycerides, and low-density lipoprotein (LDL) cholesterol in the blood, the stimulation of the secretion with bile of cholesterol and bile acid, a decrease in the level of damage to the aortic intima, and the prevention of the development of arterial calcinosis and the occurrence of myocardial infarction, and also provides, in the case of metabolic syndrome, corrective effects characterized by a normalization of arterial pressure indicators and triglyceride and glucose levels in the blood, and may be used in the case of such diseases as peripheral vascular disease, arterial hypertension, ischemic heart disease, type 2 diabetes mellitus, and a variety of other diseases associated with atherosclerosis, as well as in correcting imbalances in lipid and carbohydrate exchange in the case of metabolic syndrome.

Description

TERPENOID MEANS FOR PREVENTION AND TREATMENT

ATHEROSCLEROSIS AND METABOLIC SYNDROME CORRECTION

Technical field

The invention relates to the field of medicine and the pharmaceutical industry, and in particular a terpenoid agent for the prevention and / or treatment of atherosclerosis and correction of the metabolic syndrome, the use of the pharmaceutical substance Abisil for the prevention and / or treatment of atherosclerosis and correction of the metabolic syndrome and the use of the pharmaceutical substance Abisil for the manufacture terpenoid agents for the prevention and / or treatment of atherosclerosis and the correction of metabolic syndrome. The proposed terpenoidal agent has an anti-atherosclerotic effect, characterized by a decrease in blood total lipids, total cholesterol, triglycerides and low-density lipoprotein cholesterol (LDL), stimulation of cholesterol and bile acid excretion with bile, a decrease in the degree of damage to aortic intima, and prevention of aortic atherosclerosis and myocardial infarction , as well as the corrective effect in metabolic syndrome, characterized by normalization of blood pressure tions, triglycerides and blood glucose levels.

The proposed terpenoid agent can be used in diseases such as peripheral vascular atherosclerosis, arterial hypertension, coronary heart disease, type 2 diabetes and a number of other diseases caused by atherosclerosis, as well as in the correction of lipid and carbohydrate metabolism in metabolic syndrome.

State of the art

The basis of many existing theories of atherosclerosis is the postulate that this process is a response to damage to the endothelium of the vascular walls.

In this case, the integrity of the cell membranes of endothelial cells is violated, the products of catabolism are delayed, platelet adhesion to their surface, the synthesis of prostaglandins (thromboxanes) starts and a blood clot forms.

As agents that violate the stability of the cellular structures of the endothelium, the following are considered: chemical injuries (hypercholesterolemia and homocystinemia), mechanical stress (hypertension), immune (organ transplantation), etc.

The development and course of the atherosclerotic process is determined by the ability of the cells of the arterial wall to maintain the integrity of the endothelium, for which many complex and interconnected metabolic processes stopping destructive changes are initiated. It has been established that with atherosclerosis, the release of chemokines by almost any cells is enhanced. This occurs under the influence of cytokines, lymphokines, bacterial lipopolysaccharides and viral infections of various etiologies (Arabidze G.G., Tebloev K.I. Atherosclerosis and risk factors - M: Litterra, 2008.- 240 s).

The identification of a large number of changes in the equilibrium of organisms during the development of atherosclerosis during many years of research has contributed to the emergence of a large number of recommended methods and means for stopping these processes. But, since none of the biochemical reactions can be considered fundamental in the development of atherosclerosis, then there are no universal, sufficiently effective drugs.

For the medical treatment of atherosclerosis and the correction of the metabolic syndrome, a large number of different methods and agents have been proposed and continue to be offered. This is a series of hypocholesterolemic formulations (patent RU N ° 2164408, 2001; patent RU Jfe2192856, 2002; RU patent Ns2394587, 2009; US patent N ° 4231938, 1980), carbohydrate metabolism-normalizing agents (RU patent N ° 2158590, 2000; RU patent N ° 2185826, 2001) or blood coagulation factors (RU patent Ns2129001, 1999). The lipolipidemic agents are of primary importance in modern drug therapy for atherosclerosis: statins or inhibitors of 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMG-CoA reductase) - fluvastatin, lovastatin, pravastatin, atorvastatin, rosuvastatin and anion exchange resins (sequestrants of bile acids - cholestyramine or colestipol); nicotinic acid and its derivatives; fibrates (derivatives of fibroic acid - bezafibrate, gemfibrozil, fenofibrate); bisphenol derivative - probucol; Omega-3 polyunsaturated fatty acids and a specific inhibitor of cholesterol absorption from the intestine - ezetimibe (patent RU jY ° 4137322, 1979; patent RU N ° 2011383, 1994; patent RU M> 2372921, 2009; patent RU N ° 2372923, 2009; U.S. Patent J ° 3174901, 2000, etc.).

However, the entire existing arsenal of drugs for the treatment of atherosclerosis not only does not allow to achieve a high therapeutic effect, but also has a number of toxic side effects and, accordingly, contraindications [Guide to atherosclerosis and coronary heart disease / Ed. Acad. E.I. Chazova, Corr. RAMS V.V.Kukharchuk, prof. S.A. Boytsova - M: Media Medica, 2007. - 736 s].

For example, cholestyramine is contraindicated in obstruction of the biliary tract, nicotinic acid in case of impaired liver function, exacerbation of peptic ulcer, diabetes mellitus, gout, statins in acute liver diseases, pregnancy and lactation, infections, injuries, fibrates in pregnancy. Among the side effects, it should be noted that bile acid sequestrants often cause constipation, flatulence, anorexia, heartburn, dysphagia, hiccups, diarrhea, sometimes peptic ulcer, bowel obstruction, pancreatitis, bleeding due to vitamin K deficiency, weight gain. The main side effects of nicotinic acid: redness and itching of the skin, gastrointestinal disorders, hyperglycemia, hepatotoxicity. In the treatment with statins, myositis may develop with a significant increase in the content of creatine kinase in the blood and muscle tissue, as well as the appearance of insomnia, generalized, eczematous skin lesions, gastrointestinal disorders, hypersensitivity with the development of anaphylaxis and angioedema. Gemfibrozil can contribute to the formation of gallstones.

In diseases caused by atherosclerosis (insulin-dependent diabetes mellitus, coronary heart disease and others), starting from the 60s of the last century, clinicians began to take into account the state of metabolic and physiological factors, characterized by the concept of "metabolic syndrome". According to the criteria published in 2005. The American Association of Cardiology (AHA), together with the National Institute of Heart, Lung and Blood (NHLBI) and Russian Recommendations (2007) for the main risk factors for metabolic syndrome obesity (its abdominal form), an increase in triglycerides, a decrease in high-density lipoprotein cholesterol, an increase in blood pressure and an increase in fasting venous plasma glucose or previously diagnosed type 2 diabetes mellitus are attributed.

It is generally accepted that metabolic syndrome (MS) is a cluster of factors associated with an increased risk of developing atherosclerotic lesions of the cardiovascular system and diabetes mellitus. Numerous studies have shown that mortality from cardiovascular disease and the risk of their occurrence are higher in individuals with metabolic syndrome. World Health Organization (WHO) experts have described MS as a “pandemic of the 21st century.” Leading Russian and foreign scientists involved in the problem of MS consider it as a pre-stage of atherosclerosis and type 2 diabetes, as well as a condition in which the risk of cardiovascular complications increases. (Russian recommendations // Cardiovascular therapy and prevention, 2007, 6 (6): Appendix 2: 2-25; Grudy SM et al. Circulation, 2005, 1 12: 2735-2752; Mancia G. et al Hypertension 2007; 49: 40-47).

A number of methods and means have been proposed for the treatment of metabolic syndrome (patent RU N ° 213190, 19997; patent RU N ° 2143902, 1998; patent RU N ° 2363460, 2009; patent RU M> 2391 121, 2009; JP patent> 156320 A and others.).

However, despite the progressive direction in the treatment of atherosclerosis with the use of drugs that block the metabolic syndrome, impressive success has not been achieved. The problem remains relevant and intractable, since the disadvantages of the proposed approaches are the need for high patient adherence to such treatment methods. In addition, drugs recommended both for the treatment of atherosclerosis and the concomitant metabolic syndrome are offered taking into account the effects on individual components and links of the pathological chain. believing that they will be able to incorporate compensatory changes.

A study of the etiology, pathogenesis, and risk factors for the development of atherosclerosis at a modern level allows us to conclude that it is necessary to use many multidirectional drugs at the same time to stop this pathology, which is problematic, since with the increase in the number of drugs used funds necessary for the effective treatment of the disease and its complications, the number of side, adverse effects is increasing.

Another possible way to achieve this goal is to use drugs with a multifunctional effect. Compounds of natural origin (plant compositions, fungi, microorganisms) can meet this criterion.

Close to the present invention, the spectrum of multifunctional biological effects are statins - HMG-CoA reductase inhibitors, the positive effects of which are determined not only by a direct effect on cholesterol synthesis, but also by its pleiotropic properties such as improved endothelial function, anti-inflammatory effect, and decreased LDL oxidation processes, suppression of thrombosis, antiproliferative and other actions.

One of the known drugs from the group of natural statins is lovastatin isolated from microorganisms belonging to the genus Aspergillus (US patent No. 4231938, 1980). However, when using statins that have the necessary anti-atherogenic multifunctionality and are capable of correcting the metabolic syndrome, numerous toxic effects are noted, which causes contraindications to the appointment, and also require a long period of use for several years to achieve a therapeutic effect, which, moreover, is not sufficient.

So, against the background of the widespread use of statins, according to the American Heart Association, published in 2004, 38.5% of all deaths in 2001 in the USA were cardiovascular diseases [Ballantyne SM. - Am. Heart J., 2004, 148: 3-8].

Among the natural compounds that meet the criterion for the presence of multifunctionality of action, a special group are terpenoids (isoprenoids). For example, among terpenes and their derivatives, the number of which currently totals more than 23,000 (isolated and chemically described), a wide variety of types of biological effects are recorded b

However, an analysis of their preparation and use shows that isolated individual substances with high, but no longer multifunctional biological activity or extracts containing these substances in small or trace amounts are studied, which does not allow to achieve the desired effectiveness. Their study is mainly aimed at determining the effect on individual factors of the development of pathological processes (WO 03/007901). So, the use of extracts containing terpenes, such plants as dandelion, St. John's wort, hawthorn, valerian, mountain ash, sea buckthorn, motherwort and others, shows the presence of a mild anti-atherosclerotic effect, so they are most often prescribed in combination with other medicines [Mashkovsky M. D. Medicines, - M .: New Wave, 2005-1200 s; patents: RU N ° 2148399, RU Ko 2170099, RU> 2394587, US K "7517542).

More effective is the use of certain classes of terpenes - diterpenes (patent US N ° 7517542, 2009). A composition based on diterpenoids from Torreya nucifera showed multifunctionality, including antioxidant effects, lowering blood pressure and regulating cholesterol levels. However, a number of concomitant manifestations of destructive processes or metabolic syndrome were not corrected.

The closest, in the spectrum of multifunctional biological action, is the terpenoid pharmaceutical substance Abisil.

Previously, the pharmaceutical substance Abisil was used in the form of a dosage form (20% solution in oil) for local and external use, for which antimicrobial, anti-inflammatory, analgesic, wound healing and immunomodulating activities were identified and described [Patent RU 54945, 02.27.1996; Patent RU K ° 2198653, 02.20.2003; Patent RU JYQ2338547, 20.1 1.2008; The register of medicines of the Russian Federation, 2008, T.1: 14, 85], as well as antitumor and angiogenesis-inhibiting effects [PCT / RU 2008/000147. Published WO 2009/1 13902, September 17, 2009]. The ability of the pharmaceutical substance Abisil to activate the synthesis of humoral mediators of nerve endings in the cerebral cortex of experimental animals was also revealed. The resulting endogenous neurotransmitter pharmaceutical composition showed high antitumor activity (patent RU Na 2244928, 2005). The study the general toxic effect of the pharmaceutical substance Abisil and drugs based on it showed the safety of their use with different methods of administration.

However, despite the obvious prospect of studying complexes of various terpenoids, with a view to their possible use for the prevention and / or treatment of atherosclerosis with simultaneous correction of the metabolic syndrome, such compositions have not been identified.

Analogues of the well-known pharmaceutical substance Abisil in terms of composition and spectrum of multidirectional pharmacological action have also not been identified.

The present invention seeks to overcome the disadvantages of the state of the art.

SUMMARY OF THE INVENTION

The first aspect of the present invention provides a terpenoid agent for the prevention and / or treatment of atherosclerosis and correction of metabolic syndrome, containing from 10 to 20 mass. %, preferably 15 wt. % by weight of the pharmaceutical substance Abisil and a pharmaceutically acceptable vegetable oil.

According to the invention, vegetable oil includes sunflower oil, olive oil or soybean oil.

According to the invention, the terpenoid agent is formulated for oral administration, preferably in the form of a solution or gelatin capsules.

The proposed terpenoidal agent has an anti-atherosclerotic effect, characterized by a decrease in blood total lipids, total cholesterol, triglycerides and low-density lipoprotein cholesterol (LDL), stimulation of cholesterol and bile acid excretion with bile, a decrease in the degree of damage to aortic intima, and prevention of aortic atherosclerosis and myocardial infarction as well as corrective action for metabolic syndrome, characterized by normalization of blood pressure, triglycerides and glucose in the blood.

A second aspect of the present invention provides the use of the pharmaceutical substance Abisil for the prevention and / or treatment of atherosclerosis and the correction of metabolic syndrome.

According to the invention, the pharmaceutical substance Abisil is used in the form of from 10 to 20 mass. % solution, preferably 15 wt. % solution in pharmaceutically acceptable vegetable oil.

According to the invention, the proposed solution is administered orally, preferably in the form of gelatin capsules.

The proposed solution has an anti-atherosclerotic effect, characterized by a decrease in blood total lipids, total cholesterol, triglycerides and cholesterol low density lipoproteins (LDL), stimulating the excretion of cholesterol and bile acids with bile, reducing the degree of damage to aortic intima, preventing the development of aortic atherosclerosis and the occurrence of myocardial infarction as well as the corrective effect in metabolic syndrome, characterized by the normalization of blood pressure, level three litseridov and blood glucose.

A third aspect of the present invention provides for the use of

Abisil pharmaceutical substance for the manufacture of a terpenoid agent for the prevention and / or treatment of atherosclerosis and the correction of metabolic syndrome.

According to the invention, the terpenoid agent contains from 10 to 20 mass. % by weight of the agent, preferably 15 wt. % by weight of the pharmaceutical substance Abisil and a pharmaceutically acceptable vegetable oil.

According to the invention, vegetable oil includes sunflower oil, olive oil or soybean oil. According to the invention, the terpenoid agent is formulated for oral administration, preferably in the form of a solution or gelatin capsules.

The proposed terpenoidal agent has an anti-atherosclerotic effect, characterized by a decrease in blood total lipids, total cholesterol, triglycerides and low-density lipoprotein cholesterol (LDL), stimulation of cholesterol and bile acid excretion with bile, a decrease in the degree of damage to aortic intima, and prevention of aortic atherosclerosis and myocardial infarction , as well as the corrective effect in metabolic syndrome, characterized by normalization of blood pressure tions, triglycerides and blood glucose levels.

Disclosure of invention

The aim of the present invention is to obtain a medicinal product that has an effective effect in the prevention and / or treatment of atherosclerosis against the background of destructive processes in the vascular wall, as well as in the development of metabolic syndrome.

The goal is achieved by means of a terpenoid based on the pharmaceutical substance Abisil, containing from 10 to 20 mass. % of the pharmaceutical substance Abisil by weight of the drug in a pharmaceutically acceptable vegetable oil.

The authors of the present invention, as previously mentioned, was obtained pharmaceutical substance Abisil, for which a wide range of pharmacologically significant activities were identified.

Surprisingly, when studying the pharmacological effects of the pharmaceutical substance Abisil in laboratory animals in different concentrations, it was found that when administered orally, its 15% oil solution has a pronounced decrease in blood total lipids, total cholesterol and glucose, i.e. exhibits pharmacologically significant actions in the treatment of atherosclerosis and type 2 diabetes. This served as an incentive for further study of the pharmaceutical substance Abisil, on the basis of which a new terpenoid drug (15% oil solution in capsules, for oral administration) was developed and first proposed, with a new spectrum of pharmacological action and new possibilities for stopping pathological processes, in particular, atherosclerosis and metabolic syndrome. The use of the proposed terpenoid drug is of great clinical importance, since with appropriate therapy it is possible to eliminate or reduce the main manifestations of these pathologies, especially since the metabolic syndrome predisposes to the development of atherosclerosis, arterial hypertension and type 2 diabetes mellitus - the main causes of morbidity and mortality in the world .

The claimed pharmaceutical substance Abisil and a terpenoid preparation made on its basis for oral use (15% oil solution in capsules) differs from the above funds in that it includes natural fir terpenoids of the genus Fir (Abies) taken in a certain mass ratio, which allows to reduce the amount of pharmaceutical Abisil substances for oral use without reducing its therapeutic efficacy, by dissolving this substance in oil (sunflower, olive or soybean) to 15% concentration and and making capsules.

The composition of the claimed terpenoid agent (15% oil solution in capsules, for oral use) includes the starting components of the pharmaceutical substance Abisil in a mass fraction of%, namely: monoterpenoids (35 - 38%); sesquitepenoids (3 - 6%); neutral diterpenoids (1 1 - 15%); diterpenic acids (23 - 28%); triterpene acids (8 - 16%); unsaturated and saturated fatty acids (0, 1 - 0.3%); phenolic compounds (0, 1 - 0.2%) with a content of boronyl acetate of at least 10.0% of the total composition of terpenes dissolved in sunflower oil.

According to the invention, the pharmaceutical substance Abisil may be contained in an amount of from 10 to 20 masses. %, most preferably 15 wt. % by weight of the product.

The proposed amount of the pharmaceutical substance Abisil had specific activity and did not exert a general toxic effect on the body. animals. Even a single intragastric administration of the substance Abisil, at a dose of 10,000 mg / kg (100 times the therapeutic dose) to white mice weighing 20-22 grams, did not cause significant functional and structural changes in the organs and tissues of animals.

According to the invention, a pharmaceutically acceptable vegetable oil can be used as a pharmaceutically acceptable carrier, for example

sunflower oil, olive oil or soybean oil.

Vegetable oil is also used as a solvent for the preparation of the proposed terpenoid agent of the desired concentration, because The pharmaceutical substance Abisil, represented by fir terpenoids of the genus Abies, is by nature a lipophilic formation and is therefore preferably soluble in vegetable fatty oils.

The choice of vegetable oils as a solvent contributes to better permeability of the final product through the lipid layer of cell membranes. It has also been proven that alcohol is not an ideal solvent for capsule extract.

The amount of the pharmaceutical substance Abisil obtained in example 1, which allows to achieve the maximum therapeutic effect in the treatment and / or prevention of atherosclerosis with the correction of the concomitant metabolic syndrome, as part of the terpenoid agent used, should correspond to 15% of its concentration in a solution of sunflower (olive or soybean) oil.

According to the invention, the terpenoid agent can be formulated for oral administration, preferably in the form of a solution or gelatin capsules.

In a preferred embodiment of the invention, based on data on the general toxic effect of the pharmaceutical substance Abisil, as well as on data revealed in modeling atherosclerosis and metabolic syndrome, the claimed terpenoid agent is preferably used orally in a gelatin capsule or in the form of a 15% solution in oil, at a dose of 4-40 mg / kg body weight.

The proposed terpenoid agent, as first shown, has an antiatherosclerotic effect, characterized by a decrease in blood levels lipids, total cholesterol, low density lipoproteins and triglycerides, stimulating the secretion of bile cholesterol and bile acids, reducing the degree of damage to the aortic intima, preventing the development of aortic atherocalcinosis and the occurrence of myocardial infarction.

The preferred embodiment of the invention is the use of a terpenoid agent in the prevention and treatment of atherosclerosis, when administered orally at a dose of 10 ^ 0 mg / kg, since the most pronounced antiatherosclerotic effect was observed in rabbits atherosclerosis model at this dose.

The proposed terpenoid agent has a corrective effect in the metabolic syndrome, characterized by the normalization of blood pressure, the level of triglycerides and glucose in the blood.

A preferred embodiment of the invention is the use of a terpenoid agent for the correction of arterial hypertension, lipid and carbohydrate metabolism disorders when administered orally at a dose of 4-12 mg / kg, detected by modeling the metabolic syndrome in spontaneously hypertensive SHR rats.

All sources and publications listed in the description are an integral part of it, as if all their contents were included in the description.

The following examples are preferred, intended only to confirm the feasibility of the invention and should not be the basis for limiting the scope of claims of the Applicant. A person skilled in the art will easily find the possibilities of other embodiments of the invention, of course, subject to the claims of the Applicant, as reflected in the claims below.

Information confirming the possibility of carrying out the invention

The invention is illustrated in detail below with reference to specific examples representing its most preferred embodiments. Example 1. The preparation and composition of the terpenoid pharmaceutical substance Abisil

The preparation and composition of the terpenoid pharmaceutical substance Abisil is described in detail in patent RU No. 205494 and in patent RU Na2338547.

The initial raw material for the manufacture of the proposed pharmaceutical substance Abisil is the contents of capsules (nodules) located between the bark and bast of various representatives of the genus Fir (Abies), which is obtained by direct puncture and collection in light-proof bottles. The capsule extract obtained in this way is a thick liquid from yellow to milky white with a specific odor and contains the entire complex of natural, naturally balanced terpenoids. Moreover, this natural terpenoid composition should contain a provoked (in terms of yield and synthesis) composition for bioprotection against stress factors and be enriched with monoterpene compounds. Stress factors can be environmental (temperature fluctuations, damage by microorganisms, smoke, etc.) or specially provoked (short-term tapping of trunks).

The capsule extract thus obtained was filtered at room temperature. His physico-chemical analysis showed:

P ₀ ²⁰ 1, 51 12; d „ ²⁰ 1, 0328 g / cm ³ ,

IKS: 3400, 2900, 1740, 1690-1700, 1450, 1390, 1270, 1250, 1040, 900 words 800 words cm ^"1. PMR: 0.82; 1, 67; 4.71; 5.77 ppm (C ₆ D ₆ ). Elemental analysis data:

C 77.82; H 10, 19; S no; N no.

To obtain monoterpene compounds, a purified capsule extract is taken, placed in a flask in a water bath with an attached Klevenger apparatus, a finger cooler, and a graduated separation pipette with a drain by crane. Stripping is continued for 12 hours. The volatile fraction collected in this way is an essential oil consisting of monoterpenic compounds: tricyclene, limonene, α-pinene, santene, camphene, etc.

Physical constants of monoterpenoid fraction:

1. P _o ²⁰ = 1.4753

2. d „20 = 0.884

3. LD = -30.2 °

The terpenoid pharmaceutical substance Abisil is obtained with

mixing monoterpene compounds with capsule extract, Moreover

mixing is carried out at a temperature of 18-20 ° C and 195 ml of capsule extract are taken per 1.0 ml of monoterpene compounds.

Thus obtained pharmaceutical substance Abisil,

used in the future to obtain an oral terpenoid agent (15% oil solution in capsules), capable of exerting antiatherosclerotic therapeutic and / or prophylactic effects with simultaneous relief of the concomitant metabolic syndrome, it is characterized by the fact that it contains capsule terpenes of plants of the Pinaceae family, provoked by the release and synthesis, fir (Abies), enriched with identical monoterpenoids and includes the following components, wt.%:

- monoterpenoids (35 - 38%);

- sesquitepenoids (3 - 6%);

- neutral diterpenoids (1 1 - 15%);

- diterpenic acids (23 - 28%);

- triterpene acids (8 - 16);

- unsaturated and saturated fatty acids (0.1 - 0.3%);

- phenolic compounds (0.1 - 0.2%). Example 2. Obtaining a terpenoid agent (15% oil solution in capsules) for oral administration

To obtain a terpenoid agent according to the invention, the pharmaceutical substance Abisil obtained according to example 1 was diluted with various

solvents to a concentration of 5-40 mass. % by weight of the product. As

solvents used pharmaceutically acceptable vegetable oils, for example sunflower oil, olive oil or soybean oil, and various alcohols.

Mixing the substance with solvents is carried out at room temperature to avoid evaporation of volatile fractions and inactivation of the target product.

It turned out that alcohol is not an ideal solvent for capsule extract

In the future, this composition can be used as a solution or in the form of capsules for oral administration.

The obtained terpenoid agent is characterized by the following physical constants: density 0.85-2.50; refractive index 1, 50-1, 52; saponification number 100-130; acid number 70-90; an ether number of 10-60 and a monoterpenoid-bornyl acetate content of at least 10.0% of the total number of terpenoids.

Thus, a new terpenoid preparation based on the pharmaceutical substance Abisil was proposed, which is an oral dosage form - 15% oil solution in capsules, with a new spectrum of pharmacological action and new possibilities for stopping pathological processes, in particular, with the development of atherosclerosis and metabolic syndrome.

Example H. The study of the antiatherosclerotic effect of a terpenoid agent The study of the anti-atherosclerotic effect of the claimed terpenoid agent (15% oil solution, for oral use) was carried out in accordance with the “Methodological guidelines for the study of the lipid-lowering and anti-atherosclerotic effects of pharmacological substances” using generally accepted methods for determining changes in lipid and carbohydrate metabolism [Experimental (preclinical) manual the study of new pharmacological substances. M .: 2005: 452-461; Medical laboratory technology. St. Petersburg: Intermedica, 2002, 600 s].

To study the anti-atherosclerotic effect of a terpenoid agent (15% solution in oil), an atherosclerosis model was used, which was reproduced in rabbits by daily intragastric administration of cholesterol (manufacturer XIAMEN, China) at a dose of 0.3 g / kg. After 1 month from the start of cholesterol administration, vitamin D ₂ (ergocalciferol, manufactured by Nycomed, MerckKGaG, Germany) was additionally added to animal food to enhance aortic lipidosis over a period of 30 days. In addition, in order to enhance atherosclerotic changes and reduce the time of induction of atherosclerosis, 30 days after the start of the experiment, adrenaline was administered to rabbits twice every 15 days (producer of FSUE Moscow Endocrine Plant, Russia) at a dose of 0.04 mg / kg .

The inventive terpenoid agent (15% oil solution, for oral use) was used in doses of 10 mg / kg, 20 mg / kg and 40 mg / kg based on the active substances of the substance of this drug. As a comparison drug used the well-known lipid-lowering agent used in the treatment of atherosclerosis - lovastatin at a dose of 0.7 mg / kg. The animals were administered a terpenoid agent and lovastatin intragastrically through an atraumatic probe, once a day, on an empty stomach, for 60 days. Intact and control (model of atherosclerosis without treatment) animals were given as a placebo sunflower oil GOST 1 129-73 in a volume of 1 ml.

During the experiment, blood was taken from the marginal vein of the ear for biochemical studies. In order to assess the lipid-lowering effect of the studied drugs, the following lipid metabolism indicators were determined: total lipids (OL), phospholipids (FL), total cholesterol (OXC), triglycerides (TG), α-cholesterol - cholesterol of high density lipoproteins (HDL) and calculated indicators - cholesterol of low density lipoproteins (LDL), as well as lecithin-cholesterol index (LHI). Blood biochemical parameters were determined on an A-25 biochemical analyzer using reagents from BioSystems, Parma, Olveks, SPINREACT in serum without traces of hemolysis. The procedure was carried out on an empty stomach, after 12 hours of fasting, in the morning at 9.00 hours. These indicators were evaluated five times during the experiment, at each blood sampling (on 0, 15, 30, 45, and 60 days).

The study of the qualitative and quantitative composition of bile was performed by thin layer chromatography (TLC). Solutions of cholic (> 99%, Fluka, Italy; HC, 0.5 mg / ml), deoxycholic (> 99%, Fluka, Italy; DCA, 0.5 mg / ml), chenodeoxycholic (> 98%) were used as standards. , Fluka, Italy; HDCA, 0.5 mg / ml), lithocholic (> 99%, Fluka, Italy; LH, 0.5 mg / ml) acids and cholesterol (> 98%, Sigma, Germany; X, 0, 2 mg / ml) in methanol.

Additionally, the cholate-cholesterol index (CXI) was calculated according to the formula: CXI = EFA / OXC, where: FA - total bile acids, mg%;

OXS - total cholesterol mg /%.

On the 60th day of the experiment, after collection under bile anesthesia, euthanasia was carried out by bleeding animals from the cavities of the heart, for subsequent macroscopic and microscopic examination of organs.

At autopsy, the aorta, its thoracic and abdominal sections were completely isolated. A longitudinal section of the aorta was opened, washed physical. solution, analyzed the area of damage to the aorta. Each aorta was photographed and planimetric atheromatous changes. Expected% of the lesion area. Measurements were performed using VideoTest software - Size 5.0. In addition, a macroscopic examination of the heart, coronary, carotid and pulmonary arteries, small arteries of the liver and spleen was performed.

Fragments of the aorta and liver were fixed in a 10% solution of neutral formalin for 24 hours, then the material underwent standard processing in alcohols increasing concentration (70-95%), xylene and paraffin for the manufacture of histological and immunohistochemical preparations with a thickness of serial paraffin sections 5-7 microns.

For microscopic examination, sections were stained with hematoxylin and eosin. For histochemical studies and in order to detect lipids from fixed aortic biopsies, sections with a thickness of 7-10 μm were cut on a freezing microtome and stained with Sudan IV.

A morphological study of histological preparations and preparations stained by the immunohistochemical method was carried out using a Leica DM LS light-optical microscope with a magnification of 200 and 400.

Statistical analysis was performed using the Statistics 6.0 software. Differences were identified at 0.05 significance level.

The results of the study of the anti-atherosclerotic effect of the claimed terpenoid agent (15% oil solution) are shown in tables 1-3.

Oral administration of a terpenoid agent in the doses used in modeling atherosclerosis in rabbits had a positive effect on the level of total lipids, total cholesterol, triglycerides and low density lipoproteins (Table 1).

Table 1. Indicators of the level of total lipids, total cholesterol, triglycerides and low density lipoproteins in the peripheral blood of experimental groups of animals, after 60 days, M ± t

Note: hereinafter in the tables:

- differences are statistically significant compared with intact animals according to student criterion, at p <0.05;

** - differences are statistically significant compared with control animals according to student criterion, at p <0.05. Throughout the entire period of experimental studies, the level of total lipids in the group of intact animals remained within the physiological norm .. Against the background of exposure to atherogenic factors (control group), a sharp increase in the concentration of total lipids in the blood serum of experimental animals was observed: after 30 days, 2 times, after 45 days 3 times, by the end of the study, the level of total lipids was 3.2 times higher in comparison with the initial data.

Compared to animals that did not receive treatment (control group), the studied terpenoid agent restrained the growth of total lipids and by the 60th day their level was on average 2 times lower than the control, in the minimum dosage the difference was 2.2, in the maximum - 1, 7 times, which may indicate a tendency to inverse dose dependence.

The comparison drug lovastatin kept the level of total lipids within the physiological norm throughout the study and from day 45 had a statistically significant difference compared with the control group.

In this experiment, in all groups of animals, except for the control, the level of triglycerides was kept within the physiological norm (for rabbits, the normal value was up to 1.9 ± 0.61 mmol / L). At the same time, its level in animals using both levostatin and a terpenoid agent in all doses used, by the 60th day of the study, had statistically significant differences from the control group.

The level of total cholesterol in intact animals throughout the study did not change and fit into the range of physiological norms. In control animals and animals of the experimental groups treated with the atherogenic complex, starting from the 15th day of the study, a marked increase in the level of total cholesterol was observed. It was maximally expressed in animals that did not receive treatment by the 45th day of the study, having a value 1 1 times higher than this indicator in the intact group in the same time period. By the 60th day of the study, there was a slight decrease in this indicator, and compared with intact animals in the control group, the level of total cholesterol was 7 times higher. Against the background of the use of the comparison drug lovastatin, the cholesterol level had a maximum increase by the 45th day, with a slight tendency to decrease by the 60th study day. The studied terpenoid agent showed an inverse dose dependence with respect to the level of OXC: it turned out to be ineffective in its maximum dosage, and the minimum studied dose showed a more pronounced decrease in OXC by the final day than when using the drug comparing lovastatin.

The LDL index was calculated by the formula: LDL = (HDL-C) - (TG / 2.2). Normally, this indicator should not increase more than 3.87 mmol / l - the level of "risk". In general, the dynamics of LDL is consistent with the dynamics of total cholesterol. The level of LDL in intact animals throughout the study did not change and fit into the range of physiological norms. In control animals and animals of experimental groups receiving an atherogenic complex, starting from the 15th day of the study, a marked increase in the level of LDL was observed. The most pronounced increase in this indicator was observed in animals that did not receive treatment, and by the 60th day the excess compared to intact animals was 15.5 times. The studied terpenoid agent inhibited the growth of the studied parameter, which was more pronounced in the minimum dosage and by the 60th day was a decrease in LDL by 2.8 times compared with the control group of animals and the drug compared to lovastatin was superior in effect.

A tendency towards an increase in the level of phospholipids was observed in all groups of the studied terpenoid drug, but in comparison with the control group, the minimum dose of the drug restrained the growth of this indicator by 1.5 times by the 60th day of the study. The use of the hypolipidemic drug lovastatin did not lead to a decrease in the level of phospholipids and was 1.7 times higher at the end of the study compared to the starting point in the same group.

In response to the cholesterol load, there was a general tendency to a slight decrease in HDL, which fell within the normal range, both in animals without treatment and in experimental groups of animals treated with drugs.

The lecithin-cholesterol index is calculated as: phospholipids / OXS. Normally, this index should not fall below 0.9-1, 2. In all studied groups, the lecithin-cholesterol index, starting from day 0 of the study, had threshold values and decreased during the experiment. Neither the reference drug nor the tested terpenoid agent had a statistically significant therapeutic effect on this indicator. However, a terpenoid drug in the minimum dosage tended to normalize LHI, the index of which was 60.03 days 1, 03 ± 0.35.

The tested terpenoid drug, in all the studied doses, was effective against dyslipidemia, it positively influenced the level of total lipids, total cholesterol, LDL and triglycerides, with respect to other indicators of lipid metabolism, the positive effect was expressed only as a tendency.

The best therapeutic effect was recorded using the minimum and average doses of the test drug. A dose of 10 mg / kg terpenoid agent had a positive effect on the level of total lipids, LDL, total cholesterol, triglycerides, phospholipids, and in terms of total cholesterol, LDL, phospholipids and lecithin-cholesterol index, the effect exceeded the effect of the comparison drug lovastatin.

Under the influence of a terpenoid agent at a dose of 10 mg / kg, the amount of excreted bile increased 2.2 times, at a dose of 20 mg / kg - 2.8 times and at a dose of 40 mg / kg - 10.8 times compared with intact animals , which can characterize the presence of a choleretic effect.

During the chromatographic study of hydrolyzed samples, it was found that the qualitative set of basic bile acids (FA) in all samples is almost the same. Detected and identified by the coincidence of the values of Rf and the color of the spots after derivatization in comparison with standard samples, cholesterol (X), cholic acid (CK), deoxycholic (DCA) and lithocholic (LCC). The results of the study of the composition of bile are presented in table 2 Table 2. The content of the amount and individual FA, cholesterol and HCI, mg%

In this experiment, when cholesterol was ingested with food, there was an increase in excretion with bile of cholesterol by 45%, while the cholesterol-cholesterol index (XXI) decreased by 30%.

The use of lovastatin comparison drug against the background of pathology led to increased excretion of cholic acid, deoxycholic, while the excretion of cholesterol and bile acids increased by 22% and 43%, respectively, compared with the control group, and the cholesterol-cholesterol index increased by 17%. The terpenoid agent significantly increased the excretion of bile, as well as bile acids and cholesterol, both in comparison with intact animals and with control animals. A dose of 20 mg / kg of terpenoid agent increased the excretion of cholesterol by 1 1% and bile acids by 14% compared with the control group.

For individual bile acids, all dosages of a terpenoid agent were nearly equally effective. At the minimum and maximum dosages, an increase in the cholesterol-cholesterol index by 20% was observed in comparison with the untreated group. At a dose of 20 mg / kg, the excretion of cholesterol prevailed over the excretion of bile acids, and the cholesterol-cholesterol index changed, which increased by only 3% compared with the control group.

Morphological studies of experimental animals in modeling atherosclerosis showed that atheromatous changes were observed mainly in the abdominal aorta in the form of whitish-yellowish spots and stripes. The results of calculating the area of lesion of aortic intima in experimental groups of animals are presented in table 3.

Table 3. The area of damage to the aortic intima in the experimental groups of animals, M ± t

As can be seen from the data presented, in the control group of animals under the influence of atherogenic factors, atherosclerotic lesions of aortic intima developed (up to 51.4%). Under the influence of the studied drugs, the area of aortic lesion was reduced to 20-26%. However, it should be noted that five animals showed more pronounced aortic lesions, which can be characterized as aortic atherocalcinosis. These changes were observed in two animals of the control group (atherocalcinosis area 45-90%), one of the terpenoid group at a dose of 20 mg / kg and one of the terpenoid group at a dose of 40 mg / kg (atherocalcinosis area 12-13%) and one animal group lovastatin (area of atherocalcinosis 10-1 1%). Macroscopically, these changes were characterized as bag-shaped protrusions of the wall into the lumen of the vessel, irregular in shape, sizes from 0.2x0.3 cm to 0.3x0.5 cm. The wall in the protrusion region was thinned, whitish. Intima in the area of protrusions, as in the rest of the aorta, is smooth, shiny. Parietal blood clots were not found. Similar changes can be diagnosed as aortic microaneurysms as a result of atherocalcinosis. In the control group, these changes were observed over a large length of the aorta to 90%, while in the group of terpenoid drugs at a dose of 20 mg / kg and terpenoid drugs at a dose of 40 mg / kg, these changes were local in nature and their area did not exceed 13%.

A study of the heart and its valves showed that, against the background of atherocalcinosis of the aorta, in all cases, after a double intravenous administration of adrenaline at a dose of 0.04 mg / kg, myocardial infarction developed.

In the group of animals that received the minimum dose of a terpenoid agent, there was no atherocalcinosis of the aorta and cases of myocardial infarction. Thus, a terpenoid agent at a dose of 10 mg / kg was found to be the most effective, which prevented the development of atherocalcinosis of the aorta and myocardial infarction.

Macroscopic studies of coronary, carotid and pulmonary arteries, as well as small arteries of the liver and spleen, did not reveal pathological changes.

A histological examination revealed that when modeling atherosclerosis, changes in the aorta touched the inner and middle shell of its wall. Lipid deposition, a significant increase in the thickness of intima, and necrosis with media calcification in the control group of animals, as well as in individual animals with atherosclerosis treated with lovastatin and a terpenoid agent at doses of 20 mg / kg and 40 mg / kg, were observed. In groups of animals that were exposed to atherogenic factors, during histological examination of liver tissue, the usual arrangement of hepatocyte beams with uniformly stained cytoplasm was recorded in the hepatic lobules. Necrosis and signs of metabolic damage were absent. Slight lymphoid infiltration of the portal tracts was observed. The data obtained allow us to conclude that there are no pathological effects of drugs on liver function. The detected changes in the liver tissue are associated with the functional tension of the organ in response to an increase in the concentration of lipids in the blood.

Thus, in a model of atherosclerosis reproduced in rabbits, it was found that a terpenoid agent had a positive effect on inhibiting the growth of atherogenic lipoproteins - it prevented an increase in total lipids, total cholesterol, LDL and triglycerides. The test agent prevented the development of atherocalcinosis, reduced the area of damage to the aorta and damage to the aortic valve, thereby reducing the severity of atherosclerotic changes and the likelihood of myocardial infarction. It was revealed that the terpenoid agent has a choleretic effect, as well as the ability to stimulate the release of cholesterol and bile acids with bile, which is most likely one of its mechanisms of action, which is reflected in its effect on the blood lipid spectrum.

Example 4. Corrective action of a terpenoid agent in the development of metabolic syndrome

Modeling of metabolic syndrome (MS) was carried out using the diet "cafeteria diet" on rats - males of the breed SHR (spontaneously - hypertensive). The diet used is most recognized for the induction of a metabolic syndrome in animals close to human [Sampey B. P. et al. Obesity, 201 1, 19 (6), 1 109-1 1 17]. The products included in the nutritional composition (white bread, chocolate, potato chips, chocolate chip cookies, cheese crackers) were mixed with standard food and fed to the cage of experimental animals. The diet contained 25-35% fat and 25-30% easily digestible carbohydrates. Induction of the metabolic syndrome using the “cafeteria diet” continued for 8 weeks. The developed pathology was accompanied by arterial hypertension, glucose tolerance, impaired lipid metabolism with an increase in triglycerides and was recorded from 5 weeks. The intragastric administration of the studied terpenoid agent (15% oil solution, for oral administration) was carried out at a dose of 4 mg / kg, 8 mg / kg, 12 mg / kg and 24 mg / kg based on the active substances of the drug substance. Metformin [US Pat. No. 3,174,901], recommended by the WHO in the treatment of MS, at a dose of 200 mg / kg in a 1% solution of starch suspension was used as a comparison drug. The administration of the studied drugs was carried out according to the treatment regimen, once a day, during the last 3 weeks in a volume of 1 ml. As a placebo, GOST-1 129-73 sunflower oil was used.

To determine the lipid profile of the blood (CHS, TG, HDL) and the composition of bile in animals with metabolic syndrome, the above methods were used to model atherosclerosis.

Glucose was measured using a OneTouch Horizjn glucometer from Lifescan USA. To do this, a puncture was made along the tail vein of the animal, the device was brought up with an inserted test strip, the device automatically took 1, 5 μl of blood. The method for determining glucose is electrochemical, based on the biosensor glucose - oxidase principle. Linear measuring range 1, 1 - 33.3 mmol / L.

A glucose tolerance test (GTT) was determined - a laboratory research method used in endocrinology to diagnose impaired glucose tolerance. GTT was performed on an empty stomach (after 12 hours of fasting), the initial glucose level was estimated. Then the animals were intragastrically injected with a glucose solution prepared in such a way that 0.56 g per 100 g of animal was dissolved in 1 ml. Next, the glucose concentration was determined after 15, 30, 60 and 90 minutes using a OneTouch Horizjn glucometer (Lifescan, USA).

Blood pressure was measured at 6, 7 and 8 weeks of the experiment. To do this, an injection cuff with an integrated sensor was applied to the proximal section of the tail, and an ultrasonic pressure measurement sensor was slightly lower. The time of the most calm state of the animal, when it was in fixing house without movement. After fixing the cuff and sensors in a personal computer (PC), the LabChart 7 program window opened. Air was injected into the cuff after pressing the “Start / stop” button in the rat blood pressure measurement system (AD Instruments, Australia). The device is certified according to the rules of Good Laboratory Practice (GLP). Graphs for measuring blood pressure were displayed on a PC screen. Each animal was measured for 5-7 minutes. During this time, the device recorded blood pressure 5-7 times.

Statistical analysis was performed using the Statistics 6.0 software. Differences were identified at a 0.05 significance level.

When modeling the experimental metabolic syndrome using the ка cafeteria diet ’diet on spontaneously hypertensive rats - males of the SHR breed, we were able to induce pathology in clinical indicators similar to classical MS in humans. The pathology was accompanied by arterial hypertension, glucose tolerance, impaired lipid metabolism with an increase in triglycerides.

The baseline for assessing blood pressure was values obtained from animals kept on a standard diet of vivarium at 0 week. Changes in blood pressure were performed at 6 and 8 weeks of the experiment. The data are shown in table 4.

Table 4. The value of systolic (SBP) and diastolic (DBP)

blood pressure in experimental animals, mmHg, M ± t

Group Experiment Week

animals 0 (0) 6 (1) 8 (3)

GARDEN DAD GARDEN DAD GARDEN DAD

Intact 182 ± 5 129 ± 6 192 ± 18 143 ± 16 183 ± 6 125 ± 1 1

Control 189 ± 6 132 ± 7 174 ± 5 И0 ± 2 * 204 ± 12 133 ± 10

Metformin

187 ± 9 142 ± 10 150 ± 14 107 ± 9 212 ± 5 * 123 ± 10 200 mg / kg

Terpenoid

means according to 188 ± 6 143 ± 7 165 ± 9 1 16 ± 9 188 ± 8 123 ± 8 of the invention 24 mg / kg

Terpenoid

tool for

188 ± 6 144 ± 6 163 ± 6 106 ± 8 181 ± 1 1 129 ± 14 of the invention

12 mg / kg

Terpenoid

tool for

170 ± 8 1 15 ± 7 181 ± 9 131 ± 1 1 165 ± 7- 1 12 ± 4 of the invention

8 mg / kg

Terpenoid

tool for

185 ± 7 130 ± 4 182 ± 7 1 18 ± 15 168 ± 7 " _* 100 ± 12 of the invention

4 mg / kg

Against the background of the use of the reference drug, metformin, a slight increase in SBP was recorded by the end of the experiment.

When using a terpenoid agent in doses of 8 mg / kg and 4 mg / kg for three weeks, a statistically significant decrease in SBP was revealed in relation to the animals of the control group. No changes in DBP were detected.

Against the background of the use of the ка cafeteria diet ’diet, there were no statistically significant changes in the concentration of glucose in the peripheral blood in all experimental groups. For a more complete understanding of the nature of changes in carbohydrate metabolism against the background of the use of the ка cafeteria diet ’diet and the effect of the studied drug, GTT was performed at weeks 5 and 8 of the experiment.

In animals of the control group 90 minutes after the carbohydrate load and in the groups of animals that received the рацион cafeteria diet ’diet before drug treatment, a statistically significant increase in glucose level was observed in relation to intact animals (over 7.8 mmol / L) after 15, 30, 60 and 90 minutes after glucose loading at a dose of 0.56 g / 100 g of animal. Based on WHO clinical recommendations, it can be concluded that pronounced glucose tolerance develops in experimental animals (after 5 weeks of a high-calorie diet). After 3 weeks of systematic use of four doses of the studied terpenoid drug and the reference drug metformin, GTT was repeated to reveal the specific activity of the drugs in conditions of impaired glucose tolerance (Table 5).

Table 5. Dynamics of glucose levels in the peripheral blood of male rats after a carbohydrate load, mmol / l, M ± t

Group Time after carbohydrate load, min

ANIMALS 0 15 30 60 90

5.4 ± 0.3 6.3 ± 0.3 6.7 ± 0.3 7.3 ± 0.3 6.9 ± 0.2

Intact

n = 8 n = 8 n = 8 n = 8 n = 8

5.3 ± 0.2 8.7 ± 0.8 * 9.8 ± 0.8 * 9.5 ± 0.6 * 9.5 ± 0.6 *

The control

n = 8 n = 8 n = 8 n = 8 n = 8

Metformin 4.7 ± 0.8 7.3 ± 0.8 8.9 ± 2.1 8.6 ± 1, 1 7.9 ± 0.7 200 mg / kg p = 5 p = 5 p = 5 p = 5 n = 5

Terpenoid

means according to 4.2 ± 0.2 6.3 ± 0.9 8.4 ± 0.8 9.3 ± 0.6 * 9.2 ± 0.6 * the invention n = 9 n = 9 n = 9 n = 9 n = 9

24 mg / kg

Terpenoid

means according to 4.9 ± 0.5 6.8 ± 0.4 8.1 ± 1.0 8.9 ± 0.5 * 8.9 ± 0.7 * the invention n = 6 p = 5 p = 5 p = 5 n = 5

12 mg / kg

Terpenoid

means according to 5.2 ± 0.7 7.4 ± 0.3 * 8.7 ± 0.5 * 9.4 ± 0.4 * 8.0 ± 0.2 _{** the} invention n = 8 n = 8 n = 8 n = 8 n = 8

8 mg / kg

Terpenoid

means according to 6.0 ± 0.5 7.2 ± 0.3 7.6 ± 0.4 _** 7.7 ± 0.3 _** 8.2 ± 0.4 * the invention n = 7 n = 7 n = 7 n = 7 n = 7

4 mg / kg From the data of table 5 it is seen that in the control group of animals, 15 minutes after the carbohydrate load, the glucose level exceeded 7.8 mmol / l (which corresponds to glucose tolerance) and was statistically significantly higher than the value of this indicator in intact animals.

In the group of animals treated with the studied terpenoid drug at a dose of 8 mg / kg, a statistically significant decrease in glucose level was observed in relation to the control group at 90 minutes of GTT by 20%. A terpenoid agent at a dose of 4 mg / kg at 30 and 60 minutes of GTT reduced glucose levels by 22% and 19%, respectively. Despite a further increase in glucose level in animals of this group (by 90 minutes of GTT), we can conclude the therapeutic effect of the studied terpenoid drug, both at this dose and at a dose of 8 mg / kg, on animals with pronounced glucose tolerance. The metformin comparison drug in this study did not significantly affect glucose levels.

To evaluate the lipid spectrum, the measurement of total cholesterol, TG, and HDL was performed at weeks 5 and 8 of the experiment. After 5 weeks of using the Cafeteria Diet diet, animals observed an increase in the level of OXC. It should be noted that in the control group, this increase was statistically significant, in the rest it was of a tendency nature, and in the group taking a terpenoid drug at a dose of 4 mg / kg, the level of OXC was normal, as evidenced by the presence of a statistically significant difference from control animals.

Changes in HDL did not have clinical significance, since in none of the experimental groups there was a sharp decrease in this indicator in relation to intact animals.

To identify the specific activity of the study drug, the lipid profile was measured at 8 weeks of the experiment (table 6). Table 6. The lipid profile in the peripheral blood of male rats, mmol / l, M ± t

From the data of table 6 it can be seen that against the background of 8 weeks of use of the “cafeteria diet” diet, animals developed hypertriglyceridemia, as evidenced by a statistically significant increase in the concentration of TG in the blood of control animals in relation to the intact group. These changes correspond to the classical picture of the development of MS.

The comparison drug metformin at a dose of 200 mg / kg did not have a positive effect on lipid metabolism. The studied terpenoid agent in doses of 12 mg / kg, 8 mg / kg and 4 mg / kg contributed to a statistically significant decrease in the concentration of TG in relation to control animals.

To determine the possible mechanism of fat accumulation (due to the proliferation of adipocytes or their increase in volume), the area of fat cells from the epididymal region was studied. It was found that no statistically significant changes in the area of adipocytes occurred in any of the experimental groups. The data obtained also correspond to the absence of changes in body weight in animals.

To assess the composition of bile, the obtained samples were subjected to chromatographic studies. The data obtained are shown in table 7.

Table 7. Composition of bile and cholesterol, g%, M ± t

Indicator

Group

Deoxy-henodeoxy-

ANIMALS Sleek Cholesterol

cholic cholic

Intact

284.9 ± 15.2 13.5 ± 5.4 13.9 ± 5.1 21.6 ± 1.8 319.7 ± 22.7 n = 7

The control

307.2 ± 15.3 10.8 ± 3.9 31, 5 ± 2.4 25.9 ± 1, 3 376.5 ± 13.8 n = 7

Metfor In

200 mg / kg 247.1 ± 15.8 58.7 ± 12.0 * _** 42.4 ± 8.3 * 24.9 ± 2.0 360.5 ± 28.2 p = 4

Terpenoid

tool for

invention 202.9 ± 10, 1 * " _* 56.5 ± 16.9 * 57.6 ± 45.6 25, W, 7 336.0 ± 46.7

24 mg / kg

n = 4

Terpenoid

means according to 243.0 ± 33.5 51.9 ± 17.5 * _** 48.8 ± 9.8 * 26.5 ± 2.6 377.2 ± 57.7 of the invention 12 mg / kg

n = 4

Terpenoid

tool for

invention 248.9 ± 22.5 51, 8 ± 13.4 * _** 36.5 ± 5.5 * 21, 3 ± 0.7 _** 355, 1 ± 30.0

8 mg / kg

n = 7

Terpenoid

tool for

304.4 ± 24.3 invention 197.6 ± 15.7 * _** 28.0 ± 5.6 * _** 29.3 ± 5.6 22.5 ± 1.5

**

4 mg / kg

n = 7

From the data of table 7 it is seen that the composition of bile acids in the experimental groups differed in the content of deoxycholic and chenodeoxycholic acids. As is known, chenodeoxycholic acid is the primary bile acid that is formed in the liver from cholesterol, and deoxycholic acid is formed from primary bile acids in the intestine under the influence of enzymes of microorganisms of the intestinal microflora. An increase in the amount of deoxycholic and chenodeoxycholic acids can reduce the ability of the liver to produce cholesterol, prevent cholesterol reabsorption, and also increase cholesterol solubility. This effect can be associated with both the direct effect of the studied terpenoid drug on liver metabolism and the effect on the binding of lipoproteins and cholesterol to receptors. It can be concluded that the studied terpenoid agent has a positive effect on the synthetic function of the liver.

The use of the drug metformin against a background of experimental metabolic syndrome did not lead to the normalization of blood pressure, lipid and carbohydrate metabolism.

At the same time, the use of a terpenoid agent with systematic use led to a statistically significant decrease in the level of SBP, normalization of lipid metabolism due to a decrease in triglycerides, and a faster utilization of exogenous glucose by cells was revealed during GTT. The examples 1-4 show that the proposed terpenoid agent, in a therapeutically effective dose, when administered orally to animals with simulated signs of atherosclerosis and metabolic syndrome, leads to a decrease in blood levels of total lipids, total cholesterol, triglycerides, low density lipoprotein cholesterol (LDL) and glucose, has a choleretic effect, stimulates the secretion of bile cholesterol and bile acids, lowers blood pressure, reduces the degree of damage to the intima of the aorta and its ateroc lecinosis, prevents the development of myocardial infarction. When comparing, during the experiments, the proposed terpenoid agent with the most well-known drugs, such as lovastatin and metformin, which are widely used in the treatment of atherosclerosis and metabolic syndrome, the terpenoid agent (15% solution in oil) revealed significant advantages, as in the specificity activity and to achieve a higher therapeutic effect.

A wide range of identified pharmacological activity of the proposed terpenoid drug makes it possible to reduce the number of administered lipid-lowering, hypoglycemic, antihypertensive, anti-inflammatory, immunomodulatory and other drugs used in the complex treatment of atherosclerosis or metabolic syndrome, which also reduces the frequency of adverse reactions.

Additional advantages for the proposed terpenoid drug are that for a pharmaceutical substance a wide range of pharmacologically significant activities was previously identified: antimicrobial, anti-inflammatory, wound healing, analgesic, immunomodulating, antitumor and angiogenesis-inhibiting.

This allows us to consider the terpenoid agent as a multifunctional, promising drug for the treatment of atherosclerosis and the correction of metabolic disorders.

The use of the proposed terpenoid agent, due to the lack of manifestations of toxicity and multifunctionality of its action, with the oral administration of a therapeutically effective amount, makes it possible to carry out the task, allowing to increase the effectiveness of prevention treatment of atherosclerosis with simultaneous correction of the metabolic syndrome, as well as to overcome the disadvantages of the prior art.

USED SOURCES

1. Arabidze G.G., Tebloev K.I. Atherosclerosis and risk factors: the clinical significance of apolipoproteins in the development of IHD is a guide for doctors. - M.: Litterra, 2008.240 p.

2. Diagnosis and treatment of metabolic syndrome // Cardiovascular therapy and prevention. - 2007. T.6, 6. - Appendix 2 - e. 2-25.

3. Mashkovsky M.D. Medicines: A Manual for Doctors, 15th ed. Rev., sp., Ext. - M.: New Wave, 2005. -1200 p.

4. Medical laboratory technology. Reference // Ed. prof. Karpishenko A.I. - St. Petersburg: Intermedika, 2002 .-- 600 p.

5. Register of medicines of the Russian Federation, 2008, T.1: 14, 85

6. Guidance on the experimental (preclinical) study of new pharmacological substances / Ed. Prof. Khabrieva R.U. - M .: Medicine, 2005, - S. 452-461.

7. Guidelines for atherosclerosis and coronary heart disease / Ed. Acad. E.I. Chazova, Corr. RAMS V.V.Kuhapchuk, prof. S.A. Boytsova - M .: Media Medica, 2007 .-- 736 p.

8. Grudy S.M. Cleeman J.I., Daniels S.R. et al. Diagnosis and Management of the Metabolic Syndrome: An American Heart Association / National Heart, Lung and Blood Institute Scientific Statement // Circulation, 2005, 1 12: 2735-2752;

9. Mancia G, Bombelli M, Corrao G, Facchetti R, et al Metabolic syndrome in the Pressioni Arteriose Monitorate E Loro Associazioni (PAMELA) study: daily life bloodpressure, cardiac damage, and prognosis. Hypertension 2007; 49: 40-47.

10. Ballantyne CM Acheving greater reductions in cardivascular risk: Lessons from statin therapy on risk measures and risk reduction // Amer. Heart J. - 2004. - Vol. 148. - P. 3-8. 1 1. Sampey B. P., Vanhoose AM, Winfield H. M, Freemerman AJ et al. Cafeteria diet is a robust model of human metabolic syndrome with liver and adipose Inflammation: Comparison to High-Fat diet Obesity 2011, 19 (6), 1 109-1 117

12. Patent RU N ° 2054945 of 02.27.1996.

13. Patent RU 2198653 dated 20.02. 2003 year

14. Patent RU W 2244928 of 01.20.2005;

15. Patent RU JV22338547 of 20.11. 2008;

16. Patent RU JN ° 2129874, from 10.05. 1999 year

17. Patent RU 2148399, 10.05. 2000 year

18. Patent RU Ns 2170099, dated 10.07. 2001 year

19. Patent RU JY ° 2394587, 17.02. 2009 year

20. Patent US JSTs 7517542, 14.04. 2009 year

21. Patent US 3174901, 23.03. 1965

22. Patent US JVs 4231938, 04.1 1.1980.

23. PCT / RU 2008/000147 (priority March 14, 2008). Published by WO 2009/1 13902 09/17/2009,

Claims

Claim

1. Terpenoid agent for the prevention and / or treatment of atherosclerosis and correction of metabolic syndrome, containing from 10 to 20 mass. % by weight of the agent, preferably 15 wt. % by weight of the pharmaceutical substance Abisil and a pharmaceutically acceptable vegetable oil.

2. The terpenoid agent according to claim 1, wherein the vegetable oil comprises sunflower oil, olive oil or soybean oil.

3. The terpenoid agent according to claim 1, which is made in the form of a dosage form for oral administration.

4. The terpenoid agent according to claim 1, which is made in the form of a solution or gelatin capsules.

5. The terpenoid agent according to any one of claims 1 to 4, which has an anti-atherosclerotic effect, characterized by a decrease in blood total lipids, total cholesterol, triglycerides and low-density lipoprotein cholesterol (LDL), stimulating the secretion of bile cholesterol and bile acids, reducing the degree of damage to the aortic intima, preventing the development of atherocalcinosis of the aorta and the occurrence of myocardial infarction.

6. The terpenoid agent according to any one of items 1 to 4, which has a corrective effect in the metabolic syndrome, characterized by normalization of blood pressure, triglycerides and glucose in the blood.

7. The use of the pharmaceutical substance Abisil for the prevention and / or treatment of atherosclerosis and the correction of metabolic syndrome.

8. The use according to claim 7, where the pharmaceutical substance Abisil is used in the form of from 10 to 20 mass. % solution, preferably 15 wt. % solution in pharmaceutically acceptable vegetable oil.

9. The use of claim 7, wherein the vegetable oil includes sunflower oil, olive oil or soybean oil.

10. The use of claim 7, wherein said solution is administered orally.

1 1. The use of claim 7, wherein said solution is administered in the form of gelatin capsules.

12. The use according to any one of paragraphs. 7-1 1, where the specified solution has an anti-atherosclerotic effect, characterized by a decrease in blood total lipids, total cholesterol, triglycerides and cholesterol low density lipoproteins (LDL), stimulating the excretion of cholesterol and bile acids with bile, reducing the degree of damage to aorta intima, preventing the development of atherocalcinosis aorta and the occurrence of myocardial infarction.

13. The use according to any one of paragraphs. 7-1 1, where the specified solution has a corrective effect in the metabolic syndrome, characterized by the normalization of blood pressure, the level of triglycerides and glucose in the blood.

14. The use of the pharmaceutical substance Abisil for the manufacture of a terpenoid agent for the prevention and / or treatment of atherosclerosis and the correction of metabolic syndrome.

15. The use of claim 14, wherein said terpenoid agent contains from 10 to 20 masses. % by weight of the agent, preferably 15 wt. % by weight of the pharmaceutical substance Abisil and a pharmaceutically acceptable vegetable oil.

16. The use of claim 14, wherein the vegetable oil includes sunflower oil, olive oil or soybean oil.

17. The use of claim 14, wherein said agent is administered orally.

18. The use of claim 14, wherein said agent is administered in the form of gelatin capsules.

19. The use according to any one of paragraphs. 14-18, where the indicated agent has an anti-atherosclerotic effect, characterized by a decrease in blood total lipids, total cholesterol, triglycerides and cholesterol low density lipoproteins (LDL), stimulating the excretion of cholesterol and bile acids with bile, reducing the degree of damage to aortic intima, preventing the development of aortic atherosclerosis and the occurrence of myocardial infarction.

20. The use according to any one of p. 14-18, where the specified tool has a corrective effect in the metabolic syndrome, characterized by the normalization of blood pressure, the level of triglycerides and glucose in the blood.