RU2345761C2 - Application of kauran compounds for medicines production - Google Patents

Abstract

FIELD: medicine; pharmacology.

SUBSTANCE: kauran compounds are used for production of pharmaceutical preparations prescribed for the patients suffering from ischemic disease of tissues and organs. Herewith kauran compounds are compound A presented in the Formula 2

(isosteviol) and compound B, presented in formula 3

(steviol).

EFFECT: higher efficiency and safety in comparison with methods and medicines nowadays used for treatment of tissue and organ ischemia.

15 cl, 12 ex, 4 tbl

Description

Technical field

The present invention describes the pharmaceutical use of kauran compounds, in particular compounds A and B.

State of the art

Coronary obstruction is one of the most common heart diseases. It often leads to the development of arrhythmias, coronary heart disease, myocardial infarction and heart failure. Damage to the heart muscle can occur during reperfusion, when blocked coronary vessels suddenly open during clinical procedures such as coronary angioplasty, coronary artery bypass grafting, or the use of thrombolytic drugs. Reperfusion often leads to the development of serious arrhythmias and heart failure. Several drugs have been developed to treat cardiac muscle damage and arrhythmias caused by ischemia or reperfusion. However, the toxicity or low efficacy of these drugs severely limits their clinical use.

Ischemic or hemorrhagic stroke is one of the main causes of death in the United States. It can lead to damage to neurons and cerebral dysfunction. Cerebral ischemia also develops as a result of traumatic brain injury or hemorrhagic shock. A high level of metabolism and oxygen consumption by the brain and central nervous system leads to the fact that they are most vulnerable to damage caused by ischemia or hypoxia, each of which can lead to death or permanent injury. The development of new drugs that can protect against cerebral ischemic injury is highly desirable.

Many natural compounds are less toxic and have medicinal properties. For example, glycosides isolated from plants of the genus Digitalis play an important role in the treatment of arrhythmias and heart failure. Unfortunately, these drugs should be used very carefully, since their toxic dose is very close to the therapeutic. One of the most promising approaches to improving the medical treatment of coronary obstruction, arrhythmias, heart failure and stroke is the search for new, more active medicines among natural compounds.

The biological and pharmaceutical properties of kauran compounds (Form. 1) have been known for a long time. Most studies have focused on the effects of curanoids on metabolism (Kinghom, AD. 2002, Stevia, by Taylor & Francis Inc.). In particular, it has been shown that these compounds affect cell metabolism, carbohydrate metabolism, intestinal glucose absorption, energy metabolism in hepatocyte mitochondria, and carbohydrate and oxygen metabolism in kidney cells. It has also been shown that the above compounds cause vasodilation and hypotension. However, the effects of karanoids on heart function have not attracted sufficient scientific interest. In particular, the effects of karanoids on cerebral and cardiac ischemia, arrhythmias, or heart muscle contractions have not been published.

Where:

R ¹ is hydrogen, hydroxyl or alkoxyl;

R ² : carboxyl, carboxylate, acyl halides, aldehyde, methylhydroxyl and acylamide ester, acyl or ether group hydrolyzable to carboxyl;

R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁸ (independently): oxygen, hydroxyl, methylhydroxyl and an ether or alkyloxymethyl group hydrolyzable to methylhydroxyl;

R ⁷ : methyl, hydroxyl, as well as ether or alkyloxymethyl, hydrolyzable to methyl hydroxyl;

R ⁹ : methylene or oxygen.

Natural stevioside has a karan skeleton in the form of an aglycon; it has a sweet taste (three hundred times sweeter than sugar) and has long been used in many countries as a food supplement. It has been shown previously that stevioside can lower blood sugar (Gregersen et al., 2004) and can also lower blood pressure (Chen P et al., 2000). However, it does not affect the frequency of contraction of the heart muscle or other parameters related to the function of the heart (Hseih MH et al., 2003). In animal experiments, it was shown that stevioside has diuretic activity and is able to increase insulin secretion (Jeppeesen PB, 2000) and lower energy metabolism in mitochondria (WHO, 1999). However, there is no data on the effect of stevioside on cardiac and cerebral ischemia in the scientific literature.

Two well-known kauran compounds related to stevioside are compound A (Form 2) and compound B (Form 3).

The aglycon form of stevioside, compound B, attracted the greatest scientific interest. Its biological and pharmacological properties have been investigated in detail in various animal experiments. These properties include stimulation of insulin secretion; inhibition of absorption, transport and metabolism of carbohydrates; inhibition of energy metabolism (Jeppesen BP, 2000); as well as inhibiting the tubular transport of certain xenobiotics and stimulating the excretion of sodium and water in the kidneys (Chatsudthipong et al., 2001). Compound B has also been shown to have significant mutagenic activity (Puzzuto LM et al., 1984). However, there is no evidence in the scientific literature on the effect of Compound B on the normal function of the heart or any therapeutic properties in the treatment of cardiac ischemia and reperfusion injury, arrhythmia, and stroke.

Compound B cannot be catalyzed and clotted from stevioside by digestive enzymes, but it is exposed to intestinal microflora, as a result of which it can be absorbed. Stevioside cannot be cleaved before compound B or A after intravenous administration. Therefore, the results obtained using stevioside cannot be used to interpret the biological activity of stevioside aglycones, in particular compound B or compound A.

Compound A and compound B have relatively low biological toxicity. For compound A, the minimum lethal dose (in the case of oral administration) is 5060 mg / kg for mice and 3160 mg / kg for rats. In the case of intravenous administration, the median lethal dose (LD ₅₀ ) for rats is 503 mg / kg. The main symptoms of toxicity when receiving a lethal dose are vasodilation and acute renal failure (Zhongguo et al., 1994). The median lethal dose of compound B (in the case of oral administration) for rats is 1500 mg / kg (WHO, 1999). Compared to other commonly used drugs, compound A and compound B are relatively safe.

Based on our knowledge of the prior art, we can state that Compound A and Compound B were not used as medications.

Coronary diseases and stroke are a serious risk to human health. Natural medicines (for example, compounds derived from plants of the genus Digitalis) are available and effective in the treatment of these diseases. However, the toxicity of these drugs limits their wider clinical use. Kauran compounds (Form 1) constitute a class of natural substances that have been thoroughly studied and are widely used as food additives. The widespread use of these compounds has shown their harmlessness to health. Despite the prevalence of karanic compounds, their possible use for the treatment of coronary diseases and stroke has not been previously demonstrated. In this invention, for the first time, we have demonstrated and reported on the therapeutic use of karanic compounds represented in Form 1, as well as their most effective forms for the treatment of cardiac and cerebral ischemia. These properties were demonstrated on a generally accepted rat model of these diseases. And the most effective compounds were identified as a result of more thorough pharmacological screening.

SUMMARY OF THE INVENTION

The purpose of the invention

The aim of the present invention is to develop a new method or drug derived from less toxic karanic compounds for the treatment and prevention of coronary diseases and stroke. This method or medicine should be more effective and safer compared to the methods and medicines used in the treatment of tissue ischemia and organ ischemia at present.

The aim of this invention is also the development of karanic compounds for the treatment of heart failure.

The aim of this invention is also the development of karanic compounds for the treatment of arrhythmias.

The purpose of the invention can be achieved by the use of karanic compounds in the manufacture of pharmaceuticals for the treatment of ischemic damage to tissues and organs.

The purpose of the present invention can also be achieved by using the methods and technologies described below.

The above-described ischemic diseases of tissues and organs include cardiac and cerebral ischemia.

The above-described ischemic diseases of tissues and organs include damage or necrosis of the distal extremities, retina, optic nerve and kidneys.

The cardiac and cerebral ischemia described above include coronary disease, stroke, ischemic brain injuries, and vascular block-reperfusion injuries.

The above-described coronary diseases include angina pectoris or acute myocardial infarction.

The aforementioned stroke includes ischemic and hemorrhagic strokes.

The aforementioned cerebral ischemia can be caused by trauma, hemorrhagic shock, or poor blood supply resulting from stenosis or atherosclerosis.

The aforementioned injuries caused by blocking-reperfusion of blood vessels occur during coronary angioplasty, coronary thrombolytic therapy, when using artificial blood supply devices, cerebral thrombolytic therapy.

The purpose of the invention can be achieved by the use of karanic compounds in the manufacture of pharmaceutical preparations for the treatment of heart failure. The purpose of the present invention can also be achieved by using the methods and technologies described below.

Such heart failure includes chronic heart failure or heart failure caused by a temporary decrease in myocardial function.

The purpose of the invention can be achieved by the use of karanic compounds as pharmaceuticals for the treatment and prevention of arrhythmia.

The purpose of the present invention can also be achieved by using the methods and technologies described below.

The aforementioned arrhythmia is caused by cardiac ischemia and reperfusion.

Such arrhythmia includes ventricular, superventricular or atrial arrhythmias, depending on their origin.

Such arrhythmias include ventricular tachycardia or ventricular fibrillation.

In accordance with this invention, the aforementioned karanic compounds have the general structure shown in Formula 1.

Where:

R ¹ is hydrogen, hydroxyl or alkoxyl;

R ² : carboxyl, carboxylate, acyl halides, aldehyde, methylhydroxyl and acylamide ester, acyl or ether group hydrolyzable to carboxyl;

R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁸ (independently): oxygen, hydroxyl, methylhydroxyl and an ether or alkyloxymethyl group hydrolyzable to methylhydroxyl;

R ⁷ : methyl, hydroxyl, as well as ether or alkyloxymethyl, hydrolyzable to methyl hydroxyl;

R ⁹ : methylene or oxygen.

Also the aforementioned kauran compounds, where the substance of Form 1. is compound A, are presented in Form 2.

Also the aforementioned karanic compounds, where the substance of Form 1. is compound B, are presented in Form. 3.

As well as the aforementioned karanic compounds, where the carboxylate is replaced by an alkaline earth metal, a base metal or ammonium carboxylate.

The aforementioned karanic compounds in which the medicaments or pharmaceutical preparations described include tablets, capsules, granules, injection liquid, anal or vaginal suppositories, ointments, and any slowly soluble preparations delivered by oral route, injection, implant, catheter, etc.

The above objectives are achieved by this invention by using the best compounds described below.

Disclosure of invention

This invention describes the effects of kauran compounds (Form. 1) in the treatment of cardiac and cerebral ischemia, arrhythmias and heart failure. Compounds presented in Form. 1, are a class of natural, synthetic or semi-synthetic substances. Many of them are widely known (Kinghorn AD, 2002, p. 86-173; Sinder BB, et al., 1998; Chang FR, et al., 1998; Hsu FL, et al., 2002). These substances can have one or more asymmetric centers and they can exist in the form of various stereoisomers.

Where:

R ¹ is hydrogen, hydroxyl or alkoxyl;

R ² : carboxyl, carboxylate, acyl halides, aldehyde, methylhydroxyl, and acylamide ester, acyl or ether group hydrolyzable to carboxyl;

R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁸ (independently): oxygen, hydroxyl, methylhydroxyl and an ether or alkyloxymethyl group hydrolyzable to methylhydroxyl;

R ⁷ : methyl, hydroxyl, as well as ether or alkyloxymethyl, hydrolyzable to methyl hydroxyl;

R ⁹ : methylene or oxygen.

The group of the most active compounds is presented in Form. 1 '. These compounds have a karanic structure with substitutions in the 13th carbon region and derivatives in the carbon region 17 and 18. These compounds can have many asymmetric centers, and they exist in the form of various stereo- or diastereoisomers. The absolute configuration with respect to positions 8 and 13 is as follows - (8R, 13S) or (8S, 13R).

Formula 1' Where: R ² : carboxyl, carboxylate, aldehyde, methylhydroxyl, methyl ether, methyl acyl, acyl halides. R ⁷ : methyl, methyl hydroxyl, or methyl ether. R ⁹ : methylene or oxygen

Compound A can be obtained by acid hydrolysis of natural stevioside. Compound B is an aglycone of stevioside, which in turn is a glycoside of compound B. Compounds A and B are isomers. Compound B can be obtained from stevioside by chemical hydrolysis and oxidation, or by using a catalytic bacterial reaction in the intestines of animals.

Compound A, the chemical formula is C ₂₀ H ₃₀ O ₃ ; the chemical name is (4α, 8β, 13β) -13-methyl-16-oxo-17-norkauran-18-oic acid. It is also called isosteviol, ent-16-ketobeiran-18-oic acid. This compound is tetraditerpene with a karanic structure and the following configuration of asymmetric carbons: (4R, 5S, 8R, 9R, 10S, 13S) -, substituted methyl group at carbon 13, carbonyl group at carbon 16 and carboxyl group at carbon 18 (Rodrigues et al ., 1988).

Compound B, the chemical formula is C ₂₀ H ₃₀ O ₃ ; the chemical name is ent-13-hydroxycaura-16-en-18-oic acid. It is also called steviol. This compound is also a tetraditerpene with a karan framework and the following configuration of asymmetric carbons: (4R, 5S, 8R, 9R, 10S, 13S) -, a substituted hydroxyl group at carbon 13, a methylene group connected by a double bond at carbon 16, and a carboxyl group at carbon 18 (Rodrigues et al., 1988).

Compounds A and B may also exist in the form of carboxylates at position 18, where the carboxylate is sodium and a base metal or chloride and halogen.

Both compounds have a karanic structure and are karanic compounds. Compound A is preferred in the present invention.

This invention shows that Compound A or B have a similar therapeutic effect in the treatment and prevention of ischemic myocardial injury and arrhythmias caused by ischemia or reperfusion, as well as in improving the function of ischemic myocardium. It is understood that all other compounds presented in Forms. 1 have similar therapeutic properties. It was previously shown that large amounts of compound B, under certain conditions in vitro, can cause mutagenesis. As a result, the use of compound A in pharmaceutical preparations is preferable to the use of compound B.

This invention also describes the relationship between the activity and structure of the compounds represented in Form.1. This relationship was established by comparing the activity of compounds A and B. It is implied that their therapeutic activity is associated with the basic structure of the karanic framework. Substitutions of certain radicals (for carbons 13 and 17) or a change in the stereo configuration (for carbons 8 and 13) affect the level of activity, but not the effect of these drugs. These data show that all other compounds in Forms. 1 can have the same therapeutic effect as Compound A had in treating cardiac ischemia, cerebral ischemia, arrhythmia, and also in improving cardiac function.

This invention demonstrates methods of using compounds of Forms. 1 for the formation of salts and their use in the preparation of pharmaceutical preparations for the subsequent administration of these preparations to needy patients. Compounds A and B can form pharmaceutically acceptable salts with alkali metals (e.g., sodium) and halogen in order to increase their water solubility. Compounds A and B can be mixed with suitable pharmaceutical carriers and solvents in solid or liquid form, which include tablets, capsules, controlled or slow-dissolving forms, injections, anal or vaginal suppositories, transdermal applicators, ointments, etc. These dosages applicable for long or short use can be delivered by oral, intravenous, anal, vaginal routes, or via an intravenous or intra-arterial catheter.

This invention describes an effective dose of compound A as a dose of 0.5-4.0 mg / kg with a maximum dose of 40 mg / kg administered intravenously. Previous data showed extremely low acute toxicity of compound A; the median lethal dose is 650 mg / kg intravenously. Based on these data, we can predict a safe clinical dose between 0.1 and 0.2 mg / kg. Other compounds in Form. 1, for example compound B, also have a relatively low effective and high safe dose. In this invention, the effective dose of compound B was 2-8 mg / kg. It was previously shown that acute poisoning occurs with the introduction of 1500 mg / kg (WHO, 1999). Moreover, compound B has healing properties similar to those of compound A. These include: improving cardiac function during ischemia, protection against myocardial injury caused by ischemia and reperfusion, antiarrhythmic activity, etc. In general, more Compound B is required to achieve a comparable effect than Compound A.

This invention describes the minimum effective dose of compounds A and B. Since compound A has a high lethal dose and is relatively safe, researchers have traditionally used high doses of this substance and did not notice its effect at lower concentrations. In this invention, effective doses of compounds A and B were from 1 to 2 mg / kg for rats. The equivalent dose for pigs is from 0.2 to 0.4 mg / kg and for humans, from 0.1 to 0.2 mg / kg, based on the relative surface of the body. The effective dose of compound A and B in this invention was much lower than the previously described doses. These doses were 25 mg / kg of compound A for rats (Liu CJ, et al., 2001), 250 mg / kg of compound B for hamsters (Wasuntrawat C, 1998). For humans, 250 and 500 mg of stevioside was used three times a day for the treatment of hypertension (Chen P, et al., Hseih MH et al., 2003), which corresponds to taking 80-160 mg / kg of compound B orally, or from 1.2 up to 2.4 mg / kg. The latter was calculated based on the molecular ratio of compound B to stevioside.

Previous studies could not find that compound Forms. 1, especially compounds A and B, have therapeutic activity in the treatment of cardiac and cerebral ischemia, as shown in this invention. This circumstance may have two reasons: 1) the doses used in early studies were too high; 2) experimental animal models used in early experiments were not adequate for studying ischemia.

It has been shown previously that intraperitoneal injection of a large dose of compound A (25 mg / kg) causes vasodilation and lowering of blood pressure in rats (Lucy, et al., 2001). In this invention, the intravenous administration of compound A does not cause any hypotonic reaction in rats, but provides protection against cardio- and cerebral ischemia. This indicates a mechanistic difference between the anti-ischemic and hypotonic activity of these drugs. Moreover, the difference between effective doses cannot be explained by differences in the routes of administration of drugs (intraperitoneally or intravenously). The results of toxicological studies showed that the median of the lethal dose of compound A in rats does not depend on the route of administration. It was 503 mg / kg for intravenous and 617 mg / kg for intraperitoneal administration (Zhonggou et al., 1994). This, in turn, indicates the absence of differences in the absorption and utilization of compound A upon intravenous or intraperitoneal administration. Moreover, in the same study it was shown that the median of the lethal dose for oral administration of this drug was 3160 mg / kg, which is much higher than the two above-mentioned doses.

The therapeutic effects of compounds A and B may be due to several mechanisms. It was previously shown that the hypotonic activity of compound A may be due to the involvement of potassium channels on the membrane of myocytes (Wang KL et al., 2004). At the same time, it was shown that potassium channels are not involved in the stimulation of insulin secretion by compound A (Jeppesen PB, et al., 2000). In this invention, we have shown that the anti-ischemic activity of compounds A and B can only be partially blocked by 5-OH-decadonate (a blocker of ATP-dependent potassium channels).

The objectives of this invention are achieved through the use of a well-characterized rat model of ischemia-reperfusion, which reliably reflects the symptoms of clinical pathology. Heart failure, arrhythmia, and damage to the heart muscle were evident during rat coronary blockage / opening, which correlated well with the clinical symptoms of a heart attack and coronary reperfusion caused by thrombolytic therapy or angioplasty.

This experimental model is also suitable for studying arrhythmia. Since ischemia-reperfusion is involved in many pathological processes, the arrhythmia reproduced in this invention may correspond to clinical arrhythmia caused by various factors.

This experimental model is also suitable for studying heart failure, in which a decrease in cardiac function leads to local ischemic damage to the myocardium, which ultimately leads to cardiac arrest. These symptoms are consistent with clinical heart failure.

As indicated above, the invention describes a method for the treatment and prophylaxis of coronary diseases, heart failure, arrhythmias and stroke using karanic compounds as active components of pharmaceutical preparations. Among such compounds, compound B, a stevioside aglycon, is preferred. More desirable is compound A, the product of the acid hydrolysis of stevioside. The invention describes a method of using the above inventions to protect against damage caused by ischemia and reperfusion, improve cardiac muscle function, protect against arrhythmias and reduce the risk of ventricular tachycardia and fibrillation. The invention also demonstrates, using a model of cerebral ischemia, the use of the above compounds to protect against stroke and ischemia and to preserve brain function during ischemia.

The main advantages of this invention include the following aspects.

1. The therapeutic use of karanic compounds, including compounds A and B, for the treatment and prevention of cardiac and cerebral ischemic diseases, arrhythmias and heart failure. These compounds are derived from plants that have been and have been used by humans for many years. In particular, compounds A and B may have great clinical potential due to their relatively high efficacy and high safe dose, as well as due to their therapeutic and prophylactic nature.

2. The invention is the first to study the effects of Form compounds. 1 on the ultrastructure of cardiomyocytes. The invention, using morphological and functional methods, reproduces the pathology of myocardial ischemia in experimental animal models and demonstrates that these compounds, especially compounds A and B, are effective in protecting against ischemic damage to the heart muscle and mitochondria, reducing the area of myocardial infarction, maintaining and improving cardiac function muscle and reducing the risk of acute arrhythmias during ischemia.

3. The invention for the first time describes a significant positive ionotropic effect of karanic compounds on the myocardium, which can reduce the level of deterioration of cardiac muscle function or maintain normal myocardial function during ischemia. In the clinic, acute or chronic heart failure associated with impaired heart function can be treated with Compounds A and B. The unique advantage of Form compounds. 1 is that they reduce the risk of acute arrhythmias and prevent myocardial damage during ischemia, while improving cardiac muscle function. The therapeutic index of kauran compounds is much higher than the therapeutic index of glycosides isolated from plants of the genus Digitalis. These glycosides are widely used as a class of positive ionotropic compounds, at the same time, if the dose is exceeded, they worsen the condition of the ischemic myocardium, increase the area of myocardial infarction and cause acute arrhythmia. Given these limitations, the introduction into clinical use of karanic compounds is extremely important.

4. The invention describes a significant effect that protects against reperfusion injury. Cardiac reperfusion injury can occur during clinical procedures, including angioplasty, stent placement, operative coronary artery bypass grafting, thrombolytic therapy, medical coronary dialysis, use of a cardiopulmonary bypass, etc. The mechanisms that cause this trauma may include excess oxygen radicals, calcium overload and other factors. Reperfusion can lead to cardiac arrest, lethal arrhythmias, in particular to ventricular tachycardia, and fibrillation, which was shown in this invention. The invention indicates that the use of compounds A and B can significantly reduce myocardial damage, reduce the risk of ventricular tachycardia and fibrillation caused by reperfusion, and maintain normal heart function. Thus, the aforementioned karanic compounds, in particular compounds A and B, can be used as medicines for the treatment and prevention of reperfusion injuries caused by the previously described procedures or spontaneous coronary reperfusion caused by spasm or blood clots.

5. The present invention describes the use of kauran compounds for the treatment of arrhythmias. The incidence and duration of ventricular tachycardia and fibrillation caused by coronary ischemia and reperfusion in rats were significantly reduced by the administration of compounds A or B. Compounds of Forms. 1, in particular, compounds A and B described in this invention can be used as medicaments for the treatment of arrhythmias caused by ischemia and reperfusion, as well as for the treatment of arrhythmias caused by other factors. The last statement is based on the fact that the experimental model of reperfusion involves many pathological processes.

6. The present invention describes the use of karanic compounds for protection against stroke or brain injury. The main vital functions of the brain, such as respiratory function, are lost very quickly after a disturbance in the blood supply to the brain in mice. However, the duration of preservation of the respiratory function of the brain after a loss of blood supply to the brain in mice treated with karan compounds A or B is significantly lengthened. This result indicates that karanic compounds have significant therapeutic activity in the treatment and prevention of stroke (including ischemic and hemorrhagic strokes), cerebral ischemia or trauma caused by circulatory disorders or traumatic brain injury.

7. The present invention describes a method of using karanic compounds to protect cardiac or brain tissue from ischemia. It also describes a method of using these compounds for the treatment and prevention of ischemic necrosis of the distal extremities caused by diabetes or peripheral arterial abnormality, ischemia of the retina and optic nerve, ischemia of the kidneys (acute renal failure), etc.

8. The present invention shows a nonlinear relationship between the dose of karanic compounds and the response of the body. The introduction of compound A causes a dose-dependent increase in contractility of the heart muscle, an increase in the administered dose above a certain level does not lead to a corresponding increase in myocardial contractility. When an extremely high dose is reached, the effect of curanoids on myocardial contractility can be eliminated. In pig experiments, a catheter equipped with an inflatable balloon at the end was inserted into the left descending coronary artery using an x-ray angiograph. Coronary ischemia and reperfusion was reproduced by inflation or balloon deflation at the end of the catheter. Acute ischemia-induced arrhythmia was effectively prevented in pigs given a low dose of karan compounds (i.e., compound A). This dose, as such, does not have a noticeable effect on the heart before the onset of ischemia. The introduction through the catheter into the coronary artery of an extremely high dose of compound A (20 times the previous one) led to a significant decrease in cardiac muscle function. This result indicates that karanic compounds can affect different receptors or target cells with different affinities.

A high dose of karanic compounds can cause the opposite or undesirable effect, which can reduce or prevent the previously described therapeutic effects.

9. The present invention is also directed to the development of a method for the therapeutic and prophylactic use of karanic compounds. Based on the fact that the administration of compounds A or B to animals can effectively protect the heart and brain from ischemic and reperfusion injury and karanic compounds have a good safety profile, these compounds are suitable for the treatment and prevention of coronary and cerebral arterial diseases. In the clinic, compound A can be used for a long time as a prophylactic for patients at increased risk of developing coronary heart disease, heart attack, cerebral ischemia or embolism, acute arrhythmia, and also reperfusion injury.

In General, this invention relates to methods of using kauran compounds of Forms. 1 as active ingredients in various pharmaceutical preparations used for the treatment and prevention of coronary diseases, stroke, diseases related to ischemia of tissues or organs, arrhythmias and heart failure. Compound B and A are a preferred and more preferred compound, respectively, in this invention.

Compound Forms. 1 exist in nature or are produced by artificial synthesis. According to the invention, the compounds of Forms. 1 were screened and evaluated using experimental models of rat ischemia-reperfusion and cerebral ischemia in mice. The main therapeutic and pharmaceutical effects discovered by this invention are: a positive ionotropic effect in the ischemic heart, an increase in left ventricular systolic pressure, a maximally variable level of left ventricular pressure (dp / dt _Max mmHg / s) and heart rate. Histological examination revealed that the compounds of Forms. 1 prevent ischemic injury of the myocardium and mitochondria and reduce the size of the area of ischemic infarction, and also maintain contractile myocardial function during reperfusion. In the control group, acute arrhythmia or ventricular tachycardia and fibrillation are observed in all animals during both periods of ischemia and reperfusion. One third of the animals died as a result of continued ventricular fibrillation. In the experimental group of animals that received the compounds of Forms. 1, arrhythmia has not caused a single death. The frequency of occurrence, the number of repeated cases and the duration of ventricular fibrillation during ischemia and reperfusion were significantly reduced. The great advantage of Form compounds. 1 is their ability to simultaneously prevent myocardial ischemia and arrhythmia and at the same time improve cardiac contractile function. These compounds obviously have a better therapeutic index compared to Digitalis glycosides, which have side effects, including increased arrhythmias and worsened myocardial ischemia. In animals suffering from cerebral ischemia, Form compounds. 1 can protect the brain from ischemia and significantly extend its function. According to this invention kauranovye compounds Form. 1 can be used as therapeutic agents for the following clinical aspects: for the treatment and prevention of coronary heart diseases (coronary diseases), such as angina pectoris and acute myocardial infarction; as a positive ionotropic agent for the treatment and prevention of impaired function or heart failure (including chronic heart failure); for the treatment and prevention of arrhythmias, including ventricular tachycardia and fibrillation; for the treatment and prevention of cardio- or cerebral reperfusion injury; for the treatment and prevention of ischemic stroke, hemorrhagic stroke and other vascular diseases of the brain, as well as brain damage caused by shock or head injury; for the treatment and prevention of ischemic damage to the surface of the limbs, retina and optic nerve or kidney.

The therapeutic effect of the compounds of Form.1 is dose-dependent within certain limits. In general, compound A is more effective than compound B.

The compounds of Form 1, including compounds A and B, can be formed with pharmaceutically acceptable salts with other substances, such as base metals (sodium) and halogens. They can be combined with pharmaceutical carriers to form pharmaceutical preparations. The compounds of Form 1 and their pharmaceutical preparations can be administered orally, intravenously or by other means and delivered via a catheter intravenously or intraarterially.

Currently, there is a lack of clinically available medicines with high efficacy and low toxicity for the treatment of coronary diseases, stroke, heart failure and arrhythmia. Given this drawback, the invention provides a more desirable drug with high efficacy and low side effects, obtained from low biotoxic karanovyh compounds. These compounds are better than the compounds currently used to treat coronary disease, stroke, heart failure, and arrhythmia. Previous experiments using karanic compounds did not use similar experimental models and did not find the properties of the karanoids described in this invention. The invention for the first time demonstrates the effect of kauran compounds on the structure of cardiomyocytes. The invention describes the maximum activity and effectiveness of these compounds in vivo, which were much higher than previously described. Further, these compounds have a therapeutic index higher than that inherent in the compounds currently used to treat heart failure and arrhythmia. Obviously, this invention describes a new therapeutic use of kauran compounds, and this use was not obvious.

The above has been a general description of the invention. The methods and technologies corresponding to this invention are illustrated below in such a way that they can be reproduced by a skilled worker.

The methodologies and essence of this invention are given in the following examples.

Examples

In order to illustrate the technologies used to achieve the objective of the present invention, the methods, procedures, and special properties related to the detection and determination of the pharmaceutical and therapeutic benefits of karanic compounds are described in detail below.

The examples describe the experimental methods and results that were used to support the invention and to validate the animal models used in this invention. Appropriate control and statistical analysis were used in all experiments of this invention. The following examples are provided to illustrate and not limit the invention. These examples illustrate the methods and technologies used to screen and determine the therapeutic use of some karanic compounds from Form 1 compounds. Therapeutic use of other Formula compounds. 1 can be determined in the same way.

Materials

Animals: Sprague Dawley rats or mice of both sexes. Chemicals: Compound A (ent-17-norkauren-16-oxo-18-oic acid, molecular formula C ₂₀ H ₄₀ O ₃ , molecular weight 318.5) was obtained from stevioside by acid hydrolysis, crystallization and purification. The structure of compound A was confirmed by nuclear magnetic resonance, which corresponds to previously published data. The purity of compound A is above 99%, which was definitely by high performance liquid chromatography. Compound B (ent-13-hydroxyxyur-16-ene-18-oic acid) was obtained from stevioside by sequential oxidation, hydrolysis, acidification, extraction, purification and crystallization. The structure of compound B was confirmed by nuclear magnetic resonance, which corresponds to previously published data (Mosettig E. et al., 1963). The purity of compound B is above 99%, which was definitely by high performance liquid chromatography. The purpose of the experimental compounds: intravenous or intraperitoneal injection or oral administration. Dosage: compound A - from 0.5 mg / kg to 4 mg / kg; compound B - from 2 mg / kg to 8 mg / kg.

Methods

1. Measurement of cardiodynamic parameters.

Rats were anesthetized, tracheotomized, and the endotracheal cannula was connected to an artificial respiration apparatus. Arterial blood pressure was measured using a pressure monitor located on the femoral artery. To measure ventricular pressure, a Miller sensor was inserted into the left ventricle through the carotid artery. Pressure sensors were connected to the Power-lab, a real-time recording system. An ECG was observed and recorded using hypodermic needle electrodes located on the forelimbs of rats. The recorded parameters included: mean arterial blood pressure (SBP), left ventricular systolic pressure (LHD), the first derivative of left ventricular pressure (± dp / dt _Max ), left ventricular diastolic pressure (LVDD), left ventricular end diastolic pressure and heart rate (HR).

2. Development of an experimental model of ischemia-reperfusion in animals.

The chest was opened by a left-side thoracotomy in the fourth intercostal space. The pericardium was opened to open access to the heart. A surgical silk loop was formed around the left coronary artery (LCA). At the end of the operation, the animal was restored within 10 minutes, and then the LCA was blocked by tightening the previously formed loop. The success of the block was associated with the formation of the area of percardiocyanosis, increased T wave or CT segment on the ECG. The ischemic blocking period was maintained for 20 or 30 minutes. Cardioreperfusion was achieved by relaxing the loop and was confirmed by hyperemic overgrowth of the previously ischemic region of cyanosis and the gradual normalization of the ECG. Reperfusion was maintained for 50 or 80 minutes. ECG, CAD, and other cardiodynamic data were recorded before (control) and during periods of ischemia and reperfusion.

The experimental model described above has been well characterized and has been used for a long time in relevant studies (Liu, Y and J. Downey, 1992).

Ischemia period: blocking LCA leads to local myocardial ischemia in rats, which is similar to the symptoms and pathology of acute myocardial infarction in the clinic.

Period of reperfusion: after loosening the loop and opening the coronary artery of the heart, blood flow to the heart muscle was restored, which corresponds to ischemia-reperfusion observed in the clinic, such as opening a blocked coronary artery with a catheter, drug intervention or spontaneous thrombolysis, eliminating coronary spasm, using the apparatus cardiopulmonary bypass or coronary artery bypass grafting. The clinical situations described above can result from rapid reperfusion, and they can lead to myocardial injury and arrhythmias.

3. The protocol of the experiment and the division of animals into groups.

Groups of animals: rats were randomly divided into the following groups containing from 6 to 8 animals with an equal number of animals of different sexes.

Control group: blocking LCA / ischemia-reperfusion. Experimental group: blocking LCA / ischemia-reperfusion plus administration of compounds A or B. SHAM operation: blocking LCA / ischemia-reperfusion is absent, animals underwent only left-side thoracotomy and opening of the pericardium.

Protocol:

Control group --10 min- ----20 minutes---- ----- 50 min ----------- (saline) LCA lock opening lock, reperfusion Experimental group --10 min- ----twenty----- ------fifty-------------- (experimental compound iv) LCA lock opening lock, reperfusion

To enhance the results of ischemia-reperfusion in some experiments, the period of coronary blocking and opening were set at 30 min and 80 min, respectively.

4. Statistical analysis.

Data are presented as mean ± standard deviation. A T-test or paired T-test was used to compare two groups. A 2 × 2 dependent test was used to compare quantitative data.

Example 1

This example illustrates the effects of compound A in protecting and improving ischemic heart function and its effect on cardiodynamic data.

In the table. Figure 1 shows significant changes in the control group, before and after coronary blockage, in heart rate, -dp / dt _Max , the tendency for an increase in SBP and a decrease in LVCD was insignificant (P> 0.05). However, the LHD and + dp / dt _Max parameters, which reflect the contractile function of the heart, were significantly reduced after ischemia caused by coronary blocking (P <0.01). These results show that the contractile function of the heart in animals of the control group was significantly reduced during ischemia.

Table 1.
Cardiodynamic changes before and after ischemia caused by coronary blocking and reperfusion (control group, N = 8) Heart rate beats / min GARDEN (mm Hg) LHD (mm Hg) + dp / dt _max . Flx (mm Hg / s) Before ischemia 372 ± 17 71 ± 3 118 ± 6 8704 ± 326 Ischemia 388 ± 15 67 ± 4 98 ± 2 ** 6472 ± 219 ** Reperfusion 389 ± 11 62 ± 6 107 ± 4 * 6437 ± 395 ** * significant difference compared with data obtained before ischemia * P <0.05 ** P <0.01. + dp / dt _max . LVSP: Maximum rate of change of LHD (left ventricular systolic pressure), SBP: mean arterial pressure, heart rate: heart rate.

The results in the experimental group differ from the results in the control group. In the table. 2 shows that after ischemia caused by coronary blocking, all cardiodynamic parameters, including LPSD and + dp / dt _Max. LSD, did not change compared to pre-ischemic indicators. Comparing the results of the control and experimental groups, we can conclude that compound A has a significant protective effect on the contractile function in the ischemic heart.

Table 2.
Cardiodynamic changes before and after ischemia caused by coronary blocking and reperfusion (experimental group of compound A, N = 8) Heart rate beats / min GARDEN (mm Hg) LHD (mm Hg) + dp / dt _max . Flx (mm Hg / s) Before ischemia 367 ± 13 73 ± 3 111 ± 4 8543 ± 486 Ischemia 375 ± 10 70 ± 5 103 ± 3 7468 ± 558 Reperfusion 394 ± 7 76 ± 3 114 ± 5 8431 ± 340 + dp / dt _max . LVSP: Maximum rate of change of LHD (left ventricular systolic pressure), SBP: mean arterial pressure, heart rate: heart rate.

Example 2

This example illustrates the therapeutic effects of compound A in the treatment of arrhythmias.

Arrhythmia, ventricular tachycardia (VT) and ventricular fibrillation (VF) are the main causes of death in the clinic. During ischemia in the control group, BT was observed in all 11 animals, HF was observed in 10 of 11 animals, and 3 of 11 animals died as a result of continued HF. At the same time, in the experimental group receiving compound A, all animals survived after cardiac ischemia. The results obtained from both the control and experimental groups, including the frequency of occurrence, time of occurrence and duration of VT and WF, are presented in table. 3. These results show that during ischemia in the experimental group receiving compound A (1 mg / kg), compared with the control, the incidence and duration of BT and VF were significantly reduced (P <0.01), and the occurrence of BT and HF was significantly delayed (P <0.01), which shows the ability of compound A to protect heart function and effectively reduce the risk and severity of lethal arrhythmias, including BT and HF.

Table 3.
Therapeutic effects of compounds A and B on the treatment of arrhythmia (20 minute coronary blockage) BT Frequency (%) Duration (s) WF Frequency (%) Duration (s) VT or WF
Offensive time (s) VT + VF
score Control (n = 8) 8/8 (100) 34 + 10 7/8 (91) 37 + 11 370 +41 107 + 31 Compound A * (n = 7) (1 mg / kg) 3/7 (43) 12 + 6 1/7 (14) one 730 + 166 14 + 7 Compound B * (n = 7) (2 mg / kg) 2/5 (40) 21 + 12 1/5 (20) one 660+ 178 22 + 13 * significant difference compared with the control group (P <0.01). Account BT + VF = (duration of VT x 1) + (duration of VF × 2). VT - ventricular tachycardia; VF - ventricular fibrillation.

Example 3

This example illustrates the cardioprotective effect of compound A during reperfusion after ischemia and a change in cardiodynamics.

In the control group, after 50 minutes of postischemic reperfusion, the cardiodynamic parameters of cardiac function remain reduced, as well as during ischemia. Compared to the forerunner of the LHD and + dp / dt _Max. LHD, which reflect the contractile function of the heart, is significantly reduced (P <0.05 and P <0.01, respectively), although heart rate, LVCD and -dp / dt _Max. The LCSD has not changed. SBP was slightly reduced (P> 0.05). These results show that in the control group, reperfusion leads to a decrease in cardiac contractility, as well as during ischemia (see table 1). However, during reperfusion in the control group receiving compound A (1 mg / kg), LHD, + dp / dt _Max. LHD and other cardiodynamic parameters remained within the normal range. Compared with the ischemic period, SBP, LHD, + dp / dt _Max. LHD increased slightly (P> 0.05, see Table 2). The above results obtained from both groups show that compound A has a significant protective effect on the contractile function of the heart against reperfusion.

Example 4

This example illustrates the effect of compound A on reducing the size of a heart attack during ischemia.

Determining the size of myocardial infarction: in both the control and experimental groups after reperfusion, LCA was blocked and Evans blue dye was introduced to determine the ischemic and non-ischemic regions of the myocardium. Then the heart was removed, frozen, histological sections were prepared and washed with Tris buffer. Sections were examined under a microscope. The areas of heart attack and ischemia (risk zone) were distinguished according to the intensity of the color. After weighing the respective tissue samples, the size of the myocardial infarction was calculated and presented in accordance with the following formula: heart attack size = weight of infarct tissue / (weight of infarct tissue + weight of ischemic tissue) × 100%.

Results: The size of postischemic infarction in the control group was 58.6 ± 4.7% and in the experimental group receiving compound A (1 mg / kg) - 45.8 ± 2.9%, which significantly differs from each other (P <0.01). These data show that compound A is able to reduce the size of myocardial infarction.

Example 5

This example illustrates the protective effect of compound A on the morphology of ischemic myocardium.

At the end of the experiment, ischemic and non-ischemic myocardium were isolated from both the control and experimental (compound A 1 mg / kg) groups, as well as from the corresponding region of the heart of SHAM-operated animals. Tissues were fixed with paraformaldehyde, embedded in paraffin, cut into ultra-thin sections, dyed and studied under light or electron microscopes.

Under a light microscope, sections were examined using a 100x magnification. In non-ischemic cardiomyocyte, myocardial striate is pronounced; narrow intermyocytic space, signs of edema or inflammation are absent. In ischemic myocytes, normal striate is absent, the presence of vacuolar degradation is obvious. A large intermyocytic space was found with clear signs of edema and inflammatory cell infiltration. The destruction of myocytes was also evident. In the experimental group receiving compound A, striates were visible. No vacuolar degradation was detected. Intermiocytic space of normal size with no signs of edema and rare inflammatory cells. The destruction of myocytes is not obvious.

Ultra-thin sections were studied at 1200 × magnification using a transmission electron microscope. In non-ischemic myocyte, the membranes of myocyte and mitochondria retained their integrity. Inside the mitochondria, the crista was of high density, the distribution of the matrix granules was normal. In the ischemic control group, cell membranes were destroyed, mitochondria were swollen, mitochondrial membranes were destroyed. Loss of matrix and large vacuoles were evident. The number of mitochondrial cristae was reduced; they were irregular and destroyed. Mitochondrial lesions, compared with normal control, were obvious. In the experimental group treated with compound A, the membranes of myocytes and mitochondria retained their integrity. The density of cristae and matrix and the distribution of granules were normal. Damage to myocytes and mitochondria, compared with normal control, was not observed.

Histological data showed a significant protective effect of compound A on the myocardium, this effect was achieved due to a decrease in ischemic damage to myocytes and mitochondria.

Examples 6 to 8 illustrate the pharmaceutical and therapeutic uses of compound B.

Example 6

This example illustrates the protective effect of compound B on the morphology of ischemic myocardium caused by coronary blocking.

Compound B is an isomer of compound A. This example illustrates the protective effect of compound B on ischemic myocardium.

Analogously to example 5 except that the experimental group was treated with compound B. Myocardial tissue was isolated and prepared as described above. Myocytes and their ultrastructure were studied under light and transmission electron microscopes. The results showed that in the experimental group treated with compound B, myocytic mitochondrial membranes retained their integrity. The density of cristae and matrix looks normal. The distribution of granules within the mitochondria is normal. Damage to myocytes and mitochondria, compared with normal control, was absent. The histological results were similar to those obtained in the treatment of animals with compound A (1 mg / kg), and indicate a significant protective effect of compound B on the myocardium, achieved by reducing ischemic damage to myocytes and mitochondria.

Example 7

This example illustrates the therapeutic use of compound B for the treatment of arrhythmias during cardiac ischemia.

Analogously to example 2, the therapeutic use of compound B for the treatment of arrhythmia was studied. The following results were obtained: (also see table 2 in example 2). In the control group, ventricular tachycardia occurred in all 11 animals, 3 of 11 animals died as a result of continued ventricular fibrillation. However, in the control group (n = 5) treated with compound B (2 mg / kg), all animals experienced cardiac ischemia. Compared with the control, the incidence and duration of ventricular tachycardia and fibrillation caused by cardioischemia are significantly (P <0.01) reduced, the time of their occurrence is significantly (P <0.01) delayed. In addition, the therapeutic effect of compound B at a dose of 2 mg / kg is similar to that of compound A at a dose of 1 mg / kg (see table 3 in example 2), which indicates a higher activity of compound A compared to compound B .

Example 8

This example illustrates the therapeutic use of compound B to enhance cardiac contractile function during ischemia.

Analogously to example 1, the therapeutic use of compound B was studied. The following results were obtained. In the control (ischemic) group, left ventricular systolic pressure (LHD) and its maximum rate of change (+ dp / dt _Max. LHD) before the onset of ischemia amounted to 118 ± 6 mm Hg (mm Hg) and 8704 ± 326 mm Hg / s, respectively ; during ischemia, they amounted to 98 ± 2 mmHg and 6472 ± 219 mm Hg / s. During reperfusion, the flammable flare and + dp / dt _Max. Flax flammable flare were 107 ± 4 mm Hg and 6437 ± 395 mm Hg / s, respectively. LHD during ischemia and reperfusion were significantly (P <0.01) reduced. These data indicate that ischemia and reperfusion, in the control (ischemic) group, significantly reduces the contractile function of the heart.

On the other hand, in the experimental group of animals treated with compound B (2 mg / kg), LHD and + dp / dt _Max. Prior to the occurrence of ischemia, the flammable flare density was 112 ± 5 mm Hg and 8704 ± 326 mm Hg / s; during ischemia, they amounted to 104 ± 4 mm Hg and 7592 ± 433 mm Hg / s; during reperfusion, they amounted to 110 ± 4 mm Hg and 8362 ± 498 mm Hg / s, respectively. LHD during ischemia and reperfusion did not significantly differ (P> 0.05) from normal values. These results indicate that the contractile function of the heart during ischemia and reperfusion in animals treated with compound B does not change.

The above data show that compound B can significantly protect the heart from ischemia and improve its contractile function.

Obviously, the effects of compound B are similar to those of compound A in terms of improving the contractile function of the ischemic heart, protecting or preventing ischemic and reperfusion injuries, and reducing arrhythmias caused by ischemia and reperfusion. In general, compound A is stronger than compound B.

Example 9

This example illustrates that natural stevioside does not have the therapeutic properties described in this invention.

Analogously to example 1, the therapeutic effects of stevioside were studied in rats. In the control group treated with stevid (10 to 15 mg / kg), there is no noticeable therapeutic effect on the contractile function of the heart or arrhythmia. The results show that stevioside, as such, does not have the therapeutic properties inherent in compounds A or B.

Example 10

This example illustrates the therapeutic effect of compounds A or B in the treatment of cerebral ischemia.

An experimental model of cerebral ischemia and experimental methods.

Decapitated mouse model. In this model, the mouse is decapitated, and respiratory movements are recorded by the movement of the animal’s mouth. The frequency and duration of mouth opening was used as an indicator of cerebral function after decapitation. Animals were randomly distributed between three groups (n = 8, an equal number of animals of different sexes): control group — received only physiological saline; experimental group - compound A (4 mg / kg) or B (8 mg / kg), dissolved in physiological saline; the positive control group is Edaravone (8 mg / kg), dissolved in physiological saline. Edaravone is an antioxidant and has properties that protect against neurotrauma (Grani A. et a1., 1996).

The carrier or drugs were administered intraperitoneally 30 minutes before decapitation.

Results. In both experimental groups of compounds A or B and in the Endaravone group, the frequency and duration of mouth opening were significantly increased compared to the control group, which indicates that compounds A and B have a therapeutic effect in protecting against ischemic damage to the brain and central nervous system.

The therapeutic effects of compound A (4 mg / kg) or B (8 mg / kg) were similar to the effect of Endaravone (8 mg / kg); these results are presented in table 4.

Table 4.
Therapeutic effects of compounds A and B on the treatment of cerebral ischemia Mouth opening frequency Duration (s) The control 9.7 ± 1.6 10.5 ± 2.9 Compound A, 4 mg / kg 14.0 ± 3.2 * 22.9 ± 5 * Compound B, 8 mg / kg 12.8 ± 2.7 * 20.1 ± 4.1 * Endaravone, 8 mg / kg 13.1 ± 2.4 * 17.1 ± 4.3 * * significant difference compared with the control group (P <0.05).

Example 11

This example illustrates a method for preparing a pharmaceutically acceptable salt of compounds A or B and their liquid injection forms.

Preparation of liquid injection forms. Compounds A and B are insoluble in water, therefore, the preparation of a soluble salt of these compounds is absolutely necessary. The salt may be sodium, potassium or other inorganic salt. The use of sodium is preferred. A sodium salt solution of compound A or B is prepared by adding these compounds to 10 ml of a 0.01 molar NaOH solution and adjusting the pH to neutral. This sodium salt solution can be further diluted in sterile water or mixed with a pharmaceutical carrier at a concentration necessary for parenteral administration. The solution can be stored at room temperature.

Example 12

This example illustrates the preparation of pharmaceutical preparations containing compound A for therapeutic use.

In General, compound A can be used as an active ingredient and mixed with various acceptable pharmaceutical carriers. Kauran compounds, including compounds A and B, can be absorbed through the intestines, therefore, they can also be used in solid pharmaceutical preparations for oral use.

Preparation of pharmaceutical preparations in solid form. Kauran compounds, in particular Compounds A or B, were mixed with pharmaceutical carriers such as starch, lactose, sodium carboxyl methyl cellulose, etc. These mixtures can be used to prepare tablets, capsules, granules, etc. for oral use.

Pills. Compound A is mixed in various proportions (1-99%) with a suitable carrier (starch, lactose, sucrose, dextrin, microcrystalline cellulose); disintegrants (dry starch, sodium carboxyl starch, crospovidone, low-substituted carboxypropyl cellulose, etc.); binding agents (starches, ethanol, sodium carboxyl methyl cellulose, carboxyl propyl cellulose, methyl cellulose, carboxyl propyl methyl cellulose); lubricants (i.e. magnesium stearate). One of the preferred formulations is Compound A 2 g; starch 40 g; lactose 45 g; sodium carboxyl starch 10 g; 8% starch paste and 1% magnesium stearate in appropriate quantities. These dry materials are mixed, granulated, stained and sieved, and then the resulting mass is pressed into 1000 tablets. Each tablet contains 2 mg of compound A.

Capsules Compound A is mixed in the required amounts (1-99%) with the corresponding carriers and lubricants described above, after which it is filled into capsules. One of these formulations is Compound A 2 g, 200 g starch, mixed and filled in 1000 capsules. Each capsule contains 2 mg of compound A. For the production of soft capsules, compound A can be mixed with various solvents.

Controlled or sustained release tablets or capsules. In accordance with the above methods for preparing tablets and capsules, an extended dose delivery of compound A can be achieved by adding pharmaceutical recipients (macropolymers) to the tablets and capsules, or by coating the tablets with delivery inhibitors or semipermeable membranes, thereby forming an osmotic pump, by the use of microcapsules made of semipermeable membranes, or by the formation of liposomes. These drugs can be delivered orally to prolong the release and activity time of compound A.

Parenteral liquid preparations. The parenteral liquid preparation contains compound A (portion 1-99%) in a sterile aqueous and pharmaceutically acceptable base. The solution is adjusted to the appropriate pH, stabilized, filtered, sterilized and bottled for parenteral administration. One of these preparations contains: 2 g of compound A, 2 g of NaHCO ₃ dissolved in 1000 ml of sterile water. The solution was adjusted to the appropriate pH, filtered, sterilized and dispensed in 2 or 5 ml bottles, so each bottle contains 4 mg or 10 mg of compound A, respectively. Liquid forms of compound A are used for parenteral injection or infusion.

Other pharmaceuticals. Compound A can be prepared in the form of other pharmaceutical preparations, for example, suppositories, ointments, transdermal applicators, etc.

The above examples are preferred methods of implementing the present invention, they do not limit the invention in any way. Obviously, all modifications or changes to these examples can be made by any person skilled in the art as described in this invention. Such modifications and changes should be included in the scope of this invention.

Industrial applicability

The invention relates to the use of karanic compounds in the manufacture of pharmaceuticals for the treatment and prevention of coronary diseases, arrhythmias, etc., as has been shown in animal experiments, the invention also relates to the use of these compounds in the manufacture of pharmaceuticals to protect and preserve brain function during cerebral ischemia or stroke.

Claims (15)

1. A method of treating ischemic disease of tissues and organs, including the use of karanic compounds in the manufacture of pharmaceutical preparations for prescribing to needy patients, wherein the karanic compounds are compound A represented in formula 2 and compound B represented in formula 3

2. The method according to claim 1, characterized in that the ischemic diseases are selected from the group including cardiac and cerebral ischemia. 3. The method according to claim 1, characterized in that ischemic diseases cause damage and necrosis of the distal extremities, retina, optic nerve and kidneys. 4. The method according to claim 2, characterized in that the cardiac and cerebral ischemic diseases are selected from the group including coronary disease, stroke, cerebral ischemic injury and cerebral ischemic-reperfusion injury. 5. The method according to claim 4, characterized in that the coronary diseases are selected from the group comprising angina pectoris and acute heart attack. 6. The method according to claim 4, characterized in that the cerebral ischemic damage is selected from the group comprising a head injury, hemorrhagic shock, or ischemia caused by cerebral atherosclerosis and stenosis. 7. The method according to claim 4, characterized in that the reperfusion injury is caused by a procedure including coronary angioplasty, thrombolytic therapy, the use of coronary dilatators, the use of a cardiopulmonary bypass during surgery, cerebral thrombolytic therapy. 8. The method according to claim 1, characterized in that the pharmaceutical preparations are selected from the group comprising tablets, capsules, granules, oral or parenteral injection liquid, suppositories, ointments, sustained-release preparations for oral, parenteral or implantation use, or preparations suitable for delivery through categorical intervention. 9. A method for treating heart failure, including the use of karanic compounds in the manufacture of pharmaceuticals for use in needy patients, wherein the kauranic compounds are compound A represented by formula 2 and compound B represented by formula 3

10. The method according to claim 9, characterized in that the pharmaceutical preparations are selected from the group comprising tablets, capsules, granules, oral or parenteral injection liquid, suppositories, ointments, sustained-release preparations for oral, parenteral or implantation use, or preparations suitable for delivery using categorical intervention. 11. A method for the treatment and prevention of arrhythmias, including the use of karanic compounds in the manufacture of pharmaceutical preparations for prescribing to needy patients, wherein the kauranic compounds are compound A represented in formula 1 and compound B represented in formula 2

12. The method according to p. 12, characterized in that the arrhythmia is induced by myocardial ischemia and reperfusion injury. 13. The method according to p. 12, characterized in that the arrhythmia is selected from the group comprising ventricular, supraventricular and atrial arrhythmias in accordance with their anatomical origin. 14. The method according to p. 12, characterized in that the arrhythmia is selected from the group including ventricular tachycardia or ventricular fibrillation. 15. The method according to item 12, wherein the pharmaceutical preparations are selected from the group consisting of tablets, capsules, granules, oral or parenteral injection liquid, suppositories, ointments, sustained-release preparations for oral, parenteral or implantation use, or preparations suitable for delivery using categorical intervention.