UA122750C2 - PHARMACEUTICAL COMPOSITION BASED ON PURINE NUCLEOTIDES WITH ENHANCED EFFECT ON ADENOSINE RECEPTORS - Google Patents

Abstract

The present invention relates to medicine and pharmacy and relates to enhancing the effects of adenosine receptor agonists suitable for the treatment of neurological, cardiac and other cell proliferative diseases. What is new is that the pharmaceutical composition based on certain purine nucleotides further comprises at least one amplifier selected from the group consisting of naringin, naringenin and hesperidin, and the ratio of the mass fractions of the purine nucleotide and the mass particles of the amplifier in this composition is equal to 1: 20.

Description

This invention belongs to medicine and pharmacy, and relates to pharmaceutical compositions that allow to enhance the effects of the interaction of adenosine receptor agonists and are suitable for the treatment of neurological, cardiac and other cellular and proliferative diseases.

Adenosine is a naturally occurring nucleotide that exerts its biological effects by interacting with a family of adenosine receptors, known as AT, Aga, A2b, and AZ, which modulate important physiological processes.

Adenosine has been identified as a major local regulator of tissue function, especially in cases where cellular energy is insufficient.

Due to its ability to equalize energy intake depending on metabolic demand, it was considered a "reverse metabolite" in the 1980s (Egedpoit et al., 2011; Egedpoit, 2014).

Adenosine is ubiquitous, it is released by almost all cells and is generated in the extracellular space by the cleavage of adenosine triphosphoric acid (ATP) by ectoenzymes.

The extracellular balance of adenosine concentration is maintained by reuptake mechanisms acting under the influence of specific transporters.

Adenosine concentrations increase in metabolically unfavorable conditions.

For example, tissue hypoxia leads to increased breakdown of ATP and increased accumulation of adenosine, which later, interacting with adenosine receptors, causes a number of positive effects for the body. One of these is the effect of preconditioning, or in other words, preparing cells for energetically unfavorable conditions associated with a decrease in the level of ATP in the cell.

Because adenosine is unstable, its half-life is limited by deamination or reuptake by cells, and hypoxia-induced increases in its concentration usually affect only local signaling through adenosine receptors. Therefore, adenosine belongs to the group of autacoids, since it is not secreted by a transmitter or in a hormone-like manner.

Currently, the adenosine receptor is considered a potential therapeutic target and biological marker, given its overexpression in inflammatory and cancer cells in

Compared to the low levels found in healthy cells.

Agonists are currently considered protective agents in some therapeutic areas and drug candidates in both preclinical and clinical studies.

The main natural agonist of the adenosine receptor is adenosine itself.

However, its use in clinical practice is limited due to rapid metabolic transformation into inactive, in the receptor sense, inosine.

Introduced directly into the bloodstream within 30 seconds. adenosine is deaminated and turns into inactive, in the sense of impact on the receptor, inosine.

Deprivation of the ability of adenosine to interact with the receptor occurs by deamination of its amine group, located in the b position of the purine ring, by a specific enzyme - adenosine deaminase.

The activity of adenosine deaminase is high in all organs and tissues, it is this that limits the possibility of oral use of adenosine to achieve the necessary effects, metabolizing it in the mucous membrane of the digestive tract, starting directly in the oral cavity, since deaminated to adenosine is inosine, deprived of the ability to interact with the adenosine receptor and cause a cascade of body reactions.

Therefore, today there are two methods of drug influence on adenosine receptors: 1. Use of means protected from the action of adenosine deaminase. As a rule, this connection with

Mb-substituted groups of the purine ring, for example, Mb-cyclopentyladenosine, Mb-phenyladenosine. 2. The use of agents that prolong the time of metabolic deactivation of adenosine by suppressing the activity of adenosine deaminase, for example, dipyridamole.

However, none of these ways could fully solve the problem of optimizing the effect of adenosine on adenosine receptors.

The use of means belonging to the first option for solving the problem is still, unfortunately, experimental in nature.

Clinically available drugs are limited to adenosine itself, which is used intravenously, that is, injected directly into the blood to stop arrhythmia attacks or to limit the area of myocardial necrosis during ischemia.

The second solution to the problem is based on a wider arsenal of tools. Among 60 of them, dipyridamole, tsyparadenate, hexobendin can be singled out.

Being quite effective means, they are, however, not without drawbacks, the most important of which is the slow (gradual) onset of the effect, which limits their use in urgent situations, such as the need for quick relief of pain in ischemic heart disease.

In addition, being organic substances in themselves, they can exhibit toxic effects, which limits their wide application.

Therefore, there is a constant search for substances and their combinations that can prolong the effect of natural nucleotide on adenosine receptors.

Thus, from the state of the art, a drug called "ATP-Long" or "ATP-Forte" is widely known, which is a coordination complex of the ATP molecule with magnesium and histidine. It is believed that participation in the coordination of the sphere of the magnesium ion occurs through the nitrogen of the amino group of the purine ring in the 6th position, which complicates its availability for adenosine deaminase.

This drug is used to treat existing pain and prevent pain in coronary heart disease, cardiac arrhythmias, myocarditis and myocardial dystrophy.

To achieve the desired effect, the drug is recommended to be dissolved in the oral cavity for the fastest possible absorption into the bloodstream through the mucous membrane, in order to avoid metabolic destruction by enzymes of the gastrointestinal tract.

However, by itself, ATP-Long, having a pronounced effect on angina pain, cannot completely eliminate pain in a severe angina attack. In addition, having unpleasant organoleptic characteristics, namely - pronounced salty and bitter taste - it can cause dyspeptic disorders in some patients, especially in children, which limits its use.

In the invention of the US patent, 56425, which is taken as a prototype and which is the closest to the technical solution claimed, it is described to increase the effectiveness of ATP-Long by using it together with a weak inhibitor of adenosine deaminase - inosine, which is administered intravenously.

Indeed, this approach makes it possible to increase the clinical effectiveness of the drug.

However, this requires the need for appropriate facilities and the presence of trained personnel.

In this regard, there is a need to develop a simpler form of pharmaceutical composition, which

C0 will increase the duration of the interaction of the purine nucleotide with the receptor, and thanks to this, get a quick and long-lasting therapeutic effect.

The task is solved by a pharmaceutical composition based on purine nucleotides containing adenosine or pharmaceutically acceptable salts of adenosine triphosphate or adenosine monophosphate, which according to the invention additionally contains at least one amplifier selected from the group consisting of naringin, naringenin and hesperedin, and the ratio of purine nucleotide and amplifier is chosen equal to 1:1-20.

The proposed pharmaceutical composition is suitable for oral use. Its components can be compressed using standard means to obtain a sublingual tablet.

Next, the essence of the invention is explained by examples.

For a visual demonstration, the method of recording the electrocardiogram directly during the introduction and within 5 minutes after the introduction of purine nucleotides and the pharmaceutical composition with the amplifier, which is claimed, was used.

It is known that intravenous administration of adenosine, ATP and AMP is accompanied by short-term complete atrio-ventricular blockade of the heart, which is associated with the development of hyperpolarization of cells responsible for the generation and conduction of a nerve impulse. This effect occurs due to the fact that under the influence of adenosine receptor activation by adenosine, there is a transient activation of the release of potassium ions into the extracellular space, which leads to hyperpolarization of the outer membrane and is accompanied by its inability to generate and/or conduct a nerve impulse.

Clinically, it is characterized by the absence of cardiac complexes on the electrocardiogram, after which, as a rule, after 10-20 seconds. cardiac activity is fully restored. It is this property of purine nucleotides that is the basis of almost 10095 efficiency in supraventricular reciprocal tachycardias, since these few seconds of cardiac arrest are enough to stop the mechanism responsible for the development of atrial tachycardia. The same effect underlies the vasodilating action of adenosine. However, it is short-lived precisely according to the mechanism described above.

White, sexually mature male rats weighing 120-150 g were used as experimental animals.

All research on them was carried out in accordance with the Declaration of Helsinki on the use of laboratory animals for research purposes.

Example 1

The male rat, which is in a state of anesthesia, which was achieved by the introduction of sodium ethaminal, was subjected to ECG registration. The heart rate before the experiment was 500 beats per minute. Intravenous bolus (within 2 seconds) adenosine solution at the rate of 6 mg/kg was administered, continuously recording the ECG. Immediately after the end of the administration, a complete atrio-ventricular block (AV-block) lasting 10 seconds was recorded on the ECG, after which the normal sinus rhythm was restored. The animal was removed from the experiment.

Example 2

The male rat, which is in a state of anesthesia, which was achieved by the introduction of sodium ethaminal, was subjected to ECG registration. The heart rate before the experiment was 510 beats per minute. A solution of sodium adenosine triphosphate at the rate of 6 mg/kg (in terms of adenosine triphosphate) was injected intravenously as a bolus (within 2 seconds), continuously recording the ECG. Immediately after the end of the administration, a complete atrio-ventricular block (AV-block) lasting 12 seconds was recorded on the ECG, after which the normal sinus rhythm was restored. The animal was removed from the experiment.

Example C

The male rat, which is in a state of anesthesia, which was achieved by the introduction of sodium ethaminal, was subjected to ECG registration. The heart rate before the experiment was 500 beats. in min. An intravenous bolus (within 2 seconds) of sodium adenosine monophosphate solution at the rate of 6 mg/kg (in terms of adenosine monophosphate) was administered, continuously recording the ECG. Immediately after the end of the administration, a complete atrio-ventricular block (AV-block) lasting 10 seconds was recorded on the ECG, after which the normal sinus rhythm was restored. The animal was removed from the experiment.

Example 4

The male rat, which is in a state of anesthesia, which was achieved by the introduction of sodium ethaminal, was subjected to ECG registration. The heart rate before the experiment was 520 beats. in min. Intravenous bolus (within 2 seconds) solution of naringin at the rate of 100 mg/kg was administered, continuously recording the ECG. During 5 minutes, no changes were recorded on the ECG. The animal was removed from the experiment.

Example 5

The male rat, which is in a state of anesthesia, which was achieved by the introduction of sodium ethaminal, was subjected to ECG registration. The heart rate before the experiment was 510 beats. in min. Intravenous bolus (within 2 seconds) naringenin solution at the rate of 100 mg/kg was administered, continuously recording the ECG. During 5 minutes, no changes were recorded on the ECG. The animal was removed from the experiment.

Example 6

The male rat, which is in a state of anesthesia, which was achieved by the introduction of sodium ethaminal, was subjected to ECG registration. The heart rate before the experiment was 520 beats. in min. Hesperidin solution at the rate of 100 mg/kg was injected intravenously bolus (within 2 seconds), continuously recording the ECG. During 5 minutes, no changes were recorded on the ECG. The animal was removed from the experiment.

Example 7

The male rat, which is in a state of anesthesia, which was achieved by the introduction of sodium ethaminal, was subjected to ECG registration. The heart rate before the experiment was 500 beats per minute. A pharmaceutical composition in the form of a solution consisting of a mixture of adenosine at a dose of 6 mg/kg and naringin at a rate of 100 mg/kg was administered intravenously as a bolus (within 2 seconds), continuously recording the ECG. Immediately after the end of the administration, a complete atrio-ventricular block (AV-block) lasting 60 seconds was registered on the ECG, after which the normal sinus rhythm was restored. The animal was removed from the experiment.

Example 8

The male rat, which is in a state of anesthesia, which was achieved by the introduction of sodium ethaminal, was subjected to ECG registration. The heart rate before the experiment was 500 beats per minute. A pharmaceutical composition in the form of a solution consisting of a mixture of adenosine at a dose of 6 mg/kg and naringenin at a rate of 100 mg/kg was administered intravenously as a bolus (within 2 seconds), continuously recording the ECG. Immediately after the end of the administration, a complete atrio-ventricular block (AV-block) lasting 65 seconds was recorded on the ECG, after which the normal sinus rhythm was restored. The animal was removed from the experiment.

Example 9

A male rat, which is in a state of anesthesia, which was achieved by the introduction of ethaminal-sodium, because the ECG was recorded. The heart rate before the experiment was

500 beats per minute A pharmaceutical composition in the form of a solution consisting of a mixture of adenosine at a dose of 6 mg/kg and hesperidin at a rate of 100 mg/kg was administered intravenously as a bolus (within 2 seconds), continuously recording the ECG. Immediately after the end of the injection, complete atrio-ventricular blockade (AV-blockade) lasting 58 seconds was recorded on the ECG, after which normal sinus rhythm was restored. The animal was removed from the experiment.

The summarized data (p-b for each series of experiments) of the experimental activity of purine nucleotides and their amplifiers are shown in Table 1.

Table 1

Adenosino/////1777777771116111111111Ї111111111112

ATP /17711111171611 11111115 Her

АMF 17771116

Narynini////////17771111111151111Ї111111111101111111Ї111111111111 nn and PO SOYA PO mon ne En SPO OO nki I ET: I PO sp PO

Naringenin/

Hesperidin/////1777777711151111Ї1111111110111111111Ї111111111111 nn and PO SOYA PO mon ne En SPO OO pi ET I PO s POLYA PO

Adenosinya Naringin | 6b5 77111113 nin ST I SK E U Z PO bno 11111172 11111111 bny40 | 77777711 9b045 |Death 8th floor

Adenosin

Narinenn 6601 nin S: I PO ZE U Z PO bno Й1111111175Mo |! 11117111 b6ny40 7 | 77711111 96:28/// |Death 8 floors

Adenosin

Teseridn 10006500 nyn ST I SK E U U PO ny t t p EE I PO 11111171 6ny40 | 77119844 |death9b9

AtFeNarintin.//////7777777 65111111 30k2 | 777771 ny S: s Ts o PO bno 11111172 11111711 6y40 | 77777111 9425 |Death 8 bov

AtFeNarinenin. ///////06бя5О 77777771 3047 нин С: I PO ZE U Z PO now t El PO EE Ts I PO 11111117 6y40 | 98:48 |Death 10095

AtFeHesperidin.o 65777712 | 77777771 nin ST I SK E U U PO ny t t p EE U I PO 11111711 6ny40 | 77771111 98342 / |Death 8 floors

AMPHENarinin 17777765

AMFENArineenin.y 65711111 3047 nin S: I PO ZE U Z PO nnnnSHIIIEIVYVI TE PO EE Ts I PO 11117111 6ny40 | 7777771111190342 / |Death 8 floors

AMPHEGesperidinio |////b5 77777111 22k2 nin ST I SK E U U PO ni t t p EE U I PO 11111116 niyao | 77777171 98342// |Perished

Note - results in bold are relative to the corresponding group without an amplifier

Thus, these tables indicate the uniformity of the changes recorded on the ECG when using the pharmaceutical composition.

Based on the fact that such substances as naringin, naringenin, hesperidin do not independently affect the ECG parameters, but prolong the action of purine nucleotides on the heart (and the body as a whole) by 4-6 times, they can be called amplifiers (amplifiers) of the action of purine nucleotides nucleotides.

Based on the fact that the ratio of doses of the purine nucleotide and its amplifier, which exceeds the limits of the stated ratio - by 1 wt. part of the purine nucleotide from 1 to 20 mass parts of the amplifier, or it turns out to be ineffective or toxic, the proposed ratio of the purine nucleotide to the amplifier as 1 to 1-20 can be recognized as a significant feature.

In the future, the antianginal activity of the pharmaceutical composition of purine nucleotide and its amplifier was established, when used in the form of a dissolvable tablet under the tongue.

The tablet obtained by the technology known to the average specialist had the following composition:

Adenosine - 20 mg,

Naringin - 200 mg,

Tabletose, calcium stearate, mannitol - to obtain a tablet of average weight of 300 mg.

Patient V., 69 years old, male, with his consent, who suffers from attacks of anginal pain documented by Holter ECG monitoring, was offered to take a tablet of this composition under the tongue during the attack. In case of ineffectiveness within 5 minutes, take standard pain relievers in the form of nitroglycerin tablets under the tongue.

ECG monitoring data show that the pain attack, which was accompanied by a slight depression of segment 5 according to the ischemic type, within 3 minutes after using the tablet of the proposed composition and in the proposed manner, was stopped. No side effects were noted.

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Claims (2)

FORMULA OF THE INVENTION 1. A pharmaceutical composition based on a purine nucleotide containing adenosine or pharmaceutically acceptable salts of adenosine triphosphate or adenosine monophosphate, which is characterized by the fact that it additionally contains at least one amplifier selected from the group consisting of naringin, naringenin and hesperedin, and the ratio of the mass fractions of the purine nucleotide and the mass fraction of the amplifier in the composition is equal to 1:1-20. 2. Pharmaceutical composition according to claim 1, which differs in that it has the form of sublingual tablets.