Endogenic pharmaceutical composition prepared on basis of goal-seeking activation of humoral mediators of brain cortex nerve ending

FIELD: medicine, biotechnology, pharmacy.

SUBSTANCE: invention relates to agents used for treatment of pathological states associated with disorder of synthesis of neuromediating substances. Method involves the development of pharmaceutical composition and a method for it preparing. Pharmaceutical composition represents subcellular synaptosomal fractions: synaptic membranes, "light" synaptosomes and "heavy" synaptosomes prepared from gray matter of cerebral hemispheres from experimental animals based on the goal-seeking modification of humoral mediators of nerve endings transformed to synaptosomes in development and regression of malignant processes. The composition provides inhibiting the growth of tumor cells, to elevate span-life of patients with ascite Ehrlich's sarcoma, breast adenocarcinoma Ca-755, Wolker's carcinosarcoma-256.

EFFECT: valuable medicinal and anti-tumor properties of composition.

12 cl, 3 tbl, 3 ex

 

The present invention relates to the field of medicine and biotechnology and relates to pharmaceutical compositions on the basis of neurotransmitters, the ways of its formation and use.

Shows possible new way to obtain drugs in the example of obtaining the endogenous pharmaceutical composition, which is a complex of neurotransmitters by targeted modification of humoral mediators of nerve endings converted into synaptosome, from the cerebral cortex of experimental animals.

Background of invention:

1. In recent years significantly increased the number of detected endogenously, physiologically active compounds, engaged in the regulation of neural and humoral processes.

Along with the opening of the “classic” endogenous regulators - neurotransmitters - acetylcholine, norepinephrine, dopamine, serotonin and other identified neurotransmitter role of a large group of substances peptide - neuropeptides.

In addition to these compounds, involved in the transfer of excitation in the synapses received and widely studied endogenous growth factors of the brain, as well as a number of nutrients, sometimes related to the mediator structures (histamine, bradykinin, prostaglandins, adenosine, and others)

These groups of endogenous wishes who in recent years has been enriched detected subpopulations adrenergic (AM1, AM2, VM, WM), cholinergic (M1, M2, M3), dopaminergics (D1, D2), histamine (H1, H2, H3), serotonin (C1, C2, C3 NT, 5-HT2, 5-HT) and other receptors.

Most neurotransmitters isolated from the organism in pure form, deciphered their structure and implemented their synthesis.

Parallel to the development of revolutionary ideas on the possibility of obtaining new drugs by reproducing and modifying the natural endogenous compounds of plants, animals and humans, as well as the development of new methods (such as splicing or recombinant DNA technology, genetic engineering and so on) led to the creation of a number of effective agents (insulin, interferon, human growth hormone and many others).

Endogenous compounds identified in the process of uncovering the mechanisms of pathogenesis of various diseases.

Obtaining and practical use of new pharmaceuticals based on this knowledge, is currently in several areas:

1.1. The way the truth and the introduction of outside “similar medicines.

1.2. Use as pharmaceuticals of various tissues of humans and animals.

1.3. The path “start” in developing their own endogenous means capable of blocking pop-violation in the regulation of normal physiological processes in which organisme humans and animals.

Global development receives the first (1.1) the path of research. This contributes to the rapid development of modern research techniques and the use of emerging opportunities for experimental work in cell culture, “point” of immunohistochemistry, technology, molecular cloning, as well as the involvement of computer technology. In a short time received a huge amount of information on the structuring of different bioregulators, especially neuropeptides (trophic and regulatory).

Neuropeptides, hormones, low-molecular-weight transmitters, neurotrophins and other tissue growth factors, amyloid and apoptotic proteins, functional amino acids, proteins transductor systems is far not the full list of chemical bioregulators of the brain in normal and pathological conditions. The study of these physiologically active peptides, as well as neurotrophic growth factors with regard to the activity of the brain devoted tens of thousands of publications in the last five years.

This interest is due to the emerging new ideas, the analysis of the data of laboratory and clinical studies of higher forms of behavior of humans and animals, namely the detection material causes psychosomatic functioning of the sphere of activity of the nervous system, such as memory, fear, ILO is varovanie behavior, drug dependency, storage and processing of information by neurons in the brain, the emergence and development of diseases, etc.

Emerging new strategy underlying the creation of more effective medicines for the correction of various most significant pathologies and especially such as viral infections, AIDS, cardiovascular disease, autoimmune disorders and cancer, is of high practical interest.

The most notable achievement was the creation of neurotropic drugs, which are the natural neurotransmitters. Appeared to be an efficient and synthetic analogs of endogenous neurotransmitters, having with them their structural similarity, as well as drugs that act primarily on various subgroups of receptors.

In addition, it was found that a number of medicines (antihistamines, non-steroidal, anti-inflammatory and other) in the basis of mechanism of action have an impact on the formation and metabolism of these “mediator” substances, such as histamine, prostaglandins, bradykinin, and others.

The accumulation of experimental and practical data in these research areas has led to recognition and made it all the more evidence-based theory about the leading role of the Central nervous system, and there is but brain in the chemical regulation of various processes in humans and animals, as well as the postulate that “the development of any pathology is a violation of regulatory proportionality in the system of chemical Paltrow nervous system. And that the essence of the treatment of most pathologies is to restore proportionality functioning neurochemical systems based on specific knowledge associated, in particular, neuropeptides, growth factors, other bioregulators (Oagobmv, “Neuropeptides and growth factors in the brain,” M, 2002, s).

At the present time on the basis of the received data and study of certain aspects of the functioning of the brain serves a range of drugs.

Thus, in the known pharmaceutical compositions (application FR 94/00851 from 08.07.94, IPC 12 N 15/80, With 12 N 5/10, a 61 K 39/12) on the basis of knowledge of a number of pathological conditions with impaired synthesis of neurotransmitters suggests the introduction of the gene encoding the neurotransmitter in a defective adenovirus (devoid of pathophysiological activity), and then the use of this virus in pharmaceutical preparations for the treatment of those diseases that require the synthesis of neurotransmitters. The defective recombinant adenoviruses with therapeutic gene is invited to choose from genes encoding enzymes, blood products, hormones, lymphokines (interleukins, inter is irony, TNF and so on), growth factors, neurotransmitters or their precursors or enzymes synthesis of trophic factors (BDNF, CNTF, NGF, IGF, GMF, aFGF, bFGF, NT3, NT and so on), apolipoprotein (ApoA1, Areau, APOE, and so on), dystrophy, mini dystrophin, genes tumor suppressor or genes coding for factors involved in coagulation (factors VII, VIII, IX, and so on).

It would seem that created and proposed a series of pharmaceutical products containing at least one modified defective the recombinant adenovirus and a pharmaceutical carrier. However, the undeniable biological activity of these funds not possible to fundamentally solve the problem of effective relief of these pathological conditions, especially such as neoplastic processes, as the basis of these works were used knowledge of the individual links of the functioning of the Central nervous system, and damage to the synthesis of neurotransmitters (its mechanism and reasons) was not subject to review.

A particular case in the treatment cascade pathologies and is known altolaguirre substance recommended for treatment of pain in malignant disease, which is the modified neurotoxin of Clostridium obtained on the basis of available data on the expression of peptides in the synapses. It is able to inhibit exocytosis at least one it is mediating or neuromodulatory substances from discrete populations of neurons, while reducing the transmission of afferent pain signals from peripheral to Central pain fibers and can be used as drugs for the treatment of pain, especially chronic pain (PCT: GB 96/00916 from 16.04.1996,, C 12 N 15/31, 9/52,15/62, 07 K 19/00, 14/33, 14/48, a 61 K 38/16). However, the development of the method of regulation of the allocation of the neurotransmitter or neuromodulator from primary sensory afferent cell, or from primary nociceptive afferent cells using this substance, based on the use of the particular case, the cascade of pain, allowed accordingly to reduce pain, but not the possibility of blocking in malignant disease. Here we see an example of receiving exogenous regulatory molecules for the expression of peptides in the synapses and the creation of ineffective symptomatic agents.

Way of establishing certain aspects of truth and the introduction of outside” similar” drugs not only contributed to the development of ineffective drugs, but also created the possibility of a violation of the natural physiological processes.

So, it is known that a number of synthetic drugs has structural similarity to endogenous neurotransmitters. These synthetic analogues can by binding with receptors to inhibit the action of endogenous ligands ie to be their antagonists. Effect of synthetic neurotropic drugs may also be associated with the influence on the biosynthesis and metabolism of the natural pool of neurotransmitters and have unpredictable consequences.

In addition, synthetic natural medicines obtained on the basis of “similarity” with the natural regulators such as insulin or interferon, proved to be costly and time-consuming and a little comfortable in receiving patients, because they consist of large molecules rapidly decaying in the body, so you need to make frequent injections. There is a series of disadvantages such drugs, namely the presence of selectivity. A drug that cures some patients, can be deadly for others. The creation of such drugs has the undesirable side that perfectly rendering the effect in the laboratory, otherwise the effect on the human body.

To obtain a meaningful pharmaceutical compositions attempts are also being made to use a tissue of a human or animal (1.2) on the basis of any theoretical assumptions.

So, to create an antitumor substance is proposed to use the brain of cattle (SU # 944191, class 5 And 61 To 35/30). Or for treatment of tumors of the brain to use protein-Pat the command complex from the tissues of the brain of the embryo of the animal together with its soft tissues (RU # 2128510, CL And 61 To 35/30, 35/48; RU №2128513, CL And 61 To 35/48).

It is noted that in the clinical use of such a protein-lipid complex in neurooncology, every fourth patient revealed a significant improvement in General condition. It is clear that such patients are waiting for other results and other pharmaceuticals.

It is also proposed pharmaceutical composition of muscle cells or leukocytes (US, IL, class a 61 K 35/34; 35/16, 12 P 1/00, G 01 N 33/50//(12 P 1/00 C 12 R 1/91) for inhibiting the growth of tumor cells without affecting the growth of non-neoplastic bone marrow cells or non-neoplastic lymphocytes. It is assumed that this growth regulator, administered orally, due to the activation of the immune system will help to cure diseases such as cancer. Unfortunately, practice has shown the lack of results in the final version, immunomodulators (various) during these processes, noting always a number of positive effects.

Also known pool of proteins extracted from tissue of the umbilical cord fetal human (RU # 2055589, class a 61 K 35/44; 1996), with the ability to suppress tumor growth through an immunomodulating effect.

Attempted treatment of malignant tumors by enabling internal resources of the body, using the substrate obtained by the destruction of Nati is different malignant cells, which further acts as supressor-killer T-cells.

However, the known practical results associated with the use of methods with a detrimental impact on the tumor (radiotherapy, irradiation of the tumor infrared rays, chemotherapy, etc.)encourages researchers to continue the search for more effective means of even this direction.

Currently being tested a number of anticancer drugs, where instead of chemotherapy or radiation treatment of the tumor scientists propose to block the path of its food. It is believed that this approach is quite promising, because these new drugs, called inhibitors of angiogenesis, may suspend the development of almost any type, without toxic side effects.

Such new drugs, while experimental, known for more than ten.

The most promising are angiostatin and endostatin which were isolated from the urine of mice Folkmann and his colleague Michael O'reilly (Corporation Med Inc. from Rockville, PCs Maryland). All experiences and successes obtained in mice (60-80% reduction in melanoma).

In the direction 1.2 there are a number of studies that so far have experimental or theoretical in nature, as in the case of a proposal to use embryonic stem cell is, to update various systems and functions.

The main drawback of all these (PP and 1.2) means is that in practical use, they do not meet the urgent requirements to pharmaceutical means, namely:

- the ability to cure;

- no side effects;

- the duration of;

- no dependencies.

However, there is a clear understanding of the fact that living systems are capable of self-maintenance and self-repair, if pathological changes are not irreversible due to significant violations of the integrity of the structure or the never-ending negative impact. Even plants are able to follow this postulate. So Pinus sylvestris L. in response to the impact of stress factors: anthropogenic impact, low temperature or fungal disease, re-orienting in the synthesis of secondary metabolites. It is established that increasing the degree of unsaturation of fatty acids of lipids, increases the synthesis of low molecular weight fatty acids and the most volatile components. And in cold conditions to prevent the formation in living cells ice synthesized lipids liquid consistency, and in the rest period is the increase in the content of essential oils - reserve for future use.

It is assumed that the medicinal product received is as the basis of principle, what compounds created by the human or animal body or obtained on the basis of other natural methods of combating the disease, can act more effectively.

Promising research areas 1.1 and 1.2 based on the same principles, but using the methods of individual fragments, has not yet yielded the expected results.

Lately gaining strength in the third direction by the same principle (1.3) - targeted “start” in developing their own endogenous means capable of blocking the pathological changes. Thus, in the invention (EN, No. 2120303, C1, 6 A 61 K 39/00, g.) features obtaining specific serum that can improve the body's resistance to endogenous or exogenous factors, based on the fact that donors have in the blood programmed “memory molecule” - neuropeptides.

However, the methods and examples of treatment of pathological conditions using these serums are not listed, are not examined and the part of the brain and Central nervous system in these processes.

As reported by the employees of the Ukrainian Center of embryonic tissues “EmCell”, to treat diseases by replacement of the immune system with the introduction of embryonic cells with the given parameters from the outside.

However, it is not yet known practical results of these works, in which also the e takes into account the role of the brain and Central nervous system in the formation of endogenous resources.

Object of the invention is the creation of an effective composition for relief of pathological conditions with impaired synthesis of neurotransmitters, devoid of the above disadvantages of existing tools.

The problem was solved a new endogenous composition on the basis of neurotransmitters from the brain cortex of experimental animals. The composition according to the invention is a subcellular fraction by synaptosomes structures: SYNOPTIC membrane (CM), “light” synaptosome (LS) and “heavy” synaptosome (TC)obtained from the gray matter of cerebral hemispheres and brain of experimental animals based on the purposeful modification of humoral mediators of nerve endings converted into synaptosome, during the development and regression of the pathological process.

This fraction is preferably includes synaptosome the next time the protein content in mg/g fabric:

CM - 1,4-2,4;

LS - 1,81-2,41;

TC - 1,61-1,83.

It is assumed that the new approach is able to change the perception of drugs in the existing interpretation. It is done by targeted modification of humoral mediators of nerve endings converted into synaptosome in the cerebral cortex of experimental animals.

Effective pharmaceutical composition that meets neobhodimiye for relief of pathological processes, for example, such as the emergence and development of tumors, and targeted modification of humoral mediators of nerve endings could be obtained subject to the availability of the underlying (in this case for the development of the pathological process) a theory for the justification of the causes of these processes and on the base means, causing slow or stop the process (for example, regression of the tumor).

It is known that, periodically do a sensational reports of new effective tools to fight with malignant neoplasms.

Any "boom" could end with the victory of humanity over cancer, if it was based on the underlying cause, as with the plague, for example.

Take in as root cause "microorganisms" or "parasites" and the possible transition from infection to cancer does not allow the current practice. So, for example, found that Helicobacter pylori, affecting the mucous membrane of the stomach, promotes the development of tumors or human papillomavirus (HPV) can cause cervical cancer. But... .pylory and HPV infect millions of people, but only some of them develops cancer.

So is the situation with all the hypotheses of cancer. The real reason must explain all of the accumulated mass of information, the practice of people. To explain, not to deny the obvious.

However, integration the th process of understanding and synthesis of a vast mass of data obtained for this disease begins on the separate sections, and in particular, for the functioning of the Central nervous system and brain. This is particularly relevant to understanding the material causes of psychosomatic functioning of their sphere.

Exclusively indicative of the message in this plan can be considered a message about the cause of cancer. It is reported that the cause of uncontrolled cell growth can be only one protein, known as the ICC. The researchers believe that it is the stimulation of this protein in terms of violations of its effect on apoptosis leads to the development of tumors. Is the statement, “that cancer is not as complicated as people thought” (News. Battery.ru - Battery news, 06.05.2002,). However, the issue of feedback was not even discussed. Namely, why this and many other known processes related to the development of tumors?

In the process, we formed a “working hypothesis” development of neoplastic processes, and these processes are a consequence of the disruption of psychosomatic areas of the Central nervous system of living organisms and the brain in particular.

The aim of the present invention was not proof of this postulate. He was based research as a fact that does not require theoretical or practical research.

This “working hypothesis” was able to conclude that if the pathological process cupero the Academy of Sciences, there is a time range to the availability of the pool of regulatory substances in the Central nervous system and the brain in particular.

The aim of the present invention has been experimental substantiation of the postulate that in the brain of animals (or humans) with various pathological processes in the period of recovery is the synthesis of endogenous pharmaceutical composition, which is a complex humoral mediators of nerve endings, can deal with various deviations from the normal physiological processes.

To achieve the objective of the present invention was made to create conditions conducive to the formation of a specific set of regulatory substances in the brain of experimental animals are carriers of tumour (in the examples) or other pathology by targeted modification of humoral mediators of nerve endings converted into synaptosome, receipt and use as pharmaceutical compositions.

It was assumed that effective antitumor composition or system of products that are used in complex, must simultaneously suppressing the growth of tumor cells in their localization to change the nature of the neuroendocrine regulation. It is possible that mediators produced in the uptake in the period elimina the AI tumors, may be just such sets of regulatory substances, which will be the pharmaceutical compositions of the second order, which can be further used as self-medication.

In the first stage of the research it was necessary to choose the anticancer drug, or drugs, which could be implemented at least in some animals with inoculated tumors elimination of experimental tumor processes. These connections should have been at least in some cases, not just slow the growth of tumors or increase the lifespan of animals, but also to induce remission.

From the incredibly large number of requirements for this primary anticancer drug was separated, that he should be polyactive as endogenous elimination of tumor process is impossible without adjustments impaired antitumor immunity, which runs in the background of the modified stability of cell membranes and the pool of neuroendocrine regulation.

It was expected that the anticancer drug may be the first stage and vegetable, as it is for vegetable compositions characterized paliativa the nature of the action folded into force of the chemical composition and polyactive biological effects to the plants themselves.

WPI is local antitumor herbal remedies, found application in the clinic - vinblastine from Vinca rosea (vinca rosea L.) in Russia and vincristine from periwinkle pink in Hungary (Medmaravis, “Drugs”, M, Medicine, 1988, s). However, their use is ineffective and accompanied by a number of unwanted side effects, because they are highly toxic.

Known aqueous extract of Siberian fir needles, called “Ausib”with antitumor activity. (RU # 2104020, CL a 61 K 35/78, 29.05.95). But in the case of its use we can talk about the presence of weak biological activity, rather than therapeutic effect, because in the process of getting it is inactivated by a significant number of active ingredients even that, only the volatile fraction, which is extracted.

The most appropriate known herbal remedy is a pharmaceutical composition of Siberian fir (Abies sibiricus Led), obtained from native capsular extract by enrichment fraction monoterpenes. This drug, called “Abisil” or “Abisil-1”, meets the essential criteria, namely the presence of clinically significant spectra polyactive, including antitumor, and the absence of toxic or undesirable side effect (EN, No. 2054945, class a 61 K 35/78, 27.02.96, RU # 2002107924/14, MPK7 a 61 K /02, 35/78, 29.03.2002).

Creating conditions for the formation of a specific set of regulatory molecules in the brain of experimental animals are carriers of tumour.

As a transplantable tumor strains were used ascitic Ehrlich carcinoma (example 1), solid carcinoma, breast cancer, CA-755 (example 2), carcinosarcoma Walker - 256 (Y-256) (example 3).

Substance Abisil was obtained in a known manner (RU # 2054945, class a 61 K 35/78, 27.02.96). As a solvent used olive oil in the ratio of substance Abisil and solvent, namely 0,1:10, respectively.

Example 1.

The formation of endogenous pharmaceutical composition in the brain of experimental animals are carriers of tumour with ascitic Ehrlich carcinoma was carried out using strain BALB/c.

In the experiments was taken 4 groups of animals of 60 pieces each.

The weight of mice (males and females) is not less than 20g. The tumor, the ascitic Ehrlich carcinoma, imparted vnutriplevralno. Treatment was started 48 hours after inoculation of the tumor. As control served as the group of animals which were injected with the solvent (olive oil), but were not affected by treatment.

Mice in the control group were killed on 7-15 days on the background of the development of neoplastic pleural effusion.

In the group of mice treated with inject in the area of tumor growth prototype substance Abisil in the region of the exposure dose of 100 mg/kg of animal weight, for 9 days with an interval of 24 hours, killing 7 of 60 mice for 12 days. The remaining mice remained alive within 60 days of experience with no signs of tumor or any of toxicity.

The treatment efficiency was evaluated using generally accepted indicator of antitumor activity - UPS (increased life expectancy) in the presence of the quality control mice not treated with anti-tumour therapy.

Calculation of the increase in life expectancy was performed according to the formula

where M is the lifetime in days.

As follows from the obtained data, using the substance Abisil at a dose of 100 mg/kg of body weight of the animals - introduction to the area of tumor growth according to the described scheme causes significant regression of the tumor process (see Table 1).

It was assumed that in the brain of experimental animals during the regression of tumors by the end of this process, there must be formed a certain pool of nerve endings that have complex neurotransmitters with a given localization and specific biochemical composition, which can ensure such flow of biochemical, enzymatic and physical processes that lead to regression of tumors and survival animal carriers of tumour.

Existing methods of preparative fractionarea the ia nerve tissue was allowed to test this hypothesis.

A specific set of regulatory substances from the brain of experimental animals, inoculated with tumor strain - ascitic Ehrlich carcinoma, was obtained using the method of isolation of nerve endings, namely Sublette fractions synaptosomes structures in the wording Prokhorova M.I. (“Methods of biochemical research", Ed. by Mierkowski, Leningrad, 1982, “Express method of obtaining synaptosomes structures of the crust główny rat brain”, Shevtsov centuries Bulletin eksperimentalnoi biology and medicine, No. 1, 1972).

The method of allocation fraction synaptosomes structures.

To obtain 3 g of the gray matter of cerebral hemispheres animal deceptively, removed the brain and on the cold substrate separated the cerebral cortex. Selected bark were ground with scissors and washed away the blood with a saline solution (0.85 per cent NaCl) using decapitalise and then the environment of selection (of 0.32 M sucrose, 20 Mmol - Hcl, 1 mmol EDTA, pH 7.4 - 20°).

Then the tissue is homogenized in a glass homogenizer with a Teflon pestle in a medium discharge from the calculation: 1 g of tissue 9 ml medium, 2 g - 18 ml of medium selection (8-10 fractions gap 0.2 ml).

The obtained homogenate was centrifuged 10 min at 1500g (K-24, GDR:4000 rpm).

The precipitate was a fraction of the crude nuclei (nuclei, intact cells, shelter. vessels). The supernatant carefully on cancerous and leave.

Sediment re-resuspending in the source (18 ml) environment selection and again centrifuged 10 min at 1500g. After that, the precipitate discarded, and the supernatant liquid I and II centrifugation unite.

The combined supernatant is centrifuged in the refrigerated ultracentrifuge (VAC-60 with angle rotor 8×50) at 10,000 g for 20 min to highlight “the crude mitochondrial fraction”.

Sediment “crude mitochondrial fraction” suspended in 6 ml of cold of 0.32 M sucrose and layered on pre-cooled within 30-40 min gradient, consisting of equal volumes (6 ml) 0,8; 1,0; 1,1; 1,2; M sucrose. The separation is carried out at 130000g for 30 min in an ultracentrifuge, using a rotor Swout 3×50 (VAC-60).

As a result of ultracentrifugation get 4 layers and sediment. The top layer of 0.8 M sucrose - myelin, between 0.8 M and 1.0 M sucrose - “light”, or cholinergic synaptosomes (PA), between 1.1 and 1.2 M sucrose - “heavy”, or energicheskoi, synaptosome (TC), sediment mitochondria (MX). The obtained fractions are selected using a Pasteur pipette, diluted to 0.15 sucrose and precipitated at 20000g for 40 min (ultracentrifuge VAC-60, the angular rotor 8×50).

A specific set of regulatory substances in animal carriers of tumour enclosed in brain subcellular fractions synaptosomes structures, SFOR is aravallis in response to inoculation (control group), and subsequent regression (prototype) tumor strain - ascitic Ehrlich carcinoma, was used to treat mice as endogenous pharmaceutical compositions (EPA).

For this next group of mice as a therapeutic agent was administered EPA received from the control (untreated) groups of mice with well-developed ascites. It was noted a slight increase in life expectancy of animals, but did not reveal process regression (see table. 1.)

In the group of mice treated as treatment of EPA, isolated from the brain of animals with complete regression intertwined tumor strain - ascitic Ehrlich carcinoma, revealed a high antitumor activity. (smfbl). By the end of the experience killed 2 mouse from 60 used in the experiment. Not noted any toxic manifestations.

Example 2.

The mammary adenocarcinoma CA-755 was inoculated subcutaneously in the amount of 50 mg of tumor tissue in a mouse. In the experiments were taken from mouse C57BL×CBA, males weighing not less than 20g. The treatment in the experimental and control groups was started after 48 hours. Control mice were subcutaneously injected with the solvent (olive oil). The treatment was carried out by subcutaneous injection of a substance Abisil in the area of tumor growth in disposable dose of 100 mg/kg of animal weight. The interval between injections was 24 hours. Introduction AB the power was carried out for 9 days. A specific set of regulatory substances was also obtained analogously to example 1. The evaluation criterion was the percentage of inhibition of tumor growth, calculated by the formula:

where Vcpthe average tumor volume, which is the product of 3 measurements, expressed in cm3.

The percentage of regression developed a solid adenocarcinoma CA-755 was calculated by the formula:

where

R is the percentage of regression;

VRef- initial average tumor volume;

VLechethe average tumor volume after treatment;

The results obtained are shown in Table 2.

In mice treated with the use of the prototype (table 2), showed significant inhibition of tumor growth, but low percentage regression.

The results of the experiment using EPA from the group with the prototype indicate a high therapeutic effect from a specific set of regulatory molecules, formed in the brain of experimental animals (C57BL×CBA) with mammary adenocarcinoma CA-755, during the regression of the tumor under the influence of a prototype that allowed us to use it as a drug - endogenous pharmaceutical compositions.

Example 3.

Performed forming a specific set of regulatory substances in the brain which experimental animals are carriers of tumour intertwined with carcinoma of the Walker-256 (Y-256).

Tumor-256 transplanted subcutaneously to rats Wistar rats weighing 200-250g males and females in the groin area.

In the control was used a group of animals with the introduction of the solvent in a volume of 0.2 ml p/in the groin area

Treatment was carried out using a substance Abisil at a dose of 100 mg/kg of body weight of the animals at 48 hours after inoculation of the tumor. Abisil was injected s/C in the area of tumor growth with an interval of 24 hours for 9 days.

In control groups the mortality of rats was observed during 9-18 days. Some of the rats of the experimental group (40 of 60)who received treatment with Abesalom remained alive over the past 90 days experience. There was a gradual resorption of tumor inoculum and the absence of toxic effects.

In the group of animals treated as treatment of EPA obtained from rats of the control group, a slight increase of the life expectancy of animals and the complete absence of signs of regression of the tumor.

Animals that received as a drug of EPA, isolated from the brain of animals with complete regression of the tumor-256, remained alive within 60 days of experience with no signs of the tumor and any toxicity.

Thus, we used (examples 1, 2, 3) fashion targeted activation of synthesis of humoral mediators of nerve endings, converted what's in synaptosome, was the condition for the formation of a specific set of regulatory substances in the brain of experimental animals are carriers of tumour and allowed to make their obtain and use as endogenous pharmaceutical compositions.

Thanks this was achieved objective of the present invention.

Therefore, theoretical basis of the postulate that in the brain of animals (or humans) with various pathological processes in the period of recovery is the synthesis of endogenous pharmaceutical composition, which is a complex humoral mediators of nerve endings, can deal with various deviations from the normal physiological processes is confirmed by experimental data.

As shown (examples 1, 2, 3), various pathological processes (development of ascites carcinoma, solid tumors) is well avoided by a specific endogenous pharmaceutical composition obtained from animals with similar pathological conditions during their remission.

Thus, using this method targeted activation of humoral mediators of nerve endings of the cerebral cortex in the development and management of various pathological processes, it is possible to obtain endogenous pharmaceutical compositions from various deviations normalroutine physiological processes (ascitic cancer, solid tumors, inflammatory processes, and other dermatological diseases).

In our examples (1, 2, 3) obtaining endogenous antitumor pharmaceutical composition associated with the use of prototype - substance Abisil. In the case of any other pathological processes, there is a need for the use of appropriate medicines, can have a therapeutic effect.

This is not in receipt of endogenous pharmaceutical composition of the brain of animals with different pathological conditions in the process of healing, as their remission impossible without the formation of a specific set of regulatory substances humoral mediators of nerve endings.

Until the present invention it was not possible to obtain endogenous drugs such kind of experiment that can be used as an effective pharmaceutical compositions with the desired spectrum of biological activity.

The receipt of such therapeutic agents has become possible due to purposeful modification of humoral mediators of nerve endings converted into synaptosomes from the cerebral cortex of experimental animals.

The obtained experimental data are important not only for the principle of the social new and effective medicines, but to understand the current approaches to the treatment of various diseases, as well as for a successful evaluation of medicines among the many emerging new drugs by studying their effects on the brain, and complex biological systems.

To obtain practical results, suitable for use in universal practice, it is also possible to use transition quantummechanically level.

It is known that the mechanism of selection of mediators is the same for all structures of the nervous system. They are allocated from the synapses in the form of discrete quanta.

In the process of tumor inoculation of strains (or in spontaneous tumor development processes) changes the content of neurotransmitters in the presynaptic granule cells of the brain, changing the speed of their separation, the interaction with the membrane, etc. In the process of remission in quantummechanical field conditions are created, encoding the synthesis of certain neuropediatria, which are in the form of portions of quanta. This process of coding, you can learn to manage in order to obtain a final product (EPA), which exists in any pathology.

Our experiments (examples 1, 2, 3) showed that it is highly efficient endogenous medicine. In our examples, EPA allocated and used externally, but these connections are not necessarily the consequently to select.

It is known that the structure of neurotransmitters has the form of quantum fields. Get the right medicine from the synaptosomes structures for a person can decrypt first (with a picture of the wave form of light) resulting form at the time of remission (tumors, for example), and then artificially set the synthesis of this form. This can be achieved in different ways, but including creating a mathematical model of the set of neurotransmitters and their synthesis) specific cured at different model States (U-256, CA-755, S-180 and so on). You need to learn how to generate specific design of the electronic spins in the uptake of the brain, having a sample of the decoded created in the healing process.

In this case, the emerging endogenous pharmaceutical compositions, such as in our current understanding, can not get, but simply to record the different stages of the process of remission.

The results of the present invention determine the validity and feasibility of a new approach to control synthesis “own medicines, as well as the development of high effective radically new, therapeutically important drugs.

Table 1

Effect of endogenous pharmaceutical compositions on life expectancy, we is it with ascitic Ehrlich carcinoma in comparison with the control and prototype
StrainSubstanceThe number of animals and typeUPI, %Cure, %
Ascitic Ehrlich carcinoma (EAC)1. Control (EAC)Mouse BALB/c 60-0
 2. Prototype*Mouse BALB/c 6038188
 ARC (control)**Mouse BALB/c 60160
 4. EPA (prototype)***Mouse BALB/c 6042796
* - prototype - substance Abisil at a dose of 100mg/kg of body weight.

** - EPA(control) obtained from mice of the control group.

*** - EPA (prototype) - obtained from mice (cured) from the group with the prototype of EPA endogenous pharmaceutical composition.
Table 2

Inhibition of growth of mammary adenocarcinoma CA-755 mice endogenous pharmaceutical composition
Strain tumorSubstanceThe number of animals and typeSRW, %Cure, %
Mammary adenocarcinoma CA-7551.control (CA-755)Mouse C57BL×CBA 60 pieces 0
 2. Prototype*Mouse C57BL×CBA 60 pieces7435
 ARC (control)**Mouse C57BL×CBA 60 pieces180
 4. EPA (prototype)***Mouse C57BL×CBA 60 pieces9461
* - prototype - substance Abisil at a dose of 100mg/kg of body weight.

** - EPA(control) obtained from C57BL×CBA control group.

*** - EPA (prototype) - obtained from C57BL×CBA (cured) from the group with prototype

EPA endogenous pharmaceutical composition.

Table 3

The impact of e is doganay pharmaceutical composition on the life expectancy of rats with carcinosarcoma Walker-256 in comparison with the control and prototype
StrainSubstanceThe number of animals and typeUPI, %Cure, %
Carcinosarcoma Walker-256)1. Control (Y-256)Rats of Wistar line 60 pieces-0
 2. Prototype*Rats of Wistar line 60 pieces24793
 ARC (control)**Rats of Wistar line 60 pieces110
 ARC (prototype)***Rats of Wistar line 60 pieces268100
* - prototype - substance Abisil at a dose of 100 mg/kg of body weight.

** - EPA(control) obtained from rats of the control group.

*** - EPA (prototype) - obtained from rats (cured) from the group with the prototype of EPA endogenous pharmaceutical composition.

1. Endogenous pharmaceutical composition for the treatment of pathological States with infringement of synthesis of neurotransmitters, characterized by the fact that it is a subcellular fraction by synaptosomes structures: synaptic membrane (CM), "light" synaptosome (LS) and "heavy" synaptosome (TC)obtained from the gray matter great polular the th brain of experimental animals based on the purposeful modification of humoral mediators of nerve endings, converted into synaptosome, during the development and regression of the pathological process.

2. The composition according to claim 1, characterized in that the specified fraction includes synaptosome the next time the protein content in mg/g fabric:

CM - 1.4-2,4;

LS - 1,81-2,41;

TC - 1,61-1,83

3. The composition according to claim 1 or 2, characterized in that the targeted modification of humoral mediators of nerve endings converted into synaptosome by using a substance Abisil (in olive oil) at a dose of 100 mg/kg of animal weight.

4. The composition according to claim 1, characterized in that the targeted modification of humoral mediators of nerve endings converted into synaptosome, is carried out in experimental animals inoculated with tumor strains: ascitic Ehrlich carcinoma; mammary adenocarcinoma CA-755; carcinosarcoma Walker-256.

5. The composition according to claim 2, characterized in that the targeted modification of humoral mediators of nerve endings converted into synaptosome, is carried out in experimental animals inoculated with tumor strains: ascitic Ehrlich carcinoma; mammary adenocarcinoma CA-755; carcinosarcoma Walker-256.

6. The composition according to claim 3, characterized in that the targeted modification of humoral mediators of nerve endings, converted what's in synaptosome, is carried out in experimental animals inoculated with tumor strains: ascitic Ehrlich carcinoma; mammary adenocarcinoma CA-755; carcinosarcoma Walker-256.

7. The composition according to claim 1, characterized in that it can be enclosed in a pharmaceutically acceptable carrier, depending on the desired route of administration.

8. The composition according to claim 2, characterized in that it can be enclosed in a pharmaceutically acceptable carrier, depending on the desired route of administration.

9. The composition according to claim 3, characterized in that it can be enclosed in a pharmaceutically acceptable carrier, depending on the desired route of administration.

10. The method of obtaining endogenous pharmaceutical composition for the treatment of pathological conditions with impaired synthesis of neurotransmitters, characterized in that are purposeful modification of humoral mediators of nerve endings converted into synaptosome, through the introduction of appropriate medicines, can have a therapeutic effect in this disease, when the regression process of the gray matter of cerebral hemispheres and brain of experimental animals emit by preparative fractionation of nervous tissue subcellular fraction by synaptosomes structures, including SYNOPTIC membrane (CM), light blue is ptolomy (LS) and "heavy" synaptosome (TC).

11. The method according to claim 10, in which the targeted modification of humoral mediators of nerve endings converted into synaptosome by using a substance Abisil (in olive oil) at a dose of 100 mg/kg of animal weight.

12. The method according to claim 10 or claim 11, in which the targeted modification of humoral mediators of nerve endings converted into synaptosome, is carried out in experimental animals inoculated with tumor strains: ascitic Ehrlich carcinoma; mammary adenocarcinoma CA-755; carcinosarcoma Walker-256.



 

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