The invention and the uniqueness of the method for the production of substances used to create the key molecule “CYC-8” of the OYOX preparation relates to the field of medicine, biophysics, pharmaceuticals, and in particular to a technological method for increasing the activity of substances used in pharmaceuticals, biologically active additives, and therapeutic nutrition. The essence of the invention is a method of increasing the efficiency and, as a result, expanding the indications for the prophylactic and therapeutic use of a preparation containing cycloastragenol by treating it with a weak combined magnetic field (CMF) tuned in parametric resonance mode to targets such as atoms, ions, nuclei, or electron shells nuclei of biologically important elements, primarily metals. The finished pharmacologically active extract containing more than 10% bound water and at least 1000 μg of calcium per gram of air-dried sample is exposed for 20-30 minutes at the focus of the combined magnetic field, including an alternating magnetic field colinear to the constant magnetic field of the Earth, with an amplitude of an alternating magnetic fields in the range 1 - 200 μTesla and the amplitude / frequency ratio calculated by the Bessel function formula for the parametric resonance mode tuned to calcium ions or the corresponding Larmor frequency of the nuclear spins of other biologically important elements.
The invention presents a new practical use of parametric resonance of a combined magnetic field (CMF), which allows you to accurately establish the quantitative characteristics of an alternating magnetic field (AMF), its amplitude, directivity relative to a constant magnetic field (MF) of the Earth at the site of exposure of the treated object, focus in the working area, set the amplitude / frequency ratio in accordance with the physical nature of the primary target for the formation of the desired response of a particular biological on the object, that is, change its properties. The theoretical and experimental basis of the invention is the result of many years of studies in which it was established that CMF tuned in the parametric resonance mode to targets such as calcium ions (Ca2 + CMF), potassium (K + CMF) and hydrogen atoms significantly differently affect the activity enzyme systems of plants and animals, functional activity, regeneration rate of animal tissues, stem cell proliferation in vivo and in vitro. The activating effect was provided by extremely accurate tuning to calcium ions (Ca2 + CMF), the inhibitory effect was provided by tuning to potassium ions (K + CMF).
The novelty of this technological solution lies in the fact that for the first time using “weak” less than 0.2 mTESLA CMF tuned in the parametric resonance mode, a “canned” dry extract obtained from a biological object, which is a part of a pharmacological agent, and not an actively functioning biological object, is processed. The impact was carried out using our improved device - the “magnetotron”, the principle of the device of which allows you to configure AMF parameters strictly with the calculated requirements. The object is placed in the center of the focused CMF when controlling the magnitude of the amplitude and frequency of the AMF so that the amplitude / frequency ratio corresponds to the resonance characteristic calculated for a specific target using the formula for the Bessel function. In the present invention, the resonant nature of the amplitude-frequency dependence of the response of a biological object is most clearly manifested when using as a primary target the action of weak CMF on Ca2 + ions as a charged isotropic oscillator.
To implement the proposed method, the finished pharmaceutical form of Astragalus membranaceus extract (Cycloastragenol) containing at least 78% of the active substance was selected as follows. In this case, the extract should contain in a form associated with the substance more than 10-15% water of the total weight and about 1000 μg per gram of air-dry mass of the sample. Such a preparation preserves the aquatic environment of the substance close to the native state and the natural connection of calcium with the substance surrounded by a hydration shell. The treatment of the drug with parametric resonant CMF tuned to calcium ions lasts 20-30 minutes. After the proposed treatment, the extract preparation acquires an average of 30% higher activity, which persists for at least 2 years under normal storage conditions (outside the action of high-amplitude electromagnetic fields or a strong constant magnetic field and without additional drying of the extract). The achieved result is confirmed by experimental and clinical data indicating a steady change in the activity of the drug after treatment with Ca2 + KMF. The indicated storage conditions must be observed so that the amount of bound water in the hydration shells does not decrease below the critical level or crystallization does not occur, and that the extract is not processed in such a way that leads to the extraction of calcium ions from it.
The drug obtained after 36 hours of processing the initial ("semi-crystalline" - converted into a gel state) Ca2 + KMF extract had a statistically significant effect on laboratory animals in the form of:
- accelerating the course of the wound scarring process - epithelization and scarring occurred on average 3-4 days faster;
- an increase in the lymphocyte content by 5% and the ratio of lymphocytes/neutrophils from the initial level of 4.0-4.3 to 7-8 against the background of an unchanged total number of leukocytes (about 8-9 thousand per μl) in the peripheral blood of animals (evidence of stage development activation according to Garkavi etc and increasing the resistance of the organism as a whole;
- an increase in the ratio of CD4/CD8 by 7-9% in peripheral blood leukocytes (telomerase activation is one of the signs of a positive effect on an individual's lifespan);
- an increase of cytobiochemically determined activity of succinate dehydrogenase (a marker of mitochondria) in peripheral blood cells immobilized on a smear by an average of 70 ± 15%, which reflects an increase in the contribution of aerobic mitochondrial energy to cell function (the latter is typical for healthy blood cells not subjected to oncogenic transformation).
- The effect of treatment with Ca2+ CMF was maintained after the preparation was dried to the initial moisture content (about 15%) and was observed at the same level for a year. Drying was carried out in such a way as not to disturb the hydration shell and to preserve the altered state of calcium ions in the extract treated with CMF.
The initial extract - the finished form of the drug was processed as in example 2 and was proposed (after signing informed consent and completing additional medical insurance with the subjects) to 33 healthy volunteers: without exacerbation of any previously existing diseases, not suffering from hypertension, ischemic disease hearts with signs of rest angina, not diagnosed with type 2 diabetes) to volunteers aged 48-92 years. At the beginning of the study and at the end, the peripheral blood of volunteers, blood pressure (BP), ECG and heart rate (HR) were studied. The elevations in blood pressure and heart rate were measured every day: there were no paroxysms of rhythm disturbance or tachycardia in volunteers. The cellular composition of the peripheral blood and the standard biochemical analysis of blood did not undergo deviations beyond the range of the physiological age norm, both in the control group (15 people who received a course of the drug not treated with CMF according to the instructions) and in the “experimental” group (18 volunteers who received according to the instructions the course of the drug subjected to the processing of the CMF).
However, the following changes were observed in the “experienced” volunteers in the measured characteristics. The number of lymphocytes increased by 40-60% compared with the original. According to this parameter, people are more sensitive than animals, apparently, not only in connection with the species characteristics of rats, but also because in laboratory animals kept in a comfortable vivarium, receiving a balanced diet with rhythmic feeding, with regular cleaning of the cages, a clear regime of daylight hours, in the absence of any stressful effects, an initially much higher lymphoid status is observed than in humans. In rats, the proportion of lymphocytes is 80-85% of the total number of leukocytes, that is, the number of lymphocytes is higher than the number of neutrophilic leukocytes. Whereas in humans, the number of lymphocytes on average is 1.7-2.0 times less than the number of neutrophils circulating in the blood. Against this background, the average percentage of lymphocytes increased by 40-60% from the initial level: that is, from an average level of 25% to 35% after taking the treated extract.
In the control group of volunteers who took untreated with CMF extract, a statistically significant increase in the relative content of lymphocytes was also observed, but not more than 20%. Moreover, the shift of the CD4/CD8 ratio in a positive direction (a sign of an increase in telomerase activity) in humans is less pronounced than in animals, although in the experimental group, according to the paired Wilcoxon criterion was statistically significant.
The increase in the activity of the cytobiochemically determined activity of succinate dehydrogenase (SDH) in peripheral blood cells immobilized on a smear was much more significant in volunteers of the “experimental” group. In the control group, after a course of taking the usual extract, a pronounced tendency to increase this indicator (of the order of + 50%) was found, but it did not have statistical significance due to the large individual scatter of the results characteristic of different people. In the “experimental” group, the increase reached + 80% or more, and therefore, despite the individual scatter, it was statistically significant. Interestingly, the formazan color in the sample with malonate, which reflects the level of single-electron leaks in the respiratory chain (the formation of reactive oxygen species at the level of mitochondria), varied ambiguously in both groups depending on the initial state, but more clearly, in the “experimental group.""
In volunteers who had an initially high activity of a malonate-detectable level of ROS-dependent staining of formazan, the color intensity sharply decreased by a factor of 2–3 after a course with a treatment with CMF (inhibition of excess ROS formation).
And, on the contrary, with an initially practically zero color in the presence of malonate (also not a good situation), after a course of the drug treated with CMF, a statistically significant increase in color was observed to the level characteristic of young healthy people. Both shift lymphocytes noted at the level of mitochondria may be due to the fact that the treated preparation is more conducive to activation of mitochondrial aerobic energy and restoration of the ratio of pair and one-electron transfer in the respiratory chain. In our case, almost normalization of ROS formation was observed: an increased level decreased but normal, and, on the contrary, a sharply reduced level was restored to the level characteristic of healthy young people. The regulatory, supportive activity of mitochondrial enzymes may be due to the functioning of the known regulatory cascade, which is activated by active sirtuins 1, 3, and 6 by acting on the transcription factor PGC1α, which controls the synthesis and activity of mitochondrial oxidative phosphorylation and respiratory chain enzymes.
An analysis of the totality of data on changes in the parameters that determine the rate of aging and cell involution, as well as an increase in the activity of the mitochondrial system, found even at the level of blood corpuscles, suggests that the course of taking OYOX helps to significantly stabilize and increase the level of regulatory deacylating regulators - sirtuins. First of all, this concerns sirtuins 1, 3 and 6. Of course, to one degree or another, depending on the specific state of a person, 9 widely discussed positive effects of OYOX can be realized. Obstruction of the aging of the body, antiviral effect, regulation of the activity of the immune system, prevention of fibrotic degeneration of parenchymal organs, bacteriostatic effect, positive effect on blood glucose levels in diabetes mellitus type I and II, reduction in the likelihood of development and progression of obesity, radioprotective radiomitatory and, finally, antitumor effect.
The observed acceleration in performing the number-link test after the course of taking the treated with CMF CYC-8 in the OYOX preparation fits well with modern ideas and corresponds to data on its effect on mental abilities after a neurodegenerative lesion caused by a stroke. After 2 months of taking OYOX, human cognitive functions tend to increase the volume of short-term and long-term memory by 9-11%, respectively.
There is good reason to believe that this is due to the sirtuin activation of mitochondriogenesis, since brain structures are most sensitive to increased activity of mitochondrial energy. And it is the maintenance of mitochondrial energy that prevents the progression and development of almost all types of neurodegenerative changes in the brain, including in Parkinson's disease, Alzheimer's disease and atherosclerotic lesions of the blood vessels of the brain.
Thus, at the level of animals and humans (as a receptive system), it was shown that the CYC-8 molecule, which retains a practically native hydration shell around a biologically active plant substance and calcium cations, changes in its properties under the influence of a parametric resonant CMF tuned to calcium ions. In this case, the biological activity of the drug increases.
Typically, low sirtuin activity is associated with various age-related diseases and earlier mortality. Thus, leukocyte telomere length (LTL) and the activity of SIRT1, 3, 6 sirtuins become both biomarkers and factors contributing to age-related diseases. However, not a single clinical study examined directly the activity of sirtuins and the telomere length in different subtypes of lymphocytes isolated from the same donors, which could give an idea of the total assessment of the telomere content in leukocytes.
The first quantitative data in humans, studying both levels of sirtuin activity and telomere length in four subpopulations of lymphocytes from the same donors -CD4 +, CD8 +, CD28+, and CD8 + CD28 – T-cells and B-cells, as well as complete PBMC - in a cohort of healthy women.
We found that B-cells had the highest sirtuin activity and the longest telomere length;
CD4 + T-cells had slightly greater sirtuin activity than CD8 + CD28 + T-cells and had a similar telomere length. According to earlier reports that CD8 + CD28 T-cells are replicatively aging cells, they had the lowest telomerase activity and the shortest telomere length.
In addition, a higher percentage of CD8 + CD28-T cells correlated with shorter total PBMC TL (r = - 0.26, p = 0.05).
The sirtuin activity of CD4 + and CD8 + CD28 + T-cells of the same individual was strongly correlated (r = 0.55, r < 0.001), which indicates possible general mechanisms for regulating SIRT1; 3; 6 activity in these two cell subtypes. These data help to understand the aging of white blood cells and their relationship with human health.
