Adhesive phytoadaptogens geroprotectors in preventive oncology

O.A. Bocharova, R.V. Karpova, E.V. Bocharov, A.A. Aksyonov, T.L. Chekalina, S.V. Chulkova, N.S. Ionov, I.V. Kazeev, M.A. Lyzhen-kova, A.A. Vershinskaya, V.G. Kucheryanu, V.S. Kosorukov. Ad- hesive phytoadaptogens-geroprotectors in preventive oncology. Cardiometry; Issue No. 26; February 2023; p. 55-68; DOI: 10.18137/cardiometry.2023.26.5568; Available from: http://www.

At the current stage of medical science development there is significant knowledge about various disorders in the tumor process (expression of oncogenes and cytokines, factors of apoptosis, drug resistance, angiogenesis, deficiency of immunoreactivity, etc.). Many of the identified features of the tumor cell are trying to be used as targets for anticancer therapy. However, it is problematic to consider these numerous “details” of the characteristics of a tumor cell as targets for anticancer therapy. This is due to the fact that they do not have a key or independent significance in the appearance of a tumor cell, although they can work in experimental animal models under certain conditions and with prolonged therapy, which at times has little relation to humans.

In addition, many scientists are in favor of the fact that the future of oncological science and antitumor therapy should lie in the field of activation of the body’s defense mechanisms in general and the restoration of immunological surveillance in particular. At the same time, in 2014, IARC reported that the global war against cancer cannot be won by therapy alone, and recommended the introduction of effective prevention strategies to avert the oncological crisis. However, in this case, the globality and vagueness of these too general plans confuses.

Thus, a generalized theory of cancer still does not exist. On the one hand, none of the numerous factors listed above can be considered specific, independent, key, suppression of which can prevent or stop the tumor process. On the other hand, too general arguments about the activation of the body’s defense mechanisms do not have a clear direction. Consequently, we have incomplete knowledge of how a tumor cell coordinates and connects a huge number of events, skillfully using the “treacherous” help of its environment in the host organism to destroy it.

Disruption of cell differentiation with loss of control over proliferative processes is considered an inseparable feature of tumor cells. Close attention deserves the fact that, in this case, tumor cells lose their interaction with each other, with immune effectors, and, as a result, with the whole organism. On the basis of the research results accumulated to date, we can consider the violation of adhesive interactions as the key mechanism of neoplasia [1, 2, 3, 4].

Thus, the features of local adhesive dysregulation, which provides the main properties of the tumor: loss of tissue control of proliferation, anaplasia, invasion, metastasis, deficiency of immunological surveillance – can be controlled by central mechanisms involving the dopaminergic system, which is able to regulate the active phase using immunoadhesion interactions, immune reactions against the tumor, interfering with 56 | Cardiometry | Issue 26. February 2023

the process and thus interrupting the development of a malignant neoplasm initiated by a local mutation in a particular tissue.

At the same time, the proposed concept of the adhesion disorders key role in malignant growth in tissue and in the processes of immunoreactivity with the participation of the central dopamine loss as an adhesion-damaging factor reveals, among other things, the stress mechanism of cancer etiology. Therefore, an important conclusion of our reasoning may be a previously unknown answer to the question of the mechanism leading to the development of a malignant neoplasm as a result of chronic stress.

Finally, a significant, if not the main conclusion is that the activation of the body’s defense mechanisms and the restoration of immunological surveillance in relation to the implementation of the above approaches to the adhesive concept of local and central regulation of tumor formation creates a definite and specific perspective for increasing the effectiveness of diagnostic methods, prevention and treatment, which may be another step towards solving the problem of malignant neoplasms [5, 6.7, 8].

Preventive oncology can solve many problems of neoplasia – prevention of occurrence (primary prevention), relapses and metastasis after the main treatment (secondary prevention), complications after chemo-radiation therapy (tertiary prevention).

Innovative approaches that take into account the unique capabilities of phytoadaptogens (PhAs) natural components of antitumor and geroprotective orientation are promising in this aspect.

In traditional oriental medicine Panax ginseng, Eleutherococcus senticosus, Rhodiola rosea, etc. for more than 2000 years have been used as a means to promote longevity. The concept of adaptogens is based on Hans Selye’s theory of stress and homeostasis. Repeated low doses of stress cause an increase in the resistance of cells and organisms to subsequent stress which leads to adaptation that promotes survival. This phenomenon of adaptation to repetitive low-level stress was first described by Hans Selye in 1936.In this context adaptogens act as chronic eustress (low doses of stress) activating an adaptive response to stress, resistance and overall survival.

It is known also that there is a direct dependence of the tumors incidence on the rate of the population aging [9]. Considering that the key mechanism of aging and tumor formation can be a violation of adhesion in- teractions (and as a consequence – disdifferentiation) cancer can be considered as a rapid aging of tissue cells [10]. Therefore, the use of geroprotectors-adaptogens in oncology can be considered promising.

However, it should be emphasized that phytoad-aptogen preparations are inducers of differentiation (they are not stimulators of cell proliferation) supporting tissue systems in a highly differentiated state of mature ontogenesis homeostasis characteristics [11, 12].

That is why the interest of oncologists in these drugs is not surprising. The beginning was laid by Professor N. V. Lazarev from Naval Medical Academy in Leningrad. It was N. V. Lazarev who introduced the term adaptogen into the scientific literature. And now it is used by scientists all over the world. In vivo experiments led by his followers including Professor I. I. Brekhman showed the antitumor properties of eleutherococcus, ginseng, rhodiola rosea, etc. Antimetastatic activities of PhAs were also shown as well as intensification of chemoradiotherapy and decrease of its side effects. The pioneer of the phytoadapto-gens study in the oncological clinic was Professor V. I. Kupin from Moscow NN Blokhin Cancer center of Russian Medical Sciences Academy. The development of this direction has been received in our works. In recent years, the interest in adaptogens has increased abroad as well [11, 12, 13, 14, 15, 16, 17, 18, 19].

First of all, we have shown that extracts of adapto-gens (Panax ginseng, Rhodiola rosea) enhance the mutual adhesiveness of cells in tissue predisposed to tumor growth. Administration of Rhodiola rosea extract to high-cancer CBA mice-males in the preventive (short-term) and therapeutic (long-term) modes increased the mutual adhesiveness of target tissue (liver) cells, had immuno-, hormonemodulatory and antistress effects, as well as reduced also the frequency of hepatocarcinomas in mice-males in the second half of ontogenesis [20]. It was also demonstrated in cancer clinic that the use of Rhodiola rosea extract enhances the mutual adhesiveness of bladder urothelium cells reducing the frequency of recurrence of bladder cancer in patients [21, 22].

It should be noted that pure biologically active compounds were certainly isolated from adaptogen plants. However, their use is significantly limited for many reasons including low bioavailability, rapid excretion from the body and high toxicity. Therefore, the use of extracts remains preferable. In turn, the use of individual plants extracts is accompanied by the rap- idly advancing tolerance, that is, the loss of effects on the body. We tried to solve this problem by creating a complex preparation consisting of several extracts components [23, 24, 25].

Studies of phytotcomplexes based on the principle of the biologically active substances with structural variety rational combination and providing unique synergistic effects that cannot be obtained with any ingredient separately are relevant. More over the use of several adaptogens in one pharmaceutical composition may allow you to affect the body without causing the tolerance.

Taking into account these problems the development and research of an approach to the assessment of a complex phytoadaptogen for preventive oncology and age-related diseases seemed relevant.

Multiphytoadaptogen (MPhA) for preventive oncology. MPhA was developed by the Blokhin National Medical Research Center of Oncology. It is approved in Russia as a parapharmaceutical agent consisting of components from 40 medicinal plants including Panax ginseng , Rhodiola rosea, Eleutherococcus senticosus, Eucalyptus globules, Juniperus communis, Valeriana officinalis, Polygonum aviculare, Leonurus cardiaca , etc. [26]. MPhA has been standardized by both analytical and biological methods. With the help of NMR (nuclear magnetic resonance) and UV (ultra-violet) spectrometry the possibilities of controlling the medication by the extracts’ composition were determined. By means of HPLC (high-efficiency liquid chromatography), GC-MS (gas-liquid chromatography with mass detector), UV-HPLC (high-efficiency liquid chromatography with UV-detector) and HPLC-MS/MS (tandem mass spectrometry) analyses of the MPhA composition many compounds were quantitatively identified – triterpene glycosides, phenylpropanoids, lignans, flavonoids, essential components etc. As a result, 70 major phytocomponents have been selected in the MPhA composition. It was demonstrated that the adaptogenic activity of MPhA turned out to be higher in comparison with those of the individual PhAs. The adaptogen-ic activity was identified by the method for assessing the total adaptogenic score (TAS) – accelerated growth characteristics of baker’s yeast cells culture Saccharomy-ces cerevisiae using an energetically poor nutrient medium in the presence of the MPhA. The method TAS is important for biological standardization, making it possible to evaluate PhAs qualitatively and quantitatively, recognizing counterfeit drugs [27, 28].

Pharmacological and toxicological characteristics of MPhA have been investigated in vitro , in vivo and in clinical studies. A wide range of pharmacological effects (adaptogenic, antitumor, immunomodulatory, antimutagenic, etc.) exhibited by MPhA has been demonstrated. It was shown that MPhA corresponds to the class VI safety level according to the OECD classification (practically non-toxic) [29].

In vitro studies

The antiproliferative effect of MPhA has been shown in vitro on cell cultures of ovarian and cervical human adenocarcinomas as well as on human hypernephroma. However, MPhA did not affect the proliferation of normal kidney embryonic cells of the pig. This characterizes MPhA as a real adaptogen that provides an effect on pathology but not on the normal state. In other words, one may conclude that MPhA has a selective antitumor activity [30, 23].

The antimutagenic effect of MPhA in vitro was manifested in a decrease in the level of spontaneous and induced mutations in yeast cells Saccharomyces cerevisiae [31].

In vivo studies. In a large-scale experiment involving almost 1000 animals the preventive and therapeutic impacts of MPhA on CBA mice-males genetically predisposed to hepatocarcinomas provided a longterm effect of increasing the expression of leukocyte beta 2 integrins LFA-1 and Mac-1 on peripheral blood cells involved in contact interactions of immune effectors with target cells. At the same time a decrease in the concentration of IL-6 and IL-10 in the blood serum of animals was revealed. This leads to increased expression of beta 2 leukocyte integrins LFA-1 and Mac-1 – ligands of histo-nonspecific adhesion molecules ICAM-1, 2 on tumor cells. In this way the regulation of heterotypic adhesive interactions that cause the attachment of immune effectors to tumor cells can be restored contributing to the elimination of the latter. Prevention of dopaminergic neurons loss in the brain was accompanied by a decrease in the level of tumor formation, an increase in lifespan and maintenance of somatic status with correction of the stress hormone corticosterone serum level. Consequently, it is possible to assume control of the hepatocarcinogenesis process with increased antitumor protection and weakening of stress mechanisms including the participation of peripheral dopamine high content justifying the properties of the latter known from the literature as a stress regulator, antitumor agent and geroprotector. At the same time peripheral dopamine may play a role in the maturation of cytotoxic lymphocytes contributing to their conjugation with tumor cells. As a result, a 30% decrease in the frequency of tumor formation as well as statistically significant reduced number and size of spontaneous hepatocarcinomas were shown. The average lifespan of mice increased by 15-20% while maintaining body weight, motor (behavioral) activity and coat without signs of alopecia. With the chronobi-ological transfer of the mouse age to humans the mice of the control group lived about 62 “human years”, in the preventive mode group – about 70, in the therapeutic – about 75 years (Figure 1).

The high antimetastatic and immunomodulatory effects of MPhA were identified in male mice of the (CBAX57BL/6) F1 strain with Lewis lung carcinoma (LLC) transplanted subcutaneously. The antimetastatic properties of MPhA were revealed which correlates with the immunomodulatory effect on the T-cell immunity and increased survival of animals.

MPhA also showed radioprotective properties in mice and dogs under conditions of acute and prolonged gamma irradiation. The animals’ lifespan increased by 80% in mice (40% in the control) and 40% in dogs (0 in the control) with the best peripheral blood parameters.

In mice with MPTP-induced parkinsonian syndrome (the model of aging disease) antioxidant and neuroprotective activities of MPhA were shown. The MPhA prevented the development of hypokinesia and rigidity of animals, DA and its metabolites decreasing, suppressed the content of serotonin and malondialdehyde in the striatum of mice almost to normal values. A decrease in the activity of caspase 3 and the level of DNA fragmentation in the substantia nigra was also revealed which implies suppression of neuronal apoptosis death [24, 25].

It has been shown that the expression of β2-leuko-cyte integrins is extremely important for increasing the antitumor cytotoxicity of T-lymphocytes. It is known that some of the promising synthetic activators of antitumor T-cell immunity and, in particular, infiltration of tumors by cytotoxic T-lymphocytes (according to the international database Cortelis Drug Discovery In-telligens, 2022) are in phase 1 of clinical trials. Among them is the agonist of the adhesion molecule LFA-1 (7HP349, Hills Pharma, USA), as well as the agonist of the adhesion molecule Mac-1 (ADH-503, Leukadher-

Figure 1 .Scheme of a large-scale experiment on the effect of Multiphytoadaptogen in preventive and therapeutic modes on spontaneous hepatocarcinogenesis in high-cancer inbred mice-males CBA.

in-1cholin, GossamerBio, USA). This emphasizes the exceptional relevance and timeliness of our research. In other words, homotypic adhesion molecules of the con-tactin-cadherin type activate the expression of heterotypic adhesion molecules of beta2-leukocyte integrins, increasing the antitumor effect and lifespan.

Studies in the clinic

Clinical studies of MPhA were conducted in patients with leukoplakia of the oral mucosa (OL, an example of precancerous disease), advanced gastric cancer (GC, an example of incurable stage IV cancer), benign prostatic hyperplasia (BPH, an example of age-related hormonal disbalance disease in men), Parkinson’s disease (PD, age-related neurodegenerative disease). These studies supplemented and confirmed the results of experimental researches.

So, in the clinic and in the experimental researches the immunomodulatory effect of MPhA was revealed that was seen in the normalization of the T- and B-cell immunity as well as in the increase of NK cells level and growth of IL-2 receptors expression. At the same time the use of MPhA in PD led to the suppression of negative activation of immune system cells showing anti-inflammatory activity.

The interferon-inducing effect of MPhA was also revealed. It was expressed in the normalization of immune and interferon status indicators as well as in the induction of its own interferon (IFN-γ) production by peripheral blood lymphocytes. After the course of MPhA treatment the number of patients whose blood cells responded with IFN production increased significantly not only in relation to IFN drugs (gammapheron, reaferon, etc) but its inducers (neovir, amix-in, etc) as well. It is very important that all patients showed sensitivity to MPhA in terms of the production of their own IFN when exposed to MPhA. Thus, the fact of the individual tolerance absence to MPhA was established which favorably distinguishes it from individual phytoadaptogens.

The high interferon-inducing activity of MPhA was detected using a biological method (delayed destruction of the diploid fibroblast culture monolayer after the introduction of a test virus – vesicular stomatitis virus or mouse encephalomyocarditis virus). The results obtained allow us to conclude that MPhA has antiviral activity.

In addition, the antistress activity of MPhA was determined in these patients – a decrease in the level of the stress hormone cortisol. MPhA also showed high antioxidant activity – suppressed lipid peroxidation with a decrease in the level of malondialdehyde, activation of catalase and glutathione antiperoxide system.

The adhesive properties of MPhA are also shown on the example of precancer – OL confirming experimental studies on high-cancer CBA mice. Under the influence of MPhA on the epithelial cells of the leukoplakia lesions an increase in the expression of ICAM-1 adhesion molecules (ligands of beta2 inte-grins on immune effectors) was shown, a growth in the expression of receptors mediating apoptosis (Fas APO1) was noted as well as a decrease in the expres- 60 | Cardiometry | Issue 26. February 2023

sion of the protein-marker of immature epithelial cells (keratin 17). Modulation of keratinocyte differentiation with MPhA may be an exiting clinical possibility in the control of malignant transformation of precancers. Clinical efficacy of MPhA was twice higher than that of conventional treatment with Vitamin A preparations. Taking into account the revealed properties, the therapeutic effect of MPhA against precancerous (leukoplakia) and the preventive effect in high-cancer mice MPhA can be considered to be a medication for cancer prevention (primary prevention) [32].

The use of MPhA in advanced GC was successful. In patients who received only surgical treatment with MPhA the lifespan increased from 6.3 to 15.1 months. The combination of MPhA and surgery with chemotherapeutic drugs increased the patients’ lifespan from 5.4 to 14.3 months. At the same time in the groups using MPhA tumor markers carbohydrate antigen CA 19-9 and CEA levels were reduced. In addition, MPhA administration with conventional treatment of gastric cancer almost duplicated the number of polychemotherapy courses. So, MPhA can be used by cancer patients with conventional treatment to slow down the tumor process (secondary prevention) and decrease the side effects of chemotherapy (tertiary prevention) [23, 24].

As a result of MPhA use in patients with BPH a decrease (almost 3 times) in the age level of the chromosomal aberrations frequency in peripheral blood lymphocytes was also revealed. Administration of MPhA produced this parameter in lymphocytes of the patients lower than average population value in Russia and lower than that of the control groups without contacts with bad ecology as well as with industrial and household hazards. So, MPhA has proved itself as an adaptogen or geroprotector with an antimutagenic effect confirming in vitro studies. The hormone-modu-lating effect of MPhA with BPH was also shown which was expressed in a testosterone level increase and an estradiol level decrease in the blood serum. MPhA normalized the level of prostate-specific antigen. Urination and, consequently, the quality of patients’ life improved. So, consistent with the results obtained in high-cancer mice the effects in BPH improve the quality of patients’ life contributing to slowing down the development of age-related stress reactions, demonstrating the geroprotective effect of MPhA [33].

MPhA administration increased the efficacy of PD patients’ conventional therapy inhibiting daily and mo- tor activity violations as well as tremor, rigidity and bradykinesia according to UPDRS. Therefore, it was possible to taper gradually doses of L-DOPA-contain-ing preparations that are very toxic and provide side effects. Stress hormone cortisol level was inhibited too. So, the quality of PD patients’ life was improved. There are reasons to believe that taking into account in vivo studies MPhA has a neuroprotective effect restoring the dopamine-synthesizing function of reversibly damaged neurons and thus increasing the effectiveness of Parkinson’s disease complex pathogenetic therapy.

Thus, as a result of experimental and clinical studies it was found that MPhA has the properties of in- dividual adaptogens (with the absence of a tolerance effect) including immune (adhesive including) – and hormone-modulating, antimutagenic, antistress, neu-roprotective, antioxidant, radioprotective effects being a potential medication of preventive oncology, antitumor agent and geroprotector, showing a complex nature of action on the body [23, 24].

In silico analysis

We also carried out PASS (Prediction of Activity Spectra for Substances) predictions of biological activity spectra for each of MPhA 70 phytocomponents (Figure 2), integrated the obtained information using

Figure 2 .70 major phytocomponents selected in Multiphytoadaptogen composition.

PharmaExpert to estimate the probable additive/syn-ergistic effects, compared the predicted biological activities with known ones and determined the putative mechanisms of action caused by the observed phar-macotherapeutic effects. The results of the computer estimation correspond to the data obtained for MPhA in vitro, in vivo and in clinical studies. The in silico analysis also showed that MPhA is a potential preventive and antitumor agent against many tumor diseases as well as a geroprotector showing a complex effect on the body. In addition other activities are predicted that may be the subject of future research. The in sili-co analysis is an additional theoretical confirmation of the MPhA activity and substantiation of the relevance of adaptogenic medications for preventive oncology and aging diseases [25].

The algorithm of physicochemical, biological and computer methods (Pass2020, PharmaExpert) of analysis in quality control of phytoadaptogenic pharmaceutical compositions with a wide variety of structural compounds is an additional theoretical confirmation of the activity and justification of such drugs relevance for preventive oncology and age-related diseases.

Conclusion

Multiphytoadaptogen is approved in Russia as a parapharmaceutical agent and is consisted of 70 major components from 40 medicinal plants including Panax ginseng , Rhodiola rosea, Eleutherococcus senticos-us, Eucalyptus globules, Juniperus communis, Valeriana officinalis, Polygonum aviculare, Leonurus cardiac etc. Multiphytoadaptogen has been standardized by both analytical and biological methods. Pharmacological and toxicological characteristics of Multiphytoadapto-gen have been investigated in vitro , in vivo and in clinical studies. As a result of those investigations, a wide range of pharmacological effects (adaptogenic, immunomodulatory (adhesive including), antiviral, antitumor, antimutagenic, neuroprotective, antistress etc.) exhibited by Multiphytoadaptogen has been demonstrated.

Considering the possibility of Multiphytoadapto-gen regulating the dopaminergic neurons number in the brain with the participation of mutual adhesion of cells in the target tissue, beta2-leukocyte integrins and cytokine signaling reactivity, serum levels of the catabolic stress hormone corticosol as well as the frequency of tumors, survival and life quality of the organism it is probable to support the role of central neuronal 62 | Cardiometry | Issue 26. February 2023

processes and peripheral immune adhesion mechanisms in the control of malignant tumor formation.

Our researches open up the prospects for the creation of non-toxic regulatory medications for the effective prevention and treatment of neoplasms, age-related pathologies as well as for improving the people quality of life, increasing its duration and ameliorating the functions of body systems in conditions of multifactorial environmental influence [34].

Thus, starting from fundamental research on the violation of adhesive interactions in the target tissue during tumor formation, we came to phytoadapto-genic medications with adhesive action, which can be effective in preventive oncology and age-related pathologies.

So, fundamental oncology in essence, when it is created not for the sake of science itself, is not a brake at all, but an advance towards new prospects for effective treatment and rehabilitation of cancer and aging diseases patients.