Antitumoural Final

THE RUSSIAN ACADEMY of MEDICAL SCIENCES (RAMS) MEDICAL RADIOLOGIC SCIENTIFIC CENTRE (MRSC)

CONFIRM

Scientific Head of the company “Health Ltd”

affiliated with

                       MRCS of RAMS

Prof.  A. Konoplyannikov

REPORT

About studying of antitumoral action of the new seafood biologically active additive “Energolam Plus”

Author:

Leading scientific researcher of MRCS RAMS,

Candidate of biological sciences,

Dr. L.V.Shtein

02 March  2004

Obninsk City, Moscow Region, 2004

The given report is devoted to results of studying of possible antitumoral activity of the new, biologically active additive from seafood named «Energolam Plus». Earlier at studying of its biological activity there was applied the procedure of an assessment of radioprotective and haemostimulative action of the preparation under the test of influence for formation of endogenous splenic colonies at sublethally irradiated mice after concerning short (4-5 days) incorporation  of the given additive in a ration of laboratory animals, in this case — linear mice.

Test data received in these experience testified  a possibility of the given biologically active additive to activate a population of trunk hemopoietic cells, that can result in stimulation of hemopoietic and immune systems of animals and in some situations (at realization antitumoral therapy) to improve results of conducted therapy.

Therefore, an actual problem of studies was considered as realization of similar researches on model of animal’s treatment – tumour carriers. This approach can be most simply executed in experience on mice with subinoculated tumour. In parallel in experience on mice – tumour carriers it was necessary to verify influence of a reception of the given biologically active additive on growth of transplanted tumour cells and its ability to give metastases without realization of antitumoral therapy. At last, it was interested to receive data about a possibility of the given biologically active additive to influence on production of tumours at animals caused by influence of chemical carcinogen.

In the given report the results of experiments conducted in the described direction on studying of possible antitumoral activity of the preparation «Energolam Plus» are presented. In the first run of experience the studied additive daily per orally introduced to mice at the rate of 5 mg/kg of a body mass after slurrying in a normal saline solution and dilution up to concentration of 10 %. The preparation was introduced by a probe into a stomach daily within 7 day before a transplantation of a tumour and within 14 day after subinoculation of a tumour. In experiences mice of a line С57В1/6 were used, males in the age of 3 months, mass about 20-22 grams.

There were 15 mice in each experimental and control groups. Mice had been  intertwined (I / M) with 106 cells of (inoculum in volume 0,1 ml) intertwined tumour – Lewis carcinoma of lungs  (strain LLC) and after subinoculation during of 3 weeks regularly measured the dimensions of growing initial  tumour. In 21 days after subinoculation of a tumour the mice -tumour carriers were blocked, an initial tumour was excreted and weighed, and also lungs were fixed in the Buen’s solution for inventory under binocular magnifying glass  metastases of tumour keeping in them  (diameter more than 0,2 mm). These data allowed judging a possibility of a long influence of the biologically active additive on growth and innidiation of intertwined tumour.

The results of this run are indicated in the bellowing Table 1. As it is possible to see, on volume basis and mass of a tumour the authentic difference in experienced and control group is not revealed. Relatively weak, but statistically significant  the discrepancy between these two groups is revealed under the test of number of metastases  in lungs, accordingly, the average metastases in lungs at mice of control and experienced groups compounds 22,7 and 17,7 (р < 0,05). These interesting data can be considered as effect of immune system stimulation, which state, as it is known, can influence on effect of distant hematogenous innidiation.

At the same time similar immune system stimulation is not enough for suppression of growth of an initial tumour. The rather plausible assumption from here follows that the given biologically active additive is most perspective to use it in complex therapy of malignant neoplasm together with the agents capable to blast the basic cellular file of the initial tumour. This assumption has been verified in the second run.

Table 1

Influence of daily (21 days, 7 days of it up to a subinoculation of a tumour and 14 days after it) reception of the biologically active additive on dynamics of growth and innidiation of graft lung’s tumour Lewis (strain LLC):

In this run the subject of studies was effect of a locally conducted γ-irradiations in a doze of 15 Gr (device «Beam», γ -beams Со60, dosage rate about 50 cGr/min) on an initial tumour for 7 days after subinoculation of cells of Lewis lungs’ carcinoma (strain LLC), and including of the biologically active additive and the registration of results of experiences were the same as in the first run. There were also 15 mice in each group. The results of this series are indicated in Table 2.

As it is possible to see, the local single irradiation of a tumour in a doze of 15 Gr results in significant suppression of growth of a tumour and its innidiation in lungs. If the irradiation is carried out on a background of a reception of the biologically active additive, the effect of influence appreciably grows as on reaction of an initial tumour, and its metastases as well. These data have confirmed the stated above assumption that antitumoral effect of the given biologically active additive from seafood «Energolam Plus» is realized in the greatest measure at its incorporation in the circuit of complex therapy.

The further actual research problem in this direction undoubtedly is the receiving similar results in experiments, in which the action of the additive is combined with application of chemical preparations and operative expelling of a neoplasm or realization of the combined therapy. Besides that a number of practical questions demand the clarifications also, and in particular the questions that are connected to optimization of the schema of a combination of a reception of the biologically active additive and the basic medical influence directed on destruction of a ground mass of cells of initial locus, i.e., about those questions which now face in front of explorers who work in range of optimization of methods in an immunotherapy of tumours.

Table 2

γ-irradiation influence in a doze 15 Gr initial tumour unit of grafted  tumour LLC and daily (21 days, from them 7 days up to a subinoculation of a tumour and 14 day after it) reception of the biologically active additive on dynamics of growth   and an innidiation graft a tumour of Lewis lungs’ carcinoma (strain LLC).

The third series of experiment was devoted to studying of possible influence of the biologically active food additive on production of tumours caused by application of chemical carcinogen.  The model of a urethane carcinogenesis at mice of a genotype Swiss (on 30 mice in each group) for studying of effect of possible anticarcinogenic action of the biologically active food additive was used. Mice’s females in the age of 2 months were introduced with urethanes 5 times through each 3 days at the rate of 250 mg/kg of weight of a body.

Suspensions of the bioadditive at the rate of 5 mg/kg of mass of a body were introduced per orally to the animals of the basic experimental group within 4 months with 5 times per one week. In 6th month from a beginning of experiment animals were blocked, lungs were fixed and the number of the generated lungs adenomas was calculated. The results of this experiment are presented in the Table 3. The results of this run testify that the influence of the biologically active additive is capable to inhibit appreciably development of tumours, i.e., the given additive has expressed anticarcinogenic action. Certainly, the given conclusion further should find confirmation in experience with use of other types of chemical carcinogens.

Table 3

Studying of influence of long (5 times per one week during 4 months) per oral  reception of the biologically active additive from seafood «Energolam Plus» on quantity of lungs’ adenomas  at mice of a genotype Swiss, which were receiving and not receiving chemical carcinogen –urethane.

All received experimental data allow concluding the followings:

1. Long per oral introduction of the biologically active additive «Energolam Plus» by mice before and after transplantation of a tumour (Lewis lungs carcinomas) does not influence significantly on growth of initial tumour unit, but can inhibit a little metastatic spread of tumour cells in lungs.

2. Combination of a local irradiating of initial tumour unit and long per oral introduction of the biologically active additive «Energolam Plus» to mice up to and after a transplantation of a tumour (Lewis lungs’ carcinoma) results in amplification of medical action of ionizing radiation as on action on initial tumour unit, and on inhibition of metastatic spread of tumor cells in lungs.

3. On model of a urethane carcinogenesis at mice with a genotype Swiss it is revealed ability of long per oral introduction of the biologically active additive «Energolam Plus» to brake process of chemical carcinogenesis.

4. The received experimental data make actual the further experimental studying and clinical tests of biologically active additive «Energolam Plus» as perspective agent with properties of an adjuvant for antitumoral  therapy and agent capable to brake processes chemically sympathetic carcinogenesis.

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