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Ibragimov Shavkat Narzikulovich, Enikeeva Zulfiya Makhmudovna, Abdukhalilov Majid Makhamatkulovich, Saidkulov Bunyod Suyunovich, Narzikulova Kamola Shavkatovna, Republican Specialized Scientific-Practical Medical Center of Oncology and Radiology, Tashkent, Uzbekistan. Samarkand medical Institute, Uzbekistan E-mail: shavkat1967@inbox.ru
RADIOSENSITIZING EFFECT OF K-19 AND K-2 UPON THEIR INTRATUMORAL ADMINISTRATION TO ANIMALS WITH WALKER'S CARCINOSARCOMA STRAIN
Abstract: The efficacy of new antitumor drugs K-19 and K-2 was studied in rats with a transversal tumor strain of Walker's carcinosarcoma with intratumoral administration as substances that enhance the effect of radiation exposure. An evaluation of the efficacy showed that the combination of K-2 with irradiation causes an additive effect, Which is 22% higher than the effect of radiation, and K-19 increases the effect of irradiation by 25%.
Keywords: new anticancer drugs, K-2, K-19, radiosensitization, Walker's carcinosarcoma, intratumoral administration.
The application of antitumor drugs with properties to enhance the effect of radiation therapy seems to be the most promising direction in terms of increasing the effect of ionizing radiation [2; 3]. Antineoplastic drugs K-19 and K-2, which possess high antitumor activity [4; 5; 6], have been developed at the Republican Specialized Scientific and Practical Medical Center of Oncology and Radiology (RSSPMCOR), which are currently being studied for use in a new quality - radiosensi-tizers [7-10]. Data have been obtained that these substances have the property of enhancing the effect of irradiation when exposed to mice with transfused tumor strains of Sarcoma 180, a solid Ehrlich tumor, and rats with transfused tumor strains of Walker's carcinosarcoma (CSU) and Sarcoma 45 [7-10]. On the tumors of mice, the radiosensitizing effect of new drugs was studied and the ability of K-2 and K-19 preparations, applied for 16 hours before both single-dose irradiation at a dose of 4.5 Gy, and fractional irradiation at the same total dose (1.5 Gy x 3) The effect of irradiation is 32-42%. Also, a study of the radiosensitizing effect of drugs on rats with Sarcoma 45 tumors and Walker's carcinosarcoma revealed that new drugs applied for 16 hours before irradiation of 7Gy increase the effect of single irradiation on rats with KSU K-2 tumor by 33%, K-19 by 28%, and in rats with a Sarcoma 45 K-2 tumor at 49/56 and K-19 by 47/55%, while reducing the side effect of irradiation [10-12]. It is also necessary to evaluate the efficacy of the new antitumor drugs K-2 and K-19 as substances that enhance the effect of irradiation with intramedullary administration, since successful treatment of patients with intramedullary administration of such drugs as metronidazole
in combination treatment of rectal cancer [13] and usage of sanazol in the complex therapy of locally advanced breast cancer has been reported in the literature [1].
Aim ofresearch: Evaluation of the effectiveness of new antitumor drugs K-19 and K-2 with intratumoral administration in rats with an intertwined tumor strain ofWalker's carcinosarcoma as substances that enhance the effect of radiation exposure.
Materials and methods of research: The object of the study was the preparation K-19 and K-2, synthesized from colchamine and created in the laboratory for the development of antitumor drugs RSSPMCOR. All the experiments were carried out in accordance with the recommendations and requirements of the World Society for the Protection ofAnimals (WSPA) and the European Convention for the Protection of Experimental Animals (Strasbourg, 1986). In this work, 36 outbred rats weighing 77-90 g were used. The animals contained 4-6 individuals under the natural illumination regime with free access to water and food. Six animals were tested in experimental and control (with the infusion of physiological solution) groups. The study was performed on rats with a transversal tumor strain of Walker's carcinosarcoma, which was obtained from a tumor bank of the Russian Blokhin Cancer Research Center and passaged on donor rats, according to the strain protocol: the tumor is grafted subcutaneously with a suspension of tumor cells of 30-60 mg in 0.3-0.5 ml of nutrient medium per rat [18].
The study of the radiosensitizing effect ofthe inj ected preparations (together with irradiation) on animal tumor carriers was started on the 20th day after tumor inoculation. The drug
solution was injected into the developed tumors, the tumors were cut off from 3-4 sides, the preparations were administered at a dose of 0.3 ml per 100 g of the rat 16 hours prior to exposure to the irradiation. The irradiation was carried out on a gamma therapeutic device TENRATRON with a power of 1.25 MeV, a Co60 source in a total dose of 7 Gy, locally on the tumor. Each animal was attached to a special fixator, shielded with a lead block and a special hole in place of the tumor. Within 7 days after the drug was administered, the rats were euthanized by ether anesthesia using humane methods of working with laboratory animals [18]. The body mass of the animals was determined before and in the end of the experiment. In the course of the experiment to study the dynamics of tumor growth, tumor volumes were measured through the skin of animals in all groups (in 3 projections) at 10 and 17 days after tumor transplantation and during slaughter. At the end of the experiment, in slaughtered animals, the efficacy was determined by the volume (V) of the extracted tumor tissue, and by the tumor mass in the compared groups. The inhibition of tumor growth was calculated by the formulas [18]. The tolerability of the treatment was evaluated by the death of animals, for the indirect assessment of possible hematotoxicity in dead and killed rats, the weight of the spleen was determined. Time of observation: from the beginning of the experiment, i.e. transplantation of the tumor, to the slaughter of animals - 27 days.
Statistical processing was carried out using the program Statistica, version 6.0. p < 0.05 was taken for the level of statistical significance.
Results and discussion:
Effects on animals were evaluated in the following groups (drugs were administered intramondemally): 1 group - control received physiological solution, the 2nd group received irradiation 7Gy, the 3rd group received K-2 at a dose of 100 mg/kg once, in the 4th group the animals received K-2 at a dose of 100 mg/kg 16 hours prior to irradiation. In the 5th group, animals received K-19 in a dose of 50 mg/kg, in the 6th group, animals received K-19 at a dose of 50 mg/kg 16 hours before irradiation. An experiment on animals with the Walker's carcinosarcoma strain was started 20 days after transplantation according to the above schemes, tumor measurements were performed 10 and 7 days after transplantation, animals were slaughtered on day 7 after administration of the preparations and irradiation, the mass and volumes of tumors, and the mass of the spleen were measured. The death of animals during the experiment was in the groups receiving irradiation of 7 Gy, as well as in group 5 with the use of preparation K-19 (Table 1.).
Irradiation of 7 Gy caused an effect in 65%, (by volume and mass of tumors). When studying the effects of K-2 and K-19 preparations with a single intratumoral administration, it was found that they cause the effect in 50/45% and 65/64%, respectively. When K-2 was administered 16 hours before irradiation, the effect of suppressing tumor growth was 87/86%, which is higher than the irradiation effect of 7 Gy at 22/21%. K-19, administered 16 hours before irradiation, causes an effect of 90%, which increases the irradiation effect of 7 Gy by 25%.
Table 1.- The effect of intratumoral administration of K-19 in a dose of 50 mg / kg and K-2 in a dose of 100 mg/kg to rats with Walker's carcinosarcoma strain both independently and 16 hours prior to irradiation of 7 Gy (treatment from 20 days after tumor transplantation)
Group of animals Amount of animals/ case Mass of animals (g) Volume of tumor (sm3))) Mass of tumor (g) % inhibiton Mass of spleen (g)
Before experi-ment After experiment
On 10 day On 17 day On 27 day By volume By mass
1. control 6 88.8 ± 3.9 92.6 ± 22.4 10.3 ± 2.4 20.3 ± 6.15 57.1 ± 11.3 47.2 ± 0.73 1.7 ± 0.32
2. radiation 7gr 6/2 90.8 ± 3.3 52.2 ± 16.6 7.0 ± 1.1 17.5 ± 6.40 19.8 ± 3.2 16.3 ± 0.7 65 65 0.8 + 0.04
3. K-2100 mg/kg 6 79.2 ± 3.7 87.2 ± 18.0 4.0 ± 1.1 17.7 ± 3.45 28.6 ± 6.7 26.0 ± 6.3 50 45 1.1 +0.05
4. K-2100 mg/kg 16 hrs prior to radiation 7 gy 6 76.7 + 3.3 75.2 + 4.0 6.4 + 1.3 15.4 + 3.5 7.7 + 1.11 6.3 + 0.8 87 86 1.1 + 0.05
5. K-19 50 mg/kg 6/1 88.3 + 4.2 102.5 + 8.3 8.7 + 1.0 11.2 + 4.2 19.9 ± 6.9 16.8 ± 5.9 65 64 1.1 ± 0.13
6. K-19 50 mg/kg 16 hrs prior irradiation 7 gr 6 90.0 + 3.5 95.0 + 5.8 11.0 + 0.7 14.8 + 1.5 5.8 + 0.33 4.7 + 0.33 90 90 1.0 + 0.04
Note: * The differences are statistically significant in comparison with the control at P < 0.05. STG-inhibition of tumor growth by V/M (volume/mass)
In the experimental group with irradiation, the body weight of the animals was reduced by 42%, the spleen weight by 52%. In other experimental groups, body weight decreased only in group 4 (by 2%), and the weight of the spleen did not decrease as much as in the irradiation group: 35% in groups 3-5 and 41% in group 6, compared with control group.
70
60 50 40 30 20 10 0
The dynamics of tumor volumes is shown graphically (Fig. 1). In the control group of the tumor, from 17 to 27 days, increased by 3.8 times, in the 2-group, where irradiation at a dose of 7 Gy was applied, the tumors increased 1.13 times. The effect of K-19 reduced the tumors 2.5-fold 16 hours before irradiation. The effect of K-2 reduced the tumors 2-fold 16 hours before irradiation.
5 day
10 day
20 day
27 day
Figure 1. Dynamics of tumor growth of csu in treatment with irradiation, preparation k-19 and preparation k-19, applied 16 hours before irradiation intramammally
A study of the antitumor activity of drugs with a single in-tratumoral application on day 20 after transplantation showed that K-19, in comparison with K-2 in the transfected Walker's strain, exhibits a more significant antitumor effect with a difference of15/19%, which can be explained by a higher activity K-19, and its better solubility. However, radiosensitizing activity of K-19 with intramural injection in comparison with K-2 is only 3-4% higher in weight and volumes of treated tumors. The combination of K-2 with irradiation causes an additive effect, which is 22% higher than the effect of irradiation. K-19 increases the irradiation effect of 7Gy by 25%.
Thus, it was shown on the transfected Walker's tumor strain that both preparations possess a significant additive radiosensitizing effect upon intratumoral administration, causing a more significant (by 22-25%) antitumor effect than irradiation, while the effect of the preparations is accompanied by a reduction in the side effects caused by irradiation, such as decrease in body weight and spleen. It should be noted that intraperitoneal administration of K-19 preparation (equal in effectiveness to intravenous injection) 16 hours before irradiation causes a somewhat lesser effect 85/86% than with intu-tumoral injection, whereas in K-2, a more significant efficacy
(90%) is noted with intraperitoneal administration, therefore, the proposed intratumoral method of administration is a selection method.
Apparently, the detected ability of preparations K-2 and K-19 to suppress DNA synthesis and topoisomerase activity [7], explains their high antitumor activity and contributes to radiosensitizing activity. However, the ability to synchronize a tumor with mitotic activity and effect on DNA synthesis [9] causes the activity of new drugs as radiosensitizers. All this can contribute to their further clinical use both as cytostatics and as more effective radiosensitizers.
Conclusions:
1. The efficacy of the new antitumor drugs K-2 and K-19 was evaluated with an intratumoral single injection in rats with developed Walker's carcinosarcoma tumors, which were respectively 50/45 and 65/64%.
2. The efficacy of substances as radiosensitizers with intratumoral administration in rats with developed tumors of CSU16 hours before irradiation of 7 Gy was high and equal to 87-90%, an increase in the effect of irradiation with the introduction of K-2 by 22/21%, K-19 by 25% with a reduction in the side effects of irradiation.
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