RU2563369C1 - Method of ferrimagnetic-thermochemotherapy of malignant tumours with combinations of magnetocontrollable nanomedicines with visualisation of oncogenesis, detemination of therapy preferable in real time mode and monitoring of treatment results in experiment - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to experimental medicine and can be applied in early diagnostics and treatment of tumours, induced in experiment. For early MRT detection of tumours, invasions and metastases combinations of MRT-negative contrast nanopreparationss with positive MRT contrast preparations are introduced to animal. Diagnostics is carried out by contrast magnetic-resonance tomography (CM-RT). Ferrimagnetic-themochemotherapy (FT) of tumours with different localisation is carried out by activated combinations of magnetocontrollable nanomedicines. After-treatment of animas with metastases is carried out with combinations, including dacarbazine, decetaxel and cyclophosphane.

EFFECT: method makes it possible to improve average life expectancy to 340% with complete regression of tumours in 55% of animals.

6 ex, 7 tbl, 15 dwg

Description

The method of ferrimagnetic thermochemotherapy (FT) of malignant tumors by combinations of magnetically controlled nanopreparations (KMN) with visualization of oncogenesis, determination of therapy, preferred in real time (TP), and monitoring of treatment results in the experiment includes

- obtaining contrast magnetic resonance imaging images of oncogenesis (KM-RTI);

- definition of TP;

- suppression of tumor cell proliferation during FT;

- aftercare of infiltrative tumors with combinations of cytostatic drugs (PCC).

The invention relates to experimental medicine and can be used in the diagnosis of oncological diseases by contrast magnetic resonance imaging (MRI) using combinations of MRI negative contrast nanopreparations (MRI-NKN) with contrast MRI positive drugs and FT of tumors of different localization by activated KMN with treatment of infiltrative tumors of CC .

One of the methods for early diagnosis of tumors is to obtain CM-RTI by systemic administration of combinations of dextranferrite (DF) or citrate ferrite (CTF) with a magnevist [1] (analogue).

The disadvantages of the analogue:

- DF and CTF nanoparticles introduced systemically are concentrated in the organs of the cardiovascular system (CCC) and the reticuloendothelial system (RES);

- during the FT process, due to the high specific energy absorption (UAE), the DF and CTF nanoparticles are heated by an alternating magnetic field (PeMP) and destroy the organs of the CVS and RES, this leads to a decrease in the antitumor effect [2-8].

Known methods of treating tumors using cytostatic drugs: adenox (AO), methox (MO), melphalan (MF), sarcolysin (SL), cisplatin (CP), mitoxantrone (MK), dacarbazine (DC), docetaxel (DT) and cyclophosphamide (CF) [9-12] (analogue).

The disadvantages of the analogue:

- the lack of real-time monitoring of the distribution of PCC in the animal’s body (expectant tactics), leading to the accumulation of PCC in the CVS and RES, to a decrease in the antitumor effect [9-12];

- incomplete clearance of the body from tumor cells, leading to metastasis and relapse;

- MF and SL are hydrolyzed in aqueous media [9];

- weakly affect solid forms of tumors; carcinomas, melanomas, hepatomas, leukemia [10-12];

- non-concentrated and non-fixed in the tumor (scattered in the body) AO, MO, MF, SL, CP, MK, DK, DT and CF are concentrated in CCC and organs of RES from 20 to 40%, cause toxicosis, myelodepression and reduce the therapeutic effect [11 , 12].

A known method of induction hyperthermia of malignant tumors [5-7] (analogue).

The disadvantages of the analogue:

- uncontrolled distribution of indigenous peoples in animals, the impossibility of an objective determination of TP reduce the therapeutic effect.

A known method of thermochemotherapy of malignant tumors [4, 8] (analogue).

The disadvantages of the analogue:

- the uncontrolled distribution of indigenous peoples in animals and the impossibility of an objective determination of TP reduce the therapeutic effect.

Closest to the proposed method is a method of magnetohydrodynamic thermochemotherapy of malignant tumors with nanopreparations with magnetic resonance monitoring, in which the temperature inside the tumor rises from +47 to + 70 ° C. After heating is complete, the contents of the tumor are removed. After 6 sessions of magneto-hydrodynamic thermochemotherapy, tumors are not palpated. The use of the prototype leads to complete regression of tumors in 40% of animals with an increase in life expectancy up to 310% [2, 3] (prototype).

The disadvantages of the prototype:

- uncontrolled distribution of indigenous peoples in animals and the impossibility of objective determination of TP, reduce the therapeutic effect;

- in the tumor tissues there is an uncontrolled distribution of CMN, which leads to overheating of some and insufficient heating of other parts of the tumor and only to partial necrosis of tumor cells that penetrate deep into healthy tissues, this reduces the therapeutic effect;

- in the late stages of oncogenesis, after 2–3 months of remission, in most mice relapses of P388 lymphocytic leukemia, Ehrlich carcinoma, Lewis lung carcinoma, breast adenocarcinoma Ca 755 and melanoma B 16 are observed, as a result of invasion and metastasis of infiltrative tumors [2, 3] ;

- cytostatic agents, sarcolysin and melphalan, hydrolyze when heated under the stated conditions, therefore, DNA alkylation does not occur in all tumor cells [11];

- necrotic masses and drug residues, not completely sucked off, cause toxicosis, disrupt homeostasis, lower immunity, increase acute and chronic toxicity, this leads to a decrease in therapeutic effect, after 3-6 months of remission, death occurs as a result of impaired homeostasis, decreased immunity , invasion, metastasis and relapse of tumors [2, 3].

The objective of the invention is to remedy these disadvantages: optimizing the diagnosis, increasing the therapeutic effect and ensuring safety in the treatment of mammals.

The specified task is solved as follows:

- on 3-8 days after implantation of tumor cells in the tail vein, mice are injected with 30 to 350 μl of 0.5% sol of MRI-NKN of the General formula,

where 0 <y <1, 0 <x <1, 0 <z <1, L ³⁺ is the lanthanide [13, 14], (particle diameter 6-600 nm, citrate ions are used as a coating), specific magnetization (M _s ) up to 0.2 A · m ² / kg at a temperature of + 45 ° C in a field of 28.3 mT, with UAE 0.1-20 W / g Fe at a temperature of + 45 ° C in a field (frequency 0.01- 1.0 MHz, tension 7.2 kA / m-12.0 kA / m, power 0.15-1.5 kW) for early detection of tumors, metastases and borders of tumor cell invasion;

- the injection site is washed with warm water, wiped dry, the animal is kept at a temperature of + 25- + 28 ° C under standard conditions from 30 minutes to 30 hours, 10-20 μl of the magnevist are administered intravenously, after 6 minutes the animal is placed in BIOSPEC BC 70 biospectrotomograph / 30 USR (Bruker) and perform a KM-RT scan of the body in T ₁ -weighted modes {500/15 [repetition time, ms / echo time, ms] and T ₂ weighted (1900/80) spin echoes and T ₂ weighted gradient echo (500/15)} sequences. The signal intensity is measured and a visual analysis of the structure of the tumor and internal organs (lymphatic vessels, draining tumor, lymph nodes, spine, bones, lungs, kidneys, bladder, spleen and liver), which are usually affected by metastases of lymphocytic leukemia P388; Ehrlich carcinomas, Lewis lung carcinomas, Ca 755 adenocarcinomas of the mammary gland and B 16 melanoma. Obtain KM-RTI reflecting tumor development (Fig. 1 _{1, 2, 3, 4, 5, 6, 7, 8, 9, 10} ) and FIG. 1 ′ (a, b, c, d, e, f); 4 (a, b, c); 5 (a, b); 11 (a, b); 12 (a, b); 13 (a, b); 14 (a, b, c).

The animal is moved to the PeMP of the electronic sensor device (ESS) [17, 18]. An electronic-sensory scan of the animal’s body (ESS) is carried out, the sensor is moved along the surface of the tumor, and the whole body, from organ to organ, in the modes of sequential mutual projection of the sensor with the tumor tissues and with the tissues of the organs studied (Fig. 2, 3). In the process of scanning the body of an animal, the sensor emits a PeMP and receives electromagnetic responses in a continuous mode at combinatorial frequencies. Operators control the time taken to receive each response from a tumor, organs, body fluids and tissues. The values of the responses recorded by the computer from 2 seconds to 120 seconds determine the content of indigenous people in the tumor, organs, biological fluids and tissues in arbitrary units (ye). The results are used in formula (3) in determining TP [19].

where - _Tumor CMN - response of the KMN of the tumor;

- the _{sum of} indigenous _organs - the total response of indigenous _organs ;

- TP - the preferred therapy for this animal in real time;

- with TP≥10 - ferrimagnetic thermochemotherapy. For example, according to the results of an ESA of the animal’s body, the electromagnetic response of the tumor is ~ 9540 ye, liver ~ 42 ye, spleen ~ 45, kidney ~ 23, bladder ~ 30, the total response of the _organ CMN is 140 ye, according to formula (3), we obtain the value TP = ~ 647 , which indicates the preferable use of FT for this animal in real time, since the tumor contains KMN 647 times more than in the liver (Figs. 2, 3, 9).

To clarify the volume of the upcoming FT session, KM-RT scanning is performed in the following modes: T ₁ - weighted (TR / TE = 500/15 ms), T ₂ - weighted (TR / TE = 6000/63 ms) based on the spin echo and T ₂ - weighted gradient echo (TR / TE = 500/15 ms, angle of rotation 15 °).

The contents of the tumor are aspirated, 30 to 350 μl of 40% sol of KMN of the general formula are injected,

where 0 <y <1, 0 <x <1, 0 <z <1, L ³⁺ -Gd ³⁺ , La ³⁺ , Ce ³⁺ , Eu ³⁺ , Dy ³⁺ , Er ³⁺ , Yb ³⁺ [15, 16], (particle diameter from 5 to 900 nm, dextran of molecular weight from 10 to 70 kD, ferrite content 130 mg, M _s 8.5 kA / m, pH 6.4, ζ +15 are used as a coating mW, UAE 270-330 W / g Fe), which are obtained immediately before the introduction into the tumor by mixing the combinants DF, AO, MO, SL, MF, CP, MK, (ferrite from 2.6 to 150 mg; AO from 0.2 up to 2.0 mg, MO from 0.05 to 0.2 mg, CL or MF from 0.001 to 0.002 mg, CP from 0.01 to 0.04 mg, MK from 0.005 to 0.02 mg, M _s 40% KMN sol 8.5 kA / m, pH 7.0, ζ +13 mW, UAE 300 W / g Fe). The injection site is washed with warm water and wiped dry. Orient the animal so that the tumor is coaxial to the lines of force of an inhomogeneous constant magnetic field (NPMP) by induction of 0.15-4.0 T, and incubated for 3-6 minutes for concentration and fixation of CMN in the tumor. The tumor is oriented in PeMP (frequency 0.01–1.0 MHz, tension 7.2 kA / m –12.0 kA / m, power 0.15–1.5 kW) passing through a layer of water with a thickness of 0.01– 2.0 mm with a constant temperature, with the magnetic heating of the KMN, cytostatic drugs are activated. Simultaneously with heating, the temperature of the tumor and the body of the animal are continuously measured. After 3-15 minutes, the temperature inside the tumor rises to +44 - + 48 ° C, can withstand 30 minutes. Immediately after heating, the contents of the tumor are removed, the cavity formed at the site of the tumor is washed with a solution of antiseptic, the animal is placed in a PEMP ESSA, the magnitude of the electromagnetic responses of the formed cavity is determined, and PCC is administered .. KM-RT images of tissues that have insufficient blood flow remain for a long time light due to the slow diffusion of ferrite (Fig. 4 a, b, c; Fig. 5 a, b). The signal intensity is measured and a computer and visual analysis of tumor remnants of changes in macro- and micro-metastases of P388 lymphocytic leukemia is performed; Ehrlich carcinomas, Lewis lung carcinomas, mammary adenocarcinomas Ca 755 and melanoma B 16. The results of FT (Fig. 4 b, c) are compared with CM-RTI obtained before FT (Fig. 4a). Every 3-48 hours, depending on the somatic state of the animal (PSF), the ESA procedure, contrast magnetic resonance imaging tomography (KM-RTS) and FT are repeated from 2 to 8 sessions depending on the FSH.

The tumor at this stage of FT is usually not felt (Fig. 6). CCP is introduced into the cavity and subcutaneous metastases. When ascites and metastases of internal organs are detected, PCC is administered intraperitoneally. In parallel, a biopsy and micropreparations are done (Fig. 7 a, b, c, d) (Fig. 8).

Evaluation of the antitumor activity and the quality of aftercare is carried out in accordance with the results of an ESA and KM-RT scan, the conclusion is made taking into account the recommendations for chemotherapy of malignant tumors [12]. All the data obtained are processed statistically, the differences are taken for reliable at ρ <0.05 [20].

The claimed method is illustrated by examples 1-7

Example 1. The antitumor effect of the claimed method on the subcutaneous form of P388 lymphocytic leukemia, depending on the type and number of therapeutic procedures, expressed in complete regression of tumors and an increase in average life expectancy (VL) of mice.

A suspension of 10 ⁶ P388 lymphocytic leukemia cells is injected into 70 BDF ₁ male mice under the skin of the right scapula, 60 mice are selected and divided into 5 groups: 20 mice in group 1, 10 mice in 2, 3, 4 and 5. ESS, KM-RT scanning and FT begin when the average tumor volume reaches 30-145 mm ³ . Group 1 mice under anesthesia along the perimeter of the tumor are injected with 350 μl of a 0.9% NaCl solution and an ESA is performed. The electromagnetic tissue responses in mice of the first (control) group correspond to the background level of KMN content (~ 11 ye), according to which the number of KMN is not determined on the calibration graph (Fig. 9). On KM-RTI, with T ₁ weighted images (VI) (TR / TE = 500/15 ms), T ₂ VI (TR / TE = 6000/63 ms) based on the spin echo and T ₂ -weighted gradient echo (TR / TE = 500/15 ms, rotation angle 15 °). At the injection sites of a 0.9% NaCl solution, no images were detected.

The mice of group 2 are sucked off the contents of the tumor, along the perimeter of the tumor are injected with PCC: MK 0.01 mg, CL 0.002 mg, 0.9% NaCl up to 350 μl. The mice of group 3 are sucked off the contents of the tumor, a sol is injected along the perimeter of the tumor: DF 150 mg, water up to 350 μl. When ESA, tissue responses in mice of the third group correspond to the content of KMN 15260 ye, the sum of the responses of the liver and other organs is 150 ye, according to which the content of KMN in mg is determined on the calibration graph (Fig. 9). According to the formula 3, TP = 102 is obtained, which confirms the preference for FT for a given animal in real time. On KM-RTI of the mouse body, dark spots of KMN are revealed. Mice 4 groups sucked off the contents of the tumor. KMN is administered along the tumor perimeter: DF 150.0 mg, MK 0.01 mg, SL 0.002 mg, water up to 350 μl. When ESM mice of the fourth group receive electromagnetic responses: tumors 10890, the sum of the responses of the liver and other organs corresponds to 210. According to formula 3, get the value, TP = 52, confirming the preference for FT for this animal in real time. On KM-RTI of the mouse body, with the same scanning parameters, images of dark spots corresponding to the location of the KMN are detected.

Group 5 mice are aspirated for tumor contents. KMN is administered along the tumor perimeter: DF 150 mg, MK 0.01 mg, SL 0.002 mg, water up to 350 μl (M _s 7.5 kA / m, pH 6.6, ζ +15 mW, UAE 330 W / g Fe ) The electromagnetic responses of tissues in mice of the fifth group: tumors 15630, the sum of the responses of the liver and other organs 50 (Fig. 9). By the formula 3 get the value of TP = 313.

Mice of groups 1, 2, 3, 4 and 5 are placed in a constant magnetic field by induction of 0.2 T, a gradient of 0.015 T / cm and incubated for 5 minutes. The tumors of each mouse of groups 1, 2, 3, 4 and 5 are oriented in PeMP (frequency 0.01–1.0 MHz, intensity 12.0 kA / m, power 0.15 kW) passing through a 2.0 layer of water 2.0 mm with constant temperature (magneto-thermostating), introduced KMN is heated in PeMP. The tumor temperature in mice of groups 1 and 2 under the action of PeMP for 5 min does not rise above + 38 ° C. The temperature in the rectum and tumor of mice of groups 1 and 2 for 30 minutes is between +37 - + 38 ° C. When ESS and KM-RT scanning KMN is not detected.

In mice of groups 3 and 4, the temperature of the tumor in PeMP in 5 min rises above + 43 ° C (the temperature is measured with an alcohol thermometer). Further, the temperature inside the tumor continues to increase and after 15 minutes it reaches + 47 ° С, the temperature of the rectum + 38 ° С. On KM-RTI, images of dark spots corresponding to the places of introduction of KMN are revealed. After the heating of the tumor, its contents are removed.

Under the action of PeMP, the temperature of the tumor in mice of the 5th group rises above + 43 ° C within 5 minutes. The temperature inside the tumor continues to rise and after 15 minutes it reaches + 47 ° С, of the rectum + 38 ° С. After the heating of the tumor, its contents are removed. To remove the products of necrosis of tumor tissues and residues of CMN, the cavity formed at the site of the tumor is washed with a 0.05% formaldehyde solution in a 0.9% NaC solution, and PCC is introduced into the cavity. In groups 1, 2, 3, 4, and 5, up to 6 ESS procedures, up to 8 KM-RT scan procedures, and up to 8 FT procedures are performed (Table 1).

As a result of KM-RT scanning, the following are revealed:

- in 1 group of mice, the volume of the tumor and metastases in each mouse steadily increases without signs of regression; during 6 FT procedures, all mice die from tumor progression;

- in the second group, tumor growth is inhibited, metastases are detected in the axillary and inguinal regions of 4 mice. CCT is introduced into metastases and tumors: DT 0.05 - DK 0.1 - TF 1.0 mg in order to suppress them. Complete tumor regression 15%, increase in life expectancy of 150%;

- in the third group, the average volume of tumors is significantly reduced, metastases are revealed in the inguinal region of 3 mice, QPCs are introduced into metastases and tumors: DT 0.05 - DK 0.1 - CF 1.0 mg, complete regression of the tumor 15%, increase in life expectancy 160%

- in the fourth group, the average volume of tumors from the first day of treatment decreases sharply, metastases are revealed in the inguinal region of 2 mice, PCP is introduced into metastases and tumors: DT 0.05

- DC 0.1 - CF 1.0 mg, complete tumor regression 30%, increase in life expectancy 180%;

- in the fifth group, the average volume of tumors decreases from the first day of treatment, but metastases are detected in the inguinal region of 1 mouse, PCPs are introduced into metastases and tumors: DT 0.05 - DK 0.1 - CF 1.0 mg, complete tumor regression 40% , increase in life expectancy of 250%, (table 1).

Thus, a KM-RT scan of animals provides early detection of tumors and metastases by contrasting tumors and metastases with a combination of DF magnevist.

In ESA, the electromagnetic responses of tissues in mice of the fifth group correspond to elevated levels of CMN, which determine the content of CMN in a tumor, liver and other organs of RES in real time: in a tumor of 45,000, the sum of the responses of the liver and organs of 150, according to formula 3, the TP = 300, confirming the preference for FT for this animal in real time, on the calibration graph (Fig. 9) receive the content of KMN in mg.

Magnetic heating of the animal’s body under conditions of an increase in the rate and magnitude of heating of tumor tissues containing indigenous peoples, removal of the contents of the tumor, subsequent washing of the formed cavity, introduction of cytostatic agents into metastases as they are detected, leads to a rapid decrease in the volume of tumors, their complete regression and increase in duration the life of mammals.

KM-RT scan of the cavity formed after heating and suctioning the contents of the tumor, followed by washing with an antiseptic, helps to identify the remains of tumors and metastases, targeted controlled introduction of KMN into the remaining part of the tumor and metastases in the process of aftercare of animals.

Preferred:

- suction of the contents of the tumor to lower the pressure before the introduction of indigenous peoples and antiseptics;

- preliminary, intermediate and final magnetic resonance scans of the animal’s body for visual analysis of tumors and metastases with an individual conclusion based on FT results;

- ferrimagnetic concentration, fixation and heating of drugs in the tumor;

- FT of the tumor, depending on the FSH at + 45-48 ° C 4 × 30 min with an interval of 3 hours, with washing the cavity formed at the site of the tumor, from the products of necrosis and KMN residues with the introduction of CCC into the cavity and metastases in which the maximum therapeutic effect (table 1).

Example 2. The antitumor effect of the claimed method on the subcutaneous form of P388 lymphocytic leukemia, depending on the type and number of procedures.

A suspension of 10 ⁶ P388 lymphocytic leukemia cells is injected into 80 male BDF ₁ mice under the skin of the right scapula, 70 mice are selected and divided into 6 groups: 20 mice in group 1, 10 mice in 2, 3, 4, 5 and 6. 3-4 days after implantation, when the average tumor volume reaches 50-150 mm ^3, its contents are sucked off to lower the pressure in the tumor. Mice of group 1 along the perimeter of the tumor are injected with 350 μl of a 0.9% NaCl solution.

Mice of group 2 along the perimeter of the tumor are injected with AO 1.2 mg, 0.9% NaCl to 350 μl. Mice of group 3 along the perimeter of the tumor are injected with CCP: AO 1.2 mg, MO 0.2 mg, 0.9% NaCl up to 350 μl. Mice of the 4th group along the perimeter of the tumor are injected with 150 mg DF, water up to 350 μl. Mice of the 4th group along the perimeter of the tumor are injected with KMN: DF 150 mg, AO 1.2 mg, MO 0.2 mg, water up to 350 μl. Mice of group 5 along the perimeter of the tumor are injected with KMN: DF 150 mg, AO 1.2 mg, MO 0.2 mg, water up to 350 μl. Mice of the 6th group along the tumor perimeter are injected with KMN: DF 150 mg, AO 1.2 mg, MO 0.2 mg, water up to 350 μl (M _s 7.5 kA / m, pH 6.6, ζ + 15 mW, UAE 330 W / g Fe).

The FT procedure of mice of groups 1, 2, 3, 4, 5, and 6 is carried out similarly to Example 1: FT 1 and 2 groups are performed 6 times, 3 and 4 groups are performed 8 times, 5 groups - 6 times, 6 groups - 8 times an interval of 3 hours to 3 days, depending on the FSH (table. 2).

The results of FT mice of groups 1, 2, 3, 4, 5, and 6 are presented in Table. 2.

Thus, in accordance with Table 2, the maximum therapeutic effect is achieved by FT of the tumor at + 48 ° C for 8 × 30 min, followed by washing of the tumor from the products of necrosis and KMN residues with the introduction of PCC into the metastases detected during treatment.

Example 3. The antitumor effect of the claimed method on the subcutaneous form of Ehrlich carcinoma, depending on the number of procedures.

A suspension of 10 ⁶ Ehrlich carcinoma cells is injected into 80 male BDF1 mice under the skin of the right scapula, 70 mice are selected and divided into 6 groups: 20 mice in group 1, 10 mice in 2, 3, 4, 5 and 6. When the average tumor volume reaches 30-130 mm ³ (from 3 to 5 days), its contents are sucked off to lower the pressure in the tumor. The temperature in the rectum is measured in mice. Mice of the 1st (control) group were injected into the tumor with 350 μl of a 0.9% NaCl solution. Mice of group 2 along the perimeter of the tumor are injected with CCP: MF 0.002 mg, MK 0.02 mg, 0.9% NaCl up to 350 μl. Mice of the 3rd group along the perimeter of the tumor are injected with 150 mg DF in 350 μl of water. Mice of 4 groups along the perimeter of the tumor are injected with KMN: DF 150 mg, MF 0.002 mg, MK 0.02 mg, water up to 350 μl. Mice of group 5 along the perimeter of the tumor are injected with KMN: DF 150 mg, MF 0.002 mg, MK 0.02 mg, water up to 350 μl. Mice of the 6th group along the tumor perimeter are injected with KMN: DF 150 mg, MF 0.002 mg, MK 0.02 mg, water up to 350 μl (M _s 7.5 kA / m, pH 6.6, ζ + 15 mW, UAE 330 W / g Fe). Mice were placed for 5 minutes in a constant magnetic field by induction of 0.2 T, a gradient of 0.015 T / cm. Each mouse 2, 3, 4, and 5 groups are placed in PeMP, as in 1 example. In metastases, as they are detected, enter the PCC: DT 0.05 - DK 0.1 - TF 1.0 mg.

FT procedure is carried out:

- mice of groups 1 and 3 4 times with an interval of 3 hours to 3 days, depending on the FSH (table. 3);

- mice 4 groups 3 times with an interval of 3 hours to 3 days depending on the FSH (table. 3);

- mice of groups 2 and 5 6 times with an interval of 3 hours to 3 days, depending on the FSH (table. 3);

- mice of group 5 8 times with an interval of 3 hours to 3 days, depending on the FSH (table. 3).

The result is:

- in 1 (control) group of mice, the tumor volume in each mouse steadily increases without signs of regression;

- in the second group, the average volume of tumors decreases slowly, metastases are detected in 3 mice, PCP is administered intraperitoneally, DT 0.05 - DC 0.05 - CF 1.0 mg, an increase in life expectancy of 180%;

- in the third group, the average volume of tumors is significantly reduced, metastases are detected in 3 mice, treated with intracavitary and intraperitoneal administration of CPC, DT 0.05 - DC 0.05-TF 1.0 mg, complete tumor regression is observed in 10% of mice, an increase in duration mouse life increase in life expectancy of mice 200%;

- from the start of treatment in the fourth group of mice, a decrease in the average volume of tumors is observed, and a complete regression of tumors is observed in some mice;

- after completion of treatment, the signs of tumors do not appear for 3 months, metastases in axillary lymph nodes are detected in 3 mice, the mice are treated with intracavitary and intraperitoneal administration of CPC, DT 0.05 - DC 0.05 - CF 1.0 mg, complete tumor regression observed in 30% of mice, an increase in the life expectancy of mice 320%;

- in the fifth group, metastases are detected in 1 mouse, aftercare is carried out by intracavitary and intraperitoneal administration of CPC, DT 0.05 - DC 0.05 - CP 1.0 mg, complete regression of the tumor is observed in 30% of mice, an increase in the life expectancy of mice is 320%;

- in the sixth group, 8 FT procedures are performed, metastases are detected in 2 mice, aftercare is carried out by intracavitary and intraperitoneal administration of CPC, DT 0.05 - DC 0.05 - CF 1.0 mg, complete tumor regression is observed in 50% of mice, an increase in duration mouse life 350%;

Thus, in accordance with example 3, the tumor FT is preferable at + 48 ° C for 8 × 30 min with an interval of 3 hours to 3 days depending on the FSH, followed by washing the formed cavity with a 0.05% formaldehyde solution in 0.9% NaCl solution, and the introduction of CCP, in which receive the maximum therapeutic effect.

The experimental results (examples 1-3, tables 1-3) confirm the high antitumor effect of the claimed method, including regional administration of indigenous peoples with simultaneous heating of the body to 38 ° C with heating of the tumor predominant in speed and magnitude to + 45- + 48 ° C and subsequent removing the contents of the tumor, washing the formed cavity, treating with intracavitary and intraperitoneal administration of CPC: DT 0.05 - DC 0.05 - TF 1.0 mg.

Thus, the ES determination of TP and KM-RT monitors the results of FT of tumors and metastases with simultaneous magnetic heating of the body with heating of tumor tissues predominant in speed and magnitude containing KMN, followed by washing the cavity at the site of the tumor from necrosis products and KMN residues, with additional intracavitary and intraperitoneal administration of PCC leads to a rapid decrease in the volume of tumors, their complete regression and increase the life expectancy of mammals.

Tumor FT is preferable at + 45- + 48 ° C for 8 × 30 min with an interval of 3 hours to 3 days, depending on the FSH, with treatment with intracavitary and intraperitoneal administration of CPC, which gives the maximum therapeutic effect (Table 3).

Example 4. The antitumor effect of the claimed method on the subcutaneous form of Lewis lung carcinoma, depending on the type and number of procedures.

A suspension of 10 ⁶ Lewis lung carcinoma cells was injected into 80 muscles of C57Bl / 6j male mice in the right thigh muscle, 70 mice were selected and divided into 6 groups: 20 mice in group 1, 10, 10, 2, 3, 4, 5 and 6 mice. When the average tumor volume reaches 30-130 mm (about 8 days), its contents are sucked off to lower the pressure in the tumor. In mice, the temperature in the rectum is measured. Mice of the 1st (control) group were injected into the tumor with 350 μl of a 0.9% NaCl solution. Mice of group 2 along the perimeter of the tumor are injected with CPC: CP 0.03 mg, MK 0.01 mg, 0.9% NaCl up to 350 μl. Mice of the 3rd group along the perimeter of the tumor are injected with 150 mg DF in 350 μl of water. Mice of the 4th group along the perimeter of the tumor are injected with KMN: DF 150 mg, CP 0.03 mg, MK 0.01 mg, water up to 350 μl. Mice of group 5 along the perimeter of the tumor are injected with KMN: DF 150 mg, CP 0.03 mg, MK 0.01 mg, water up to 350 μl. Mice of the 6th group along the tumor perimeter are injected with KMN: DF 150 mg, CP 0.03 mg, MK 0.01 mg, water up to 350 μl (M _s 7.5 kA / m, pH 6.6, ζ + 15 mW, UAE 330 W / g Fe). Mice were placed for 5 minutes in a constant magnetic field by induction of 0.2 T, a gradient of 0.015 T / cm. Each mouse of groups 2, 3, 4, 5, and 6 was placed in PeMP, as in 1 example. It is treated with intracavitary and intraperitoneal administration of CCP: DT 0.05 - DK 0.1 - TF 1.0 mg.

FT procedure is carried out

- mice of groups 1, 2, 3 and 5 6 times with an interval of 3 hours to 3 days, depending on the FSH (table. 4);

- mice 4 groups 4 times with an interval of 3 hours to 3 days depending on the FSH (table. 4);

- mice of group 5 8 times with an interval of 3 hours to 3 days, depending on the FSH (table. 4).

The result is

- in 1 (control) group of mice, the tumor volume in each mouse steadily increases without signs of regression;

- in the second group, the average volume of tumors decreases slowly, metastases are detected in 3 mice, treated with intracavitary and intraperitoneal administration of CPC, DT 0.05 - DC 0.1 - CF 1.0 mg, an increase in life expectancy of 180%;

- in the third group, the average volume of tumors is significantly reduced, metastases are detected in 2 mice, treated with intracavitary and intraperitoneal administration of CPC, DT 0.05 - DC 0.1 - CF 1.0 mg, complete tumor regression is observed in 10% of mice, increase in duration mouse life; an increase in mouse life expectancy of 160%;

- in the fourth group, the average volume of tumors decreases from the first day of treatment and their symptoms do not appear for 3 months, metastases in axillary lymph nodes are detected in 3 mice, they are treated with intracavitary and intraperitoneal administration of CPC, DT 0.05 - DC 0.1 - CF 1.0 mg. Complete tumor regression is observed in 10% of mice; an increase in the life span of mice is 220%;

- in the fifth group, metastases are detected in 1 mouse, treated with intracavitary and intraperitoneal administration of CPC, DT 0.05 - DC 0.05 - CF 1.0 mg, complete regression of the tumor is observed in 20% of mice, an increase in the life expectancy of mice is 240%;

- in the sixth group, 8 FT procedures are performed, metastases are detected in 2 mice, treated with intracavitary and intraperitoneal administration of CPC, DT 0.05 - DC 0.05 - CF 1.0 mg, complete tumor regression is observed in 25% of mice, an increase in life expectancy mice 260%;

Thus, in accordance with example 4, the tumor FT is preferable at + 48 ° C for 8 × 30 min with an interval of 3 hours to 3 days depending on the FSH, followed by washing the formed cavity with a 0.05% formaldehyde solution in 0.9% NaCl solution, and the introduction of CCP, in which receive the maximum therapeutic effect.

The experimental results (examples 1-4, tables 1-4) confirm the high antitumor effect of the claimed method, including regional administration of indigenous peoples with simultaneous heating of the body to 38 ° C with heating of the tumor predominant in speed and magnitude to + 45- + 48 ° C and subsequent removing the contents of the tumor, washing the formed cavity, treating with intracavitary and intraperitoneal administration of CPC: DT 0.05 - DC 0.05 - TF 1.0 mg.

Thus, the ES determination of TP and KM-RT monitors the results of FT of tumors and metastases with simultaneous magnetic heating of the body with heating of tumor tissues predominant in speed and magnitude containing KMN, followed by washing the cavity at the site of the tumor from necrosis products and KMN residues, with additional intracavitary and intraperitoneal administration of PCC leads to a rapid decrease in the volume of tumors, their complete regression and increase the life expectancy of mammals.

Tumor FT is preferable at + 45- + 48 ° C for 8 × 30 min with an interval of 3 hours to 3 days, depending on the FSH, with treatment with intracavitary and intraperitoneal administration of CPC, which gives the maximum therapeutic effect (Table 4).

Example 5. The antitumor effect of the claimed method on the subcutaneous form of breast adenocarcinoma Ca 755, depending on the type and number of procedures.

A suspension of 10 ⁶ cells of Ca 755 breast adenocarcinoma is injected into 80 female C57Bl / 6j female mice under the skin of the right axillary region, 70 mice are selected and divided into 6 groups: 20 mice in group 1, 10, 10, 2, 3, 4, 5 and 6 mice . When the average tumor volume reaches 30-130 mm ³ (about 4 days), the temperature in the rectum is measured in mice. To lower the pressure in the tumor, its contents are aspirated. Mice of the 1st (control) group were injected into the tumor with 350 μl of a 0.9% NaCl solution. Mice of group 2 along the perimeter of the tumor are injected with melphalan (MF) 0.002 mg and mitoxantrone (MK) 0.02 in 350 μl of 0.9% NaCl. Mice of the 3rd group along the perimeter of the tumor are injected with dextranferritis (DF) 150 mg, water up to 350 μl. Mice of 4 groups along the perimeter of the tumor are injected with KMN: DF 150 mg, MF 0.002, MK 0.02 mg, water up to 350 μl. Mice of group 5 along the perimeter of the tumor are injected with KMN: DF 150 mg, MF 0.002, MK 0.02 mg, water up to 350 μl. Mice of group 6 along the perimeter of the tumor are injected with KMN: DF 150 mg, MF 0.002, MK 0.02 mg, water up to 350 μl. Mice were placed for 5 minutes in a constant magnetic field by induction of 0.2 T, a gradient of 0.015 T / cm. Each mouse of groups 2, 3, 4, and 5 is placed in PeMP, as in Example 1. The tumor temperature in mice of groups 2, 3, 4, and 5, rises above + 42 ° C for 5 minutes under the influence of PeMP, after 15 min the tumor temperature reaches + 48 ° C, after 30 minutes the heating is stopped. At the same time, the temperature in the rectum and tumor of group 1 mice remains between + 37- + 38 ° C. After heating, remove its contents. The cavity formed at the site of the tumor is washed with a 0.3% formaldehyde solution in a 0.9% NaCl solution.

FT procedure is carried out

- mice of groups 1 and 5 6 times with an interval of 3 hours to 3 days, depending on the FSH (table. 5);

- mice of groups 2, 3 and 4 4 times with an interval of 3 hours to 3 days, depending on the FSH (table. 5);

- group 5 mice 6 times with an interval of 3 hours to 3 days, depending on the FSH (table. 5).

- mice of group 6 8 times with an interval of 3 hours to 3 days, depending on the FSH (table. 5).

The result is

- in 1 (control) group of mice, the tumor volume in each mouse steadily increases without signs of regression;

- in the second group, the average tumor volume decreases slowly, metastases are detected in 3 mice, treated with intracavitary and intraperitoneal administration of CPC, DT 0.05 - DC 0.05 - CF 1.0 mg, an increase in life expectancy of 160%;

- in the third group, the average volume of tumors is significantly reduced, metastases are detected in 3 mice, treated with intracavitary and intraperitoneal administration of CPC, DT 0.05 - DC 0.05 - CF 1.0 mg, an increase in the life expectancy of mice is 200%;

- in the fourth group, the average volume of tumors decreases from the first day of treatment, their signs do not appear for 3 months, metastases in axillary lymph nodes are detected in 2 mice, they are treated with intracavitary and intraperitoneal administration of CPC, DT 0.05 - DC 0.05 - CF 1.0 mg, complete tumor regression is observed in 30% of mice, an increase in the life expectancy of mice is 220%;

- in the fifth group, in one mouse, tumor metastasis is detected in the right groin, treated with intracavitary and intraperitoneal administration of CPC, DT 0.05 - DC 0.05 - CF 1.0 mg, complete tumor regression is observed in 30% of mice, an increase in the life span of mice 280%;

- in the sixth group, metastases in the right groin are detected in one mouse, treated with intracavitary and intraperitoneal administration of CPC, DT 0.05 - DC 0.05 - CF 1.0 mg, complete regression of the tumor is observed in 50% of mice, the increase in life expectancy of mice 350 %

Thus, in accordance with example 5, determining the type of therapy preferable in real time, preliminary, intermediate and final magnetic resonance scans of tumors and metastases with an individual conclusion according to the results of FT, simultaneous magnetic heating of the body with predominant heating and speed of tumor tissues containing KMN, removal of necrotic masses and washing of the cavity formed at the site of the tumor, the introduction of a combination of cytostatic agents into metastases leads to fast a decrease in the volume of infiltrative tumors, their complete regression, and an increase in the lifespan of mice. In breast adenocarcinoma Ca 755, FT is preferable at + 47- + 48 ° C for 8 × 30 min with an interval of 3 hours to 3 days depending on the FSH, with the introduction of metastases and PCC cavity in which they obtain the maximum therapeutic effect (Table 5) .

Example 6. The antitumor effect of the claimed method on the subcutaneous form of melanoma B 16 depending on the type and number of procedures.

80 female C57Bl / 6j mice were injected with a suspension of 10 ⁶ melanoma cells B 16 in the right axillary region, 70 mice were selected and divided into 6 groups: 20 mice in 1 group, 10 mice in 2, 3, 4, 5 and 6. When the average tumor volume reaches 30-130 mm ³ (about 6 days) in mice, the temperature in the rectum is measured and the contents of the tumor are aspirated. Mice of the 1st (control) group were injected into the tumor with 350 μl of a 0.9% NaCl solution. In the mice of the 2nd group, the contents of the tumor are aspirated, along with the perimeter of the tumor, bis (L-malato) IV oxovanadium (BMOV) 6.0 mg in 350 μl of water is injected. In the mice of the 3rd group, the contents of the tumor are sucked off; along the perimeter of the tumor, PCCs are administered: BMOV 6.0 mg and MK 0.02 mg in 350 μl of water. In mice of the 4th group, the contents of the tumor are aspirated, 150 mg of DF in 350 μl of water are injected along the perimeter of the tumor. In the mice of group 5, the contents of the tumor are aspirated, KMN is administered along the tumor perimeter: DF 150 mg, BMOV 6.0 mg and MK 0.02 mg in 350 μl of water. In the mice of group 6, the contents of the tumor are sucked off, along with the perimeter of the tumor are injected KMN: DF 150 mg, BMOV 6.0 mg and MK 0.02 mg in 350 μl of water.

Mice were placed for 5 minutes in a constant magnetic field by induction of 0.2 T, a gradient of 0.015 T / cm. Each mouse of groups 2, 3, 4, 5, and 6 is placed in PeMP, as in Example 1. The tumor temperature in mice of groups 2, 3, 4, and 5, rises above + 42 ° C for 5 min under the action of PeMP 15 minutes the tumor temperature reaches + 48 ° C, after 30 minutes the heating is stopped. At the same time, the temperature in the rectum and tumor of group 1 mice remains between + 37- + 38 ° C. After heating, the contents of the tumor are removed. The cavity formed at the site of the tumor is washed with a 0.3% formaldehyde solution in a 0.9% NaCl solution.

FT procedure is carried out

- mice of groups 1 and 5 6 times with an interval of 3 hours to 3 days, depending on the FSH (table. 6);

- mice of 2, 3, 4 and 8 groups 8 times with an interval of 3 hours to 3 days, depending on the FSH (table. 6);

The result is

- in 1 (control) group of mice, the tumor volume in each mouse steadily increases without signs of regression;

- in the second group, the average volume of tumors decreases slowly, metastases are detected in 3 mice, treated with intracavitary and intraperitoneal administration of CPC, DT 0.05 - DC 0.05 - CF 1.0 mg, complete tumor regression is observed in 10% of mice, increase in duration mouse life 160%;

- in the third group, the average volume of tumors decreases slowly, metastases are detected in 1 mouse, treated with intracavitary and intraperitoneal administration of CPC, DT 0.05 - DC 0.05 - CF 1.0 mg, complete tumor regression is observed in 20% of mice, an increase in duration mouse life 200%;

- in the fourth group, the average volume of tumors decreases from the first day of treatment and, finally, their symptoms do not appear for 3 months,

- in the fifth group, metastases in axillary lymph nodes are detected in 2 mice, treated with intracavitary and intraperitoneal administration of CPC, DT 0.05 - DC 0.05 - CF 1.0 mg, complete tumor regression is observed in 30% of mice, an increase in the life span of mice 240%;

- in the sixth group, metastases in axillary lymph nodes are detected in 2 mice, treated with intracavitary and intraperitoneal administration of CPC, DT 0.05 - DC 0.05 - CF 1.0 mg, complete regression of the tumor is observed in 40% of mice, an increase in the life span of mice 250%;

Thus, in accordance with example 6, determining the type of therapy preferable in real time, preliminary, intermediate and final magnetic resonance scans of tumors and metastases with an individual conclusion according to the results of FT, simultaneous magnetic heating of the body with predominant heating and speed of tumor tissues containing KMN, removal of necrotic masses and washing of the cavity formed at the site of the tumor, aftercare with intracavitary and intraperitoneal administration of CPC: DT 0.05 - DK 0.05 - CF 1.0 mg, leads to a rapid decrease in the volume of infiltrative tumors, their complete regression and increase the life expectancy of mice. For melanoma B 16, FT with KMN is preferable: DF 150 mg, BMOV 6.0 mg and MK 0.02 mg in 350 μl of water at + 47- + 48 ° C 8 × 30 min with an interval of 3 hours to 3 days with intracavitary and intraperitoneal administration of CCP, DT 0.05 - DC 0.05 - TF 1.0 mg, at which the maximum therapeutic effect is obtained (Table 6).

Example 7. The antitumor effect of the claimed method on the subcutaneous form of spontaneous mammary adenocarcinoma in female Wistar rats, depending on the type and number of procedures. Out of 10 female Wistar rats with spontaneous tumors that arise as they age, 8 rats with infiltrative mammary carcinoma are selected and divided into 4 equal groups: 2 rats in each group. Age from 14 to 26 months, weight from 250 to 450 g, tumor volume from 40 to 450 mm ³ . In rats, the temperature in the rectum is measured. The rats of the 1 (control) group are sucked off the contents of the tumor, 4000 μl of a 0.9% NaCl solution is injected into the tumor. In rats of group 2, the contents of the tumor are sucked off, along the perimeter of the tumor are administered PCP: MF 0.02 mg and MK 0.2 mg in 4000 μl of water. In rats of group 3, the contents of the tumor are sucked off, along with the perimeter of the tumor, a DP of 1500 mg is administered in 4000 μl of water. In rats of group 4, the contents of the tumor are aspirated, KMN is administered along the tumor perimeter: DF 1500 mg MF 0.02 and MK 0.2 mg in 4000 μl of water.

Tumors of all rats withstand 5 minutes in a constant magnetic field by induction of 0.2 T, a gradient of 0.015 T / cm and placed in PeMP. The temperature of the tumor in rats of groups 3 and 4, under the action of PEMP, rises above + 42 ° C for 5 minutes, after 15 minutes the temperature of the tumor reaches + 48 ° C, and after 30 minutes the heating is stopped. At the same time, the temperature in the rectum and tumor of group 1 rats remains between + 37- + 38 ° C. After heating, the contents of the tumors are removed. The cavity formed at the site of the tumor is washed with a 0.3% formaldehyde solution in a 0.9% NaCl solution.

FT procedure is carried out

- to rats of groups 1, 2, 3 and 4 8 times with an interval of 3 hours to 3 days, depending on the FSH (table. 7).

The result is:

- in 1 (control) group, the tumor volume in rats is steadily increasing without signs of regression;

- in the second group, the average tumor volume decreases slowly, metastases are detected in rats, treated with intracavitary and intraperitoneal administration of CPC: DT 0.5 - DC 0.5 - CF 10.0 mg, increase in life expectancy of rats by 150%;

- in the third group, the average volume of tumors decreases rapidly, metastases are detected in mice, treated with intracavitary and intraperitoneal administration of CPC: DT 0.05 - DK 0.05 - CF 1.0 mg, increase in life expectancy of rats 200%;

- in the fourth group, the average volume of tumors decreases from the first day of treatment and, finally, their symptoms do not appear for 3 months,

Thus, in accordance with example 7, determining the type of therapy preferable in real time, preliminary, intermediate and final magnetic resonance scans of tumors and metastases with an individual conclusion according to the results of FT, predominant in speed and magnitude heating of tumor tissues containing indigenous people, removal of necrotic masses and washing the cavity formed at the site of the tumor, treating with intracavitary and intraperitoneal administration of CPC, DT 0.5 - DC 0.5 - CF 5.0 mg, leads to a rapid decrease in ema infiltrative tumors, their complete regression and increased survival in rats. In spontaneous infiltrative tumors of rat mammary glands, FT with intratumoral administration of KMN is preferable: DF 1500 mg, MF 0.02, MK 0.2 mg in 4000 μl of water at + 47- + 48 ° C 8 × 30 min with an interval from 3 hours to 3 days with intracavitary and intraperitoneal administration of CCP: DT 0.5 - DC 0.5 - CF 5.0 mg, and the maximum therapeutic effect is obtained (Table 7).

The experimental results are as follows: Examples 1-7, Tables 1-7, and FIG. 4 (a, b, c); 5 (a, b); 6; 7 (a, b, c, d) confirm the high antitumor effect of the claimed method, which includes the regional administration of CMN containing CF while heating the body to 38 ° C with heating the tumor predominantly in speed and magnitude to + 45- + 48 ° C, removing the contents of the tumor and washing the formed cavity from the products of necrosis. After-treatment with intracavitary and intraperitoneal administration of CPC: DT 0.05 - DK 0.05 - TF 1.0 mg, leads to a rapid decrease in the volume of infiltrative tumors, their complete regression and an increase in the life expectancy of mice.

The claimed method is illustrated in tables: 1, 2, 3, 4, 5, 6, 7.

Table 1

The antitumor effect of the claimed method on the subcutaneous form of P388 lymphocytic leukemia in female BDF ₁ mice, depending on the type and number of therapeutic procedures, expressed in complete regression of tumors and an increase in the average life expectancy of the mice

Table 2.

The antitumor effect of the claimed method on the subcutaneous form of P388 lymphocytic leukemia in female BDF1 mice, depending on the type and number of therapeutic procedures, expressed in the complete regression of tumors and VL of mice.

Table 3.

The antitumor effect of the claimed method on the subcutaneous form of Ehrlich carcinoma in female BDF1 mice, depending on the type and number of therapeutic procedures, expressed in complete regression of tumors and VL of mice.

Table 4.

The antitumor effect of the claimed method on the subcutaneous form of Lewis lung carcinoma in female C57Bl / 6j mice, depending on the type and number of therapeutic procedures, expressed in the complete regression of tumors and OLC of mice.

Table 5.

The antitumor effect of the claimed method on the subcutaneous form of adenocarcinoma of the mammary gland Ca 755 in female C57Bl / 6j mice, depending on the type and number of therapeutic procedures, expressed in complete regression of tumors and VL of mice.

Table 6

The antitumor effect of the claimed method on B 16 melanoma in female C57Bl / 6j mice, depending on the type and number of therapeutic procedures, expressed in the complete regression of tumors and OLC of mice.

Table 7

The antitumor effect of the claimed method on the subcutaneous form of spontaneous adenocarcinoma of the mammary glands in female Wistar rats, depending on the type of therapeutic procedure, expressed in complete regression of tumors and VL of rats.

The claimed method is illustrated by figures 1 ( _{1, 2, -3, 4, 5, 6, 7, 8, 9, 10} ) and 1 ′ (a, b, c, d, e, f); 2, 3, 4 (a, b, c); 5 (a, b); 6, 7, 8, 9, 10, 11 (a, b), 12 (a, b), 13 (a, b), 14 (a, b, c); fifteen.

FIG. 1 ( _{1, 2, -3, 4, 5, 6, 7, 8, 9, 10} ). Contrast magnetic resonance imaging tomography (KM-RT) monitoring of oncogenesis of the tubular form of Ca 755 breast adenocarcinoma implanted under the skin of the right axillary region of a female mouse C57Bl / 6j: ( ₁ ) implantation channel (arrow) 4-24 hours after tumor implantation; ( ₂ ) negative; ( ₃ ) after 24-48 hours, the size of the transplantation channel increases and a network of branching lateral tubules appears; ( ₄ ) negative; ( ₅ ) after 48-96 hours, the tubules increase, the boundaries of the invasion of tumor cells into healthy tissues appear as a veil, the growth of large tubules, their thickening and fusion are directed mainly towards the nipple; ( ₆ ) negative; ( _7, ) in 96-192 hours the volume of the tubule network increases and they merge into nodules, which turn into large nodes; ( ₈ ) negative; ( ₉ ) after 192-384 hours, the fusion of individual large nodes and tubules into dense tuberous formations that merge with the formation of a solid tumor, the invasion of tumor cells into healthy tissues, the vessels of the lymphatic and circulatory system, and the growth of collateral vessels in proliferation areas increase; ( ₁₀ ) negative. FIG. 1 ′ (a, b, c, d, e, f). Contrast magnetic resonance imaging (KM-RT) monitoring of oncogenesis of the lobular form of Ca 755 breast adenocarcinoma implanted under the skin of the right axillary region of a female mouse C57Bl / 6J: (a) implantation channel (arrow) 4-24 hours after tumor implantation; (b) after 24-48 hours, the size of the channel increases; (c) after 48-96 hours, the tumor grows, the boundaries of the invasion of tumor cells into healthy tissues appear as separate outgrowths (thin arrows); (d) after 96-192 hours, the tumor volume increases, invasions into healthy tissues (thin arrows) appear; (f) after 192-384 hours, an increase in the volume of a solid tumor is observed, numerous invasions of the tumor into healthy tissues (thin arrows) are visible; (f) partial tumor necrosis and invasion of tumor cells into healthy tissues (thin arrows). FIG. 2. A Wistar rat weighing 450 g with a subcutaneous form of spontaneous mammary adenocarcinoma, anesthesia: zoletil 100, 10 mg / kg, (Zoletil 100, Virbac), to determine the TP by electron-sensor scanning, the content of CMN in the tumor and in the organs of RES is measured, directions scan (arrows). FIG. 3. A Wistar rat weighing 350 g with a subcutaneous form of spontaneous mammary adenocarcinoma: intravenous anesthesia, “Zolet 100”, 20 mg / kg; to determine TP electron-sensory scanning measures the content of CMN in the tumor and in the organs of the RES, the scanning direction (arrows). FIG. 4 (a, b, c), (a, b, c) MP tomograms of the mouse body with mammary adenocarcinoma Ca 755: (a) tumor tissue before FT procedures (between the arrows); (b) tumor tissue after the first FT procedure (between the arrows); (c) tumor tissue after the third FT procedure (between the arrows). FIG. 5 (a, b), (a, b) MP tomograms of a portion of the mouse body with mammary adenocarcinoma Ca 755 during FT. (a) tumor tissue after the fourth FT procedure (between the arrows); (b) tumor tissue after the sixth FT procedure (between the arrows). FIG. 6. Female BDF ₁ mouse with P388 lymphocytic leukemia 3 weeks after receiving 8 FT procedures, as a result of which the primary tumor is not palpable, lymph node metastases are not detected. After exfoliation of the crust at the location of the primary tumor, alopecia is formed, which gradually overgrows with gray hair. FIG. 7 (a, b, c, d). Microphotographs (magnification × 200) of Ca 755 mammary adenocarcinoma tissue: (a) tumor cells endocytosed KMN aggregates (nanoparticle aggregates are stained with Berlin blue in blue color); (b) ferrite aggregates concentrate and fix NPMP in tumor tissues; (c) 3 days after the first FT procedure, partial replacement of tumor tissues with elements of connective tissue is observed (light areas); (d) 6 days after the sixth FT procedure, the replacement of tumor tissues with elements of connective tissue is observed (light areas). FIG. 8. Digital photo image of pathological and anatomical changes in the body of a female C57Bl / 6J female mouse on the 15th day after implantation of breast adenocarcinoma Ca 755: (a) primary tuberous tumor Ca 755 with a volume of more than 500 mm ³ (between four long black arrows); (b) overflow of the bladder and kidneys caused by metastases of the bladder blocking the urethra (short white arrows); (c) metastases of sentinel lymph nodes of the right groin (short thick white arrow), (d) metastases in the lungs (three short black arrows); (e) the spleen is enlarged (long white arrow); (g) the intestines are full and swollen due to overlapping of the anus with metastases (short white arrows). FIG. 9. The calibration graph. FIG. 10. Image of a C57Bl / 6j mouse in which 90 days after completion of FT, palpation reveals a secondary Lewis lung carcinoma tumor developing from metastases of the lymph nodes of the neck (arrow). FIG. 11 (a, b). KM-RTI of female C57Bl / 6J female mouse with Lewis lung carcinoma with a volume of more than 600 mm amplify MRI-NKN at T ₂ weighted gradient echo (500/15) longitudinal scanning of the mouse body: (a) 1 hour after intravenous administration of MRI-NKN (a long arrow and thick arrows indicate proliferating areas of the tumor), the spread of the tumor through the internal organs, macro-metastases (short arrows); (b) negative. FIG. 12 (a, b). KM-RTI of a female mouse C57Bl / 6J with Ca 755 breast adenocarcinoma with a volume of more than 800 mm ³ is enhanced by the intravenous administration of negative MRI of a contrast agent with the addition of a magnevist, which increases the brightness of KM-RTI with T ₂ weighted gradient echo (500/15) longitudinal scan of the mouse body: (a) 6 minutes after intravenous administration of the drug, the structure of the primary tumor and the area of tumor invasion through the chest into the liver (large arrow), metastases (small arrow) are visible; (b) negative. FIG. 13 (a, b). KM-RTI of a female mouse C57Bl / 6J with Ca 755 breast adenocarcinoma with a volume of more than 500 mm ³ is enhanced by the intravenous administration of MRI-NKN with the addition of a magnevist, which increases the brightness of KM-RTI at T ₂ weighted gradient echo (500/15) longitudinal scan of the mouse body: (a) KM-RT image of the primary tumor (short arrow); areas of proliferation, tumor spread through the lymphatic vessels, neovascularization, macro-metastases in healthy tissues (long arrows); (b) negative. FIG. 14 (a, b, c). KM-RTI is obtained by longitudinal scanning the body of a mouse with mammary adenocarcinoma Ca 755 (tumor diameter> 500 mm ³ ), 20 hours after the intravenous administration of MRI-NKN (citrate ferrite), an additional magnevist is added, which increases the brightness of KM-RTI: (a) invasion tumors in the lung (long arrow), metastases in the kidneys and other internal organs (short arrows); (b) negative; (c) invasion of the tourniquet consisting of twisted pathogenic tubules into healthy tissues, the head of the tourniquet destroys the capsule wall and penetrates into healthy tissues through the hole (long black arrows), the tourniquet opens, the tubules grow in all directions (long white arrows), merge form metastatic nodules in healthy tissues (short arrows). FIG. 15. Female C57Bl / 6j mouse after 8 procedures of FT 75 breast adenocarcinoma Ca 755: the primary tumor is not palpable, metastases to the lymph nodes are not detected, after detachment of the cutaneous crust, alopecia is formed, gradually overgrowing with gray hair (arrow).

Technical result

The claimed method is new and effective in the treatment of malignant tumors in the experiment and includes:

- early detection of tumors, metastases and borders of invasion of tumor cells by combinations of MRI of negative contrast nanopreparations of the general formula (1) with MRI-positive drugs, for example, magnevist, which increase the brightness of CM-RTI under the stated conditions;

- the combined antitumor effect of KMN containing compounds of the general formula (2) and cytostatic agents (ferrite from 26 to 150 mg; AO from 0.2 to 2.0 mg, MO from 0.05 to 0.2 mg, SL or MF from 0.001 to 0.002 mg, CP from 0.01 to 0.04 mg, MK from 0.005 to 0.02 mg) with retreating infiltrative tumors with combinations of cytostatic drugs, dacarbazine-docetaxel-cyclophosphamide (DC-DT-CF);

- lowering the pressure in the tumor before injections of KMN or antiseptic, by aspirating its contents;

- determination of the content of KMN in the tumor and internal organs in real time before each heating procedure;

- determination of therapy preferable for this animal in real time according to the formula (3),

where: - KMN of a _tumor - response of KMN ye a tumor;

- _{the amount of} KMN _bodies - total response KMN ye _bodies;

- TP - the preferred therapy for this animal in real time, (with TP≥10 - ferrimagnetic thermochemotherapy with TP≤1 - other types of therapy);

- assessment of the completeness of the removal of indigenous people after each session of FT from the cavity formed at the site of the tumor, the rapid determination of the size of the remaining part of the tumor, the content of indigenous people in the tumor and in the liver, and individual planning of the next session of FT;

- individual control of the concentration of KMN in the tumor, allowing you to quickly evaluate the results of FT, plan the volume of subsequent FT sessions and enhance the therapeutic effect, which is manifested in a rapid decrease in the volume of malignant tumors in the absence of toxic effects and a reduction in the number of therapeutic procedures.

The application of the claimed method leads to complete regression of tumors in 55% of mice with an increase in their life expectancy up to 350% and ensures the safety of mammals.

The proposed method is illustrated by examples 1-7, tables 1-7 and figures 1, 1′-15.

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19. Brusentsov HA, Nikitin P.I., Pirogov Yu.A., Brusentsova T.N., Nikitin M.P., Yuryev M.V., Kupriyanov D.A., Dubina A.I., Uchevatkin A. A., Ivanov A.V., METHOD FOR NON-INVASIVE QUALITATIVE AND QUANTITATIVE DETERMINATION OF MAGNETO-CONTROLLED NANO DRUGS AND ASSESSMENT OF THEIR REAL TIME FUNCTIONS AT EXPERIMENTAL ANIMALS, RF Patent 2427390, 08.06.2009. fig. 24, 2011.

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Claims (1)

A method of ferrimagnetic thermochemotherapy (FT) of malignant tumors in an experiment, including the introduction of combinations of magnetically controlled nanopreparations (KMN), monitoring of treatment results and determining indications for FT in real time, characterized in that animals are administered intravenously magnetic resonance tomographic negative contrast nanopreparations of the general formula

where 0 <y <1, 0 <x <1, 0 <z <1, L ³⁺ is a lanthanide in combination with a magnevist who visualize contrast magnetic resonance imaging images (CMRTI) reflecting the development of tumors, invasions and metastases in dynamics ;
- the content of indigenous people in the tumor, liver, spleen, kidneys and lungs is determined by electron-sensory scanning (ES) of the body of experimental animals;
- indications for FT for this animal in real time are the results of determining the preferred therapy (TP), which is determined from the ratio of the content of CMN in the tumor to the sum of the CMN contained in the liver, spleen, kidneys and lungs, according to the formula

Where
- TP _{therapy is preferred} - therapy, preferred for this animal in real time;
- KMN of a _tumor - response of KMN of a tumor;
- the _{sum of} indigenous _organs - the total response of indigenous _organs ;
- with TP≥10 conduct ferrimagnetic thermochemotherapy;
- the contents of the tumor are aspirated;
- combinations are introduced along the perimeter of the tumor, including at least two of the listed cytostatic drugs, adenox, methox, melphalan, mitoxantrone, cisplatin and bis (L-malato) IV oxovanadium with magnetically controlled nanopreparations (KMN) of the general formula

where 0 <y <1, 0 <x <1, 0 <z <1, L ³⁺ is a lanthanide, and it is heated by magnetization reversal in an alternating magnetic field for 30 minutes at a temperature of +45 - + 48 ° С;
- after heating is completed, the contents of the tumor are removed, the remaining part of the tumor, invasions and metastases are determined by contrast MP tomographic scanning;
- before carrying out the next FT procedure, the content of CMN in the tumor, liver, kidneys and spleen is determined;
- aftercare of animals with infiltrative tumors is carried out by compositions comprising dacarbazine, docetaxel and cyclophosphamide.

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