Method of predicting efficiency of local chemical therapy in treatment of patients with malignant tumours of brain

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to oncology and can be used for predicting efficiency of local chemical therapy of brain in patients with malignant tumours of central nervous system. For this purpose during surgical intervention for ablation of brain tumour from perifocal zone of tumour taken is brain tissue, which is divided into samples, 100 mg each. Number of samples is limited by number of studied chemical preparations. Chemical preparationbs are added to samples in dose 0.2 mg. Incubation at temperature 37°C for 30 minutes is performed. Activity of alpha-2-macroglobulin is determined before and after incubation. If index increases by 40% and more, in comparison with the value before incubation, efficiency of chemical preparations is predicted.

EFFECT: method ensures high specificity, possibility of objective evaluation of efficiency of chemical preparation impact in performing analysis on operation day, which makes it possible to start adequate therapeutic measures, including cancelling or replacement of chemical preparation in each specific case.

2 ex

 

The invention relates to medicine, namely to Oncology, and can be used to assess the effectiveness of local chemotherapy of brain in patients with malignant tumors of the Central nervous system.

There is a method of assessing response to treatment with chemotherapy drugs, which are used biochemical indicators, such as

the concentration of TNF-α, IL-1, IL-6 in serum (Orlov V.N., Gusev S.A. Goncharov, AP Influence of chemotherapy on the concentration in the serum of patients with multiple myeloma interleukin-1, interleukin-6 and tumor necrosis factor. RBM. W. hematol. transposal. 2007, N 3, p.17-20.).

However, this analysis was not conducted in malignant tumors of the brain, and the determination of TNF-α and interleukins in serum informative, but is not specific to the operation of the authority and does not allow to predict therapeutic effect for intracranial gliomas.

A known method for determining the expression of P-glycoprotein in normal, okolokrugovykh and cancerous tissues (Chen Chun-Yan, Zhu Zhao-Hua Shiyie huaren xiaohua zazhi=World Chin. J. Dig. - 2003. - 11, N 1. - P.36-38;). The level of P-glycoprotein was different in normal, allopurinol and cancerous tissues in cancer of the stomach: 13, 22, and 43%from the norm and correlated with the number of surviving patients.

The authors use this pokazatel is to confirm the effectiveness of chemotherapy, but the main point of reference become clinical signs of the patient, the duration and quality of their lives.

Playback of the following methods is not used for predicting the efficacy of chemotherapy when choosing chemotherapy drugs during the operation.

There is a method of assessing the efficacy of chemotherapy (Cirak O.V., Lisovskaya POSTGRADUATE performance Criteria endolymphatic chemotherapy in patients with advanced breast cancer and ovarian cancer. RBM. chemotherapist. J., 2000. N 4, P.17-21.), selected as a prototype. The essence of the method lies in the fact that prior to treatment from the peripheral blood of cancer patients produce lymphocytes and leukocytes, register in lymphocytes admonitive DNA fragments in the gel electrophoresis, and then examine the index change in the affinity of cells to antigens of the primary tumor and placenta in LAI-test. Check admonitive DNA fragments in peripheral blood mononuclear prior to treatment and decrease the affinity of cells to the antigen of the primary tumor with simultaneous rising to the antigen of the placenta in the period of treatment was correlated with a marked clinical effect of chemotherapy.

However, the disadvantages of this method are, on the one hand, its duration (2-3 days), associated with the separation of lymphocytes and leukocytes, the implementation of the PE the work with antigens and the need for repeat studies after the introduction of chemotherapy drugs, that slows down the production and analysis of results, on the other hand, the inability of a cancellation or replacement of a chemical in the case of resistance of the investigated tissues to the previously selected before the introduction of the drug to the patient. In addition, this method is not applicable in patients with malignant brain neoplasms.

The aim of the invention is to be able to predict the efficacy of chemotherapy drugs directly to the target organ malignant process.

This objective is achieved in that during the surgical intervention, removal of brain tumors from perifocal zone tumor take the tissue of the brain, divided into samples of 100 mg each; the number of samples is limited to the number of chemotherapy plus one sample, which leave intact; other samples add chemotherapy at a dose of 0.2 mg for each sample, incubated for 30 min at 37°C. after incubation, determine the activity of the inhibitor alpha-2-macroglobulin in all samples, including intact (before incubation); increase to 40% and more compared with the value prior to incubation predict the efficacy of chemotherapy.

The invention of "a Method for predicting the effectiveness of regional chemotherapy in the treatment of patients with malignant brain tumors" is but the YM, as it is unknown the level of achievements of medicine in the field of predicting the effectiveness of chemotherapy and resistance to chemotherapy drugs.

The novelty of the invention lies in the fact that in the perifocal tissue determine the activity of the universal inhibitor of proteolysis of alpha-2-macroglobulin before and after incubation with chemotherapy and with its increase by 40% or more after incubation predict a positive clinical effect. If the level of alpha-2-macroglobulin less than 40% predicted resistance fabric perifocal zone of the tumor to the chemotherapy drugs, which gives the possibility of making appropriate decisions about the choice of chemotherapy drugs during the operation. This method of forecasting is new and allows for individual selection of chemotherapy drugs during surgery, as well as the timely adoption of measures to prevent adverse consequences.

In open sources information of Russia, CIS and abroad indications for a similar method for predicting the effectiveness of chemotherapy drugs us not detected.

The invention is industrially applicable as it can be reproduced and re-reproduced in any clinical laboratory medical health care institutions.

The method is as follows. During the op the radio removed the tumor and the required number of perifocal brain tissue, which was used for the determination. Fabric perifocal zone was divided into several samples of 100 mg each (2-3, depending on the number of tested drugs). One of the samples was left intact, others were added to chemotherapy in a dose of 0.2 mg each, and incubated 30 minutes at 37°C. After incubation in each sample were used to define the alpha-2-macroglobulin. The tissue was ground in a homogenizer with 1 ml of physiological solution. Osmotic shock caused by the addition of 1 ml of bidistilled water and after 10 minutes of exposure of the obtained homogenate was centrifuged 15 min at 3000 Rev/min the Precipitate was discarded, and the supernatant was determined by the activity of alpha-2-macroglobulin. The measurement was repeated unified kinetic method for speed cleavage of the synthetic substrate BAEE (Artikova V.F., Pashina T.S., J. The matters. the honey. chemistry, 1979, V.25, no 4). The activity of alpha-2-macroglobulin was calculated as the increase in optical density of the investigated solution in the cuvette of a spectrophotometer for 10 min vs. control for the reagents. Recalculation in Sg tissue was performed using the conversion factor proposed by the authors of the method, adjusted to 1 g of fresh tissue. The duration of definition 2 hours, which allows analysis to be performed on the same day.

An example of the application method can be the issue the ski records.

1. Patient K., 1958 born., Case history No.-726/kg, was admitted to the Department of tumours of the Central nervous system FGU RNII and gynecology 15.10.2007 and was in the hospital on 30.10.2007.

Diagnosis (primary, secondary, complications): glioblastoma left fronto-parietal region of the brain; the spasms.

G/a No. 974384-389 - isomerisation glioblastoma with diffuse infiltrative growth, extensive hemorrhage.

17.10.2007 the patient underwent surgery decompressive craniotomy with removal of the tumor of the left fronto-parietal region of the brain. During the operation, after removal of the tumor was preparing 4 sample perifocal brain tissue 100 mg One sample was left for comparison as the original, others were added 0.2 mg of chemotherapy in each (1 - cyclophosphamide, 2 methotrexate, 3 - carboplatin), and incubated at 37°C for 30 minutes. Alpha-2-macroglobulin was determined in the original sample and in the samples after incubation. The increase in the activity of alpha-2-macroglobulin after incubation with carboplatinum was 62.8% from the original. The drug is considered effective, which was confirmed by the subsequent dynamics of the patient. Currently (within 2 years) the patient is under observation, data in favor of relapse no.

2. Patient B., 1939 born., Case history No.-8235/C, was dropped at the re with 19.06.2008 on 04.07.2008.

Diagnosis (primary, secondary, complications): glioblastoma right temporal lobe of the brain; the spasms.

G/a No. 36957-962 - glioblastoma with infiltrative growth, foci of necrosis.

10.12.2007 the patient underwent surgery osteoplastic craniotomy with removal of the tumor on the right temporal region of the brain. During the operation, after removal of the tumor was preparing 4 sample perifocal brain tissue 100 mg One sample was left for comparison as the original, others were added 0.2 mg of chemotherapy in each (1 - cyclophosphamide, 2 methotrexate, 3 - carboplatin), and incubated at 37°C for 30 minutes. Alpha-2-macroglobulin was determined in the original sample and in the samples after incubation. The increase in the activity of alpha-2-macroglobulin after incubation with drugs was less than 27% in all samples. Drugs considered ineffective, which was confirmed by the subsequent dynamics of the patient's condition.

In may 2008 (5 months after chemotherapy and determining the activity of the inhibitor), the patient showed a relapse of the tumor, his condition deteriorated; 23.06.08 re - operation-recruitme with Subtotal removal of the tumor. The patient died in August 2008.

Were examined 7 patients with brain tumors who were treated in the Department of tumours of the NS RNII and subjected to operation with local chemotherapy.

Found that 5 patients with a favorable clinical effect after intraoperative injection of chemotherapy drugs value of alpha-2-macroglobulin in the tissue of the perifocal zone tumors, incubated with the cancer, varied 13,65 inhibitory units (S), tissue - 19,11 Sg tissue, which exceeded the original value on 44-76%%. In 2 patients developed a relapse, and died subsequently, the activity of alpha-2-macroglobulin, defined in the same tissue in the same conditions ranged from 10,37 Sg tissue to 13,65 Sg tissue (increased to 27% from the original)that is clearly linked to the resistance of the fabric perifocal zone of the tumor to the chemotherapy. The source believed the value of the activity of alpha-2-macroglobulin in the same perifocal tissue prior to incubation. In all patients, the initial activity of alpha-2-macroglobulin ranged from 8,19 Sg tissue to 13,92 Sg tissue.

Technical and economic efficiency of the method is its high specificity, being able to objectively evaluate the impact of chemotherapy drugs directly to the affected organ, analysis on the same day that allows for timely assign adequate therapeutic measures, cancel or replace the chemo.

A method for predicting the effectiveness of local himit is rapie in the treatment of patients with malignant brain tumors, including operation, spectrophotometric studies, characterized in that during the surgical intervention - remove a brain tumor from perifocal zone tumor take the tissue of the brain, divided into samples of 100 mg each, the number of samples limit the number of investigational drugs, one sample is left intact to other samples add chemotherapy at a dose of 0.2 mg for each sample, incubated for 30 min at 37°C, before and after incubation to determine the activity of the inhibitor alpha-2-macroglobulin, when the increase by 40% or more compared with the value before incubation predict the effectiveness of chemotherapy drugs.



 

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3 ex

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2 ex, 3 tbl

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