RU2166944C2 - Plasma substitute solution with hemocorrecting and immunomodulating properties - Google Patents

Abstract

FIELD: medicine, pharmacy. SUBSTANCE: invention relates to pharmaceutical agents used for treatment of patients with severe acute and chronic damages resulting to blood loss or depletion of its functional properties and accompanying immunodeficiency state. Plasma substitute solution has isotonic solution and DNA sodium salt additionally obtained from animal raw with mean molecular mass (1.5-550) x 10³ Da and hyperchromic effect 28-46% taken in the amount 5 x 10^-5 g/1 l of isotonic solution. Except for the basic curative effect this agent stimulates hemopoiesis and myelopoiesis being without adverse effects. EFFECT: increase percent of viability, decreased time of patients treatment. 5 tbl, 1 ex

Description

 The invention relates to medicine, in particular to the field of pharmaceuticals used to treat severe acute and chronic lesions of the human body, leading to loss of blood or loss of its functional properties with a concomitant state of immunodeficiency.

 The purpose of the invention is to increase the percentage of survival and reduce the treatment time for patients for whom a quick restoration of the quantity or functional properties of blood is necessary for health reasons.

 Severe acute injuries of the human body are known, such as extensive burns with deep penetration into tissues, blood poisoning from tissue decay products as a result of severe injuries, frostbite, penetrating wounds, and so on.

 Severe chronic lesions are known, such as various types of anemia, leuko- and thrombocytopenia, resulting from irradiation or poisoning during radiological or environmental disasters, specific chemotherapy for cancer patients.

 As a rule, all of the above diseases are accompanied by a state of immunodeficiency.

 Severe acute and chronic infectious diseases such as AIDS, hepatitis B, genital herpes II, etc., which entail a persistent state of immunodeficiency, are also known.

 A key component of the treatment of such diseases, in addition to concomitant therapy of lesions, is transfusion therapy with plasma-replacing solutions (PZR).

 Modern plasma-substituting solutions are conventionally divided into four main groups according to their purpose and functional properties (1, 2), namely: hemodynamic (anti-shock), the main representatives of which are polyglyukin and reopoliglyukin; detoxification, the main representatives are hemodez, polydez; solutions for protein parenteral nutrition and regulators of water-salt and acid-base conditions.

 Of the hemodynamic plasma-substituting solutions, the most popular is polyglucin, which is a medium-molecular fraction of partially hydrolyzed dextran dissolved in an isotonic sodium chloride solution freed from toxicity and pyrogenicity (prototype).

 Casein hydrolyzate, hydrolysin, aminokrovin are most known as solutions for protein parenteral nutrition; each of which is a mixture of amino acids and simple peptides in an isotonic solution of sodium chloride or glucose (prototype).

 Among the regulators of the water-salt and acid-base conditions are known solutions of disol, trisol, as well as a water-salt infusion solution of lactasol containing a balanced salt composition and lactic acid.

 Plasma-replacing solutions are known that simulate the respiratory functions of blood (blood gas carriers) based on a concentrated hemoglobin solution purified from stromal impurities and procoagulants (2, 7). A model of oxygen transfer based on modified hemoglobin was developed (8, 9), among which emulsions of perfluorane and perfucol (3) constitute a special group. A new plasma-substituting hemodynamic action solution based on starch has been developed (5, 4).

 The need for the simultaneous use of various medical manipulations related to filling the bloodstream, restoring and maintaining constant blood pressure (BP), freeing the body of toxins, normalizing blood stasis, providing nutrients and oxygen, accompanying, as a rule, massive blood loss and severe shock, predetermined the creation of a number of complex multifunctional plasma-substituting solutions, for example, such as a solution of hemodynamic and hematopoietic action - Polifer or diuretic and rheological - reoglyuman. In addition, in these cases, plasma-substituting solutions have been widely used, in which polyglucin, which increases blood pressure and keeps it at a stable level, reopoliglyukin, normalizing microcirculation, eliminating blood stasis, leading to the re-deposition of red blood cells from capillaries; compounds that promote excretion of metabolites and lactasol from the body with urine, which eliminates tissue acidosis and increases the overall therapeutic effect.

 However, none of the known plasma substituting solutions, including multifunctional action, does not directly affect the immune system in order to normalize its functioning.

The essence of the invention lies in the fact that to overcome the above disadvantages and achieve the purpose of the invention, it is proposed to add to the composition of existing PZR a composition in an amount of 0.5 • 10 ^-5 - 0.5% by weight, consisting of the main component in the form of the sodium salt of the DNA derivative, obtained from animal raw materials, with an average molecular weight of 1.5 - 550.0) • 10 ³ Daltons and a hyperchromic effect of 28 - 46% and additional components: protein - not more than 1.5% by weight; polysaccharides - not more than 2% by weight; RNA - not more than 6% by weight
The essential features of the proposal should be considered the quantitative composition, physical characteristics of the main component, as well as its quantitative content in the PZR, which makes it possible to obtain the optimal therapeutic effect in the absence of complications (Tables 1, 2).

 The preparation of the claimed funds is carried out in the factory by the traditional volumetric weight method. In this case, a solvent is added to a certain weight amount of the composition in the form of a powder to obtain the required volume of solution with a concentration of more than 0.5% by weight. As a solvent, distilled water or saline is used. Next, the solution is filtered, sterilized and added in the required amount to the blood substitute until the desired concentration of the composition is obtained in it. The resulting solution is a clear, colorless liquid, does not contain mechanical or other impurities, is stable. It can be subjected to long-term storage of at least 2 years. With chemical and bacteriological control, there are no contaminants. Meets the requirements of GFH.

In experimental clinical trials, the hemocorrecting properties of the claimed drug were revealed under conditions of cytopenia caused by a cytostatic agent (immunodeficiency model due to chronic poisoning caused by environmental factors) or gamma radiation (immunodeficiency model caused by radiological contamination). Studies were conducted on male mice line F ₁ (CBA x C ₅₇ B1), male rats F1 (AMCY x Wistar) and dogs breed "English Beagle."

 As cytotoxic drugs used: cyclophosphamide domestic production, farmarubicin manufactured by Farm-Italy, cis-platinum (DDR) manufactured in the USA. Irradiation was performed using the installation "Stem-3a". Cytostatics were used for the intraperitoneal route of administration. When conducting experiments on the “Stebel-3a” installation - the source of gamma irradiation - mice were placed in glass chambers of 3-4 animals and introduced into the irradiation zone of the installation with the location of the sources as a “squirrel wheel”. Dosimetric control recorded a dose difference within the container of not more than 10%. The radiation dose was 6 Gy. The claimed tool was administered intravenously at the rate of 2 mg of an immunomodulating composition per 1 kg of animal weight.

 When analyzing the morphological composition of peripheral blood after the use of cytostatics, it was revealed that the claimed agent stimulated the release of both lymphocytes and granulocytes into peripheral blood. On the 6th day, the number of granulocytes in mice that received the claimed drug exceeded 3 times this figure in the control group (table. 3).

 The effect of the claimed funds on the dynamics of the restoration of blood parameters after total gamma radiation is shown in table. 4. There is a statistically significant excess of the number of leukocytes in the peripheral blood several times in comparison with the control group.

 Pharmacokinetic studies were conducted in mice, for which conditions were developed for the preparation and purification of an immunomodulatory additive labeled with tritium. Several routes of administration of a pharmacological agent have been used, including intravenous. The concentration and time dependences of the distribution of the labeled drug and its metabolic products in blood, plasma, blood cells, kidneys, liver, lymph nodes, spleen, thymus, stomach, small intestine, brain, bone and bone marrow were studied. All pharmacokinetic curves describing a change in the concentration of ID and metabolites in the studied organs and tissues were characterized by a fast phase of a decrease in concentrations in the time interval of 3-6 days. Lymph nodes, spleen, stomach and kidneys had the greatest affinity for ID. The average retention time for organs and tissues was 60 + 2.9 hours. Excretion of ID occurred mainly with urine. The main amount of ID was excreted on the first day after administration of the drug: 37% and 3.3% of the dose, respectively, with urine and feces. Further, the rate of the elimination process slowed down and by the 6th day after the drug was administered, no more than 10-15% of the administered dose remained in the animal organism. Completely eliminates ID from organs and tissues in at least 10 days.

 The toxicological properties of the drug were studied in the field of acute toxicity in mice, rats, dogs, subchronic toxicity in rats and dogs, allergenic and immunological effects in guinea pigs, embryotoxic and teratogenic effects in rats, pyrogenic properties in rabbits, harmlessness of the drug with investigation of potential mutagenicity and prognosis for carcinogenesis. Based on the results obtained, it can be concluded that the drug is a low-toxic substance, pathomorphological studies have shown that changes in organs and tissues correlate with the magnitude of the applied dose of the drug and are reversible. The drug does not have teratogenic, carcinogenic and embryotoxic effects, does not affect the morphological and biochemical composition of peripheral blood, does not have gastrointestinal, cardio, hepato- and nephrotoxicity, does not have a locally irritating effect, and is pyrogen-free. The main types of toxicity of the drug were revealed, which appear only when it is administered intravenously in high doses and concentrations (see tab. 1, 2). The drug is not anaphylactogenic and does not cause sensitization with repeated use.

In general, the analysis of experimental data allows us to draw the following main conclusions:
1. In contrast to the prototype of the claimed tool can dramatically accelerate the restoration of the immune status of the body.

 2. When using the tool, no side effects and complications were detected.

 The clinic conducted studies of the claimed invention in patients with chronic anemia and in cancer patients with inhibition of hematopoiesis due to the use of chemoradiotherapy (permission of the Pharmacological Committee is attached).

In studies of patients with chronic anemia, patients over the age of 18 years with anemia were included (men - the number of red blood cells less than 4.0 • 10 ¹² / l; women - the number of red blood cells less than 3.5 • 10 ¹² / l) at the time of inclusion in a study that either lasts for at least 6 months, or periodically recurs after treatment for 1 year. Before the start of the study, the patients underwent a complete examination to obtain clinical, hematological and biochemical data confirming the diagnosis: anamnesis, the number of leukocytes, red blood cells, reticulocytes, hemoglobin, hematocrit, leukocyte blood count, ESR, iron and iron binding capacity of serum, transferrin, ferritin, copper zinc, ceruloplasmin. Clinical and immunological data characterizing the state of the immune system were also determined: CD3, CD4, CD8, CD16, CD72 - lymphocytes, serum immunoglobulins, phagocytic number and index, complement hemolytic activity, rheumatoid factor titer.

The course of treatment was carried out by intravenous daily administration of 100-200 ml of polyglucin with a BMI content of (0.5-1.5) • 10 ^-3 % for 3 days.

 At the end of treatment, the results were evaluated according to the following criteria: condition and well-being of the patient, dynamics of hematological parameters, dynamics of immunological parameters, dynamics of biochemical parameters, the presence or absence of relapse of the disease during the observation period.

As a result of summarizing the data for 18 patients, a statistically significantly positive dynamics was determined with the subsequent normalization of hematological, biochemical and immunological parameters in 83% of patients, the remaining patients showed a positive dynamics of indicators with unexpressed relapse of the disease 40-60 days after the end of the course of treatment. In 22% of patients, a hyperthermic reaction was observed with an increase in temperature to 38-38.5 ^o , which disappeared spontaneously on the 2nd - 3rd day without the use of symptomatic agents. No other adverse effects were observed.

 In studies in cancer patients, a group of 14 people was selected who received combined chemotherapy in intensive CAM regimen for small cell lung cancer. Prior to the study, patients underwent a complete clinical examination: radiography of the lungs, CT of the liver, adrenal glands, brain, skeleton scan, biochemical blood test, myelogram.

Patients with morphologically proven localized MRL on the first day of treatment received: cyclophosphamide 1.5 g / m ² iv, adriamycin 60 mg / m ² iv, methotrexate 30 mg / m ² iv.

The course of treatment was carried out daily by intravenous administration of 100 - 200 ml of polyglucin with a BMI content of (0.5 - 1.5) • 10 ^-3 %, starting from the second day of treatment for 5 to 10 days. Blood tests were performed daily from the 2nd to the 20th days, on the 6th and 10th days a control myelogram was performed. During the study period, patients did not receive other hemostatic stimulants, vitamins, corticosteroids, blood transfusions.

As a result of the generalization of the data, the use of the claimed agent in patients with MRI during chemotherapy allowed to reduce the depth of leukemia and neutropenia, to shorten their duration, to conduct chemotherapy without septic complications, and in 64% of patients to start repeated chemotherapy courses with a 20-30% reduction in the interval between them. . All patients noted satisfactory tolerability of the drug, in 21% of patients there was a temperature increase to 38 ^o , which disappeared spontaneously, did not require the use of symptomatic agents and did not prevent further administration of the drug.

The invention is illustrated by the following example:
Example 1

Patient K.A.U., born in 1935, chronic anemia, was observed for 6 months, the therapeutic effect of the use of known drugs is short-lived or absent. I received a course of treatment for PZR with BMI in the form of daily intravenous administration of 200 ml of polyglucin with a BMI of 0.5 • 10 ^-3 % for 3 days. The results are presented in table. 5.

 In the given example, a characteristic is a steady increase in the number of red blood cells and white blood cells during the 50 days of observation, as well as a change in other indicators in the direction of approaching them to normal. There was a persistent improvement in the patient's well-being, the disappearance of symptoms of dizziness, weakness.

 Complications of a general nature, as well as side effects, were not observed.

 Thus, the example illustrates the effectiveness of the proposed tool in the field of the invention.

 All this allows us to recommend it for widespread use as a PZR with immunomodulating properties.

 List of references.

 1. Mashkovsky MD, Medicines, 1983, 4.2, p. 122-135.

 2. Handbook of blood transfusion and blood substitutes, Moscow, Medicine, 1982, p. 155-178.

 3. Bulletin of the USSR Academy of Sciences, 1989, N 6, p. 55-64.

 4. E.R. 059633, 04/27/94, A 61 K 31/72.

 5. U.S. 5470841, 11.25.95, NCI 514/60.

 6. E.P. 0083469, 07/13/83, A 61 K 37/02, 37/12, 45/06.

 7. J.P. Application 3-59883, 09/12/91, A 61 K 37/14.

 8. U.S. 5610137, 11.03.97, NKI 514/12.

 9. U.S. 5449759, 09/12/95, NKI 530/375.

Claims (1)

Plasma-substituting solution, including an isotonic solution, characterized in that it additionally contains a sodium salt of a DNA derivative obtained from animal raw materials having an average molecular weight (1.5 - 550.0) • 10 ³ Daltons and a hyperchromic effect of 28 - 46%, in the amount of 5 • 10 ^-5 - 5 g per 1 liter of isotonic solution.

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