RU2251432C2 - Method for preventing hypoxia - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: the present innovation deals with preventing the development of hypoxia due to introducing ceruloplasmin 1 d before hypoxia. The method enables to exclude the development of allergic reactions.

EFFECT: higher efficiency of hypoxia prevention.

2 ex, 3 tbl

Description

The invention relates to medicine, namely to the use as a antihypoxic drug of human blood - ceruloplasmin, and can be used in clinical practice.

A number of drugs with antihypoxic properties are known that provide normalization of energy metabolism and contribute to adaptation to oxygen deficiency conditions. These include amtisol, trimetazidine, oliphene and cytochrome C (Abramchenko V.V. “Antioxidants and antihypoxants in obstetrics”, St. Petersburg, Publishing House DEAN, 2001, p. 220-236). These are mainly synthetic compounds, sometimes foreign to the body, with their overdose, the development of a prooxidant action is possible, i.e. effect opposite to expected.

The closest to the claimed in essence and the result obtained is the antihypoxic drug Energostim, which includes the enzyme cytochrome C, nicotindiimide dinucleotide and inosine, described in US patent 5439883, A 61 K 31/455, published on 08.08.95. The proposed drug helped to normalize energy metabolism and reduce oxygen deficiency in the tissues of the heart, brain and liver. The disadvantage of this combination is that one of its components, cytochrome C, is isolated from animal tissue. Intravenous and intramuscular administration of such a drug can cause the development of allergic reactions in recipients.

The problem solved by the invention is the expansion of the range of drugs with antihypoxic effect.

The technical result from the use of the invention is the exclusion of the development of allergic reactions due to the fact that the proposed drug is obtained from human blood and does not contain foreign components.

The specified result is achieved by the use of ceruloplasmin as an antihypoxic drug.

The antihypoxant we offer is a drug released from human blood plasma. This is ceruloplasmin, a copper-containing enzyme belonging to the alpha-2-globulin fraction of blood plasma. Due to the ferrooxidase action, ceruloplasmin is able to stimulate the absorption of iron by the body. It is the main antioxidant in blood plasma and the main reserve of copper in the body. The antioxidant, radioprotective and antianemic properties of ceruloplasmin have long been known, and this has allowed its use in clinical practice in the treatment of anemia, critical conditions of various origins, in the treatment of tumors, etc. (Kraineva T.A., Efremova L.M. “Ceruloplasmin - biological properties and clinical application.” Nizhny Novgorod, publ. NGMA, 2000, 19-24). The neuroprotective properties of ceruloplasmin are known (Patent US 2002/0094949, A 61 K 38/41, published July 18, 2002). However, the authors of these inventions used non-commercial preparations of ceruloplasmin when exposed to isolated brain cell cultures. Antihypoxic properties of the drug can be proved only when conducting experiments on animals. Were not proposed methods of introducing the drug into the body.

The drug “Ceruloplasmin” is administered intravenously or intramuscularly in pathological conditions accompanied by hypoxia. The antihypoxic effect of the drug was studied on the basis of the Central Scientific Research Laboratory of the Nizhny Novgorod State Medical Academy. The effect of the drug is demonstrated in the examples below.

Example 1

1. The study was carried out on sexually mature white outbred male rats. Statistical groups of experimental animals consisted of 13 individuals. A model of acute hypobaric hypoxia (OGBG) was used - the "death pad" test. Using a vacuum pressure chamber, the animal was placed at a "height" of 11500 meters at a speed of 183 m / s. Recorded such indicators as the time of loss of posture (runway) after lifting, life time (Tg) - the time interval from the completion of the "lifting" to the second agonal inhalation. The number of animals that recovered posture (VP) from the start of the "descent" within three minutes. By the time before the appearance of the second agonal inhalation, the animals were divided into three groups:

1) low resistance (NU) - the life time (TJ) is less than three minutes;

2) medium stable (SU) - life time (TJ) from three to ten minutes;

3) highly stable (WU) - the life time (TJ) is more than ten minutes.

Due to the need to study the metabolic parameters during hypoxia, the death of animals, especially low resistance animals, was not expected at the "death site". Immediately after the second agonal inhalation, the pressure in the pressure chamber was reduced to the initial values. After the experiment, the blood of animals was taken for biochemical studies. Quantitative determination of pyruvate and lactate was carried out according to the method of Asatiani B.C. “New methods of biochemical photometry”, - M., Nauka, 1965, p.541. The intensity of free radical oxidation was determined by the method of induced chemiluminescence (EI Kuzmina et al. Interuniversity collection of biochemistry and biophysics. Gorky, 1983. - p. 179-183). The study of lipid peroxidation (lipid peroxidation) and plasma antioxidant activity (AOA) was carried out using the methods of induced biochemiluminescence (IBCL), while Imax was determined - the maximum luminescence intensity of the test sample, measured in mV, reflecting the free radical activity of the sample, indicator S is the CL light sum for a certain time inversely proportional to the antioxidant activity (AOA) of the sample, the ratio Lmax / S was calculated, which characterizes the total AOA of blood plasma. When UV spectroscopy of the primary products of lipid peroxidation (Lankin V.Z. et al. Cardiology, 1979, No. 6, pp. 71-75), the content of diene conjugates (DC), triene conjugates (TC) and double bonds (DS) were determined. The determination of superoxide dismutase activity was carried out according to the method of M. Nishicimi, A. Roo, K. Xagi, Biochem. Biophys. Res. Commun., 146 (2), 849-854, (1972). Ceruloplasmin was dissolved in physiological saline and administered to animals intraperitoneally (analogous to intravenous administration in small rodents) at a dose of 50 mg a day before hypobaric hypoxia was created. In the control series used sterile saline in appropriate quantities.

The experiments showed that the preventive use of ceruloplasmin affects the survival of animals in modeling acute hypobaric hypoxia (Table 1). With the introduction of ceruloplasmin, the lifetime (Tg) in conditions of acute hypoxic hypoxia exceeds the control almost three times. Among the population of rats subjected to hypoxia, the number of highly resistant animals increases and the number of low-resistant animals decreases. The time of loss of posture (VPP) under conditions of hypobaric hypoxia is significantly longer than in the control. After the hypoxic period, rats of this group normalize their neurological status faster, by 3 minutes most animals restore their posture, unlike the control group, without administering drugs, and indicators of neurological status in the posthypoxic period are restored faster. According to biochemical studies of indicators of carbohydrate metabolism in blood plasma, we can conclude about a decrease in lactic acidosis in the blood during hypoxia. Significantly lower than the control, the lactate content in the blood plasma and the ratio of lactate / pyruvate (table 2). Data on the state of tissue hypoxia in the body (lactate / pyruvate) during the development of acute hypoxic hypoxia indicate a protective antihypoxic effect of ceruloplasmin. Ceruloplasmin caused activation of the antioxidant enzyme SOD (Table 3). According to other biochemical parameters, free radical activity, the content of primary lipid peroxidation products (DC and TC) were significantly lower when using ceruloplasmin compared to the control.

Thus, the administration of ceruloplasmin significantly increased the antioxidant defense of the body, as a result of which, with hypoxia, there was no pathological increase in free radical oxidation and accumulation of lipid peroxidation products, in contrast to the control group without the use of the drug.

Table 1.
Indicators of survival and resistance to hypoxic hypoxia of white outbred rats. Indicator Control, n = 13 Intramuscular injection n = 13 Intraperitoneal administration n = 13 T _w, min 16.5 ± 0.5 55 ± 1.5 * 50 ± 1.5 * Number of WU 0 3 (25%) 2 (16%) NU 9 (75%) 6 (50%) 5 (42%) Number of SU 3 (25%) 3 (25%) 5 (42%) Runway, min 6.5 ± 0.3 10 ± 0.4 * 10.5 ± 0.2 * VP count 18%) 7 (58%) 9 (75%) Note: * - significance of differences with the control series, p <0.05. Table 2.
The effect of ceruloplasmin on blood carbohydrate metabolism in experimental animals Indicator Control n = 13 Intramuscular administration of ceruloplasmin n = 13 Intraperitoneal administration of ceruloplasmin n = 13 Pyruvate 0.33 ± 0.05 0.29 ± 0.02 0.29 ± 0.04 Lactate 1.43 ± 0.07 1.1 ± 0.03 * 1.2 ± 0.08 * L / R 4.3 ± 0.02 3.8 ± 0.013 * 4.1 ± 0.02 *

Table 3.
The effect of the drug on the performance of the pro- and antioxidant systems of the body of experimental animals Indicator Control n = 13 Intramuscular injection, n = 13 Intraperitoneal administration, n = 13 Imax 4.3 ± 0.2 3.5 ± 0.3 * 3.8 ± 0.2 * 1 / S 0.018 ± 0.005 0.031 ± 0.004 * 0.030 ± 0.003 * DK 0.075 ± 0.005 0.059 ± 0.005 * 0.060 ± 0.004 * TC 0.034 ± 0.002 0.028 ± 0.001 * 0.029 ± 0.001 * SOD 183.7 ± 9.6 284.3 ± 12.4 * 276.0 ± 10.2 * Note: * - significance of differences with the control series, p <0.05.

Example 2

Ceruloplasmin was administered to animals intramuscularly one day before hypoxia. The same analyzes were performed as in Example 1. Similar results were obtained (Table 1-3). The use of ceruloplasmin improved the survival of animals in conditions of acute hypoxia. There was no difference between the parameters obtained by comparing groups of animals with intramuscular and intravenous administration of the drug.

Thus, the studies obtained have confirmed that ceruloplasmin has antihypoxic properties and can be administered both intravenously and intramuscularly in pathological conditions accompanied by hypoxia.

Claims (1)

A method of preventing hypoxia, characterized in that ceruloplasmin is administered one day before hypoxia is created.