LT4369B - Use of beta-endorphin and prostaglandins e1, e2, f2alfa, i2 for the preparing of infusions which can compensate disorders of vitaly important functions during anaphylactic shock - Google Patents

Abstract

Invention covers biologically active beta-endorphin (β-lipotropin 61-91) and E1, E2, F2α, I2 prostaglandins (derivatives of arachidine, linolenic poly-non-saturated lipid acids) use for production of intravenous microdropping infusions, which may be used for vital function disorder compensation during anaphylaxis. Afore mentioned substances are being dissolved in ethyl alcohol at 1/1,000 ratio, and afterwards preparing an infusion, taking effective dose it is diluted by isotonic solution at 1/130 ratio. Dropping is done as follows: β-endorphin by the method of intravenous microdropping infusions in 0.5 μg/kg/min, E1, E2, F2α prostaglandins in 10 μg/kg/min, and I2 prostaglandin in 100 μg/kg/min.

Description

The invention relates to medicine and can be used in the preventive therapy of anaphylactic shock (IOP), other extreme conditions, in the treatment of cardiac arrhythmias, cardiac muscle ischemia, haemocoagulation and other vital functions. It can also be used in scientific experimental research.

β-endorphin (β-lipotropin 61-91), isolated from camel hypolysis, known as the biologically active neuropeptide, is involved in the regulation of neural processes. The powdered solution thereof is usually injected into the cerebral ventricles (Clin. Exp. Pharmacol. Physiol. 17, 1990, 17, 1, 33-37; Biomed. Pharmacothcr. 47, 8, 311-320, 1993; Neuropsyehologia, 1993). 31.7, 655-660).

Hey, E ₂ , F _2a , I ₂ prostaglandins (arachidic derivatives of linoleic polyunsaturated lipid acids) isolated from animal prostate and other tissues are known as biologically active lipids that regulate vital cardiovascular, respiratory, haemocoagulatory and other functions. Their solutions are injected intravenously or locally into the damaged wire. When injected into an artery, prostaglandins are more potent than intravenous as they prevent the destruction of their lung and liver enzymes (Ažgichin I-, Prostaglandiny. - M., 1978 - 565, Russian). Although β-endorphin and prostaglandins are more rapidly degraded and act more potently when administered to the artery, they can only be used for a short period of time because they cannot compensate for the long-term effects of IGs because they are shorter than IG pathologies. In addition, β-endorphin or prostaglandin injected into the artery have a relatively local effect and therefore do not affect the function of the whole body, and long-term arterial infusion is practically inappropriate due to high blood pressure. Continuous microvascular intravenous infusions of βendorphine and prostaglandins greatly reduce the catastrophic (lethal) effects of IOP. Their performance is determined by recording and tracking the parameters of the specified malfunctions.

Both β-endorphin and EH, E ₂ , F _2a , I ₂ prostaglandins are rapidly degradable (within seconds). Therefore, when used in stressful states (anaphylactic shock), where severe and long-lasting damage to functional systems occurs, only short-term, ineffective functioning is achieved.

For a more consistent effect, the present invention provides methods and dosages of intravenous microvascular infusions of β-endorphin, E _b E ₂ , F _2a , I ₂ prostaglandins, as well as their concentrations, for compensation of vital dysfunction during anaphylaxis.

Examples of preparation of intravenous microvascular infusions of β-endorphin and E _t E ₂ , F _2a , I ₂ prostaglandins.

1.Example

The stock solution of β-endorphin was obtained by dissolving its crystalline powder (Heidelberg Servo firm) in 98% ethyl alcohol in a ratio of 1: 100. From this (stock solution), we took aliquots of saline (isotonic 0.9% sodium chloride) at a ratio of 1: 130, after taking the active dose (125 pg).

2. Example

E ₂ , F _2ct or I ₂ prostaglandin solutions (individually) were prepared by dissolving their powder (Tallinn-made) in 98% ethyl alcohol at a ratio of 1: 100 and taking the effective dose (8 mg) of E ₂ and F ₂ prostaglandins and prostaglandin I ₂ active dose - 80 mg infusions were finalizing them atskiesdami physiological (isotonic 0.9% sodium chloride) solution in a 1: 130th

Biological challenge

Following the use of intravenous microvascular infusions of these biologically active substances with adaptive-compensatory and immunomodulatory activity, we performed tests by inducing IOP and recording functional blood pressure, cardiac electrocardiographic (ECG), hemocoagulographic and spirometric parameters.

The aim of this study was to determine the composition and concentrations of β-endorphin and Εμ E ₂ , F _2a , I ₂ prostaglandin solutions by the preparation of intravenous microvascular infusions that allow dosing and constant compensation for impaired vital functions (arterial blood pressure, respiration, heart function, coronary blood flow and haemocoagulation). We have found that this method of preparation and administration is more effective than single intravenous or intraperitoneal injections, since it helps to permanently compensate for these functional disorders during anaphylaxis. The steady state of intravenous infusions is determined by recording and monitoring the parameters of the reported functional disorders.

67 rabbits of both sexes, weighing 2.0 to 3.5 kg, were tested. They were divided into 6 experimental groups receiving β-endorphin or Εμ E ₂ , F _2cc , I ₂ prostaglandins by intravenous microvascular infusion, with anaphylactic shock, and recording functional blood pressure, cardiac electrocardiographic, hemocoagulographic and spirometric measurements. Rabbits were sensitized with native chicken egg protein diluted to 20% in isotonic 0.9% sodium chloride solution for 1 ml by subcutaneous injection 4 times every 5 days. 25-30 days after the first sensitization injection, rabbits were challenged with anaphylactic shock by intravenous injection (1 ml / kg) of the same antigen. Blood pressure in the carotid artery was measured with a mercury pressure gauge, external breathing with a pneumograph, cardiac activity with an electrocardiograph and blood coagulation with a hemocoagulograph.

The test data are shown in Tables 1-5.

1. Experiment Group

After intravenous microvascular infusions of 0.5 µg / min (0.2 µg / kg / min, mean 2.5 kg rabbit) for 10 min in sensitized rabbits, we found that this dose of β-endorphin significantly reduced rabbit blood pressure, worsens cardiac intravascular conduction and impairs the flow of IOP (several rabbits killed).

2. Experiment group

After administration of 0.15 µg / kg / min intravenous microvascular infusions of 0.15 µg / kg / min to sensitized rabbits and subsequent induction of anaphylactic shock, we found that this amount of β-endorphin had little effect on arterial blood pressure in sensitized rabbits and sinus 236.5 rpm to 215.5 rpm (Table 3). In the first I minutes, 50% of rabbits recorded single ventricular and one rabbit atrial extrasystoles, and after 6-10 minutes they disappeared or moved to S-T segment dislocation. In the first minutes of HR, arterial blood pressure increased (from 117.6 mm Hg to 121.0 mm Hg) and decreased to 87.1 mm Hg in 2-3 min. At 10-15 min, arterial blood pressure stabilized (106.0 mm Hg) (Table 2).

3. Experiment group

Cause sensibilizuotiems rabbits routine I the native hen egg white (specific antigen) by intravenous injection (1 mL / kg) and is not used than β-endorphin than Εμ E2, F2 _a, I2 prostaglandins were registered anafilaksiniam shock typical blood pressure and lung dynamics: the initial transient increases in blood pressure, with a mean reduction in peak IOP of 3 minutes (p <0.01), increased respiratory rate (p <0.05) and worsening (Table 1-2). Extrasystole (30%), cardiac conduction abnormalities (30%), and signs of myocardial ischemia (35%) were reported, which in some cases resulted in a myocardial infarction (sheet 3). Blood collected at the second to third minute of anaphylactic shock did not coagulate in most rabbits within 10 minutes (Table 4). 50% of rabbits were killed by anaphylactic shock.

4. Experimental group

Following intravenous microvascular infusions of 10 pg / kg / min Εμ E ₂ , F _2ct prostaglandins (individually) 10 minutes prior to rabbit anaphylactic shock and during anaphylactic shock, we observed a mean increase in arterial blood pressure of 12, 1 minute after anaphylactic injection. 7%, 12.8%, and 20.1%, respectively, for the indicated individual prostaglandins, and external respiration was not significantly different from normal anaphylactic shock (Tables 1, 2). After anaphylactic injection, 30% of rabbits receiving Ei prostaglandin, 30% of rabbits; we received extrasystoles and ventricular fibrillation during E2 prostaglandin and 50% - F _2a prostaglandin shock, and in others (10%, 10% and 15%) cardiac conduction abnormalities and signs of ischemia (10% each) (Table 3). Extrasystoles (except in two cases) and ventricular fibrillation resolved within 5-10 min.

5. Experiment group

Following the administration of 100 µg / kg / min I2 prostaglandin microinjections prior to anaphylactic shock and anaphylactic shock, two rabbits had atrial extrasystoles and two had ischemic events (Table 3). The thromboelastographic (blood coagulation) indices of rabbits sensitized to prostaglandin I ₂ by H. Hartert (Clin. Wschr., 1948, 26, 577-583) were changed by: "r" (blood prothrombin response time), "k" (clot formation time). and “t” (specific coagulation time constant) was prolonged, whereas “ma” (maximum amplitude reflecting clot elasticity) decreased (p <0.01, p <0.05, p <0.05, and p <0.05). (Fig. 3, Table 5). The blood of these rabbits underwent coagulation within 10 minutes of the action of I-prostaglandin (Table 4). During normal IG, blood does not coagulate due to the development of intravascular coagulopathy (Barkagan Z., Gemoragičeskije zabolevanija i syndrome. M. 1980, p. 5, Russian), with the formation of multiple microthrombi in capillaries. This disrupts microcirculation and causes tissue hypoxia. Therefore, it is important to prevent the development of primary hypercoagulation by slowing down blood coagulation prior to induction of anaphylactic shock by infusion of I ₂ prostaglandin or β-endorphin (Fig. 1, Table 5). Consequently, from I killed only 6% of rabbits received the prostaglandin I _2, 8% - When β-endorphin, 10% received E _b prostaglandin E2, 16% F ₂ acting "Prostaglandins and 50% of untreated (Fig. 2).

Figure 1 Rabbit haemocoagulograms: a - sensitized, b - second to third minute anaphylactic shock, c - receiving I ₂ prostaglandin second to third minute anaphylactic shock.

Figure 2 Rabbit blood pressure tendency and outcome of anaphylactic shock: percentage of deaths due to β-endorphin (β-End.), E _b E ^, F _2a and I ₂ prostaglandins, K to prostaglandins and βendorphin.

Figure 3 Hemocoagulogram parameters (according to H. Hartert): r - Blood prothrombin response time (from start of recording to 1 mm amplitude); k - time of clot formation (amplitude from 1 mm to 20 mm); ma is the maximum amplitude representing the elasticity of the clot (platelet and fibrinogen clotting activity); t - specific coagulation constant (duration min, sec).

Rabbit Respiratory Rate (rpm) Table of βendorphin (β-End) and E _b E2, F _2a , I ₂ Prostaglandins (Pg) Received in Anaphylactic Shock (HR)

Time of investigation Expel · ^ nature Sensitized (up to IU), cpm 1st min, rpm On the 10th min, rpm Number of rabbits (n) At the moment Received 115.6 * 13.2 130.0 * 16.5 p <0.05 110-14.8 10th β-End Received 113.5-10.5 132.5 * 12.5 p <0.05 110.3 * 12.5 12th Pgi2 114.3 12.2 131.5 * 14.3 p <0.05 115.3 * 13.5 15th Received Pg 1 120.0 * 12.5 145.0 * 23.5 p <0.05 105.0 * 11.2 10th Received Pg ^ -2 121.5 * 10.8 142.3 * 20.5 p <0.05 106.5 * 10.3 10th Received Pg ^h 2a 123.3 * 14.8 148.0 * 25.2 108.5 * 13.2 10th

p <0.05

Changes in rabbit arterial blood pressure (mmHg) during anaphylactic shock (AS) and table of β-endorphin (β-End) or hey, E _2> F _2a and L prostaglandins (Pg)

Time of investigation Sensitizing. (up to I) mm Hg 1st O min, mm Hg 2-3 O min, mm Hg 10th O min, mm Hg Fatalities number in) -Exper. character ^ At the moment 120.0 * 2.5 128.0 * 2.0 68.5 * 10.8 p <0.01 82.0 * 21.5 p <0.05 5 Gavisius β-End 117.6 * 5.5 121.0 * 5.6 87.1 * 14.3 p <0.01 106.0 * 18.5 p <0.02 1 Receiving Pgl ₂ 100.7-6.0 121.0 * 3.6 67.8 * 30.7 p <0.01 79.5 * 15.8 p <0.05 1 Received PG No. 99.4 * 5.6 112.0 * 9.5 68.4 * 17.2 p <0.01 75.6 * 19.5 p <0.05 1 After receiving Pg ^E 2 100.7 * 6.0 113.5 * 3.6 67.8 * 20.7 p <0.01 74.7 * 18.5 p <0.01 1 Received Pg ^F 2 « 94.0 * 5.3 112.9 * 6.0 60.5 * Ί4.2 p <0.01 60.8 * 3.7 p <0.01 2

Heart rate (rpm) and ECG data of rabbits during anaphylactic shock (HR) and Table of prostaglandins (Pg) receiving β-endorphin (β-End) or ^E 1, ^E 2, ^E 2a, h

ECG lesions Lkspt ^. nature Sensitive lysed, rpm At I, rpm Excitement disorders, (cases number) Conductivity lesions, (cases number) Ischemia phenomena, (cases number) At the moment 255.0 * 5.0 261.0 * 7.0 Ms. Ex-1 Sk.Ex-2 7th 5 Received β-End 236.5 * 5.6 215.5 ^ 6.2 Pr.Ex-l Sk.Ex-3 2 2 Received 260.0 * 7.8 277.5 * 9.6 Pr.Ex-2 1 2

Received PgEl 210.0 * 10.5 211.0 * 10.2 Pr.Ex-3 1 1 Received Pg ^E 2 208.0 * 9.5 210.5 * 9.3 Pr.Ex-3 1 1 Receiving Pg ^E 2cc 245.0 * 7.8 237.0 * 22.0 See Ex-4 2 1

Note: Pr.Ex - Atrial Extrasystoles Sk. Ex - ventricular extrasystoles

Rabbit haemocoagulogram data during anaphylactic shock (HR) and Table of prostaglandins (Pg) receiving β-endorphin (β-End) or Ei, E2, F _2a> I2

Hemocoagulographic indices Expert> \ nature r min, sec k min, sec ma mm t min, sec Obvious rabbit number (n) Sensitizing 2'50 * Γ05 5Ί5 * Γ40 50.2 * 3.0 12Ό-4Ί5 - At the moment 18'54 * 6'40 21'54 * 8'30 19.0 * 3.8 - 6th Received β-End 7Ό8 * Γ45 9'1O * 1'2O 17.5 * 3.3 - 1 Received Pg 16'30 * 3'20 25'40 * 7'30 22.0 * 3.5 - - Receiving Pg ^E i 18Ί5 * 6’20 20'51 * 5'40 20.3 * 3.8 - 4 After receiving PgE ₂ 18:30. 54θ 2Γ10 * 4’35 20.614.1 - 3 Receiving ^P 8 ^F 2a 17'40t5'15 19'30t6'20 18.4 * 3.2 - 5

r - Blood prothrombin response time (from recording to 1 mm amplitude);

k - time of clot formation (amplitude from 1 mm to 20 mm);

ma is the maximum amplitude representing the elasticity of the clot (platelet and fibrinogen clotting activity);

t - specific coagulation constant (duration min, sec).

Hemocoagulogram data for rabbits receiving β-endorphin (β-End) or E ^ E ₂ , F _2a , I ₂ prostaglandins (Pg)

Hemocoagulum lyograms indicators Expert \ character ^ r min, sec k min, sec ma mm t min, sec Sensitizing 2'48 * 0'30 5'24 ^ l'20 52.4 * 3.9 12'40 * 4'30 ^3, 15 ^ l'3O " 6'40 * 2'15 1630 ^ 530 ¹¹ p <0.05 Received 5'45 * 1'15 6'36 * 2'30 37.0¾ 15'51T5'45 ^p gi ₂ p <0.01 p <0.05 p <0.05 p <0.05 Receiving Pg ^E i 4'30 * T20 p <0.02 5'30 ri-40 50,313.6 13'20 * 4'15 Receiving P8E ₂ 3'20 * O'45 5'40 ^r l'3O 51.5 - * 3.2 14Ί0 * 4’30 After receiving PgF ⁶ 2a 2'40 ^t 0'30 5'30 * Γ45 52.0 * 3.5 13Ί0 - ⁺ 4’15

r is the blood prothrombin response time (from the start of recording to Ϊ mm amplitude);

k - time of clot formation (amplitude from 1 mm to 20 mm);

ma is the maximum amplitude representing the elasticity of the clot (platelet and fibrinogen clotting activity); t - specific coagulation constant (duration min, sec).

SUMMARY OF RESULTS

1. Microvascular intravenous infusions of β-endorphin (0.15 pg / kg / min) stabilized the dynamics of blood pressure changes in anaphylactic shock, reduced cardiac dysfunctions and fatal outcome of anaphylactic shock.

2. Shortly before anaphylactic injection in rabbits given an epaphylactic shock of Ep E ₂ or F _2a prostaglandins (10 pg / kg / min), a slight increase in arterial blood pressure was observed, but the dynamics of drop in blood pressure and external respiration subsequently did not differ significantly from the more favorable anaphylactic shock.

3. Hey, E ₂ , F _2a and I ₂ prostaglandins were antiarrhythmic during anaphylactic shock and I ₂ and anti-ischemic.

4. Prostaglandins Ei, E ₂ , F _{2a were} unaffected by the anaphylactic shock-related retardation of blood coagulation, whereas I ₂ prostaglandin and β-endorphin stabilized blood coagulation and apparently resulted in the survival of most rabbits.

Studies have shown that biologically active substances such as β-endorphin and Ej, E ₂ , F ₂ a, I ₂ prostaglandins would be appropriate for infusions.

An advantage of the present invention is that the use of solutions of this concentration by intravenous micro-infusion can attenuate the effects of anaphylactic shock and compensate for impaired functions such as blood pressure, cardiac function, and coagulation. In this case, β-endorphin and Ei, E ₂ , F _2a , I ₂ prostaglandins act as anti-anaphylactic (anti-stress) factor.

Claims (4)

DEFINITION OF INVENTION 1. Use of biologically active substances β-endorphin (β-lipotropin 61-91) and Ei, E ₂ , F _2a , I ₂ prostaglandins (arachidic, linoleic polyunsaturated lipid acid derivatives) by intravenous microvascular infusion with compensation of vital dysfunction in anaphylaxis during the preparation. Use according to claim 1, characterized in that β-endorphin and E ₁ , E ₂ , F _2a , I ₂ prostaglandins are used in the form of intravenous micro-infusions obtained by dissolving their crystalline powder in 98% ethanol at a ratio of 1: 100 and isotonic dilution. in a 1: 130 solution. 3. An intravenous micro-infusion, characterized in that it contains β-endorphin and Ei, E ₂ , F _2a .I ₂ prostaglandins as active ingredient for use in compensation of vital dysfunction during anaphylaxis. The infusion according to claim 3, characterized in that the active ingredient is used as follows: β-endorphin in the form of a solution for intravenous microvascular infusions of 0.15 pg / kg / min, E ₁ , E ₂ , F _{2ct for} prostaglandins in solution for intravenous microvascular infusions. pg / kg / min and I ₂ prostaglandin at 100 µg / kg / min.