RU2403911C1 - Agent for increasing survival rate in acute blood loss - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, particularly to biologically active peptides capable to reduce death rate caused by acute blood loss. The application of octapeptide H-Phe-Nle-Phe-Gln-Gly-Gln-Arg-Phe-NH₂ (I) amide and its pharmaceutically acceptable salts as an agent for increasing survival rate after acute blood loss.

EFFECT: peptide described above effectively increases survival rate after acute blood loss.

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Description

The invention relates to biologically active peptides that can reduce mortality after acute blood loss, and can find application in various fields of medicine, in particular in ambulance, in disaster medicine and in field medicine.

The total mortality from injuries in the world is more than 2.5 million annually (46 people per 100 thousand of the population) [1]. Most injured die within the first hour as a result of acute blood loss [2]. Despite numerous studies, the optimal strategy for stabilizing the post-traumatic condition has not yet been developed [3]. The main causes of death in acute blood loss are insufficient tissue perfusion, cellular hypoxia and the subsequent development of inflammation. Therapy is aimed mainly at stopping bleeding and restoring intravascular blood volume. However, massive and rapid infusion of fluids to maintain blood pressure does not always have the desired effect due to the possible increase in blood loss and cell damage [1-4]. In addition, the use of blood replacement therapy is not always possible at the site of injury or on the battlefield. Therefore, researchers pay particular attention to ways to improve survival in acute blood loss based on the introduction of substances that counteract the development of damaging changes in organs. Some of them reduce the intensity of oxidative stress, while others have an adrenomodulating effect.

The use of antioxidant substances in a number of experiments on animals made it possible to improve survival after acute blood loss. However, these results are only of theoretical, but not practical value, since synthetic free radical traps, as a rule, are toxic for cell metabolism and, therefore, cannot be used for practical use in humans [5].

A more promising direction is the use of various natural neurotransmitters, including neuropeptides. Most of them modulate the activity of the sympathoadrenal system, the main emergency defense mechanism in blood loss. The use of catecholamines alone [6] or in combination with vasopressin [7] during fluid replacement therapy requires very accurate dosing, as they can have both protective and damaging effects [8]. More promising is the targeted use of substances that modulate natural mechanisms.

The positive effect on the survival of animal peptides containing the sequence-His-Phe-Arg-Trp-; such peptides are secreted in large quantities in acute blood loss [9]. This is adrenocorticotropic hormone (ACTH), melanocyte-stimulating hormone (α-MSH) and its analogues - melanocortins. The action of these peptides is carried out by activating the MC4 receptors of the medulla oblongata [10-12] and is accompanied by increased secretion of glucocorticoids, normalization of the level of nitric oxide, and inhibition of the formation of free radicals [9]. Jiuliam et al. [12] showed that as specific Ac-Nle-c [Asp-Pro-D-Phe-Arg-Trp-Lys] Pro-Val-NH ₂ (PG-931), Butir-His-DPhe-Arg-Trp- Sar-NH ₂ (RO27-3225) and non-specific Ac-Ser-Tyr-Ser-Nle-Glu-His-D-Phe-Arg-Trp-Gly-Lys-Pro-Val-NH ₂ (NDP-α- MSH) MC4 receptor agonists reduced the level of reactive oxygen species in the blood, reduced the degree of organ damage, improved circulatory and respiratory functions, and also the survival of rats (of both sexes) during lethal blood loss. High doses of these peptide preparations ensured the survival of all animals in a day. However, these experiments are not free from disadvantages. First, the authors used rats of both sexes, and it is well known that females are much more resistant to blood loss than males [4]. The uncontrolled distribution of females and males in groups could be one of the reasons for the extremely high mortality in the control group, when all rats died within 35 minutes. Secondly, the blood loss from the femoral vein in these experiments occurred gradually over 25 minutes, and such a duration is more characteristic of venous bleeding, while bleeding occurs much faster with damage to the arterial bed. Thirdly, the experiments were carried out under urethane anesthesia, which suppresses natural compensatory mechanisms.

The objective of the invention is to find a means that can prevent mortality within 1 hour after an injury and increase survival in acute blood loss. The urgency of the task is dictated by the need to provide practical medicine with non-toxic drugs for the treatment of acute blood loss, as well as expanding the arsenal of existing means [7].

The problem is solved using the octapeptide amide H-Phe-Nle-Phe-Gln-Gly-Gln-Arg-Phe-NH ₂ (I) and its pharmaceutically acceptable salts (RF patent No. 2346001 [13]) to prevent acute mortality for 1 hours and increase animal survival in acute blood loss. This peptide belongs to the group of neuropeptides, does not contain the -His-Phe-Arg-Trp- sequence in its structure, the presence of which is a distinctive feature of melanocortins and determines their activity, therefore its use to prevent animal mortality from acute blood loss is not obvious. The ability of octapeptide (I) amide to increase blood pressure (BP) and heart rate frequently for a short time is known [13]. However, this ability is not decisive for ensuring better survival in acute blood loss, since many data indicate the absence of a direct relationship between them. Thus, the increase in blood pressure observed upon administration of a hypertonic NaCl solution with dextran [14] or vasopressin with adrenaline [7] was not accompanied by an improvement in survival.

Example. Animal survival in acute blood loss

Male Wistar rats weighing 354 ± 13 g under ketamine anesthesia (100 mg / kg) were inserted with catheters in the femoral and carotid arteries, and in the jugular vein for continuous recording of mean arterial pressure (BP), heart rate (HR) with computer recording, blood sampling and the introduction of saline. Acute blood loss was caused by the selection of arterial blood from the carotid artery with a volume of 2.2-2.5% of the animal’s body weight in 10 minutes according to the method developed by N. Ya. Kovalenko et al. [4]. 3 minutes after the cessation of blood loss, an octapeptide (I) amide in a dose of 0.2 mg dissolved in 0.5 ml of physiological saline was injected into the jugular vein three times, with a break of 5 minutes. Control animals were injected only with physiological saline. An hour after acute blood loss, the catheters were removed, the wounds were sutured, treated with an antiseptic, and the animals were returned to the cells for observation during the day. The control group consisted of 17 animals, the experimental group of 15 rats.

Acute mortality was assessed by the death of animals within 1 hour after acute blood loss. Long-term mortality was estimated by the number of animals that died within 24 hours after acute blood loss. In the control series, 29% of the animals died within an hour after acute blood loss and another 59% died during the next 23 hours, that is, after 24 hours only 12% of the animals survived (drawing). In the group of animals that received peptide (I) after acute blood loss, no one died during the first hour after acute blood loss, 33% of the animals died in the next 23 hours, that is, after 24 hours, 67% of the animals remained alive. Thus, the relative number of animals surviving after 24 hours was 5 times greater in the experimental group than in the control.

The data presented indicate that the peptide (I) completely prevented mortality within 1 hour after acute blood loss and 5 times increased animal survival in the next day.

The proposed dosage form based on the claimed compound may be ampoules for intravenous administration with a substance dissolved in physiological saline. It is assumed that the tool will be used in bolus, which makes the procedure convenient (equipment for transfusion of blood-replacing drugs is not required) and can significantly reduce the amount of the drug used.

Bibliography

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9. Jochem J. & al. The role of melanocortin peptides in the cardiovascular regulation in hemorrhagic shock. Folia Med. Cracov. 2005; 46 (3-4); 13-21.

10. Versteeg D.H., Van Bergen P., Adan R.A., De Wildt D.J. Melanocortins and cardiovascular regulation. Eur. J. Pharmacol. 1998; 360 (1): 1-14.

11. Guarini S., Cainazzo M.M., Giuliani D., et al. Adrenocorticotropin reverses hemorrhagic shock in anesthetized rats through the rapid activation of a vagal anti-inflammatory pathway. Cardiovasc. Res. 2004; 63 (2): 357-65.

12. Giuliani D., Mioni C., Bazzani C., et al. Selective melanocortin MC4 receptor agonists reverse haemorrhagic shock and prevent multiple organ damage. Br. J. Pharmacol. 2007; 150 (5): 595-603.

13. Chazov E.I., Bespalova Zh.D., Sidorova M.V., Palkeeva M.E., Azmuko A.A. Kapelko V.I., Lakomkin V.L. RF patent No. 2346001. 10/18/2007. Publ. 02/10/2009, bull. Number 4.

14. Rocha E, Silva M. Hypertonic saline resuscitation. Medicina (B Aires). 1998; 58 (4): 393-402.

Claims (1)

The use of the octapeptide amide H-Phe-Nle-Phe-Gln-Gly-Gln-Arg-Phe-NH ₂ (I) and its pharmaceutically acceptable salts as a means to increase survival in acute blood loss.

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