WO2020027686A1 - Method of preventing entero-entero anastomotic leaks - Google Patents

Abstract

This invention relates to experimental medicine, and specifically to experimental gastroenterology, and may be used to prevent entero-entero anastomotic leaks. A method of preventing entero-entero anastomotic leaks comprising action using a medicinal preparation, characterized in that the action using a medicinal preparation is realized by irrigating the surface of an entero-entero anastomosis with a peptide of structural formula Tyr-D-Ala-Gly-Phe-Leu-Glu.

Description

 METHOD FOR PREVENTING INSOLVENCY

 INTESTINAL ANASTOMOSIS

The present invention relates to experimental medicine, and in particular, to experimental gastroenterology and can be used to prevent interintestinal anastomosis insolvency.

Failure of the intestinal anastomosis is a deadly complication in various operations on the organs of the abdominal cavity.

Various methods are known in the art for the prophylaxis of interintestinal anastomosis insolvency, including drug exposure [http://www.rlsnet.ru/tn_index_id_3097.htm], [http://t-pacient.ru/articles/6022/], [Patent RF 2475247]. Despite the methods used, the likelihood of developing an intestinal anastomosis failure during abdominal surgery remains high.

The invention is aimed at creating an effective and safe method for the prevention of interintestinal anastomosis insolvency, including exposure to a drug.

The solution to this problem is provided by the fact that in the method for preventing insolvency of the intestinal anastomosis, including exposure to a drug according to the invention, the drug is exposed by irrigating the surface of the intestinal anastomosis with a peptide of the structural formula Tyr-D-Ala-Gly-Phe-Leu-Glu.

The applicant experimentally established the effectiveness of the method, including irrigation of the surface of the intestinal anastomosis with a peptide

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SUBSTITUTE SHEET (RULE 26) structural formula Tyr-D-Ala-Gly-Phe-Leu-Glu for the prevention of interintestinal anastomosis insolvency.

The above peptide has multifunctional properties. It accelerates the regeneration of blood vessels and nerves, has anti-inflammatory and anti-ischemic activity. Activation of the lymphatic flow helps to accelerate detoxification mechanisms, increase interstitial transport in tissues, regulate water-salt metabolism, and stimulate the immune system as a result of an increase in the release of lymphocytes and lymphopeptides from lymph nodes into the lymph and into the bloodstream.

An example embodiment of the invention is given below.

Example 1

Evaluation of the effectiveness of the method for preventing insolvency of the intestinal anastomosis during operations on the abdominal cavity, including exposure to the peptide, was studied by the survival of animals in which the obstruction of the small intestine was simulated, followed by resection of the non-viable part of the small intestine and restoration of intestinal patency by applying an interintestinal anastomosis. At the same time, microcirculation of the walls of the small intestine and surrounding tissues was studied in vivo. A histological study of small intestine preparations confirmed the positive results obtained by using the peptide for the prevention of interintestinal anastomosis failure in operations on the abdominal cavity.

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SUBSTITUTE SHEET (RULE 26) Table 1.

Rat survival in various models of acute intestinal obstruction (OKN) and peptide exposure.

 n is the initial number of animals; nl is the number of surviving animals at

OKN in units, n 2 - in%. “+” - one effect, “+++” - three effects (in the table, the ligature on the 3 arteries of the mesentery).

Table 1 presents the survival results of animals with various models of intestinal obstruction. We studied 9 groups of animals, a total of 210 outbred white rats of males weighing 200-260 g. All animals were anesthetized with intramuscular injection of 8% chloral hydrate solution at the rate of 0.5 ml per 100 g of weight.

The first group of animals after anesthesia was subjected to laparatomy;

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SUBSTITUTE SHEET (RULE 26) Group 2 - in addition to laparatomy, a ligature was placed on a small intestine located at a distance of 7-10 cm proximal to the ileocecal transition by 7-10 cm;

Group 3 was ligated on one segmental arterial vessel of the mesentery of the small intestine;

In the 4th group, a ligature was placed on three mesenteric vessels;

The 5th group underwent a torsion of a section of the small intestine with a feeding vessel using ligature;

The 6th group crossed the small intestine with the subsequent restoration of its patency, stitching the ends of the intestine;

The 7th group performed a resection of a fragment of the intestine followed by the application of an inter-intestinal anastomosis;

The 8th group caused intestinal obstruction by ligation 7-10 cm proximal to the ileocecal transition, after 24 hours a non-viable small intestine section was resected, then intestinal patency was restored by application of an inter-intestinal anastomosis. Before suturing the wound, the surface of the intestinal anastomosis was irrigated with 1 ml of physiological saline.

The 9th group differed from the 8th by dissolving the peptide at a dose of 40 μg / kg in 1 ml of physiological saline.

An analysis of the results of Table 1 indicates that the operation of laparatomy, ligation of one mesenteric artery, as well as only the intersection or removal of a fragment of an intact small intestine with subsequent application of an inter-intestinal anastomosis, ends in 90-

100% of cases are favorable. Other consequences (100% death of animals) occur with a significant violation of the blood supply to the walls of the small intestine as a result of ligation of the 3 mesenteric arteries, in violation

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SUBSTITUTE SHEET (RULE 26) patency of the intestine in conditions of its torsion or ligature. Resection of a non-viable part of the small intestine with subsequent application of an inter-intestinal anastomosis increases the survival of animals from 0% to 43%. The cause of the death of animals in groups 8 and 9 was the failure of the intestinal anastomosis. The highest survival rate of 66% was obtained using the studied peptide (see Table 1 gr. 9).

During the operation, antibiotics and other antimicrobial and anti-inflammatory drugs were not used. An increase in the survival of animals of the 9th group by 23% compared with the control (group 8) was achieved only through local single use of a small dose (40 μg / kg) of the peptide during the operation.

Table 2 presents the results illustrating the survival of animals with different weights without the use of a peptide in the control (group 8) and when exposed to the peptide (group 9). The results of a comparative analysis of the survival of rats indicate greater survival of animals treated with the peptide (an average of 42% more in all subgroups). Based on the results of table 2, we can conclude: the peptide is effective for the prevention of insolvency of the intestinal anastomosis in various age subgroups of the studied animals.

Table 2.

Comparative characteristics of the survival of animals with acute intestinal obstruction of various weights in the control (group 8) * and when exposed to the peptide (group 9) **.

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SUBSTITUTE SHEET (RULE 26) Note: n is the number of animals. * - control included the following actions: 1) laparatomy; 2) ligature application to the rat small intestine proximal to the ileocecal transition by 7-10 cm; 3) resection of a fragment of the small intestine above and below the ligature after 24 hours; 4) the imposition of an inter-intestinal anastomosis. ** - peptide treatment: the inter-intestinal anastomosis at the end of the operation was irrigated with a peptide at a dose of 40 μg / kg in 1 ml of physiological saline.

Table 3 presents the results of a pathomorphological study of the fragment of the intestinal anastomosis on the 7th day after application in rats of the 8th (control) and 9th (peptide) groups.

Table 3.

The effect of the peptide on the restoration of the intestinal wall in the area of the intestinal anastomosis (histological examination).

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SUBSTITUTE SHEET (RULE 26)

The results obtained indicate not only an improvement, but also a restoration of morphological examination indicators, indicating restoration of impaired blood circulation and lymph circulation in the small intestine wall with restoration of lymphatic drainage, regression of interstitial edema, structural damage to the suction epithelium and smooth muscle cells of the muscle layers responsible for contractile function and intestinal motility.

The results of an intravital study of microcirculation in blood and lymphatic microvessels of the small intestine wall are presented in Table 4 and Table 5. The diameters of small veins and the frequency of contractile activity of lymphatic microvessels in rats of the 8th (control) and 9th (peptide) groups are compared.

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SUBSTITUTE SHEET (RULE 26) Table 4.

The diameter of the small veins on the surface of the small intestine of the surviving rats in the control * and on the 7th day after the resection of the intestine and the creation of an intestinal anastomosis in surviving animals of the 8th and 9th groups.

 Note: * - the initial diameter of the small veins before applying the ligature.

Table 5.

The frequency of contraction of the wall of the lymphatic microvessels of the mesentery of the small intestine of surviving rats in the control * and on the 7th day after resection of a fragment of the intestine, and the creation of an intestinal anastomosis in surviving animals of the 8th and 9th groups.

 Note: Control * is the initial frequency of contraction. Significant differences with control - *, differences between subgroups - **.

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SUBSTITUTE SHEET (RULE 26) On the 7th day after resection of the damaged part of the intestine and application of an intestinal anastomosis, the plethora of venules and small veins in all surviving animals decreases: in control, the diameter of small veins decreases on average by 15%, when using the peptide, the diameter of the veins decreases to a greater extent - by 25% .

 In similar groups of rats, the contraction rate of lymphatic microvessels when using the peptide was 28% higher compared to the control, despite the long period (7 days) after a single dose of a small dose (40 μg / kg) of the peptide was used, which also indicates the high efficiency of the peptide.

All the obtained biomicroscopic and histological studies of the intestinal anastomosis on the 7th day after the operation correspond to indicators of increased survival of animals of the general group and age subgroups when using the peptide.

The data obtained as a result of the study confirm the effectiveness of the method for the prevention of interintestinal anastomosis insolvency, including exposure to a drug, in which the drug is exposed by irrigation of the surface of the intestinal anastomosis with a peptide of the structural formula Tyr-D-Ala-Gly-Phe-Leu-Glu.

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 SUBSTITUTE SHEET (RULE 26)

Claims

 CLAIM A method for the prevention of interintestinal anastomosis insolvency, including exposure to a drug, characterized in that the exposure to the drug is carried out by irrigating the surface of the intestinal anastomosis with a peptide of the structural formula Tug-D-Ala-Gly-Phe-Leu-Glu. 10  SUBSTITUTE SHEET (RULE 26)