Method for preventing renal microcirculation disorders in cisplatin-induced nephropathy using 11-membered peptide simulating the α-helix of b erythropoietin

Abstract

FIELD: experimental pharmacology; urology.SUBSTANCE: rats are injected intraperitoneally daily with an 11-membered peptide that simulate α-helix of B erythropoietin (pHBSP), which does not have an erythropoietic effect, at a dosage of 25 mcg / kg for 14 days. On the first and eighth days of the experiment, 30 minutes after the introduction of the 11-membered peptide simulating α-helix of B erythropoietin (pHBSP), cisplatin is administered intraperitoneally at a dosage of 5 mg / kg.EFFECT: more effective prevention of microcirculation disorders in the kidney in cisplatin-induced nephropathy.1 cl, 1 tbl, 1 ex

Description

[0001] The invention relates to medicine, in particular to experimental pharmacology and urology.

[0002] Every year in the Russian Federation more than 600 thousand cases of malignant neoplasms are detected [S.M. Okladnikov, S.Yu. Nikitina. Healthcare in Russia. 2023: Stat. collection / Rosstat. - M., 3-46 2023. - 179 p.]. Despite all the modern methods of laboratory and instrumental diagnostics, in 18.9% of cases the diagnosis of cancer was first established when the process was already at stage IV [A.D. Kaprin et al. Malignant neoplasms in Russia in 2023 (morbidity and mortality) - M .: P.A. Herzen Moscow Oncology Research Institute - branch of the Federal State Budgetary Institution "NMITs Radiology" of the Ministry of Health of the Russian Federation, 2024. - ill. - 276 p.]. According to current recommendations, drug-based antitumor therapy is the gold standard upon which all treatment of patients with advanced and metastatic forms of cancer is based.Cisplatin is recommended for systemic therapy of a large number of malignant neoplasms; however, it has pronounced nephrotoxicity, which in some cases leads to dose reduction or even complete discontinuation of the drug [Burnasheva E.V., Shatokhin Yu.V., Snezhko I.V., et al. Kidney damage during antitumor therapy / / Nephrology. - 2018. - Vol. 22. - No. 5. - Pp. 17-24]. Given this fact, one of the main tasks facing doctors and scientists around the world is the prevention of cisplatin-induced nephropathy caused by damage to nephron structures due to the use of cisplatin [Altındag F, Ergen H. Sinapic acid alleviates cisplatin-induced acute kidney injury by mitigating oxidative stress and apoptosis. Environ Sci Pollut Res Int. 2023 Jan;30(5):12402-12411].

[0003] The prototype of the claimed invention is a method for correcting microcirculatory disorders in the kidneys in an experiment, the model of which includes a single intraperitoneal administration of cisplatin at a dosage of 6 mg / kg [Bagnis C, Beaufils H, Jacquiaud C, Adabra Y, Jouanneau C, Le Nahour G, Jaudon MC, Bourbouze R, Jacobs C, Deray G. Erythropoietin enhances recovery after cisplatin-induced acute renal failure in the rat. Nephrol Dial Transplant. 2001 May;16(5):932-8]. In order to prevent microcirculatory disorders in the kidneys, erythropoietin was administered intraperitoneally once at a dosage of 100 IU / kg before the cisplatin injection. To assess renal blood flow, para-aminohippuric acid and inulin clearance were measured 4 and 9 days after cisplatin administration. This resulted in improved microcirculation in kidney tissue.

[0004] The disadvantage of this method is the erythropoietic effect exerted by erythropoietin, which is undesirable and may contribute to an increased risk of thrombotic events in the first year after kidney transplantation (erythropoietin 24.4% compared with placebo 6.4%) [Aydin Z, Mallat MJ, Schaapherder AF. et al. Randomized trial of short-course high-dose erythropoietin in donation after cardiac death kidney transplant recipients. Am J Transplant. 2012;12(7):1793-1800]. The risk of adverse cardiovascular events was also significantly increased with erythropoietin treatment in end-stage chronic kidney disease [Singh AK, Szczech L, Tang KL, Barnhart H, Sapp S, Wolfson M, Reddan D; CHOIR Investigators. Correction of anemia with epoetin alfa in chronic kidney disease. N Engl J Med. 2006 Nov 16; 355(20):2085-98].It is worth mentioning such important side effects of erythropoietin use as arterial hypertension and stimulation of growth and progression of malignant neoplasms [Phrommintikul A, Haas SJ, Elsik M, Krum H. Mortality and target haemoglobin concentrations in anaemic patients with chronic kidney disease treated with erythropoietin: a meta-analysis. Lancet. 2007 Feb 3;369(9559):381-8]. Therefore, the results of correction of microcirculation in the kidney in modeling cisplatin-induced nephropathy in animals using erythropoietin are unsatisfactory.

[0005] The objective of the invention is to create a more effective method for preventing microcirculation disorders in the kidney in cisplatin-induced nephropathy, including the use of an 11-membered peptide that imitates the α-helix B of erythropoietin, which does not have an erythropoietic effect.

[0006] The technical result of the invention is a method for preventing microcirculation disorders in the kidney in cisplatin-induced nephropathy by daily intraperitoneal administration of an 11-membered peptide that imitates the α-helix B of erythropoietin (pHBSP) at a dosage of 25 μg / kg for 14 days, wherein on the first and eighth days of the experiment, 30 minutes after the administration of the 11-membered peptide that imitates the α-helix B of erythropoietin (pHBSP), cisplatin is administered at a dosage of 5 mg / kg.

[0007] The method is as follows: the study is carried out on white male Wistar laboratory rats weighing 320-360 g. Rats without external signs of disease, which had undergone a quarantine regime (14 days), were selected for the study. For the experiment, the rats are divided into 3 groups: the first group simulated pathology; the second group of intact rats; the third study group. In the first group, cisplatin-induced acute kidney injury was modeled by intraperitoneal administration of cisplatin at a dosage of 5 mg / kg on the first and eighth days of the experiment. In the second group, intact rats were intraperitoneally injected with 2 ml of 0.9% NaCl solution on the first and eighth days of the experiment as a placebo.In the third study group, rats were administered an 11-membered peptide mimicking the α-helix B of erythropoietin (pHBSP) intraperitoneally daily at a dosage of 25 μg / kg for 14 days, and on the first and eighth days of the experiment, 30 minutes after the administration of the 11-membered peptide mimicking the α-helix B of erythropoietin (pHBSP), cisplatin was administered at a dosage of 5 mg / kg.

[0008] The results are statistically processed by calculating the arithmetic mean (M) and standard error of the mean (±m). The statistical significance of differences in intergroup comparisons is assessed using a two-tailed Student's t-test for independent groups. Differences are considered statistically significant at p values ​​< 0.05.

[0009] Example 1

[0010] Rats that had undergone a 14-day quarantine at the vivarium of Belgorod State National Research University and showed no signs of acute or chronic diseases were selected for participation in the study. Housing conditions complied with the standards of the Decree of the Chief State Sanitary Doctor of the Russian Federation dated August 29, 2014, No. 51 and GOST 33044-2014: the animals were housed in plastic cages equipped with special metal mesh lids and a feeding section, with 5 animals per cage, free access to food and water, and fed a balanced diet for this species. Environmental parameters were maintained at the following levels: temperature was 20-25°C and relative humidity was 60-65%. Wood sawdust, pre-sterilized with UV light, was used as bedding.

[0011] For the experiment, rats were divided into three groups. In the first group, cisplatin-induced acute kidney injury was modeled by intraperitoneal administration of cisplatin at a dose of 5 mg / kg on the first and eighth days of the experiment. In the second group, intact rats were intraperitoneally administered 2 ml of a 0.9% NaCl solution as a placebo on the first and eighth days of the experiment. In the study group, rats were administered an 11-membered peptide mimicking the α-helix B of erythropoietin (pHBSP) intraperitoneally daily at a dosage of 25 μg / kg for 14 days, and on the first and eighth days of the experiment, 30 minutes after the administration of the 11-membered peptide mimicking the α-helix B of erythropoietin (pHBSP), cisplatin was administered at a dosage of 5 mg / kg.

[0012] Fourteen days after the initial administration of cisplatin, rats are sedated with general anesthesia using intraperitoneal administration of chloral hydrate (Sigma-Aldrich) at a dose of 300 mg / kg body weight. After anesthesia onset, rats are transferred to a room with an ambient temperature of at least 25°C and secured in a supine position on a specially equipped heated veterinary table. After preliminary preparation of the surgical site, which consists of mechanical removal of hair from the skin of the anterior abdominal wall and subsequent treatment of the access area with an antiseptic solution twice, a midline laparotomy is performed with an incision along the linea alba. Next, the intestinal loops are carefully mobilized, the retroperitoneal space is opened, and the renal body and elements of the renal pedicle are isolated on both sides.Renal cortex microcirculation is measured using the LAZMA ST laser diagnostic system (LAZMA Research and Production Enterprise, Russia), which includes a LAZMA-D peripheral blood flow, lymph flow, and tissue coenzyme analyzer. The peripheral sensor is applied to the mid-portion of the kidney, sparing the hilum. Results are recorded and processed using AcqKnowledge software version 3.8.1. Values ​​are expressed in perfusion units (PU).

[0013] The level of microcirculation in the kidney in the group of intact animals was 25.1±0.7 PU. The development of cisplatin-induced nephropathy led to a deterioration in microcirculation in the kidney, as evidenced by a drop in the level of microcirculation to 13.4 ± 0.9 PU (p < 0.05) 14 days after the first administration of cisplatin. Daily intraperitoneal administration of an 11-membered peptide mimicking the α-helix B of erythropoietin (pHBSP) at a dosage of 25 μg / kg against the background of modeling cisplatin-induced kidney injury in rats led to a reliable increase in the level of microcirculation in the kidney on the 14th day to 18.7 ± 0.6 PU, which is statistically higher (p < 0.05) than in the group of "untreated" rats.

[0014] Thus, the obtained results convincingly indicate the pronounced prevention of microcirculation disorders in the kidney when using an 11-membered peptide that imitates the α-helix B of erythropoietin (pHBSP), which does not have an erythropoietic effect, under the conditions of a reproducible model of cisplatin-induced kidney injury.

[0015] Table 1

[0016] Effect of an 11-membered peptide mimicking α-helix B of erythropoietin on microcirculation in a model of cisplatin-induced renal injury in rats

[0017]

[0018]

[0019] Notes: PU - perfusion units; x - p<0.05 in comparison with the group of intact animals; y - p<0.05 compared to the pathology modeling group.

Claims

A method for preventing microcirculation disorders in the kidney in cisplatin-induced nephropathy, including intraperitoneal administration of cisplatin, characterized in that rats are administered daily an 11-membered peptide that imitates the α-helix B of erythropoietin (pHBSP), which does not have an erythropoietic effect, intraperitoneally at a dosage of 25 μg / kg for 14 days, while on the first and eighth days of the experiment, 30 minutes after the administration of the 11-membered peptide that imitates the α-helix B of erythropoietin (pHBSP), cisplatin is administered at a dosage of 5 mg / kg.

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