RU2519727C2 - Antiproliferative agent - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, namely to an agent for treating benign prostatic hyperplasia. The antiproliferative agent for treating benign prostatic hyperplasia which contains red deer's antler lipids prepared by extraction of the red deer's antlers in a mixture of the solvents ethanol: ethyl acetate: water in the specific environment.

EFFECT: agent is effective for treating benign prostatic hyperplasia.

2 tbl

Description

The invention relates to medicine, namely to the creation of medicines of natural origin and can be used to treat benign prostatic hyperplasia (BPH).

BPH is one of the most common diseases of elderly men [Alyaev Yu.G., Vinarov A.Z., Lokshin K.L., Spivak L.G. New in the conservative treatment of patients with prostatic hyperplasia // Consilium medicum. - 2008. - No. 4. - S. 45-49]. Histological signs of BPH are found in more than 40% of men over 50 and 90% of men over 80. Clinical symptoms of the disease are diagnosed in most men older than 50 years [Dhingra N., Bhagwat D. Benign prostatic hyperplasia: an overview of existing treatment // Ind. J. Pharmacol. - 2011. - Vol. 43, N 1. - P.6-12]. According to epidemiological studies conducted in Russia, the frequency of BPH has increased from 11.3% among 40-50-year-olds to 81.4% in 80-year-old men [Rational pharmacotherapy in urology / Ed. ON. Lopatkina, T.S. Pepanova. - M .: Litterra, 2006. - S.30-42].

The use of the homeopathic preparation rhododendron for the treatment of benign tumors is known [Pat. RF №2207868]. Its disadvantage is the need for individual dose selection for each patient, which makes it difficult to use in outpatient practice.

It is known the use of decoction of licorice root naked for the treatment of chronic prostatitis [Pat. RF №2058152]. The disadvantages of this tool are unstable during storage and inconvenient to prepare the dosage form, weak symptomatic effect in chronic prostatitis without therapeutic effect on the pathogenesis of the disease, which does not exclude the occurrence of relapses.

It is known to use a herbal preparation for the treatment of prostatitis and prostate adenoma in the form of complex therapy, including oral and rectal agents [Pat. RF №2275929]. This tool has an anti-inflammatory effect, but does not inhibit proliferative processes in the prostate gland. The therapeutic effect with benign prostatic hyperplasia occurs only with combined oral administration and rectal administration in the form of enemas, which creates inconvenience for outpatients.

Known therapeutic and prophylactic tool "Prostanorm" for the treatment of prostatitis, urethritis and other inflammatory diseases of the genitourinary sphere, including licorice root, St. John's wort grass, Canadian goldenrod grass, roots with rhizomes of Echinacea purpurea, taken in equal proportions [Pat. RF No. 209811] This remedy also has only an anti-inflammatory effect and does not have an antiproliferative effect in benign prostatic hyperplasia.

More than 10 groups of drugs are used for the treatment and prevention of BPH, although only the α-adrenergic blocking agents, finasteride 5α-reductase inhibitor and herbal remedies - extracts of the American fanifolia palm Serenoa repens (permixone, prostamol) and Cameroonian plum Pygeum africanum (trianol) have been proven to be therapeutic [ Rational pharmacotherapy in urology / Ed. ON. Lopatkina, T.S. Pepanova. - M .: Litterra, 2006. - S.30-42].

In light of the foregoing, it is relevant to search for drugs that have a complex effect on regulatory mechanisms that trigger a cascade of physiological changes that lead to the emergence of such a serious pathology as BPH.

Closest to the proposed solution is the tool according to patent CN101884759, taken as a prototype.

The objective of the invention is the expansion of the arsenal of funds of natural origin based on domestic raw materials effective in the treatment of BPH.

The technical result is achieved by the fact that as the antiproliferative use lipids antlers of maral.

The extraction of lipids from maral antlers is carried out with a mixture of ethanol: ethyl acetate: water solvents at their ratio, parts by weight, 1: 1: 1 and the ratio of raw materials: extractant 1:10 parts by weight. three consecutive macerations of 4 hours each. The extract obtained is concentrated in a vacuum evaporator and dried in an evaporator.

The experiments were performed on 120 outbred white rats of males of late reproductive age (8-10 months) weighing 500-550 g, grown under conventional conditions in a laboratory of laboratory animals of the Scientific Research Institute of Pharmacology SB RAMS. Antiproliferative effect of maral antler lipids was evaluated on a BPH model induced by daily intraperitoneal administration of sulpiride (eglonil, ampoule solution, Sanofi-Winthrop Industrie, France) at a dose of 40 mg / kg for 60 days. Throughout the experiment, rats were injected with maral antlers lipids in the form of a suspension of 1% starch mucus in doses of 10, 30, and 60 mg / kg. An extract of Serenoa repens permixon (Pierre Fabre Medicament Production, France) at a dose of 50 mg / kg (according to Serenoa repens extract) was used as a reference preparation. Control animals received 1% starch mucus. After 60 days, the rats were decapitated under ether anesthesia. The anterior, lateral and posterior prostate glands were weighed. For histological examination, the lateral and posterior lobes were fixed with 10% neutral formalin, dewaxed sections were stained with hematoxylin-eosin. In the homogenates of the lateral and posterior prostate glands, the content of protein, diene conjugates, the rate of formation of malondialdehyde, the amount of reduced glutathione, the activity of selenium-dependent glutathione peroxidase and superoxide dismutase were determined. In the blood plasma, the content of prolactin, testosterone and 5α-dihydrotestosterone was analyzed by the immunofluorescence method (DRG International kits, USA).

Statistical processing of the results was carried out by the method of pairwise comparisons using the nonparametric Wilcoxon-Mann-Whitney criterion with the probability of an erroneous conclusion not exceeding 5% (p≤0.05).

Sulpiride, by blocking dopamine D ₂ receptors in the tuberoinfundibular system, significantly increases the secretion of the prolactin proliferation proliferation stimulator with the development of experimental BPH. With the introduction of sulpiride for 60 days, the mass of the anterior lobe of the prostate gland did not change, the mass of the lateral and posterior lobes and weight coefficients (the ratio of the mass of the gland to the body weight of the animals) became 1.8-2.1 times more than in intact animals (table .one). Histological examination of the prostate gland determined an increased number of acini with papillary proliferation of the papillary epithelium. The stroma between the acini contained a large number of hypertrophied smooth muscles, coarse collagen fibers, was infiltrated by neutrophils, lymphocytes, macrophages, mast cells. The expanded lumen of the acini was filled with acidophilic mass and neutrophils. In the homogenates of the lateral and posterior prostate glands, the protein content increased 1.6 times and lipid peroxidation was activated against the background of inhibition of antiperoxidation. The content of diene conjugates increased 2.1 times compared with their normal amount, the production of malondialdehyde in the presence of ascorbate accelerated 2.2 times, after the addition of NADP · H - 2.7 times. The content of reduced glutathione, the activity of glutathione peroxidase and superoxide dismutase decreased by 1.7-2.2 times. In the blood serum of rats with experimental BPH, the prolactin content increased 5.8 times, 5α-dihydrotestosterone - 1.5 times, the amount of testosterone decreased 3.2 times (Table 2).

Maral antlers lipids at doses of 10, 30 and 60 mg / kg and Serenoa repens extract at a dose of 50 mg / kg effectively prevented the progression of experimental BPH caused by injections of sulpiride in male rats of late reproductive age. At a dose of 10 mg / kg, maral antler lipids and Serenoa repens extract reduced the mass of the lateral and posterior prostate glands by 1.4-1.5 times, although it remained 1.3-1.4 times more than normal. At doses of 30 and 60 mg / kg, lipids of maral antlers reduced the mass of the lateral and posterior lobes by 1.8-2.1 times, while the mass became the same as in intact animals. Weighting factors decreased by 1.8-2.1 times. The antiproliferative effect of maral antler lipids at doses of 30 and 60 mg / kg exceeded the effect of Serenoa repens extract (Table 1).

For an in-depth study of the antiproliferative effect of the BPH model, maral antlers lipids were administered at a dose of 30 mg / kg. Under the influence of experimental therapy with maral antler lipids, the area of epithelial structures was reduced in the tissue of the lateral and posterior lobes of the prostate against the background of the expansion of the lumen of acini. The tissue of the prostate gland consisted of individual glands separated by narrow bundles of connective tissue with smooth muscle cells, collagen and elastic fibers of normal structure. Cellular infiltration of interstitium was absent; leukocytes were not detected in the acini. With the introduction of Serenoa repens extract, moderate lymphocytic macrophage infiltration persisted. In the homogenates of the lateral and posterior prostate glands, experimental therapy with maral antler lipids was accompanied by a 1.5-fold decrease in protein content, and by a significantly lesser extent by 1.3 times when Serenoa repens extract was used (Table 2).

Maral antlers lipids had a pronounced antioxidant effect. In the homogenates of the lateral and posterior lobes of the prostate, the content of lipid peroxidation products and anti-peroxidation indicators — the amount of reduced glutathione, the activity of glutathione peroxidase and superoxide dismutase returned to normal. The content of diene conjugates decreased by 2.2 times, the production of ascorbate and NADP · H-dependent malondialdehyde slowed down by 1.8–2.1 times, the content of glutathione, the activity of glutathione peroxidase and superoxide dismutase increased by 1.5–2 times compared to indicators for BPH model. With the introduction of Serenoa repens extract, the intensity of lipid peroxidation decreased by 1.4-1.8 times, the activity of antiperoxidation increased by 1.3-1.5 times (differences with the effects of lipids of antlers of maral are significant only for the content of diene conjugates). The production rate of malondialdehyde exceeded the normal rate by 1.4-1.5 times, the content of reduced glutathione, the activity of glutathione peroxidase and superoxide dismutase remained reduced by 1.4 times (Table 2).

In the blood of rats treated with maral antler lipids, the prolactin content decreased threefold compared with the hormone level in rats that were administered only sulpiride, but it was 1.9 times higher than normal. With the introduction of the extract of Serenoa repens, the prolactin content in the blood decreased to a lesser extent - 1.5 times, remaining 3.8 times higher than normal and twice as much as with the introduction of maral antlers lipids. The content of male sex hormones in the blood of rats protected by maral antlers lipids and Serenoa repens extract changed to the same extent and became close to the hormones in intact animals - the testosterone content increased 2.2-2.6 times, 5α-dihydrotestosterone decreased 1.5-1.7 times compared with the amount of these hormones, measured with the introduction of one sulpiride (table 2).

Thus, in the model of BPH caused by the introduction of a prolactin secretion stimulator by sulpiride to rats of males of late reproductive age, maral lipids in an effective dose of 30 mg / kg and Serenoa repens extract in a dose of 50 mg / kg prevented the development of lateral and posterior prostate hyperplasia, proliferation of its epithelium and inflammatory infiltration. Both drugs had a pronounced antioxidant effect and changed the hormonal balance towards normal. In experimental BPH, antler lipids, to a greater extent than Serenoa repens extract, restrained the increase in the mass of the lateral and posterior prostate glands, the production of primary lipid peroxidation products - diene conjugates and the development of hyperprolactinemia.

Judging by the increase in testosterone levels and the decrease in the amount of 5α-dihydrotestosterone in the blood of rats treated with maral lipids from sulpiride injections, this drug inhibits 5α-reductase and inhibits the recovery of testosterone in 5α-dihydrotestosterone, which stimulates the proliferation of prostate tissue. The antioxidant effect is of great importance in the mechanism of the antiproliferative effect of lipids of antlers of maral. Lipoperoxidation products stimulate the proliferation of epithelium and connective tissue in the prostate gland.

Table 1

; n=10)The effect of maral antler lipids in comparison with the action of Serenoa repens extract (permixon) on the mass of the prostate gland in a model of its benign hyperplasia ( $$\overline{\text{X}}\pm \text{S}\overline{\text{x}}$$

; n = 10) Indicator Intact animals Sulpiride Antler antlers lipids Serenoa repens extract 10 mg / kg 30 mg / kg 60 mg / kg + sulpiride The mass of rats, g 510.6 ± 7.5 523.4 ± 8.7 525.7 ± 8.4 512.8 ± 17.6 529.4 ± 19.6 529.6 ± 11.5 Anterior lobe of the prostate weight mg 191.8 ± 12.7 194.7 ± 10.5 187.1 ± 11.3 184.8 ± 12.2 192.0 ± 18.2 190.9 ± 12.0 weight coefficient, mg / g 0.38 ± 0.05 0.37 ± 0.04 0.36 ± 0.03 0.36 ± 0.05 0.35 ± 0.04 0.36 ± 0.02 Lateral Prostate weight mg 43.6 ± 2.9 87.9 ± 7.1 * 59.2 ± 3.0 * ⁺ 42.4 ± 4.5 ⁺ 43.9 ± 4.3 ⁺ 64.5 ± 3.5 * ^{+ #} weight coefficient, mg / kg 0.08 ± 0.01 0.17 ± 0.02 * 0.11 ± 0.02 0.08 ± 0.01 ⁺ 0.08 ± 0.02 ⁺ 0.12 ± 0.02 Posterior prostate weight mg 49.6 ± 3.8 96.7 ± 4.8 * 62.6 ± 3.7 * ⁺ 48.3 ± 5.2 ⁺ 52.6 ± 3.4 ⁺ 70.2 ± 4.0 * ^{+ #} weight coefficient, mg / kg 0.10 ± 0.02 0.18 ± 0.02 * 0.12 ± 0.03 0.09 ± 0.02 ⁺ 0.10 ± 0.02 ⁺ 0.13 ± 0.02 Note. P <0.05: compared with the indicator * for intact animals, ⁺ - with the introduction of sulpiride, ^# - with the introduction of lipids of antlers of maral in doses of 30 and 60 mg / kg.

table 2

; n=10)The effect of maral antler lipids on lipoperoxidation and hormone content compared with the action of Serenoa repens extract (permixon) in a model of benign prostatic hyperplasia ( $$\overline{\text{X}}\pm \text{S}\overline{\text{x}}$$

; n = 10) Indicator Intact animals Sulpiride Antler antlers lipids Serenoa repens extract + sulpiride Prostate homogenate Protein, mg / g of tissue 120.7 ± 7.9 197.4 ± 10.3 * 129.0 ± 5.9 ⁺ 156.8 ± 7.3 * ^{+ #} Diene conjugates, µmol / g tissue 18.6 ± 1.5 38.7 ± 2.7 * 17.9 ± l, 6 ⁺ 26.8 ± 3.4 ^{+ #} Malondialdehyde, nmol / mg protein · min: ascendant dependent 0.44 ± 0.03 0.98 ± 0.07 * 0.55 ± 0.06 ⁺ 0.63 ± 0.07 * ⁺ NADP · H-dependent 0.57 ± 0.06 1.54 ± 0.06 * 0.75 ± 0.08 ⁺ 0.88 ± 0.09 * ⁺ Reduced glutathione, nmol / mg protein 5.2 ± 0.4 2.4 ± 0.3 * 4.8 ± 0.7 ⁺ 3.5 ± 0.5 * ⁺ Glutathione peroxidase, nmol NADP · H / g tissue · min 2528 ± 123 1478 ± 96 * 2198 ± 119 ⁺ 1865 ± 114 * ⁺ Superoxide dismutase, E / mg protein 0.87 ± 0.07 0.41 ± 0.05 * 0.69 ± 0.08 ⁺ 0.61 ± 0.06 * ⁺ Blood serum Prolactin, ng / ml 10.9 ± 1.8 62.7 ± 3.3 * 21, l ± 2.5 * ⁺ 41.7 ± 5.3 * ^{+ #} Testosterone, pg / ml 2187 ± 312 673 ± 57 * 1765 ± 164 ⁺ 1460 ± 97 ⁺ 5α-dihydrotestosterone, pg / ml 179.7 ± 10.9 265.9 ± 23.4 * 171.6 ± 13.5 ⁺ 158.7 ± 17.9 ⁺ Note. P <0.05: compared with the indicator * - of intact animals, ⁺ - with the introduction of sulpiride, ^# - with the introduction of lipids of antlers of maral.

Claims (1)

Antiproliferative agent for the treatment of benign prostatic hyperplasia based on extracts from raw materials of animal origin, characterized in that it contains maral antlers lipids obtained by extraction of a mixture of solvents ethanol: ethyl acetate: water in their ratio, parts by weight, 1: 1: 1 and the ratio of raw materials: extractant 1:10.