RU2739491C1 - Agent for preventing and treating asthenozoospermia and teratozoospermia - Google Patents

Abstract

FIELD: medicine; pharmaceuticals.

SUBSTANCE: invention refers to medicine and pharmaceutics and can be used for prevention and treatment of asthenozoospermia and teratozoospermia. For this purpose, it is proposed to apply Verva fir extract in an effective dose once a day daily for 5 days before and 5 days after cytostatic exposure as an agent for prevention and treatment of asthenozoospermia and teratozoospermia.

EFFECT: invention provides higher sperm cell mobility and reduced number of pathological forms in asthenozoospermia and teratozoospermia.

1 cl, 1 ex, 2 dwg

Description

The invention relates to medicine, specifically to pharmacology, and relates to a means of preventing and treating asthenozoospermia and teratozoospermia.

The problem of male infertility has acquired not only medical, but also social significance [1]. The most common forms of male infertility are asthenozoospermia (occurs in 41% of cases of male infertility) and teratozoospermia (occurs in 26% of cases) [1]. Asthenozoospermia is characterized by low rates of sperm motility, teratozoospermia - by an increased number of their pathological forms [2].

For the treatment of male infertility, the drug Speman is recommended [3]. This drug is recommended primarily for the treatment of asthenospermia. Speman is a multicomponent composition of medicinal herbs that grow in different countries of the world. The drug has a complex composition of medicinal herbs that grow in different countries of the world. Their raw materials are limited. As a disadvantage of Speman, it should be noted the possibility of allergic reactions, and its effectiveness is not always high, the drug does not reduce the severity of teratozoospermia. In addition, Speman is a rather expensive drug.

Among the drugs that reduce the severity of asthenozoospermia and teratozoospermia is the drug Dibornol [4]. This drug is a semi-synthetic terpenophenol, in the structure of which there are two terpene substituents (2,6-diisobornyl-4-methylphenol). Dibornol exhibits the properties of an effective antioxidant.

However, Dibornol is insufficiently effective, since its use in an experimental model of pathospermia does not allow reaching the values of the percentage of motile spermatozoa of the intact control level (background) [4].

Known remedy - fir extract "Verva", which increases the yield of high-quality sperm production of healthy bulls-producers [5]. It was also found that this extract helps to increase the milk yield of cows [6]. The natural preparation Verva is an economically available product, it is obtained from the woody greenery of fir. The active substance of the biological product "Verva" is triterpenic acids (5 g / l), and there are also natural phenolic compounds with antioxidant properties. We have shown for the first time its ability to significantly increase sperm motility and reduce the number of their pathological forms in conditions of asthenozoospermia and teratozoozoospermia.

The problem solved by this invention is to expand the arsenal of agents that increase sperm motility and reduce the number of pathological forms of spermatozoa used for the prevention and treatment of asthenozoospermia and teratozoospermia. The technical result of the proposed invention is to obtain a therapeutic effect in conditions of asthenozoospermia and teratozoospermia.

The technical result is achieved through the use of fir extract "Verva" for the prevention and treatment of asthenozoospermia and teratozoospermia, taken in an effective dose, providing sperm motility, once a day every day for 5 days before and 5 days after cytostatic exposure.

The “Verva” fir extract agent used in the invention was developed by the Institute of Chemistry of the Komi Scientific Center of the Ural Branch of the Russian Academy of Sciences on the basis of an emulsion extract of fir tree greenery (state registration certificate No. PVR-2-5.0 / 0260). It has been established that fir extract is a means of increasing the physiological reserves of the body [7].

The claimed essential features in the aggregate have shown new properties that do not follow explicitly from the prior art in this field and are not obvious to a specialist. The fact that the Verva extract leads to an increase in sperm production in sire bulls does not yet indicate that this agent will be effective in asthenozoospermia and teratozoospermia. Special scientific research is required to establish the effect of fir extract on sperm motility and the number of pathological forms of spermatozoa. This is evidenced by the data obtained in the experimental study of the effectiveness of a number of adaptogens (pantocrine, ginseng, eleutherococcus) in various models of asthenozoospermia and teratozoospermia in rats [8]. The results of this study indicate that the introduction to healthy male rats (immature, elderly), not characterized by signs of asthenozoospermia and tetrazoospermia (in other words, without signs of pathology), adaptogens (pantocrine, ginseng, eleutherococcus) leads to an increase in sperm motility and a decrease in the number of pathological forms of sperm. Testing these compounds in four different models of asthenozoospermia and tetrazoospermia (folliculin, cyproterone, androcuric, fenosteride) does not lead to an increase in the percentage of motile forms and a decrease in the number of pathological forms of sperm. All these adaptogens were found to be effective only in one model of pathospermia (metaminyl). These data indicate that the positive effects of the drug on the number of motile forms of sperm, the percentage of their pathological forms that are detected in healthy animals may not be confirmed in a specific model of tetrazoospermia asthenospermia. This is due to the fact that the pathogenesis of asthenozoospermia and tetrazoospermia may not be associated with the mechanism of the stimulating effect of the drug on these parameters. It is no coincidence that the use of models of pathological processes is a prerequisite for the search for new drugs [9].

The revealed new properties of Verva extract - to prevent the development of asthenozoospermia and teratozoospermia and increase sperm motility - have not been found in the patent and scientific medical literature.

The proposed invention can be used in experimental medicine with access to practical health care.

Based on the foregoing, the claimed technical solution should be considered the corresponding criteria: "Novelty", "Inventive level", "Industrial applicability".

The invention is illustrated by the experimental diagrams in FIGS. 1 and 2.

FIG. 1 shows the data on the percentage of mobile forms of germ cells in animals receiving etoposide (1 - control); etoposide in combination with fir extract "Verva" (2 - etoposide + "Verva" 100 mg / kg; 3 - etoposide + "Verva" 220 mg / kg; 4 - etoposide + "Verva" 500 mg / kg); etoposide in combination with Dibornol (5 - etoposide + Dibornol 10 mg / kg).

FIG. 2 shows the data on the percentage of pathological forms of sperm in animals treated with etoposide (1 - control); etoposide in combination with fir extract "Verva" (2 - etoposide + "Verva" 100 mg / kg; 3 - etoposide + "Verva" 220 mg / kg; 4 - etoposide + "Verva" 500 mg / kg); etoposide in combination with Dibornol (5 - etoposide + Dibornol 10 mg / kg).

The method is carried out as follows.

To laboratory animals (male rats) fir extract "Verva" in doses of 100, 220 or 500 mg / kg is administered once a day every day for 5 days before and 5 days after a single intravenous injection of the cytostatic drug etoposide in the maximum tolerated dose of 30 mg / kg.

The proposed method was studied in experiments on male rats of the Wistar population (n = 60) of reproductive age (4 months) weighing 250-300 g. Animals were obtained from the laboratory of biological modeling of the Research Institute of Physics and Mathematics named after. E. D. Goldberg. The rats were kept in accordance with the rules adopted by the European Convention for the Protection of Vertebrate Animals used for Experimental and Other Scientific Purposes (Strasbourg, 1986). Damage to the gonads was simulated by a single intravenous injection at the maximum tolerated dose (30 mg / kg) of the cytostatic drug etoposide (Teva, Israel). The choice of the drug was due to the fact that etoposide causes significant suppression of spermatogenesis and various types of asthenozoospermia and teratozoospermia [10].

Of the 60 rats used, 10 made up the group of intact animals (background). Control males (n = 10) were injected with etoposide (Teva, Israel) once, intravenously at the maximum tolerated dose of 30 mg / kg. Rats of three experimental groups (10 in each) received Verva fir extract intragastrically using a probe at doses of 100, 220 or 500 mg / kg once a day every day for 5 days before and 5 days after the administration of etoposide. The rats of the 6th group (n = 10) were injected with Dibornol intragastrically at a dose of 10 mg / kg of body weight once a day for 5 days before and 5 days after the administration of etoposide. Dibornol was used as a reference drug. Upon completion of administration, the animals were euthanized in a CO ₂ chamber after opening the tail portion appendages was isolated testes. It was cut open and placed in warm saline solution. In a drop of the obtained cell suspension on a glass slide, the percentage of motile forms of spermatozoa was calculated. Then, a smear was made from the same suspension, it was fixed and the percentage of degenerative forms of mature spermatozoa was calculated [11].

The results were processed by the method of variation statistics using the nonparametric Mann-Whitney U test.

Example.

The number of motile sperm forms 6 days after the administration of etoposide significantly decreased and amounted to 62.2% of the background (Fig. 1), which indicates a pronounced asthenozoospermia. In rats that were injected with fir extract at a dose of 100 mg / kg (2), this indicator slightly increased (compared to the control ((1) P≥0.05) and amounted to 81.3% of the background. However, its values were significantly different from those in the background (Fig. 2). When the drug was administered at a dose of 220 mg / kg (3), the number of motile forms of sperm increased in comparison with the control (P≤0.05) and fully corresponded to the background values (100% of the background) When using fir extract "Verva" at a dose of 500 mg / kg (4), this indicator significantly increased compared to control (1), and did not statistically significantly differ from the background values (89.9% of the background). who received the reference drug (5), the number of motile sperm forms significantly increased compared to the control (1), but did not reach the background values (78.9% of the background). Thus, the Verva plant extract prevented the development of asthenozoospermia at doses of 220 and 500 mg / kg The most pronounced therapeutic effect of the drug the agent was detected at a dose of 220 mg / kg. The efficacy of Dibornol was lower than that of the herbal extract, which was administered at doses of 220 and 500 mg / kg (Fig. 2 (3 and 4)).

The number of pathological forms of sperm 6 days after the introduction of etoposide was statistically significantly higher compared to the background and amounted to 138.9% of the background (Fig. 2), which indicates pronounced teratozoospermia. In rats that were injected with Verva fir extract at a dose of 100 mg / kg (2), this indicator decreased compared to the control (P≤0.05) and amounted to 72% of the background (P≤0.05). With the introduction of the fir extract "Verva" at a dose of 220 mg / kg (3), the percentage of pathological forms of spermatozoa also decreased compared to the control (P≤0.05), but to a greater extent (up to 60% of the background values; P≤0, 05). When using the plant extract "Verva" at a dose of 500 mg / kg (4), this figure was up to 71.4% of the background (P≤0.05), and was reduced compared to the control (1). In the group of rats receiving the reference drug, the number of pathological forms significantly decreased compared to the control (P≤0.05), and the baseline values did not differ statistically significantly (90.6% from the background). Thus, the plant extract prevented the development of teratozoospermia when used in all studied doses. Noteworthy is the fact that the percentage of pathological spermatozoa in all rats receiving fir extract was statistically significantly lower than the background values. The most significant pharmacological activity of the "Verva" extract was detected when administered at a dose of 220 mg / kg. The therapeutic effect of the reference drug was less pronounced.

The presented data indicate that the fir extract "Verva" in doses of 100, 220 and 500 mg / kg has pharmacological activity under the conditions of the model of experimental asthenozoospermia and teratozoospermia caused by etoposide. The most effective dose was 220 mg / kg. Against the background of the use of the extract in this dose, the normalization of the number of motile and pathological forms of spermatozoa was noted. The values of the percentage of mobile forms reached the level of intact animals (background), and the percentage of pathological forms was even lower than the background values. The comparison drug Dibornol also led to a decrease in the severity of asthenozoospermia and teratozoospermia. However, after its use, such an indicator as the percentage of mobile forms did not reach the level of intact animals. Thus, the advantage of “Verva” fir extract over Dibornol is its greater efficiency, namely, the ability to completely prevent the development of asthenozoospermia and teratozoospermia. In addition, the number of pathological forms of spermatozoa against the background of the introduction of the plant extract decreased so much that it turned out to be even lower than background values.

The proposed application of the fir extract "Verva" at a dose of 220 mg / kg once a day for 5 days before and 5 days after cytostatic exposure to male rats allows for highly effective correction of the morphological and functional state of spermatozoa in conditions of asthenozoospermia and teratozoospermia.

The revealed properties of the proposed agent indicate the promise of its use in male infertility caused by asthenozoospermia and teratozoospermia.

Information sources

1. Andreeva M.V., Khayat S.Sh., Sorokina T.M., Chernykh V.B., Shileiko L.V., Shtaut M.I., Mashina E.V., Dobrodeeva L.T., Sedova A.O., Kurils L.F. Forms of pathozoospermia in men with infertility in marriage and / or with disorders of the reproductive system // Andrology and Genital Surgery. 2017. No. 2. S. 33-38.

2. Showell M., Mackenzie-Proctor R., Brown J. et al. Antioxidants for male subfertility // Cochrane database syst. Rev. 2014.170.

3. Directory Vidal. Medicines in Russia. M., 1995. AstraFarmService. P. 49.

4. Borovskaya TG, Kamalova VA, Vychuzhanina AV, Poluektova ME, Kuchin AV, Chukicheva I.Yu. Means for the prevention and treatment of pathozoospermia // Patent (RU) 2619330, publ. 05/15/2017. Bul. No. 14, prior. dated 03.24.2016. Federal State Budgetary Scientific Institution "Research Institute of Pharmacology and Regenerative Medicine named after E.D. Goldberg ".

5. Ryaposova M.V., Shkuratova I.A., Mymrin V.S., Busygina O.A., Busygin P.O., Tarasenko M.N., Belousov A.I., Mymrin S.V., A.V. Kuchin, T.V. Khurshkainen The method of increasing the yield of sperm production from bulls // Patent 2601156. 2016. BI No. 30.

6. Zharikov Ya.A., Khurshkainen T.V. Influence of fir feed additives on the productivity of dairy cows // Zootechnia. 2011. No. 5. S. 9-12.

7. Garnov I.O., Kuchin A.V., Mazina N.K., Karpova E.M., Boyko E.R. Fir extracts as a means of increasing the physiological reserves of the body // Khimicheskie nauki. 2014. Issue. 3, No. 19. S. 44-52.

8. Lisakovskaya O.V. Influence of the Knotweed and some adaptogens on the functional parameters of the reproductive system: abstract of Ph.D. dis. … Candidate of Biological Sciences: 19.03.99 / Olga Vladimirovna Lisakovskaya. Tomsk, 1999.S. 15, 19, 22, 23.

9. Chereshnev V.A., Shilov Yu.I., Chereshneva M.V., Samodelkin E.I., Gavrilova T.V., Gusev E.Yu., Gulyaeva I.L. Experimental models in pathology. Perm. state un-t. Permian. 2011.267 p.

10. Goldberg E. D. Borovskaya T.G. Timina E.A. et al. State of spermatogenesis in rats after administration of the antitumor drug Vepeside // Byull. experimental biol. and honey. 2005. T. 140, No. 9.S. 301.

11. Guidelines for experimental (preclinical) study of new pharmacological substances / Ed. Mironova N.A. Part 1. M .: Medicine. 2005.S. 92.

Claims (1)

The use of an effective dose of fir extract "Verva", which provides an increase in sperm motility and a decrease in the number of pathological forms of spermatozoa, once a day, every day for 5 days before and 5 days after cytostatic exposure as a means for the prevention and treatment of asthenozoospermia and teratozoospermia.

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